Biodiversitas vol. 11, no. 1, January 2010 by Biodiversitas Unsjournals

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic)

Journal of Biological Diversity V o l u m e

1 1

–

N u m b e r

–

J a n u a r y

2 0 1 0

FIRST PUBLISHED: 2000

ISSN: 1412-033X (printed edition) 2085-4722 (electronic)

EDITORIAL BOARD (COMMUNICATING EDITORS): Abdel Fattah N.A. Rabou (Palestine), Dato A. Latiff Mohamad (Malaysia), Alan J. Lymbery (Australia), Ali Saad Mohamed (Sudan), Bambang H. Saharjo (Indonesia), Charles H. Cannon Jr. (USA), Edi Rudi (Indonesia), Guofan Shao (USA), Hassan Poorbabaei (Iran), Hwan Su Yoon (USA), John Morgan (Australia), Joko R. Witono (Indonesia), Katsuhiro Kondo (Japan), Mahendra K. Rai (India), María La Torre Cuadros (Peru), Mochamad A. Soendjoto (Indonesia), Peter Green (Australia), Salvador Carranza (Spain), Shahabuddin (Indonesia), Sonia Malik (Brazil), Sugiyarto (Indonesia), Thaweesakdi Boonkerd (Thailand)

EDITOR-IN-CHIEF: Sutarno

EDITORIAL MEMBERS: English Literary Editor: I Made Sudiana (sudianai@yahoo.com) Technical Editor & Banking: Solichatun (solichatun_s@yahoo.com) Distribution & Marketing: Rita Rakhmawati (oktia@yahoo.com) Webmaster: Ari Pitoyo (aripitoyo@yahoo.com)

MANAGING EDITORS: Ahmad Dwi Setyawan (unsjournals@gmail.com)

PUBLISHER: Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University, Surakarta and The Society for Indonesian Biodiversity

ADDRESS: Jl. Ir. Sutami 36A Surakarta 57126. Tel. +62-271-7994097, Tel. & Fax.: +62-271-663375, Email: unsjournals@yahoo.com

BANKING: Solichatun, BNI KC Sebelas Maret, Acc. No. 0033691646

ONLINE: www.unsjournals.com

ACCREDITED BY DECREE OF THE DIRECTORATE GENERAL OF HIGHER EDUCATION, THE MINISTRY OF NATIONAL EDUCATION, REPUBLIC OF INDONESIA No. 65a/DIKTI/Kep/2008 (valid until October 2011)

EXPERTATION AND CORRESPONDING EMAIL OF THE COMMUNICATING EDITORS: GENETIC DIVERSITY: Alan J. Lymbery (a.lymbery@murdoch.edu.au), Hwan Su Yoon (hsyoon@bigelow.org), Mahendra K. Rai (pmkrai@hotmail.com), Salvador Carranza (salvicarranza@gmail.com), Sonia Malik (sonia.unicamp@gmail.com). SPECIES DIVERSITY: Dato A. Latiff Mohamad (latiff@ukm.my), Joko R. Witono (jrwitono@yahoo.com), Katsuhiro Kondo (k3kondo@nodai.ac.jp), Thaweesakdi Boonkerd (Thaweesakdi.B@chula.ac.th). ECOSYSTEM DIVERSITY: Abdel Fattah N.A. Rabou (arabou@iugaza.edu), Ali Saad Mohamed (alisaad48@yahoo.com), Bambang H. Saharjo (bhsaharjo@gmail.com), Charles H. Cannon Jr. (chuck@xtbg.ac.cn), Edi Rudi (edirudi@yahoo.com), Guofan Shao (shao@purdue.edu), Hassan Poorbabaei (hassan_pourbabaei@yahoo.com), John Morgan (morgan@latrobe.edu.au), Mochamad A. Soendjoto (asoendjoto@telkom.net), Peter Green (p.green@latrobe.edu.au), Shahabuddin (shahabsaleh@gmail.com), Sugiyarto (sugiyarto_ys@yahoo.com). ETHNOBIOLOGY: María La Torre Cuadros (angeleslatorre@lamolina.edu.pe).

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 1-4

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110101

Genetic variations among Indonesian native cattle breeds based on polymorphisms analysis in the growth hormone loci and mitochondrial DNA SUTARNO♥ Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University, Jl. Ir. Sutami 36A Surakarta 57126, Tel./fax. +62-271663375, email: nnsutarno@yahoo.com Manuscript received: 20 October 2009. Revision accepted: 26 December 2009.

ABSTRACT Sutarno (2010) Genetic variations among Indonesian native cattle breeds based on polymorphisms analysis in the growth hormone loci and mitochondrial DNA. Biodiversitas 11: 1-5. Genetic variation within breeds is important and its study has become a subject of interest in livestock species, as it has many applications in animal breeding and genetics, such as the identification of animals and parentage testing, gene mapping and identifying markers for performance traits. Two loci of bovine growth hormone genes, and two regions of mitochondrial DNA, D-loop and ND-5 were characterized using polymerase chain reaction – restriction fragment length polymorphism (PCR-RFLP) involving 120 Indonesian native cattle of Bali, Madura, PO and West Sumatra breeds. The results indicated that sequence variations were detected both in the growth hormone loci and mitochondrial DNA. Key words: marker assisted selection, local Indonesian cattle, PCR-RFLP, GH gene, mtDNA.

INTRODUCTION Advanced techniques of molecular biology have provided the opportunity to study genetic diversity within and among breeds at gene level. Candidate QTLs, such as the growth hormone gene, BoLA gene and casein gene have been extensively studied. Variation in the growth hormone gene has been reported in many cattle breeds (Lucy et al. 1993; Sutarno 1998, 2001; Beauchemin et al. 2006). Schlee et al. (1994b) reported different concentrations of plasma growth hormone related to different growth hormone genotypes of cattle. Mitochondrial DNA polymorphism has been reported within and between breeds of mostly European cattle (Ron et al. 1990; Sutarno and Lymbery 1997). Loftus et al. (1994) used mtDNA polymorphisms to study the phylogeny of different breeds of cattle from Europe, Asia and Africa, however there was not any report for Indonesian native cattle. In cattle, the growth hormone gene (bovine somatotropin gene) has been cloned and completely sequenced (Woychik et al. 1982). It is 1793 bp long, consisting of five exons separated by four intervening sequences (introns) of 248 bp (A), 227 bp (B), 227 bp (C) and 274 bp (D) in length (Gordon et al. 1983). The bovine growth hormone gene has been assigned to chromosome region 19q26-qtr (Hediger et al. 1990). Variation in the growth hormone gene has been reported in many livestock animals, including cattle (Sutarno 2004, 2005; Beauchemin et al. 2006; Thomas et al. 2006), pigs (Nielsen and Larsen 1991), and sheep (Gootwine et al. 1990). Harvey (1995) suggested that deletions result in growth hormone deficiency or the synthesis of defective growth hormone variants. PCR

procedures followed by restriction endonuclease digestions were recently used for typing further growth hormone gene variations, such as a MspI polymorphism (Mitra et al. 1995; Sutarno 2004), and an AluI polymorphism (Mitra et al. 1995) in the bovine growth hormone gene. Mammalian mtDNA is a double stranded, covalently closed-circular molecule of approximately 16500 nucleotides located within the inner mitochondrial membrane (Shoffner and Wallace 1990). Bovine mitochondrial DNA has been completely sequenced and is 16.338 kb long (Anderson et al. 1982). Mammalian mitochondria carry multiple copies of mtDNA which are replicated and expressed in the mitochondria, and inherited maternally (Wallace 1993). The mitochondrial genome contains genes that encode 13 polypeptides involved in the oxidative phosphorylation (OXPHOS) system, together with the 12S and 16S ribosomal RNAs and the 22 mitochondrial transfer RNAs necessary for mRNA expression. Loftus et al. (1994) described the organization of bovine mitochondrial DNA with locations of known restriction sites. Because of their involvement in the energy generating pathway (ATP synthesis), mitochondria and mitochondrial DNA are found in all animal species. Enzymes encoded by mtDNA participate in mitochondrial oxidative phosphorylation of ADP to ATP. In this process, both nuclear and mitochondrial DNA are required to cooperatively perform the respiratory function (Shadel and Clayton 1997). Variation of mitochondrial DNA has also been demonstrated within species. Brown and Vinograd (1974) were the first to demonstrate intraspecific mtDNA variation using restriction endonuclease analysis. This technique was then applied in pedigree analysis, the estimation of

B I O D I V E R S IT A S

evolutionary relationships, determination of mtDNA involvement in disease, intraspecific migration rates, and involvement of mtDNA in affecting production traits. The aims of the study were therefore to investigate polymorphisms in the growth hormone gene and mitochondrial DNA of many Indonesian native cattle breeds such as Bali, Madura, PO and West Sumatra coastal area

MATERIALS AND METHODS Experimental cattle The cattle used in this study were of Indonesian local cattle comprising of 4 different breeds, Bali, Madura, Ongole derived (PO) and Western Sumatra coastal area cattles. The samples were consist of 30 cattle of each breed. Blood collection Blood was collected by venepuncture into a 10 mL Venoject tube containing heparin as anticoagulant. White blood cells were then isolated from the remaining 10 mL of blood. Isolation of white blood cells Whole blood was dispensed into centrifuge tubes, and then spun at about 1500g for 15-20 minutes. The buffy coat was removed with a pipette, transferred to 10 mL centrifuge tubes, topped up with TE-1 buffer and centrifuged at 2000g for 10-15 minutes. The pellet was resuspended in 1 mL of TE-2 buffer, transferred to 1 mL Nunc storage tube, and frozen at -84oC. Extraction of genomic DNA and mtDNA from white blood cells The genomic DNA was extracted using Wizard genomic DNA purification system from Promega as instructed by the manufacturer, while the mitochondrial DNA was extracted using the Wizard Minipreps DNA Purification System (Promega, Madison, USA). Genotyping Polymerase chain reaction and restriction fragment length polymorphisms (PCR-RFLP) were used to detect polymorphisms in the growth hormone gene and mitochondrial DNA.

11 (1): 1-4, January 2010

D-loop primers: D-L: 5’-TAG TGC TAATACCAACGGCC-3' D-R: 5’-AGGCAT TTTCAG TGCCTTGC-3' ND-5 primers: ND-L: 5'-ATCCGTTGGTCTTAGGAACC-3' ND-R: 5'-TTGCGGTTACAAGGATGAGC-3' All amplification reactions were performed in a 25 uL reaction mix consisting of 200 ng of template DNA, 0.15 uM each of the oligonucleotide primers 200 uM each dNTPs, 2 mM MgCl2, 10x buffer and 1.5 units Taq DNA polymerase (Promega) in 0.2 mL PCR reaction tube. RFLP Analysis The PCR products were used directly in restriction enzyme digestion reactions. Amplified DNA of growth hormone locus 1 was digested with restriction endonuclease AluI (Promega) to identify the AluI site polymorphism, and amplified DNA of growth hormone locus 2 was digested with restriction endonuclease MspI (Promega) to identify the MspI site polymorphism. The Amplified products of D-loop and ND-5 fragments were digested using restriction enzymes of SspI and HindIII respectively. Electrophoresis was performed using horizontal gels, in electrophoretic cells (Bio-Rad, Richmond, U.S.A) for 90 minutes at 55 volts. Ethidium bromide was included in the gel at a final concentration of 0.12g/mL. After electrophoresis, DNA was visualized under UV-illumination and photographed using Polaroid type 57 film with a red filter.

RESULTS AND DISCUSSION Results The 223 bp fragment of locus 1 spanning intron IV and exon V of the growth hormone gene, amplified using primers GH-1 and GH-2, and the 329 bp locus 2 spanning exon III and exon IV of the growth hormone gene were amplified using primers GH5 and GH6. An example of gel photograph showing the polymorphisms of the amplified product cut by AluI restriction enzyme were shown in Figure 1.

PCR amplification of growth hormone gene Growth hormone locus 1 (GHL1), a 223 bp region spanning intron IV and exon V, and growth hormone locus 2 (GH-L2), a 329 bp region spanning exon III and exon IV of the growth hormone gene were amplified by PCR using primers GH1/GH2 and GH5/GH6 respectively. The primers used to amplify these fragments were: GH1: 5’-GCTGCTCCTGAGGGCCCTTCG-3’ GH2: 5’-GCGGCGGCACTTCATGACCCT-3’ GH5: 5’-CCCACGGGCAAGAATGAGGC-3’ GH6: 5’-TGAGGAACTGCAGGGGCCCA-3’ PCR amplification of D-loop and ND-5 of mtDNA The non-coding D-loop region and part of the gene coding for NADH dehydrogenase sub-unit 5 (ND-5) were amplified by PCR, using primers D-L/D-R and ND-L/NDR (Suzuki et al. 1993 cit. Sutarno, 2008):

100bp marker

600bp 223bp 171bp 52bp

100bp

Figure 1. Gel photographs showing growth hormone gene polymorphisms detected by PCR-RFLP using AluI in locus 1 fragment. Lane 1 = LV, Lane 2 = VV, Lane 3 = LL, and lane 4 = Uncut.

SUTARNO â&#x20AC;&#x201C; DNA polymorphism of Indonesian local cattle

The patterns of restriction enzymes digestion to those loci were described in Table 1 for AluI and MspI in the growth hormone loci, and the D-loop and ND-5 digestion were presented in Table 2. The frequencies of allele and genotype resulted based on the PCR-RFLP analysis at growth hormone loci by employing AluI and MspI restriction enzymes were shown in Table 3 and 4 subsequently. Table 1. Restriction sites for AluI in the 223bp locus 1 (GH L1) fragment, and for MspI in the 453 bp locus 2 (GH L2) fragment of the growth hormone gene. Enzyme Alu I Msp I

No of restriction sites 1 0 1 0

Allele L V Msp I + Msp I -

Fragment size (bp) 171, 52 223 224, 105 329

Table 2. Restriction sites for SspI in D-loop, and HindIII for ND5 regions of mitochondrial DNA. Enzymes D-loop SspI ND-5 HindIII

Alleles

Number of sites

Fragment size (kb)

A B

1 2

0.85, 0.29 0.66, 0.29, 0.15

A B

1 0

0.33, 0.13 0.46

Table 3. Alleles frequency of MspI (+), MspI (-), L, V from PO, Bali, Madura, and Pesisir cattle population Breeds

MspI (+)

MspI (-)

PO Bali Madura Pesisir

0.74 0.36 0.28 0.38

0.26 0.54 0.72 0.52

0.9 0.8 0.9 0.8

0.1 0.2 0.1 0.2

Table 4. Genotype frequency of PO, Bali, Madura, and Pesisir cattle population Breeds PO Bali Madura Pesisir

MspI (++) 0.50 0.53 0.40 0.47

MspI (--) 0.07 0.07 0.03 0.03

MspI (+-) 0.43 0.40 0.57 0.50

1.00 0.94 1.00 0.74

0 0 0 0.06

0 0.06 0 0.20

Discussion As demonstrated in this study, recent developments in molecular techniques have resulted in an abundance of data on genetic polymorphisms from DNA analysis. These data will provide us with a better understanding of the nature of genetic variation within and between cattle breeds. PCRRFLP analysis can detect the same type of polymorphisms as traditional RFLP analysis, but without the need for Southern blotting (Cushwa and Medrano 1996), thus decreasing the time taken and increasing sensitivity. This

method is therefore very useful for the study of genetic variation. Genetic variation within breeds is important and its study has become a subject of interest in livestock species, as it has many applications in animal breeding and genetics, such as the identification of animals and parentage testing, gene mapping and identifying markers for performance traits. Since all phenotypic characters are influenced by the genetic information carried by DNA, DNA variation may be correlated with variation in performance traits. This idea is the basis for marker assisted selection (MAS), which has aroused much interest in recent years (Soller 1994; Schwerin et al. 1995). Genetic variation, measured at the DNA level, can also be used as a check on the level of genetic variation in quantitative traits maintained within breeds. The genotype frequencies of MspI (++), MspI (+-) and MspI (--) were relatively similar in the four breeds of the Indonesian cattle. However, the genotype frequencies of LL, VV and LV were quite different between breeds. The genotype of VV was not detected in PO, Bali and Madura cattle breeds, and very rare (0.06) in Pesisir cattle breed. The genotype LV was also not detected in PO and Madura cattle breeds, and quite rare (between 0.06 and 0.2) found in Bali and Pesisir cattle breeds. It thus both VV and LV genotypes were quite rare in Indonesian native cattle breeds. Increasing the number of sample for analysis in each breed possibly could reveal or increase the rare frequency of both genotypes. The AluI and MspI restriction site polymorphisms in the locus 1 and locus 2 fragments of the growth hormone gene found in the study have also been previously reported in dairy cattle (Hoj et al. 1993a; Lucy et al. 1993), Bavarian Simmental (Schlee et al. 1994a), Indian cattle (Mitra et al. 1995) and Hereford and composite cattle (Sutarno 1998). Polymorphisms were found in the mitochondrial D-loop region with PstI and SspI, and in the mitochondrial ND-5 region using HindIII and SpeI. As previously reported by Sutarno and Lymbery (1997), the new polymorphisms in the D-loop by PCR-RFLP analysis using SspI and in the ND-5 region using SpeI were also found in the Indonesian local cattle. Compared to the standard bovine sequence of mitochondrial DNA (Anderson et al. 1982), these variant sequences have an extra AvaII and SspI site in the D-loop and have lost an SpeI site in mitochondrial ND-5. Sutarno and Lymbery (Sutarno and Lymbery 1997) suggested that the most likely scenarios for restriction site gains in the Dloop are a T to C transition at position 16273 to create a new AvaII site, and a C to T transition at position 245 to create a new SspI site. This was confirmed with the sequence data presented in by Sutarno (1998). For conservation purposes, study of the genetic variation between livestock breeds is very important. Adaptation to their different environmental challenges may have resulted in a unique combination of alleles specific to certain breeds, and this would be difficult to recreate (Hall and Bradley 1995). Breeds which are very different to others may need to be conserved, since the genes and gene combinations that they carry may be useful to agriculture in the future.

B I O D I V E R S IT A S

Bali cattle (Bos javanicus javanicus), more popularly called Banteng, have been domesticated largely in Indonesia, especially in Java, Bali, Sumbawa and Borneo. Cattle of this type superficially resemble Zebu, as they possess a hump, but the bone structure of the head is quite different. Copland (1996) suggested that Bali cattle are more similar to ancestral cattle than other modern types. According to the assessment done by AWCSG (Asian Wild Cattle Specialist Group) in 1995, Bos javanicus has been categorized as endangered, due to disease, hunting, hybridization or trade. Indeed, the introduction of modern cattle to Indonesia in the last few decades has partly caused a reduction in the diversity of Bali cattle. This is unfortunate because they are considered an original species with several economic advantages such as high fertility rate, adaptability and carcass percentage. Another important application of genetic variation between breeds is to predict the crosses between breeds that will produce crossbreed offspring with maximum heterosis. Much more attention has been paid in recent years to the utilization of heterosis in beef cattle and other livestock species. However, because there are so many breeds that could be used for crossbreeding, it is impossible to experimentally cross and compare all breeds.

CONCLUSION Based on these results, and previous studies, indicated that both of growth hormone gene and mitochondrial DNA are vary between breeds of Indonesian native cattle, and it is reasonable to presume that genetic variation in production traits is partly caused by variation in the genetic constitution of the genes such as growth hormone gene that coding for hormones and receptors in the growth hormone axis, and mitochondrial DNA that responsible for oxidative phosphorylation for energy production.

REFERENCES Anderson S, De Bruijn MHL, Coulson AR, Eperon IC, Sanger, F, Young, IG (1982) Complete sequence on bovine mitochondrial DNA: conserved features of the mammalian mitochondrial genome. J Mol Biol 156: 683-717. Beauchemin VR, Thomas MG, Franke DE, Silver GA (2006) Evaluation of DNA polymorphisms involving growth hormone relative to growth and carcaas characteristics in Brahman steers. Genet Mol Res 5 (3): 438-447. Beckmann JS, Kashi Y, Hallerman EM, Nave A, Soller M (1986) Restriction fragment length polymorphism among Israeli HolsteinFriesian dairy bulls. Anim Genet 17: 25-38. Copland JW (1996) Bali cattle: origins in Indonesia. ACIAR Proceedings 75: 29-33. Corden J, Wasylyk B, Buchwalder A, Sassone-Corsi P, Kedinger C, Chambon P (1980) Promoter sequences of eukaryotic protein-coding genes. Science 209: 1406-1414. Cowan CM, Dentine MR, Ax RL, Schuler LA (1989) Restriction fragment length polymorphisms associated with growth hormone and prolactin genes in Holstein bulls: Evidence for a novel growth hormone allele. Anim Genet 20: 25-38. Cushwa WT, Medrano JF (1996) Applications of the random amplified polymorphic DNA (RAPD) assay for genetic analysis of livestock species. Anim Biotechnol 7: 11-31.

11 (1): 1-4, January 2010 Gootwine E, Valinsky A, Shani M (1990) Restriction fragment length polymorphism at the growth hormone gene in sheep, goats and cattle. Proceedings of The 4th World Congress on Genetics Applied to Livestock Production XIII, Edinburgh 13: 83-85. Gordon DF, Quick DP, Erwin CR, Donelson JE, Maure RA (1983) Nucleotide sequence of the bovine growth hormone chromosomal gene. Mol Cell Endocrinol 33: 81-95. Hall SJG, Bradley DG (1995) Conserving livestock breed biodiversity [Review]. Trends Ecol Evolut 10: 267-270. Hallerman EM, Nave A, Kashi Y, Halzer Z, Soller M, Beckmann JS (1987) Restriction fragment length polymorphisms in dairy and beef cattle at the growth hormone and prolactin loci. Anim Genet 18: 213222. Harvey S (1995) Growth hormone synthesis. In: Harvey S, Scanes CG, Daughaday WH (ed.) Growth hormone. CRC Press, Boca Raton. Hediger R, Johnson SE, Barendse W, Drinkwater RD, Moore SS, Hetzel J (1990) Assignment of the growth hormone gene locus to 19q26-qter in cattle and to 11q25-qter in sheep by in situ hybridization. Genomics 8: 171-174. Hilbert P, Marcotte A, Schwers A, Hanset R, Vassart G, Georges M (1989) Analysis of genetic variation in the Belgian Blue cattle breed using DNA sequence polymorphism at the growth hormone low density lipoprotein receptor, a-subunit of glycoprotein hormones and thyroglobulin loci. Anim Genet 20: 383-394. Hoj S, Fredholm M, Larsen NJ, Nielsen VH (1993) Growth hormone gene polymorphism associated with selection for milk fat production in lines of cattle. Anim Genet 24: 91-96. Loftus RT, Machugh DE, Ngere LO, Balain DS, Badi AM, et al. (1994) Mitochondrial genetic variation in European, African and Indian cattle populations. Anim Genet 25: 265-271. Lucy MC, Hauser SD, Eppard PJ, Krivi GG, Clark JH, et al. (1993) Variants of somatotropin in cattle - gene frequencies in major dairy breeds and associated milk production. Domest Anim Endocrinol 10: 325-333. Mitra A, Schlee P, Balakrishnan CR, Pirchner F (1995) Polymorphisms at growth-hormone and prolactin loci in Indian cattle and buffalo. J Anim Breed Genet 112: 71-74. Nielsen VH, Larsen NJ (1991) Restriction fragment length polymorphisms at the growth hormone gene in pigs. Animal genetics 22 (3): 291-294. Schlee P, Graml R, Rottmann O, Pirchner F (1994a) Influence of growth hormone genotypes on breeding values of simmental bulls. J Anim Breed Genet 111: 253-256. Schlee P, Graml R, Schallenberger E, Schams D, Rottmann O, et al. (1994b) Growth hormone and insulin like growth factor I concentrations in bulls of various growth hormone genotypes. Theor Appl Genet 88: 497-500. Schwerin M, Brockmann G, Vanselow J, Seyfert HM (1995) Perspectives of molecular genome analysis in livestock improvement. Arch Anim Breed 38: 21-31. Shadel GS, Clayton DA (1997) Mitochondrial DNA maintenance in vertebrates. Annu Rev Biochem 66: 409-435 Shoffner JM, Wallace DC (1990) Oxidative phosphorylation diseases: disorders of two genomes. Adv Hum Genet 19: 267-380. Soller M (1994) Marker assisted selection - an overview. Anim Biotechnol 5: 193-207. Sutarno (1998) Candidate gene marker for production traits in beef cattle. In: Vet Biology. Murdoch University, Perth. Sutarno, Lymbery AJ (1997) New RFLPs in the mitochondrial genome of cattle. Anim Genet 28: 240-241. Suzuki R, Kemp SJ, Teale AJ (1993) Polymerase chain reaction analysis of mitochondrial DNA polymorphism in Ndama and Zebu cattle. Anim Genet 24: 339-343. Thomas MG, Silver GA, Enns RM (2006) Relationships of DNA polymorphisms in growth hormone (GH) to growth and carcass traits of Brangus. Bull Int Plant Anim Genome 14: P526. Wallace DC (1993) Mitochondrial diseases: genotype versus phenotype. Trends Genet 9: 128-133. Welter C, Dooley S, Blin N (l989) A rapid protocol for the purification of mitochondrial DNA suitable for studying restriction fragment length polymorphism (RFLP). Gene 83: 169-172. Woychik RP, Camper SA, Lyons RH, Horowitz S, Goodwin EC, Rottman FM (1982) Cloning and nucleotide sequencing of the bovine growth hormone gene. Nucleic Acids Res 10: 7197-7210.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 5-8

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110102

The genetics of glutamate oxaloacetate transaminase (GOT) in Pinus merkusii Jungh. et de Vriese ISKANDAR ZULKARNAEN SIREGARâ&#x2122;Ľ, TEDI YUNANTO Department of Silviculture, Faculty of Forestry, Bogor Agricultural University, Kampus IPB Darmaga, P.O. Box 168, Bogor 16680, West Java, Indonesia, Tel. +62-251-8626806, Fax. +62-251-8626886, ď&#x201A;Šemail: siregar@ipb.ac.id Manuscript received: 10 November 2009. Revision accepted: 29 December 2009.

ABSTRACT Siregar IZ, Yunanto T (2010) The genetics of glutamate oxaloacetate transaminase (GOT) in Pinus merkusii Jungh. et de Vriese. Biodiversitas 11: 5-8. Inheritance and linkage analysis of glutamate oxalacetate transaminase (GOT) in P. merkusii were performed using megagametophyte tissues of single tree seed-lots. One monomorphic locus (GOT-A) and three polymorphic loci (GOT-B, GOT-C and GOT-D) were identified, based on the allelic 1: 1 segregation of putatively heterozygous trees. Linkage analysis revealed that GOTC and GOT-D were closely linked loci with an average recombination rate (R) of about 5%. This result which is in contrast to the GOT genetics in other pine species was discussed with regard to the probably occurring gene duplication of one GOT locus. Key words: Pinus merkusii, GOT, isozymes, inheritance and linkage.

INTRODUCTION The glutamate oxaloacetate transaminase (GOT) which is identical to aspartate amino transferase (AAT, E.C. 2.6.1.1) is one of the enzyme systems often used in isozyme studies on plant and animal genetics. It catalyzes the reversible reaction of glutamate and oxaloacetate to 2oxoglutarate and aspartate, thus having an important role in nitrogen metabolism by distributing this nutrient originally assimilated into glutamate to other compounds (Ireland and Joy 1985). In surveys of higher plant isozymes, three to four electrophoretic GOT zones have been reported (for review, see Gottlieb 1982). In maize, three different isozymes associated with the mitochondria (mGOT), with the glyoxysomes (gGOT), and with the soluble fraction of the cytosol (sGOT) were described, however, in later studies it was established that GOT occurs also in plastids (Weeden and Gottlieb 1980). In genetic studies on forest tree species, GOT has frequently been assayed to determine genetic diversity and differentiation (Aguirre-Planter et al 2000; Ledig 2000; Ledig et al. 2001; Shea et al. 2002), since its controlling gene loci were found to be largely polymorphic. In conifer, GOT is generally encoded by three gene loci, one of which (GOT-C or GOT-3) specifies an isozyme consisting of double- or triple-banded variants near the cathode in zymograms. If the separation buffer in electrophoretic analysis has a pH value of 8.0 or 8.1, one or two bands of GOT-C migrate to the anode and one band migrates to the cathode, but all bands of this isozyme reveal a comigration, so that they were believed to be encoded by the same gene locus. In pine species, GOT has also been found to be encoded by three gene loci, i.e. GOT-A, GOT-B and GOTC (Mejnartowicz and Bergmann 1985). However, Chung

(1981) reported a fourth locus of GOT in P. sylvestris and classified it as GOT-D, although this additional isozyme may correspond to one of the above-mentioned subbands comigrating with bands of GOT-C (Mejnartowicz and Bergmann 1985). Similarly, four isozyme zones were found in seed samples of Pinus merkusii from Thailand, however, one of these zones near the origin was invariant and was assigned to GOT-C which controls another variable zone (Changtragoon and Finkeldey 1995). However, new data with P. merkusii single tree seed-lots from Indonesia revealed the existence of two separate gene loci, i.e. GOT-C and GOT-D, controlling these multiband configurations in GOT zymograms (Siregar and Yunanto 2008). The studies on inheritance and linkage were carried previously with objective to confirm the number of GOT loci in P. merkusii. Output of the studies would be of significant importance in future biodiversity assessment, i.e. genetic diversity, of the genus Pinus.

MATERIALS AND METHODS Seeds Bulk seed-lots of P. merkusii from Aceh and Java (Indonesia) were primarily used to optimise the experimental methods for GOT resolution. In the second step haploid megagametophytes (endosperms) of 18 single tree seed-lots were analysed according to the procedure largely used by Changtragoon and Finkeldey (1995). Seeds were immersed overnight in water, dissected and the embryo carefully separated from the megagametophytes. The megagametophytes were ground in two drops of homogenising buffer (0.1 m Tris-HCl pH 7.3, containing 0.03% DTT and 2.5 % PVP).

B I O D I V E R S IT A S

11 (1): 5-8, January 2010

Electrophoresis These crude homogenates were then subjected to horizontal starch gel electrophoresis, using the buffer system of Ashton and Bradon, pH 8.6. Further details of electrophoretic and staining procedures were given by Liengsiri et al. (1990). Segregation analysis Inheritance of isozyme variants was confirmed through analysis of segregation ratios of haploid megagametophytes from putatively heterozygous mother trees. A test for goodness of fit to the 1: 1 ratio, chisquare test (2), was performed separately for each parent tree. In this analysis, all statistical tests were set up at 5% level of significance.

Zone A + Zone B Zone C

ORIGIN

Zone D B3 B3 B3 B2 B2 B2 C2 C2 C2 C2 C2 C2 D1 D1 D2 D1 D2 D1

B2 B3 B2 B2 B3 B3 C2 C2 C2 C1 C2 C2 D1 D1 D1 D1 D1 D1

Linkage analysis Chi-square tests (2 test) of linkage were performed for each pair of segregating loci (Weir 1990). Figure 1. The GOT banding patterns of megagametophytes (1n) from two trees. The Two chi-square tests (2) were indication of independent GOT-C and GOT-D zones is shown on the left zymogram. calculated: the first one tests Mendelian segregation of alleles at each individual locus in the pair, and Four allozymes were identified in the GOT-C zone. The the last one tests for linkage between loci. A linkage same number was also observed by Na’iem and Indrioko analysis was performed for each individual parent tree (1996). The segregation at GOT-C was regular for several when its progeny was found to be heterogenous at a given conifers species (Sousa et al. 2001). Nine out of twelve pair of loci. When linkage between loci was detected, the seed lots tested for segregation showed Mendelian 1: 1 estimated frequency of recombination (r) was calculated ratio, confirming that GOT-C is controlled by a single gene using the formula r = l/n, where l is the number of locus with four alleles (Table 1). Two heterozygous trees recombinants and n is the total number of (J54 and J47) with genotype C C and C C respectively, 2 3 2 4, megagametophytes. indicated a very strong segregation distortion. RESULTS AND DISCUSSION The phenotypic variation and inheritance of GOT After electrophoretic separation of extracts from seed megagametophytes, the gels stained for GOT showed four zones where one zone migrated towards the cathode. Figure 1 shows the four GOT zones in zymograms as well as variations in zone B and zone D. Contrary to this study, Zhelev et al. (2002) reported a monomorphic GOT enzyme system on Pinus peuce. Estimates of the statistics used to test the conformity to the Mendelian mode of inheritance for each polymorphic locus of GOT zone are presented in Table 1. The zone GOT-A was invariant in the material used and supposed to be controlled by a single gene locus. Contrary to P. merkusii in Thailand, three allozymes were observed in the GOT-B zone and genetic analysis conducted with 10 single tree seed-lots showed that this zone is controlled by one single gene locus (Table 1).

The GOT-D zone was highly polymorphic, possessing two allozymes and genetic analysis conducted with 18 single tree seed-lots showed that this zone is controlled by a separate gene locus. Two seed lots showed segregation distortion and the allele D2 seemed to be in favour. Linkage relationships As was shown in Figure 1, the zones GOT-C and GOTD in zymograms appear to be independent isozymes which are encoded by two separate gene loci, however, these loci may be linked. In the following analysis, linkage among the three variable GOT loci will be assessed. Three pairs of heterozygous GOT loci were tested in different tree samples and the results of linkage analysis and statistical tests are presented in Table 2. According to the observed recombination values for significantly linked loci, only one linkage group can be constructed. GOT-C/GOT-D was closely linked with an average recombination value of 0.046. Other possible linkages were GOT-B/GOT-D (R=0.167) based on only one significant case of linkage, however, the allelic distribution in the other three samples

SIREGAR & YUNANTO – The genetics of GOT in Pinus merkusii

also tends to some loose linkage. Significant linkage in one but not all trees has also been reported for other conifer species (Fallour et al. 2001; Pastorino and Gallo 2001). Table 1. Observed segregation of allozymes from megagametophytes trees of P. merkusii at gene loci of GOT and goodness of fit to the 1: 1 expected ratio. Tree

Genotype

Sample size (Nij)

Observed segregation (Ni) (Nj)

2test

GOT-B J04 J09 J14 J29 J33 J34 J37 J54 AI40 A04 Pooled

B2B3 B2B3 B2B3 B2B3 B2B3 B2B3 B2B3 B2B3 B2B3 B2B3 B2B3

12 12 15 22 16 29 16 43 18 20 203

7 6 10 13 4 15 9 24 11 8 107

5 6 5 9 12 14 7 19 7 12 96

0.333 0.000 1.667 0.727 4.000* 0.034 0.250 0.581 0.889 0.800 0,596

GOT-C J01 J02 J36 Pooled

C1C2 C1C2 C1C2 C1C2

27 34 40 101

12 21 25 58

15 13 15 43

0.333 1.882 2.500 4.715

J16 J26 J54 Pooled

C2C3 C2C3 C2C3 C2C3

36 62 43 141

16 39 39 94

20 23 4 47

0.444 4.129* 28.488*** 282***

J27 J38 J47 J48 J52 AI41 Pooled

C2C4 C2C4 C2C4 C2C4 C2C4 C2C4 C1C2

57 16 36 20 43 25 197

31 9 26 12 18 12 108

26 7 10 8 25 13 89

0.439 0.250 7.111** 0.800 1.140 0.040 1.980

GOT-D J01 D1D2 27 16 11 0.926 J02 D1D2 34 14 20 1.059 J04 D1D2 12 7 5 0.333 J06 D1D2 12 6 6 0.000 J09 D1D2 12 6 6 0.000 J11 D1D2 22 9 13 0.727 J12 D1D2 39 6 33 18.692*** J27 D1D2 57 48 9 26.684*** J29 D1D2 21 10 11 0.048 J31 D1D2 22 12 10 0.182 J34 D1D2 29 14 15 0.034 J36 D1D2 40 14 26 3.600 J47 D1D2 36 14 22 1.778 J51 D1D2 24 10 14 0.667 J52 D1D2 40 20 20 0.000 J54 D1D2 43 20 23 0.209 AI41 D1D2 25 13 12 0.040 A39 D1D2 17 11 6 1.471 Pooled D1D2 512 250 262 0.281 Note: Significant levels are 0.05 (*), 0.01 (**), 0.001 (***) Table 2. Pairs of loci for which significant levels of linkage were detected in at least one tree.

Tree no.

Observed number by allelic combination AiBi AiBj AjBi AjBj

7 Recombination Fraction

2 Test (df=1) locus A

locus B

Linkage

GOT-B/GOT-C J54 20 4 19 0 0.581 28.488*** 0.209 GOT-B/GOT-D J54 14 10 6 13 0.581 0.209 2.814 J04 6 1 1 4 0.333 0.333 5.333* J12 8 7 6 8 0.034 0.034 0.310 J22 14 10 6 13 0.581 0.209 2.814 GOT-C/GOT-D AI41 2 10 11 2 0.040 0.040 11.560*** J01 0 13 12 0 0.040 0.040 25.000*** J02 0 11 7 0 0.889 0.889 18.000*** J36 0 25 14 1 2.500 3.600 36.100*** Note: Significant levels are 0.05(*), 0.001(***)

(R) 0.535 0.372 0.167 0.448 0.372 0.160 0.000 0.000 0.025

Discussion The genetic analysis conducted in this study has been based on megagametophytes of seeds from putatively heterozygous trees, since progenies from controlled crosses were not available. In coniferous tree species, isozyme analysis of haploid megagametophytes, which genetically represent the female gametes, allows observation of regular meiotic segregation at individual loci (Bergmann 1973). This method therefore allows for both, the identification of the number of loci controlling an enzyme system and the independent transmission or linkage of alleles among these different loci. The study on the genetic control of observed isozyme phenotypes in P. merkusii was mainly carried out to confirm the results of previous works (Changtragoon and Finkeldey 1995; Szmidt et al. 1996). However, in contrast to these studies it was noticed that the enzyme system GOT turned out to be controlled by four gene loci. This result is surprising because it is much in contrast to all other pine species where only three GOT loci were identified. Furthermore, it was established that the allelic segregation at two loci did not occur independently. However, Mejnartowicz and Bergmann (2002) and Lewandowski et al. (2002) reported that four AAT or GOT activity zones were found in Abies alba and Picea abies, respectively. The close linkage between the loci GOT-C and GOT-D has not been reported previously because the two loci are often regarded as one locus. In many conifers, the GOT-C and GOT-D are actually two widely separated, always comigrating bands and one of these bands is often moving towards the cathode. This band configuration has led to the one-locus hypothesis. This interpretation may be correct if we assume a gene duplication which has occurred just very recently. In this case, recombinants between adjacent gene loci can only be detected among thousands of seed megagametophytes, which were not included in the former inheritance studies. Later on, the distance between the duplicated GOT loci may have become larger (e.g. by some insertions) and, hence, recombinants between the two loci can be detected with smaller seed samples, as is the case with P. merkusii. The various recombination frequencies between GOT-C and GOT-D observed in four trees, i.e.

B I O D I V E R S IT A S

from 0% to 16%, might be attributed to the genotype at the GOT-C locus. The highest recombination rate (R=16%) was observed in tree AI41 which is C2C4, while the other three trees were C1C2. Indication of possible influence of temperature during meiosis has been mentioned by Rudin and Ekberg (1978) to explain the different levels of recombination frequency observed among trees of P. sylvestris. The later reason should be excluded from our results because of the absence of such high temperature fluctuation in the tropics. Additionally, it must be emphasized that the linkage analysis was only based on female gametes. In spite of the general observation that gamete selection is less pronounced on the female than on the male side in flowering plants (Rudin and Ekberg 1978), female gamete selection may have influenced recombination data. In P. radiata, it was found that the rate of recombination is 43% greater in male gametes than in female gametes. In addition, significant linkage in one but not in all trees has also been reported for other conifer species. The possible reason for different recombination values among trees is related to environmental effects or modification of chromosome structure, i.e. insertion, translocation, deletion, reversion, etc. A high frequency of events leading to such meiotic irregularities has been observed in conifers (Andersson et al. 1969). Furthermore, the history of colonization of a species might contribute also to slightly different chromosomal arrangement resulting in a deviating linkage pattern as shown in P. sylvestris from southern and northern Sweden (Rudin and Ekberg 1978). The gene duplication followed by mutation is a major mechanism of evolution. Duplication results in two identical copies, each of which can retain original function, becomes functionless (formation of pseudo-gene) or develops a new function. The GOT loci has often been reported to be tightly linked with PGI loci in pines and other coniferous species and this linkage has become an indication of a striking conservatism of chromosomal structure in this species (Ledig 1998). Using P. merkusii samples from Thailand and Vietnam, Szmidt et al. (1996) have found polymorphism in PGI and this opens up the possibility for future work to confirm the finding of such conservatism of linkage groups.

CONCLUSION In P. merkusii, GOT was identified to have one monomorphic locus (GOT-A) and three polymorphic loci (GOT-B, GOT-C and GOT-D). Linkage analysis confirmed that GOT-C and GOT-D were closely linked loci with an average recombination rate (R) of about 5%. ACKNOWLEDGEMENT The authors would like to thank Perum Perhutani for providing sample materials.

11 (1): 5-8, January 2010

REFERENCES Aguirre-Planter E, Furnier GR, Eguiarte LE (2000) Low levels of genetic variation within and high levels of genetic differentiation among populations of species of Abies from southern Mexico and Guatemala. Am J Bot 87: 362-371. Andersson E, Ekberg I, Eriksson G (1969) A summary of meiotic investigations in conifers. Stud For Suec 70: 1-20 Bergmann F (1973) Genetische untersuchungen bei Picea abies mit hilfe der isoenzym-identifizierung. II. Genetische kontrolle von esteraseund leucinamino-peptidase-isoenzymen im haploiden endosperm ruhender samen. Theor Appl Genet 43: 222-225. Changtragoon S, Finkeldey R (1995) Inheritance of isozyme phenotypes of Pinus merkusii. J Trop For Sci 8 (2): 167-177. Chung MS (1981) Biochemical methods for determining population structure in Pinus sylvestris L. Act For Fenn 173: 28. Fallour D, Fady B, Lefevre F (2001) Evidence of variation in segregation patterns within a Cedrus population. J Hered 92: 260-266 Gottlieb LD (1982) Conservation and duplication of isozymes in plants. Science 216: 373-380. Ireland RJ, Joy KW (1985) Plant transaminases. In Christen P, Metzler DE (eds). Transaminases. John Wiley & Son, New York. Ledig FT (1998) Genetic Variation in Pinus. In Richardson DM (ed) Ecology and biogeography of Pinus. Cambridge University Press, Cambridge, UK. Ledig FT (2000) Founder Effect and the Genetic Structure of Coulter Pine. American Genetic Association 91: 307-315. Ledig FT, Capó-Arteaga MA, Hodgskiss PD, Sbay H, Flores-López C, Conkle MT, Bermejo-Velázquez B (2001) Genetic diversity and the mating system of a rare Mexican piñon, Pinus pinceana, and a comparison with Pinus maximartinezii (Pinaceae). Am J Bot 88: 1977-1987. Lewandowski A, Samocko J, Burczyk J (2002) Inheritance of AAT in Picea abies – some old and new facts. Silvae Genetica 51: 161-163 Liengsiri C, Piewluang C, Boyle TJB (1990) Starch gel electrophoresis of tropical trees. A Manual. ASEAN-Canada Forest Tree Seed Centre and Petawawa National Forestry Institute, Muak Lek, Saraburi, Thailand. Mejnartowicz L, Bergmann F (1985) Genetic differentiation among Scots pine populations from the lowlands and the mountains in Poland. In: Gregorius, H.-R. (ed.) Population genetics in forestry. Springer, Berlin. Mejnartowicz L, Bergmann F (2002) Mode of Inheritance of Aspartate Aminotransferase in Silver-Fir (Abies alba Mill.). Silvae Genetica 52: 15-17 Na’iem M, Indrioko S (1996) Inheritance of isozyme variants of Tusam (Pinus merkusii) artificial stands in Java. In: Rimbawanto A, Widyatmoko AYPBC, Shuaendi H, Furukoshi T (eds). Proceeding of International Seminar on Tropical Plantation Establishment: improving productivity through genetic practices. FTIRDI and JICA Yogyakarta. Pastorino MJ, Gallo LA (2001) Linkage relationships as a useful tool to state interspecific gene homology: case study with isozyme loci in Austrocedrus chilensis (Cupressaceae). Silvae Genetica 50: 233-239. Rudin D, Ekberg I (1978) Linkage studies in Pinus sylvestris L. using macro-gametophyte allozymes. Silvae Genetica 27: 1-12. Shea KL, Furnier GR (2002) Genetic variation and population structure in central and isolated populations of balsam fir, Abies balsamea (Pinaceae). Am J Bot 89: 783-791. Siregar IZ, Yunanto T (2008) Inference on the possible causes of segregation distortion from open pollination progenies of merkus pine (Pinus merkusii). Hayati J Biosci 2008 15: 155-158. Sousa VA, Hattemer HH, Robinson IP (2001) Inheritance and linkage relationships of isozyme variants of Araucaria angustifolia (BERT.) O. KTZE. Silvae Genetica 51: 5–6. Szmidt AE, Wang XR, Changtragoon S (1996) Contrasting patterns of genetic diversity in two tropical pines: Pinus kesiya (Royle ex Gordon) and P. merkusii (Jungh. et De Vriese). Theor Appl Genet 92: 436-441. Weeden NF, Gottlieb LD (1980) The genetics of chloroplast enzymes. J Hered 71: 392-396. Weir BS (1990) Genetic data analysis. Sinauer, Sunderland, M.A. Zhelev P, Gomory D, Paule L (2002) Inheritance and linkage of allozymes in a Balkan endemic Pinus peuce Griseb. J Hered 93: 60-63.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 9-14

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110103

Systematic studies of Australian stipoid grasses (Austrostipa) based on micro-morphological and molecular characteristics BETTY MAULIYA BUSTAMâ&#x2122;Ľ Biology Department, Faculty of Mathematics and Natural Sciences, Syiah Kuala University, Jl. Syech Abdur Rauf 3, Darussalam, Banda Aceh 23111, Aceh, Indonesia, Tel./fax. +62-651-7428212, +62-651-7410248, ď&#x201A;Šemail for correspondence: liya1304@yahoo.com. Manuscript received: 25 October 2009. Revision accepted: 26 December 2009.

ABSTRACT Bustam BM (2010) Systematic studies of Australian stipoid grasses (Austrostipa) based on micro-morphological and molecular characteristics. Biodiversitas 11: 9-14. This research is one of many studies on stipoid grasses organized by the International Stipeae Working Group (ISWG). This research tested the subgeneric classification of Austrostipa proposed by Jacobs and Everett (1996) and tested how informative the micro morphological characters used. Data were collected from herbarium specimens of 36 species (33 species of Austrostipa, two species of Hesperostipa and one species of Anemanthele) at Royal Botanic Gardens, Sydney. Twenty eight micro morphological characters were used. The data were collected from both adaxial and abaxial surfaces of leaves, and from the lemma epidermis using a scanning electron microscope (SEM). ISWG provided the molecular data. Parsimony analysis and a distance method (Unweighteic Pair Group with Arithmatic Mean: UPGMA) were used to analyze micro-morphological and molecular data separately. Only UPGMA analysis was used to analyze the combined data. The results support the monophyly of Austrostipa. However, there is a little support for the subgeneric classification of Austrostipa proposed by Jacobs and Everett (1996), other than for the consistent recognition of Falcatae. The characters for comparisons between genera are too homoplasious at this level and do not contain enough information for analyses at subgeneric level, a problem apparently shared with the DNA sequences. Key words: Austrostipa, stipoid grasses, micro morphological, molecular

INTRODUCTION The tribe Stipeae was first formulated by Dumortier in 1823, based on the genus Stipa L. English names applied to the tribe include Speargrass, Feathergrass and Needlegrass (Townrow 1978). This is a cosmopolitan tribe comprising approximately 500 species (Barkworth 1993). These grasses grow in temperate Australia, North and South America, Europe and Central Asia (Barkworth and Everett 1986; Hsiao et al. 1999). This tribe has been variously placed. The treatment most widely accepted at present is to regard the Stipeae as a tribe of the subfamily Pooideae (Hsiao et al. 1999; Jacobs et al. 2000; GPWG 2000; Wheeler et al. 2002). At present, the relationships within the stipoid grasses are poorly understood, with different data sets suggesting different relationships (Ariaga and Barkworth 2000; Cialdella and Giussani 2002; Connor and Edgar 2002; Maze et al. 2002; Vasques and Barkworth 2004; Ariaga and Barkworth 2006; Ariaga and Jacobs 2006; Barkworth et al. 2008). Understanding the relationships of these grasses allows more effective and efficient resource management (Garden et al. 2000; Clarke 2003; Landberg et al. 2003; de Lange et al. 2004; Huxtable et al. 2005). Bentham (1878) was the first person to provide treatment of Australian species of Stipa. After that, some studies have been conducted to get better understanding of those Australian species of Stipa (Hughes 1921, 1922; Everett and Jacobs 1983; Barkworth and Everett 1987; Vickery et al. 1986; Everett 1990; Jacobs and Everett

1996). Based on those studies and the fact that Australian species are more closely related to each other than to any non-Australian species, Jacobs and Everett (1996) decided the best option was to place all the Australian species formerly included in Stipa in a new genus, Austrostipa. However, the relationships among subgenera in Austrostipa still need to be tested. Jacobs et al. (2000) have DNA sequences for several species. While these DNA sequences strongly supported some groupings or sub genera, other groupings were either poorly supported or not supported at all. The most reliable way of testing any data set would be to compile other data sets based on different characters and look for corroboration. In an attempt to find the relationships of the subgenera within genus Austrostipa, it was decided to compile micro morphological data sets for comparison with DNA sequences. The objectives of this study were: (i) testing whether the subgenera in Austrostipa are natural or monophyletic groups, (ii) how informative the micro morphological characters are. This study was conducted at Royal Botanic Gardens, Sydney, Australia, from August 2002 to March 2006.

MATERIALS AND METHODS All micro morphological data were collected from herbarium specimens of 36 species (33 species of Austrostipa, two species of Hesperostipa and one species

B I O D I V E R S IT A S

of Anemanthele). Data were collected from leaves and lemma. Leaves are elongated structures made up of a basal cylindrical sheath and an upper blade or lamina. For this study, only the blade (lamina) of the leaf was used. The lamina was cut approximately 4 cm from the blade/sheath junction and, for both abaxial and adaxial surfaces; a segment of approximately 3 mm of both abaxial and adaxial surfaces was taken for examination. The lemma is the outer bract subtending the floret and is on the side of the spikelet axis away from the main inflorescence axis (Wheeler et al. 2002). For every species, approximately six lemmas were taken, particularly those that were mature and loose from the glumes. Micro morphological data were collected using a Cambridge S360 Scanning Electron Microscope (SEM). Theoretically, all specimens were examined with the same magnification. However, due to different structures of each specimen, the magnification is changed to suit the specimens. All micro morphological characters were scored. In determining the scoring characters, the suggestions of Ellis (1979) were followed, except for the stomata and subsidiary cell size characters, where the scoring was based on the result of preliminary analyses. The parameters of micro morphological data were: A. Leaf epidermis characters (adaxial and abaxial surfaces) 1. Difference over (costal) and between veins (intercostal): 0 - absent (the costal and intercostals areas are indistinguishable); 1 - present (the costal and intercostals areas are distinguishable) 2. Silica bodies presence: 0 - absent ; 1 - present 3. Longitudinal cell wall: 0 - smooth; 1 - sinuous 4. Stomata abundance: 0 - few stomata = < 10 stomata per unit area exposed on the SEM (magnification of 200x); 1 - sinuous = ≥ 10 stomata per unit area exposed on the SEM (magnification of 200x). 5. Stomata size, this character measured the length of guard cell approximately (μm): 0 - (≤ 20); 1- (>20 23); 2 - (>23 - 26); 3 - (>26 - 29); 4 - (>29 - 32); 5 (>32 - 35); 6 - (>35). 6. Stomata subsidiary cell size: The scoring is the same as in stomata size (μm) 7. Stomatal shape: 0 - dome-shaped; 1 - parallel sided 8. Macrohairs presence: 0 - absent; 1 - present 9. Macrohairs position: - missing (inapplicable); 0 both in costal and intercostals areas; 1 - in costal areas only; 2 - in intercostal areas only 10. Prickles presence: 0 - absent; 1 - present 11. Prickles position: - missing (inapplicable); 0 - both in costal and intercostals areas; 1 - in costal areas only; 2 - in intercostal areas only B. Lemma characters of fundamental cells 12. Length of fundamental cells compared to short cells: - inapplicable; 0 - fundamental cells longer than short cells; 2 - fundamental cells shorther than short cells 13. Sidewall shape: 0 - straight; 1 - wavy; 2 - dentate 14. Sidewall thickness: 1 - not conspicuously thickened; 2 - conspicuously thickened 15. Endwall shape: 1 - straight; 2 - wavy 16. Short cells hooks presence: 0 - absent; 1 - present 17. Lemma silica bodies: 0 - absent; 1 - present

11 (1): 9-14, January 2010

There were 28 micro morphological characters used in this research, 22 characters from leaves (eleven characters for abaxial and eleven characters for adaxial) and six characters for lemmas. Sequences for the ITS region (molecular data) were obtained for the same taxa. The ITS sequences were done by Dr. Randall Bayer, a member of International Stipeae Working Group (ISWG) from Australia, and Dr. Catherine Hsaio from USDA, Longan. All data, micro morphological and molecular were entered into a computer programmed MacClade 4.05 (Madison and Madison 2002) and were then analyzed with PAUP (Phylogenetic Analysis Using Parsimony) version 4.0b10 (Swofford 2002). Parsimony analysis and a distance method (Unweighted Pair Group with Arithmetic Mean: UPGMA) were used to analyze micro morphological and molecular data separately. However only UPGMA analysis was used to analyze the combined data, since preliminary analyses of separated data retrieved some similar groups.

RESULTS AND DISCUSSION Micro morphological analyses There are several groups resolved from the micro morphological analyses in both parsimony and UPGMA. However, most of the groups have no bootstrap support and have no support from the molecular analyses. In addition, most of the groups are not the same as groups in recent classification of Austrostipa by Jacobs and Everett (1996). For example, the group consisting of three species: Austrostipa acrociliata, A. bigeniculata and A. aristiglumis retrieved in the parsimony analysis of micro morphological data (Figure 1), were placed in two groups in the recent classification (Jacobs and Everett 1996). A. acrociliata is in subgenus Arbuscula whereas A. bigeniculata and A. aristiglumis are in subgenus Ceres. However, the analysis placed A. acrociliata together with A. bigeniculata rather than A. bigeniculata with A. aristiglumis. Morphologically, there are many similarities between A. bigeniculata and A. aristiglumis. For instance, both species have open and spreading panicles and nonbranching culms. Both A. bigeniculata and A. aristiglumis have a lemma with a coma, non-tuberculate surface and a short strongly-angled callus while A. acrociliata does not have a coma and the lemma has a tuberculate surface and a blunt callus. There are two groups retrieved from the micro morphological analyses that are consistent with the subgenera in the recent classification by Jacobs and Everett (1996). The first group is retrieved from the parsimony analysis (Figure 1) and consists of two species: A. nitida and A. nodosa. This group is also retrieved in the UPGMA analysis of micro morphological data (Figure 3), with the addition of one species, A. scabra subsp. falcata. All three species: A. nitida, A. nodosa, A. scabra subsp. falcata are placed in subgenus Falcatae in Jacobs and Everett (1996). The other group that is retrieved in the UPGMA analysis of micro morphological data (Figure 3) consists of two species: A. geoffreyi and A. juncifolia, which placed in subgenus Lobatae (Jacobs and Everett 1996). While there

BUSTAM â&#x20AC;&#x201C; Systematic studies of Australian stipoid grasses (Austrostipa)

Figure 2

Figure 3

Figure 4

Figure 5

Figure 1. Parsimony analysis of mico morphological data. A is the outgroup. Strict consensus of 25,698 equally parsimonious tress (140 steps, CI = 0.32, RI = 0.57, RC = 0.19). Bootstrap percentage values (1000 replicates) are shown above the branches for all values > 50. Figure 2. Parsimony analysis of the molecular data. A is the outgroup. Strict consensus of 2, 808 equally parsimonious tress (292 steps, CI = 0.49, RI = 0.61, RC = 0.3). Thicker lines indicate the group was also identified in the UPGMA analyses. Groups in bold case indicate the groups identified in the UPGMA analysis of the molecular data. Bootstrap percentage values (1000 replicates) are shown above the branches for all values > 50.

BUSTAM â&#x20AC;&#x201C; Systematic studies of Australian stipoid grasses (Austrostipa)

Figure 1

Figure 3. UPGMA analysis of the micro morphological data. Thicker lines indicate the group also was identified in parsimony analysis of the molecular data and other UPGMA analyses. Groups in bold case indicate the groups also identified in the UPGMA analysis of combined data. Figure 4. UPGMA analysis of the molecular data. Thicker lines indicate the group also was identified in other analyses. Bold case indicates the groups identified in the parsimony analysis of the molecular data. Figure 5. UPGMA analysis of the combined data. Thicker lines indicate the group also was identified in parsimony analysis of the molecular data and other UPGMA analyses. Bold case indicates the groups identified in UPGMA analyses of both the micro morphological and molecular data.

B I O D I V E R S IT A S

is a little support for the subgeneric classification of Austrostipa proposed by Jacobs and Everett (1996) by the micro morphological analyses, the monophyly of Austrostipa received strong support (91%) from the parsimony analysis (Figure 1). Molecular analyses Unlike micro morphological analyses, most groups retrieved in the parsimony analysis of molecular data were also retrieved in the UPGMA analysis. Except for the outgroups, there are five groups that are resolved in the parsimony analysis and four groups in the UPGMA analysis (Figure 2 and Figure 4.). Moreover, most groups retrieved in the parsimony analysis of molecular data have quite strong bootstrap support (>70%). The first group retrieved in the parsimony analysis consists of four species: A. drummondii, A. nitida, A. nodosa and A. scabra subsp. falcata. All four species in the group are placed in the subgenus Falcatae by Jacobs and Everett (1996). This group received 92% bootstrap support and also is supported by UPGMA analyses of the micro morphological and molecular data (Figure 2, Figure 3 and Figure 4). In the UPGMA analysis of the molecular data, the group contains four species, while in the UPGMA analysis of the micro morphological and molecular data the group contains three species, without A. drummondii (Figure 3 and Figure 4). In addition, in the UPGMA analysis of the molecular data, A. nitida placed together with A. nodosa, as in the UPGMA analysis of the micro morphological data, while in the parsimony analysis, A. nitida placed with A. drummondii and this clade receives weak bootstrap support (53%). Morphologically, all Falcatae species are caespitose (growing in tufts), have culms that only branch in the inflorescence and, in addition, all have a falcate bristle on the awn, and hairy lemmas (Jacobs and Everett 1996). The spikelets are all so similar that vegetative characters often play an important role in distinguishing taxa in the Falcatae. It is often very difficult to tell A. nitida and A. nodosa apart. In spite the similarity, there are some differences among species. Austrostipa scabra is characterized by having very fine narrow leaves, much finer than the others in the subgenus. A. drummondii has hairy awns and leaves. A. nitida and A. nodosa are very similar, differing only in minor characteristics of the inflorescence but also in the basal formation of new culms, A. nitida being intravaginal and A. nodosa being extravaginal. The species also tend to grow in different habitats. A. nitida grows particularly on (mostly red) sandy soils in all mainland States, while A. nodosa grows on heavier soils than A. nitida in all regions in Australia except the North Coast and South Coast of New South Wales, Victoria, South Australia and Western Australia, often associated with mallee. Austrostipa scabra subsp. falcata grows mainly in the woodlands on the Tablelands and southern areas of New South Wales, Queensland, Victoria and South Australia (Vickery et al. 1986). The second group that is resolved in the parsimony analysis of molecular data is the group that contains four species: A. elegantissima, A. acrociliata, A. platychaeta and A. ramosissima. This group was also resolved in the

11 (1): 9-14, January 2010

UPGMA analysis of molecular data and received 80% bootstrap support (Figure 2 and Figure 4). Although, A. acrociliata and A. platychaeta have been placed in the same subgenus (subgenus Arbuscula, Jacobs and Everett 1996), the analyses do not put those two as sister species. The analysis supported the grouping of A. acrociliata with A. elegantissima (Figure 2) but only with low bootstrap support (67%). Morphologically, there are many similarities between A. acrociliata and A. platychaeta. For example, both species have linear-cylindrical to ovate-cylindrical panicles and glabrous, scabrous or pubescent branches and pedicels. Moreover, both species have branched culms, a blunt callus and the lemma longer than palea. They also share an almost-falcate awn. There has been some speculation as to their relationships with the well-defined Falcatae (S. Jacobs pers. comm.), though there is no other link and none of the analyses has supported any such connection. It is understandable that these species have been placed in the same subgenus but perhaps the characteristic growth form, which may relate to habitat and appears to be a homoplasious character, disguises significant differences. In the recent classification of Austrostipa (Jacobs and Everett 1996), A. elegantissima is placed in subgenus Petaurista along with A. tuckeri. A. elegantissima is more similar to A. tuckeri than to A. acrociliata in terms of morphology. For example, both A. elegantissima and A. tuckeri have pyramidal panicles when mature and spreading, with whorled branches, and the whole unit detaches and acts as the diaspore, while A. acrociliata has linear-cylindrical to ovate- cylindrical panicles that remain attached and the florets break off separately. Moreover, both A. elegantissima and A. tuckeri have characteristic fine hairs on the branches and pedicels, while A. acrociliata has glabrous, scabrous or pubescent branches and pedicels. These hairs and the whole inflorescence detaching are characteristic of subgenus Petaurista (Jacobs and Everett 1996). A. acrociliata has glabrous, scabrous or pubescent branches and pedicels, which is the usual situation in Austrostipa. Despite the similarity between A. elegantissima and A. tuckeri, there are some differences. For example, hairs on the branches of A. elegantissima are longer than in A. tuckeri. A. elegantissima has hairs up to 2 mm long whereas A. tuckeri hairs up to only 0.5 mm long. A. elegantissima has glabrous nodes while A. tuckeri has nodes with sericeous hairs 0.6 mm long. While the inflorescence characters seem quite highly derived and suggest close relationship, any analyses that concentrate on other characteristics seem to consistently suggest that they are not particularly closely related (Jacobs et al. 2000). Wind dispersal does seem to have been derived several times in the Stipoid grasses (Jacobs and Everett 1997; Jacobs et al. 2000), and several obviously different syndromes have evolved. In every other case so far, species with the same syndrome have been shown to form a closely related group (Jacobs et al. 2000). Of all these syndromes, that developed in subgenus Petaurista is the most divergent (the whole inflorescence acting as a diaspore and the characteristic long-hairy branches and/or pedicels) and is the most difficult to imagine as being an example of

BUSTAM – Systematic studies of Australian stipoid grasses (Austrostipa)

parallel evolution. The relationships here are still not clear and can probably only be solved by exploring other even more variables sites for genes sequences. The third group that was recognized in both parsimony and UPGMA analysis of molecular data and received 81% bootstrap support is the group that consists of two species: A. densiflora and A. rudis subsp. nervosa. These species s are placed in two subgenera in the recent classification (Jacobs and Everett 1996). A. densiflora is in subgenus Austrostipa while A. rudis subsp. nervosa is in subgenus Tuberculatae. There are a lot of differences between the two species in term of gross morphology and habitat. For example, A. densiflora has a lemma that is hairy/scabrous near the apex and a characteristic long-hairy awn (one of the characteristics of subgenus Austrostipa (Jacobs and Everett 1996)), while A. rudis subsp. nervosa has a lemma that is glabrous for varying lengths below the lemma apex and which is covered with characteristic silicious crystalline blunt tubercles (characteristics of subgenus Tuberculatae (Jacobs and Everett 1996)). Both species also differ in their habitats. A. densiflora grows in low fertility soils and is more common after disturbance, in drier regions away from the coast of New South Wales, Queensland, Victoria and South Australia. A. rudis subsp. nervosa grows on sandstone, mostly in undisturbed, higherrainfall coastal areas of New South Wales, Queensland and Victoria (Vickery et al. 1986). Another group recognized in the parsimony analysis of molecular data (with 75% bootstrap support) is a group consisting two species: A. mollis and A. tuckeri. However, this group is not supported by the UPGMA analysis. Like most groups retrieved in the parsimony analysis of molecular data, the species in this group are placed in different subgenera in the recent classification (Jacobs and Everett 1996). A. mollis is in subgenus Austrostipa while A. tuckeri is in subgenus Petaurista. The last group retrieved in the parsimony analysis of molecular data (with 71% bootstrap support) consists of two species: A. muelleri and A. breviglumis, supported by UPGMA analysis of molecular data (Figure 2 and Figure 4). This group again comprises two species that have been placed in different subgenera by Jacobs and Everett (1996). A. muelleri was placed in subgenus Tuberculatae while A. breviglumis was in subgenus Arbuscula. There are several differences between the two species in term of gross morphology. For instance, A. breviglumis has sturdy simply-branched culms, while A. muelleri has spreading, scrambling or decumbent much-branched culms. The parsimony analysis of molecular data also supports the monophyly of Austrostipa with 100% bootstrap support (Figure 2). This research has not supported the subgenera of Austrostipa (Jacobs and Everett 1996) with the exception of subgenus Falcatae. While there are occasional hints of further relationships, there is nothing substantial. It is possible that the subgenera (Jacobs and Everett 1996) do not adequately reflect relationships within the genus, but then the analyses do not produce strong evidence for improving that classification.

As shown in the Consistency Index (Table 1), most micro morphological characters are considered to be highly homoplasious, the exception being stomata size (μm) and stomata subsidiary cell size (μm) abaxial surface, which made it difficult to get meaningful result at this level of relationships. There are 26 micro morphological characters in the Table 1 whereas the complete data were collected for 28 characters. The two uninformative but variable characters with CI = 1 are: (i) Leaf adaxial surface different over and between veins. This character consists of two states, absent and present. With the exception of A. muelleri, all remaining species that were examined scored as ‘present’, (ii) Leaf adaxial stomata abundance. This character consist of three states, no stomata present, few stomata and abundant. As for the previous character, A. muelleri has only few stomata, while the reminder of species that were examined have abundant stomata. Table 1. The consistency index (CI) value of all micromorphological characters used can be seen in the table below. ad = adaxial, ab = abaxial No

Characters

1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26.

Leaf ad. silica bodies Leaf ad. long cell wall morphology Leaf ad. stomata size (μm) Leaf ad. stomata subsidiary cell size (μm) Leaf ad. stomata shape Leaf ad. macrohairs presence Leaf ad. macrohairs position Leaf ad. prickles presence Leaf ad. prickles position Leaf ab. different over and between veins Leaf ab. silica bodies presence Leaf ab. silica bodies shape Leaf ab. Long cell wall morphology Leaf ab. Stomata abundance Leaf ab. stomata size (μm) Leaf ab. stomata subsidiary cell size (μm) Leaf ab. macrohairs presence Leaf ab. macrohairs position Leaf ab. prickles presence Leaf ab. prickles position Lemma fundamental cell lenght Lemma sidewall shape Lemma sidewall thickness Lemma endwall shape Lemma short cell hooks Lemma silica bodies presence

CI 0.125 0.500 0.316 0.308 0.333 0.125 0.500 0.333 0.500 0.143 0.333 0.465 0.333 0.500 0.833 0.800 0.333 0.316 0.167 0.125 0.300 0.333 0.250 0.250 0.167 0.333

Although in all analyses, Anemanthele lessoniana is consistently included within a monophyletic of Austrostipa, admittedly at several different positions, there is no suggestion that the genera be combined. There are some obvious gross morphological characters that can be used to distinguish Anemanthele from Austrostipa, including stamen number, hilum shape and lemma nerves and length (Jacobs and Everett 1996).

B I O D I V E R S IT A S

14 CONCLUSION

Parsimony analysis highly supported the monophyly of Austrostipa (>90%). However, there is a little support for the generic classification of Austrostipa proposed by Jacobs and Everett (1996). Only the subgenus Falcatae is supported by all analyses. The micro morphological characters used are uninformative (too homoplasious), consider the consistency index value less than 1.

ACKNOWLEDGMENTS I would like to thank Surrey Jacobs, Joy Everett and Paul Adam as my supervisors. I also would like to thank all staff at Royal Botanic Gardens Sydney and all member of International Stipeae Working Group.

REFERENCES Ariaga MO, Barkworth ME (2000) Leaf anatomy in Nassella and other South American Stipoid Grasses. Am J Bot 87 (6): 33-36. Ariaga MO, Barkworth ME (2006) Amelichloa: a new genus in the Stipeae (Poaceae). SIDA 22 (1): 145-149. Ariaga MO, Jacobs SWL (2006) An anatomy-ecological experiment in Austrostipa aristiglumis, a lowland Stipoid species. Telopea 11 (2) 161-170. Barkworth ME (1993) North American Stipeae (Gramineae): taxonomic changes and other comments. Phytologia 74: 1-25. Barkworth ME, Ariaga MO, Smith JF, Jacobs SWL, Reyna JV, Bushman BS (2008) Molecules and morphology in South American Stipeae (Poaceae). Syst Botany 33 (4): 719-731. Barkworth ME, Everett J (1987) Evolution in the Stipae: Identification and relationships of its monophyletic taxa. In Soderstrom TR, Hilu KW, Campbell CS, Barkworth ME (eds.) Grass systematics and evolution. Smithsonian Institution Press, Washington D.C. Bentham G (1878) Gramineae. In Flora Australiensis 7: 564-571. Lovell, Reeve & Co. London. Cialdella MA, Giussani LM (2002) Phylogenetic relationships of the genus Piptochaetium (Poaceae, Pooideae, Stipeae) evidence from morphological data. Ann Miss Bot Gard 89 (3): 305-336. Clarke PJ (2003) Composition of grazed and cleared temperate grassy woodland in Eastern Australia: patterns in space and inferences in time. J Veget Sci 14: 5-14. Connor HE, Edgar E (2002) History of the taxonomy of the New Zealand native grass flora. J R Soc N Z 32 (1): 89-112. Dumortier BCJ (1823 [1824]) Observations sur les GraminĂŠes de la Flore Belgique. J Casterman, Tournay.

11 (1): 9-14, January 2010 de Lange PJ, Norton DA, Heenan PB, Courtney SP, Molloy BPJ, Ogle CC, Rance BD, Johnson PN, Hitchmough R (2004) Threatened and uncommon plants of New Zealand. N Z J Bot 42: 45-76. Ellis RP (1979) A procedure for standardizing comparative leaf anatomy in the Poaceae. II. The epidermis as seen in surface view. Bothalia 12 (4): 641-671. Everett J (1990) Systematics relationships of the Australian Stipeae (Poaceae). [Thesis]. University of Sydney, Australia. Everett J, Jacobs SWL (1983) Studies in Australian Stipa (Poaceae). Telopea 2: 11-15. Garden DL, Logde GM, Friend DA, Dowling PM, Orchard BA (2000) Effects of grazing management on botanical composition of native grass-based pastures in temperate South-East Australia. Aust J Exp Agric 40 (2) 225-245. GPWG (Grass Phylogeny Working Group) (2000) A phylogeny of the grass family (Poaceae), as inferred from eight character sets. In Jacobs SWL, Everett J (eds.) Grasses: systematics and evolution. CSIRO, Melbourne. Hsiao C, Jacobs SWL, Chatterton NJ, Assay KH (1999) A molecular phylogeny of the grass family (Poaceae) based on the sequences of nuclear ribosomal DNA (ITS). Aust Syst Bot 11: 667-668. Hughes DK (1921) A revision of the Australian Species of Stipa. Kew Bull 1921: 1-30. Hughes DK (1922) Further notes on the Australian Species of Stipa. Kew Bull 1922: 15-22. Huxtable CHA, Koen TB, Waterhouse D (2005) Establishment of native and exotic grasses on mine overburden and topsoil in the Hunter Valley, New South Wales. Rangeland J 27 (2): 73-88. Jacobs SWL, Everett J (1996) Austrostipa, a new genus, and new names for Australian species formerly included in Stipa (Gramineae). Telopea 6: 579-595. Jacobs SWL, Everett J (1997) Jarava plumosa (Gramineae), a new combination for the species formerly known as Stipa papposa. Telopea 7: 301-302. Jacobs SWL, Everett J, Barkworth ME, Hsiao C (2000) Relationships within the Stipoid Grasses (Gramineae). In Jacobs SWL, Everett J (eds). Grasses, systematics and evolution. CSIRO, Melbourne. Landberg J, James CD, Morton SR, Mueller WJ, Stol J (2003) Abundance and composition of plant species along grazing gradients in Australian rangelands. J Appl Ecol 40: 1008-1024. Madison DR, Madison WP (2002) MacClade 4 Release Version 4.05. Sinauer, Sunderland, Massachusetts. Maze J, Kathleen AR, Benerjee S (2002) Studies into abstract properties of individuals VII. Emergence in Hesperostipa comata and three species of Achnatherum (Poaceae). Int J Plant Sci 163 (3): 379-385. Swofford DL (2002) PAUP (Phylogenetic Analysis Using Parsimony) and other methods. Version 4.0b10. Sinauer, Sunderland, Massachusetts. Townrow JES (1978) The genus Stipa L. in Tasmania. Part 3-Revised Taxonomy. Papers and Proceedings of the Royal Society of Tasmania 112: 227-287. Vasques FM, Barkworth ME (2004) Resurrection and emendation of Macrochloa (Gramineae: Stipeae). Bot J Linn Soc 144 (4): 483-495. Vickery JW, Jacobs SWL, Everett J (1986) Taxonomic studies in Stipa (Poaceae) in Australia. Telopea 2: 391-400. Wheeler DJB, Jacobs SWL, Whalley RDB (2002) Grasses of New South Wales. University of New England Press, Armidale.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 15-18

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110104

Begonia “Tuti Siregar” (Begonia listada Smith &Wasshausen x Begonia acetosa Vellozo): a new cultivar from Bali Botanic Garden, Indonesia HARTUTININGSIH-M. SIREGAR1,2,♥, I MADE ARDAKA1, MUSTAID SIREGAR1,2 1

Bali Botanic Garden, Indonesian Institute of Sciences (LIPI), Candikuning, Baturiti, Tabanan 8219, Bali, Indonesia. 2 Centre for Plant Conservation-Bogor Botanic Gardens, Indonesian Institute of Sciences LIPI, Jl. H. Juanda 13, Bogor 16122, West Java, Indonesia, Tel. +62-251-8322187; Fax. +62-251 8322187,email: hartutiningsih@yahoo.co.id Manuscript received: 9 September 2009. Revision accepted: 28 December 2009.

ABSTRACT Siregar HM, Ardaka IM, Siregar M (2010) Begonia “Tuti Siregar” (Begonia listada Smith &Wasshausen x Begonia acetosa Vellozo): a new cultivar from Bali Botanic Garden, Indonesia. Biodiversitas 11: 15-18. A new Begonia cultivar was generated by crossing Begonia listada Smith & Wasshausen with Begonia acetosa Vellozo at Bali Botanic Garden began in 2005. The female flowers of B. listada were pollinated with pollen grains from B. acetosa. Begonia listada is characterized by the following characters: erect, non rhizomatous, branched subshrub to 1 m tall; leaves: blade above bronze, beneath purple with two pale green veins. B. acetosa has the following characters: creeping rhizomatous perennial to about 1 m tall; leaves: thick, blade above olive green, beneath bright red, both surfaces with a dense covering of short stiff white hairs. The results of crossing is, herb, erect, non-rhizomatous, 40-75 cm height, stem brownish green, hairy, stipules in pairs, marcescent, ascending, membranous. Leaves, beautiful asymmetrical leaves intermediate between both parents; petiole deeply red, covered with wooly hairs, veins yellow like B. listada. The new cultivar, Begonia ‘Tuti Siregar’, has the American Begonia Society registration number 1001 (Salisbury 2008). Key words: Begonia “Tuti Siregar”, Bali Botanic Garden, Indonesia, new cultivar.

INTRODUCTION Wild begonias are very abundant in Indonesia, especially in the herb layers of mountain rainforests (Tebbitt 2005). There are more than 200 indigenous Indonesian begonias, which are distributed in Java, Sumatra, Kalimantan, Sulawesi and Papua. Papua is the most species-rich region with 70 species. More than 1600 species of Begonia, which show a pantropical distribution, have been described (Kiew 2005). In the tropical zone, begonias can grow and adapt well from the lowland up to the highland. At 1400 m above sea level (m a.s.l), these plants need light intensities between 30-60%. Begonias can be quickly propagated and grown at Bali Botanic Garden in Indonesia. This Garden encompasses 154 ha at an altitude of 1250 m a.s.l. The propagation of begonias commonly requires well-lit conditions, the light usually filtered by netting which absorbs 70% of the light intensity. Researchers of Bali Botanic Garden have conducted floristic explorations of the entire Indonesian archipelago collecting begonias for ex-situ conservation. More over, seed exchange with both national and international conservation institutions and seed donations have increased the species number in the collection. In 2009 the collection comprised 81 indigenous Indonesian species and 213 exotic. These collections have a great potential for the generation of cultivars, and make Bali Botanic Garden as a center for conservation of Begonia in Indonesia (Hoover et al. 2006). The objectives are to save

Begonia species from the threat of extinction, and to cultivate and propagate them. In addition to the conservation research, the potential of begonias as ornamental plants, and the generation of cultivars, either directly or through a cross-breeding approach, have been investigated. These cultivars can be used for commercial purposes. The aims of these studies were to obtain a new hybrid of Begonia amalgamating the characters of both parents and to create unique character combinations including erect, ovate leaves with yellow veins and a bright red lower leaf surface.

MATERIALS AND METHODS Crossbreeding research on two species of Begonia (B. listada and B. acetosa) has been conducted at the nursery of Bali Botanic Garden since 2005. The garden is located at 1250 m a.s.l, with day time temperatures of 24-26oC, 1820oC in the night, and 70-90% of humidity. The female flowers of B. listada were pollinated with pollen grains from B. acetosa. Begonia listada is characterized by the following characters: erect, non rhizomatous, branched subshrub to 1 m tall; leaves: blade above bronze, beneath purple with two pale green veins. Begonia acetosa has the following characters: creeping rhizomatous perennial to about 1 m tall; leaves: thick, blade above olive green, beneath bright red, both surfaces with a dense covering of short stiff white hairs.

B I O D I V E R S IT A S 11 (1): 15-18, January 2010

The Begonia crossbreeding was conducted in the morning at 09.00-10.00 a.m. The crossing method followed Hooley (1999) and Siregar (2008a,b). Pollen was taken using a cotton bud and smeared directly on the stigma of B. listada flowers. The flowers were then wrapped with clear plastic bags. After fruit production the plastic bags were opened and the fruits left to ripen. A label was put on the plants, which contained information about the female and male parents, the crossing time and the breeder’s name. The crossing was repeated six times. The fruit ripening time, the picking time, and the time it took to raise the seedlings were documented. The seedlings were raised following Siregar et al. (2007). Sterile organic fertilizer (Kompenit, trade mark of organic fertilizer, produced by Bali Botanic Garden) was used. The pots were placed into container containing water to maintain high humidity. The pots were sealed and only opened after the seeds had germinated. After the development of two leaves the plants were transferred into trays with a substrate containing 75% organic fertilizer. After they developed four leaves they were planted in pots or poly bags. RESULTS AND DISCUSSIONS Crossing time There was a high rate of successful crosses. From six repeats, 50% of the hand-pollinated flowers developed into mature fruits. The reasons for this are the good growing conditions at Bali Botanic Garden, especially the glass house temperatures. Optimum temperatures for most begonias are in the range of 13-29oC. The vast majority of them require a relative humidity between 40 and 60% (Tebbitt 2005). Some studies indicate that pollination is more successful in the morning than in the early evening. Sanusie et al. (2008) showed that pollination of Anthurium reached a success rate of 90% in the morning, while the success rate in the early evening was only 70%. Successful pollination depends on several factors such as cleanliness of the tools to avoid contamination, the timing, and the crossing protocol. Failure of pollination is indicated by wilting and falling of the flowers, and the lack of fruit set. Fruits ripened in 30-45 days after fertilization. The color of the fruit changed from green to brownish. The mature brownish fruit was picked and placed into an envelope to avoid seed release. The next step was to open the fruits and to separate the seeds from the pericarp. Time after harvest and cultivation of seedling Fruit harvesting was conducted when the fruits were physiologically ripe, as indicated by a brownish or yellow color and wilted peduncles, but before the seeds were released. The usage of specific materials helps to accelerate the seed germination. The seed germinated about 11-40 days after they were sown. A flexible watering regime and fertilization are essential for the care of the seedlings. Numerous factors including the climate, and the growing medium influence how much water a plant needs at a particular time. Over watering and a lack of adequate atmospheric humidity are probably the two most common reasons why begonias and other cultivated tropical plants

are lost. The most common symptoms of over watering among begonias are dropping of leaves, root rot, and wilted stems. Hybrid morphology The morphology of about 60 seedlings has been documented and four different groups can be recognized: (i) Group with asymmetrical leaves. The leaf color is intermediate between both parents, the blade is ovate like in B. acetosa and fused with yellow veins like in B. listada. Blade green above, hairy, ovate, 15-17 x 10-11 cm, venation palmate, primary veins 7-8, bright yellow; beneath deeply red with pink hairs. This group shows characters which are intermediate between the two parents. Seedlings of this group grew in thirty pots. (ii) Group with symmetrical leaves. Crossing of the two species resulted in plants with symmetrical, orbicular leaves, which are unusual in Begonia. Blade green above, veins pale. The habit in this group is cane-like, erect, with flat stems. Seedlings of this group grew in ten pots. (iii) Group with leaf asymmetry resembling B. acetosa. Blade green above, hairy, beneath bright red, veins invisible. Seedlings of this group grew in ten pots. (iv) Group with not uniform leaves. Blade green above, hairy, beneath bright red, veins invisible. Seedlings of this group grew in ten pots. To avoid the generation of hybrids which are not of interest for the breeder, groups two, three, and four were destroyed, while the first group was maintained and continually propagated. Table 1. shows the characters of the new cultivar and the characters of the two parents. Characters of the new cultivar are: shrub-like habit (SL), erect, non-rhizomatous, height 40-75 cm, internodes 3-4 cm. These are the dominant characters of B. listada. Thomson (2007a,b) showed that a cross using the cane-like type (CL) as either the female or the male parent with the rhizomatous type (Rh) will always result in a new plant with the CL type. The new cultivar at Bali Botanic Garden has been propagated using cuttings, and more than 200 pots were available at the time of writing. The next step was to register the cultivar with the American Begonia Society, providing data like the name of the hybrid plant, data of the parents, the breeder’s data, the time of pollination, first flowering data, the date of request, the plant description, photographs, illustrations, and herbarium voucher details. The cultivar name “Tuti Siregar” was chosen as reference to the breeder. Crossbreeding of Begonia is common abroad, but for me and Bali Botanic Garden, this was the first time to obtain a new hybrid name from a legal institution. There are some necessary requirements for designating a name for a hybrid in accordance with the regulations of the American Begonia Society: the hybrid plant name must be original, it the prefix of the, and some other requirements. At the time of writing, the name of Begonia “Tuti Siregar” was registered less than a year ago and it is now legitimately registered with the American Begonia Society, USA, as a new cultivar with the registration number 1001 (Salisbury, 2008). Herbarium specimen collector Hartutiningsih no. HK 866, kept in Herbarium of Bali Botanic Garden as a reference this new cultivar.

SIREGAR et al. – A new Begonia cultivar from Bali Botanic Garden

D 3 cm

F G

2 cm

5 mm

Figure 1. Begonia “Tuti Siregar”. A. General appearance, B. Entire leaf, C. Leaf cross section, D. Leaf margin, E. Stipule, F. Pistillate flower, G. Staminate flower.

5 cm

Figure 2. Begonia “Tuti Siregar”. A. leaf, B. flower, C and D. habit.

25 cm

50 cm

B I O D I V E R S IT A S 11 (1): 15-18, January 2010

Table 1. Morphological comparison of B.listada, B. acetosa and F1 Begonia “Tuti Siregar”. Characters Plant shape Plant height Internodus Stipules Leaves: Blade

B. listada CL = cane-like 30-75 cm tall 3 cm White, small, 1.5x1 cm

B. acetosa Rh = rhizomatous 35-40 cm tall 1-2 cm Dry, small

F1 = B. “Tuti Siregar” (B. listadaxB. acetosa) SL = shurb-like 40-75 cm tall 3-4 cm Reddish, pairs, white hairy, 3.8-2.7 cm.

Above bronze beneath purple Hastate, 6-10x3.5-5 cm, asymmetry

Above olive green, beneath bright red Ovate to almost orbicular, 418x3.3-13 cm, asymmetry Minutely toothed

Above green, beneath deep red Ovate to almost orbicular, 15-17x10-11 cm, asymmetry Serrulate

Cordate Abruptly short acuminate Rust-brown to red, covered with wooly hairs, 6-30 cm long Axillary, many-flowered Bisexual Cymose

Very obliquely cordate Truncate Deep red, covered with wooly hairs, lanate, 12-15 cm long Axillary, many-flowered Bisexual Cymose

Four, white Outer pair elliptic; 10-12x58 mm, inner pair narrowly elliptic, 7-14x1.5-3 mm

Four, white Outer pair ovate; 1.2x1.3 cm, inner pair obovate 0.8x0.4 cm

Margin

Very shallowly toothed and ciliate Base Very obliquely cordate Apex Acuminate Petiole Pale green to pink with short white hairs, 4.5-15 cm long Inflorence Axillary , few-flowered Bisexual Cymose Male flowers: Tepals Four, white Outer pair ovate-cordate to almost orbicular; 1.11.5x1.1-1,5 inner pair narrowly obovate 11.2x0.3-0.4 cm Stamens About 25-35, arranged symmetrically, anther connectives projecting Female flowers: Tepals Five, white sometimes tinged red Narrowly to broadly elliptic, slightly unequal 0.7-1.5x0.4-1 cm Ovary

Green with a pink tinge, hairy Unequally three-winged, 0.5-1x0.25-0.48 cm Peduncle 20-25 cm tall

About 20-30, arranged in a Many, yellow, arranged symmetric mass on top of a symmetrically column, anther connectives projecting Five, white

Four, white

Outer two narrowly elliptic 0.6-10x0.15-2 cm, inner three elliptic 1-1.3x0.4-0.5 cm White, hairy

Outer pair ovate , inner 0.6x0.3 cm, inner pair narrowly elliptic 0.4x0.2 cm White, hairy

Unequally three-winged, 0.6-1.1x0.3-0.5 cm 28-57 cm tall

Unequally three-winged 0.9x0.5 cm 16-24 cm tall

CONCLUSIONS Description of Begonia “Tuti Siregar” (♀ B. listada Smith & Wasshausen collector Hartutiningsih, no HK 674 x ♂ Begonia acetosa Vellozo collector Hartutiningsih, no HK 676) Herb, erect, non-rhizomatous, 40-75 cm heigh. Stem brownish green, hairy, 1-2 times branched. Stipules in pairs, marcescent, ascending, membranous, 3.8x2.7 cm. Leaves: beautiful asymmetrical leaves intermediate between both parents; 20-24 per stem; petiole deeply red, covered with wooly hairs, lanate, 12-15 cm long; blade almost circular like B. acetosa, veins yellow like B. listada, ovate, 15-17x10-11 cm, base very obliquely cordate, apex truncate, above green, with wooly hairs, beneath deep red with pink wooly hairs, venation palmate with 7-8 primary veins, veins bright yellow. Margin serrulate; Inflorescence: axillary, 6-8, many-flowered with 16-27 flowers, bisexual, cymose, peduncle 16-24 cm long, bracteoles 1.1x0.4 cm, soon falling. Male flowers: tepals 4, white, outer pair covered with short pink hairs, ovate, 1.2x1.3 cm, inner pair

obovate, 0.8x0.4 cm; stamens yellow, numerous. Female flowers: tepals 4, white, outer pair ovate, 0.6x0.3 cm, glabrous, inner pair narrowly elliptic, 0.4x0.2 cm; ovary white, glabrous, unequally three-winged, 0.9x0.5 cm. Flowering season: DecemberFebruary. The propagation methods most commonly used for Begonias are stem and leaf cuttings, and division of a large plant into two or more smaller ones (Figure 1 and 2).

ACKNOWLEDGEMENTS We are deeply indebted to Dr. Harry Wiriadinata, Herbarium Bogoriense, The Research Centre of Biology, The Indonesian Institute of Sciences; Dr. Scott W. Hoover, New England Conservatory, Vermont, USA, for their expertise, advice, and their essential support of our research. We are also very grateful to Daniel C. Thomas, Royal Botanic Garden Edinburgh, UK and Bramantyo T.A. Nugroho who had helped us to edit this paper.

REFERENCES Holley F (1999) Setting and saving seed for species conservation. A Seminar for SWR-American Begonia Society-USA, May 1999. Hoover SW, Hunter JM, Wiriadinata H, Girmansyah D (2006) Begonias at Bali Botanic Gardens, Indonesia. Begonian. November-December 2006: 224-225. Siregar HM, Ardaka IM, Siregar M (2007) Masa berbunga 22 jenis Begonia alam di Kebun Raya “Eka Karya” Bali. Biodiversitas 8: 188-192 Siregar HM (2008a) Mengenal dan merawat Begonia. PT Agromedia Pustaka, Jakarta. Siregar HM (2008b) Begonia ”Tuti Siregar”, the new hybrid from Bali Botanic Garden-Indonesia. Begonian. September/October 2008: 214-217. Kiew R (2005) Begonias of Peninsular Malaysia. Natural History Publications (Borneo) Sdn. Bhd, Kota Kinabalu, Sabah, Malaysia. Salisbury G (2008) New cultivar Begonia “Tuti Siregar” (♀ Begonia listada Smith & Wasshausen x ♂ Begonia acetosa Vellozo). Official International Registration number 1001. Begonian. September/ October 2008: 212-213. Tebbitt MC (2005) Begonias: cultivation, identification, and natural history. Brooklyn Botanic Garden, New York. Thompson B (2007a) Begonia registration handbook. www.begonias.org/ registered/registered AF.htm. [19 December 2007]. Thompson B (2007b) Hybridizing page, hints on hybridizing. www. bradsbegoniaworld.com/hybrid.htm. [19 December 2007]. Sanusie I, Qodriyah L (2008) Teknik penyerbukan silang dan pembibitan Anthurium. www.kebonkembang.com/mod.php?mod=publisher&op= viewarticle&artid=164. [1 January 2008].

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 19-23

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110105

Population dynamic of the swallowtail butterfly, Papilio polytes (Lepidoptera: Papilionidae) in dry and wet seasons SUWARNO♥ 1

Department of Biology, Faculty of Mathematics and Natural Sciences, University of Syiah Kuala, Jl. Syekh Abdur Rauf No. 3, Darussalam, Banda Aceh 23111, Aceh, Indonesia, Tel.: +62-651-742821, Fax: +62-651-7552291, email: j_suwarno@yahoo.com Manuscript received: 17 November 2009. Revision accepted: 9 December 2009.

ABSTRACT Suwarno (2010) Population dynamic of the swallowtail butterfly, Papilio polytes (Lepidoptera: Papilionidae) in dry and wet seasons. Biodiversitas 11: 19-23. The population dynamic of Papilio polytes L. (Lepidoptera: Papilionidae) in dry and wet seasons was investigated in the citrus orchard in Tasek Gelugor, Pulau Pinang, Malaysia. Population of immature stages of P. polytes was observed alternate day from January to March 2006 (dry season, DS), from April to July 2006 (secondary wet season, SWS), and from October to December 2006 (primary wet season, PWS). The population dynamics of the immature stages of P. polytes varied between seasons. The immature stages of P. polytes are more abundance and significantly different in the PWS than those of the DS and the SWS. The larval densities in all seasons decreased with progressive development of the instar stages. Predators and parasitoids are the main factor in regulating the population abundance of immature stages of P. polytes. There were positive correlations between the abundance of immature stages of P. polytes and their natural enemies abundance in each season. Ooencyrtus papilioni Ashmead (Hymenoptera: Encyrtidae) is the most egg parasitoid. Oxyopes quadrifasciatus L. Koch. and O. elegans L. Koch. (Araneae: Oxyopidae) are the main predators in the young larvae, meanwhile Sycanus dichotomus Stal. (Heteroptera: Reduviidae), Calotes versicolor Fitzinger (Squamata: Agamidae), birds and praying mantis attacked the older larvae. Key words: population dynamic, seasons, Papilio polytes, immature stages, natural enemies.

INTRODUCTION Most studies on population dynamics of tropical butterflies were focused on the adult populations (Ehrlich and Gilbert 1973; Hayes 1981; Koh et al. 2004); only few reported on the larval population and their relationship with the environmental factors, particularly the biological factors (Hirosie et al. 1980; Watanabe 1981). Hirosie et al. (1980) and Watanabe (1981) reported that the population of immature Papilio xuthus L. (Lepidoptera: Papilionidae) varied throughout the year with the population of young larval instars fluctuated differently from the older instars. Meanwhile, Diez et al. (2006) reported that the population of Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae) in lemon orchard as well as its parasitoids increased from spring to fall and declined during the winter. Usually, the butterflies display different growth performances (Rafi 1999a; Baguette and Schtickzelle 2003; Munir et al. 2007), fecundity (Davies et al. 2006) and population density in different weather conditions (Davies et al. 2006; Woods et al. 2008; Tiple et al. 2009). Papilio polytes L. (Lepidoptera: Papilionidae) is a tropical or subtropical papilionid butterfly distributed from Southeast Asia to the Southwestern Island of Japan (Corbet and Pendlebury 1992; Nakayama and Honda 2004). They have a wide ecological tolerance enabling them to thrive in a broad variety of climatic conditions (Rafi et al. 1999a). However there are no studies or reports on the growth performances of P. polytes in different seasons in tropical areas.

Generally, in tropical regions, there are two seasons, dry and wet seasons. The temperature is usually higher in the dry season than in the wet season; meanwhile humidity and rainfall are lower in the dry than in the wet season. The weather parameters such as temperature, humidity, and rainfall are environmental factors that affect the development, growth and abundance or population dynamics of insects (Bryant et al. 2000; Warren et al. 2001; Perdikis et al. 2003; van Nouhuys and Lei 2004; Freitas et al. 2005; Pickens 2007). Therefore, the objectives of this research were first, to study the population dynamics of the immature stages of P. polytes in different seasons. Secondly, to determines the relationship between the immature stages of P. polytes and their natural enemies.

MATERIALS AND METHODS The study of population dynamic of the immature stages of P. polytes was conducted in the Taman Firdauce orchard, Tasek Gelugor, Pulau Pinang, Malaysia located at 5o 28’ 60 north latitude, and 100o 30’ 0 east longitude, 34 m above sea level. Taman Firdauce is an orchard planted with several tropical fruits, such as star fruit (Averrhoa carambola L.), citrus (Citrus microcarpha Bunge, C. maxima (Burm.) Merr.), mango (Mangifera indica L.), guava (Psidium guajava L.), avocado (Persea americana Mill.), jackfruit (Artocarpus integra Merr.), longan

B I O D I V E R S IT A S

11 (1): 19-23, January 2010

(Dimocarpus longan Lour.), rambutan (Nephelium lappaceum L.), sour sop (Annona muricata L.) and water apple (Syzygium aqueum Merr.). In the Taman Firdauce orchard, 216 seedlings of Citrus reticulata Blanco were planted in six rows, with each row containing 36 seedlings that were planted in “triangles” of 3.0 m apart and 4.5 m between the rows. Regular farming practices, such as fertilization, irrigation, and insecticidal applications were carried out to ensure a proper and healthy growth. A month prior to observation, insecticide applications were terminated. Unfortunately, six seedlings died during the preparation. The experimental observations were initiated after six months of planting and the plants attained a height of 50-75 cm. The natural population of P. polytes was observed on 210 young plants of C. reticulata (50-75 cm in high) in orchard. Based on the data of climatic factors (temperature, humidity and rainfall) during six years (2000-2005) (Figure 1), the observations were carried out in three different seasons, whereby three months (January – March 2006) was the dry season (DS), four months (April – July 2007) was the secondary wet season (SWS) and three months (October – December 2006) was the primary wet season (PWS). The weather data were taken from Bayan Lepas Meteorological station, in Penang, Malaysia. The population levels of all stages of growth from egg to adult were recorded. Before starting the observation in each season, all immature stages of P. polytes was removed from the host plants to prevent overlapping of seasons. The eggs of P. polytes was attached to the citrus plant (leaves and stems) were counted and labelled. The newly hatched larvae were recorded and their development was labelled and monitored in the alternate day until the adult emerged. The observations were made on specific seasonal cohorts. The data on abundance of immature stages and natural enemies of P. polytes in all seasons were not normally distributed (Smirnov-Kolmogorov, P < 0.05) hence they

were all analyzed using the non-parametric procedures. The abundance of the immature stages in various seasons was analysed using the Friedman test (Dytham 2003; Pallant 2005). The relationships of the abundance of immature stages of P. polytes with their natural enemies were evaluated by simple correlation and simple linear regression analyses. The data of eggs, larvae, parasitized and predated egg and predated larvae population in each season were pooled and transformed using log(x), log(x+1) or √(x), √(x+1) whenever appropriate (Dytham 2003; Pallant 2005), to normalize their distributions before data use subjected to ANOVA. All the above comparisons and relationships were analyzed using the SPSS software version 12 (Dytham 2003; Pallant 2005).

RESULTS AND DISCUSSION The abundance of the immature stages of P. polytes was investigated in three seasons; dry season, secondary and primary wet seasons. Each stage fluctuated differently and significantly at P < 0.05 in all seasons (Table 1). Among the three seasons, the abundance of P. polytes was very low in the dry season and the highest in the primary wet season. Predators and parasitoid are the main natural enemies of the immature stages of P. polytes. The increased and decreased in number of immature stage of P. polytes closely related to the increase and decrease of their natural enemies. A strong positive correlation between the immature stages density of P. polytes and their natural enemies was recorded in the dry season (r = 0.94), in the secondary wet season (r = 0.89) and in the primary wet season (r = 0.96). Furthermore, the regression analysis showed that the density of immature stage of P. polytes in dry season, secondary and primary wet seasons were significantly affected on their natural enemies (Figure 2).

Figure 1. The distribution abiotic factors in Tasek Gelugor, Pulau Pinang, Malaysia from 2000-2005 (average) and 2006. (a) Temperature (oC), (b) Relative humidity (%), (c) Rainfall (mm).

1.6 y = 0.697x + 0.052 R2 = 0.844

1.2 0.8 0.4

Log (x+1) total natural enemies

2.5

2.0

1.6 y = 0.900x - 0.243 R2 = 0.859

1.2 0.8 0.4 0.0

0.0 0.0

0.5

1.0

1.5

Log (x+1) total immature stage

2.0

2.5

2.0 y = 0.777x - 0.061 R 2 = 0.926

1.5

1.0

0.5

0.0

0.5

1.0

1.5

Log (x+1) total immature stage

2.0

2.5

0.0

0.5

1.0

Log

(x+1)

1.5

2.0

2.5

3.0

total immature stage

Figure 2. Relationship of total number of immature stage (egg to larva instar 5) of P. polytes and their natural enemies on Citrus reticulata, in Taman Firdauce orchard, Tasek Gelugor, Pulau Pinang, Malaysia in different season from January to December 2006. A. DS = Dry Season, B. SWS = Secondary Wet Season, C. PWS = Primary Wet Season.

SUWARNO â&#x20AC;&#x201C; Population dynamic of Papilio polytes

Generally, fluctuation of natural enemies density of the immature stage of P. polytes in each season depended on the immature stage density. During the dry season egg predated and egg parasitized of P. polytes increased significantly positive correlated to increasing of egg density (Table 2). Regression analysis showed that the increased of egg density significantly affected (P < 0.01) on the predators of egg (Table 2). The larval density was related to predators of larval density. The positive correlation was recorded between the density of P. polytes larval and their natural enemies in dry season (Table 2). Furthermore, the regression analyses showed that the predated larval density on each instar was significantly affected (P < 0.01) by each larval density of P. polytes (Table 2). A modest to strong positive correlation was observed between immature stages density of P. polytes and its natural enemies in the secondary wet season. Furthermore, a simple linear regression analyses showed that the increased in the egg and larvae density of P. polytes in secondary wet season significantly increased (P < 0.01) the number of the natural enemies (Table 3). In the primary wet season, fluctuation density of the egg of P. polytes closely related to their predated and parasitized egg. A strong positive correlation was recorded between the egg density of P. polytes and the predated and parasitized egg in this season (Table 4). The similar phenomenon was observed on the larvae whereby, fluctuation of larvae in each instar was closely related to their predated larvae fluctuation. The strong positively correlations were observed between the larval density of P. polytes and predated larvae in each instar (Table 4). The simple linear regression analyses showed the egg and larvae density of P. polytes in primary wet season significantly affected (P < 0.01) on predated egg and larvae with the equation as shown on Table 4. Population dynamics of the immature stages of P. polytes varied between seasons. It might be affected by several factors such as climate, nutrient and natural enemies. Natural enemies are the important component in the population dynamics of this butterfly. The natural enemies have influence on total insect number and some-times their impact is related to the density of their prey (Hirosie et al. 1980; Watanabe 1981; Feeny et al. 1985; Stefanescu 2000; Zalucki et al. 2002; Pearce et al. 2004; Nava et al. 2005; Stefanescu et al. 2006).

Table 1. The abundance of immature stage of P. polytes on C. reticulata in all seasons in Taman Firdauce orchard, Tasek Gelugor, Pulau Pinang, Malaysia, from January 2006 to December 2007. Stages Chi-Square df sig. Egg 82.88 2 0.00* L1 129.97 2 0.00* L2 79.02 2 0.00* L3 84.63 2 0.00* L4 71.16 2 0.00* L5 42.67 2 0.00* Pupa 18.09 2 0.00* Note: L1 = first instar larvae; L2 = second instar larvae; L3 = third instar larvae; L4 = fourth instar larvae; L5 = fifth instar larvae; Significant values marked with an asterisk â&#x20AC;?*â&#x20AC;? are significantly different (Friedman test, P < 0.05)

Table 2. Correlation and regression analyses between the number of each stage of P. polytes and they had attacked by natural enemies on C. reticulata in Taman Firdauce orchard, Kampong Paya Tok Akil, Tasek Gelugor, Pulau Pinang, Malaysia in dry season from January to March 2006. r = The Pearson correlation (P = 0.01). Relationships n r F value y = a + bx R2 P value EggVs.Parasitoids 31 0.49# Egg Vs. Predators 31 0.69 15.985 y = 0.127+0.341x 0.60 < 0.01 L1 Vs. Predators 32 0.86 87.582 y=-0.381+0.759x 0.87 < 0.01 L2 Vs. Predators 32 0.80 40.765 y = 0.097+0.588x 0.76 < 0.01 L3 Vs. Predators 30 0.62 14.521 y=-0.449+1.005x 0.60 < 0.01 L4 Vs. Predators 30 0.60 14.758 y = 0.282+0.560x 0.60 < 0.01 L5 Vs. Predators 32 0.81 51.872 y = 0.122+0.992x 0.80 < 0.01 # ) Data not normally distribution, L1 = first instar larva, L2 = second instar larva, L3 = third instar larva, L4 = fourth instar larva, L5 = fifth instar larva.

Table 3. Correlation and regression analyses between the number of immature stage of P. polytes and they had attacked by natural enemies on C. reticulata in Taman Firdauce orchard, Tasek Gelugor, Pulau Pinang, Malaysia in SWS from April to July 2006. r = The Pearson correlation (P = 0.01). Relationships n r F value y = a + bx R2 P value # EggVs.Parasitoids 46 0.654 Egg Vs. Predators 46 0.677 70.431 y = 0.285+0.861x 0.788 < 0.01 L1 Vs. Predators 48 0.645 30.954 y = 0.050+0.655x 0.638 < 0.01 L2 Vs. Predators 47 0.447 13.964 y = 0.068+0.487x 0.483 < 0.01 L3 Vs. Predators 48 0.861 79.342 y = 0.013+0.677x 0.799 < 0.01 L4 Vs. Predators 49 0.776 65.454 y = 0.069+0.723x 0.767 < 0.01 L5 Vs. Predators 50 0.887 65.597 y = 0.103+0.891x 0.763 < 0.01 # ) Data not normally distribution, L1 = first instar larva, L2 = second instar larva, L3 = third instar larva, L4 = fourth instar larva, L5 = fifth instar larva.

Table 4. Correlation and regression analyses between the number of immature stage of P. polytes and they had attacked by natural enemies on C. reticulata in Taman Firdauce orchard, Tasek Gelugor, Pulau Pinang, Malaysia in PWS from October to December 2006. r = The Pearson correlation (P = 0.01). Relationships n r F value EggVs.Parasitoids 39 0.72 50.57 Egg Vs. Predators 39 0.86 77.26 L1 Vs. Predators 40 0.74 60.11 L2 Vs. Predators 39 0.87 137.00 L3 Vs. Predators 39 0.86 97.62 L4 Vs. Predators 40 0.88 151.55 L5 Vs. Predators 39 0.85 97.13 L1 = first instar larva, L2 = second instar fourth instar larva, L5 = fifth instar larva.

y = a + bx R2 P value y=-0.558+0.678x 0.76 < 0.01 y = 0.154+0.656x 0.83 < 0.01 y = 0.041+0.617x 0.79 < 0.01 y = 0.192+0.584x 0.89 < 0.01 y = 0.036+0.667x 0.86 < 0.01 y = 0.033+0.619x 0.90 < 0.01 y = 0.007+0.653x 0.86 < 0.01 larva, L3 = third instar larva, L4 =

B I O D I V E R S IT A S

11 (1): 19-23, January 2010

The lower abundance of eggs and larvae of P. polytes in dry season was probably due to a lesser amount of young leaves and poorer quality of leaves of C. reticulata. Females of Papilio species (Ono et al. 2000; Saljoqi et al. 2006) and Hypna clytemnestra Cramer (Lepidoptera: Nymphalidae) (Gomes-Filho 2003) have great precision to lay their eggs on the healthiest young leaves which subsequently prefer to lay their eggs on its as preparation for their offsprings. This statement supported the results in the present P. polytes study. In the secondary and primary wet seasons, flushes of the young leaves after the rains attracted more females to oviposit on them. Barone (2000) found that the spatial and temporal distributions of Bassaris gonerilla Fabr. (Lepidoptera: Nymphalidae) are correlated to the availability of young leaves. Furthermore, Ba et al. (2008) stated that the adult populations of Chilo spp. (Lepidoptera: Pyralidae) were more abundant during the wet than the dry cropping seasons. When more eggs of P. polytes were laid by females in the secondary and primary wet seasons, the larval abundance were subsequently higher in those seasons. Rafi et al. (1999b) reported that the egg density of Papilio demoleus L. (Lepidoptera: Papilionidae) was correlated to their larval density. Since the larvae were growing into the adulthood, many factors affected their population. The larval densities of P. polytes in all seasons decreased with progressive development of the instar stages. The abundance of larvae (Instar 1-5) of P. polytes, were generally significantly different in the various seasons. The variation in the number of eggs determined the larval abundance in each season. The abundance certain stage of the life cycle was determined by the abundance and mortality of the previous stage. Since the mortality agents in the larval stage of P. polytes were living together on the same host plant, it would strongly affect the butterfliesâ&#x20AC;&#x2122; population. As a result constant mortality rate occurred in that population. In the Firdauce orchard, natural enemies were found to be the most important factor regulating the population of P. polytes. Among all stages, the eggs suffered the most attack especially by the predators and parasitoids. Spiders, such as Oxyopes spp. were the most important predators. Others include the red ant, Solenopsis sp. Parasitoid species was mostly Ooencyrtus papilioni Ashmead (Hymenoptera: Encyrtidae). At the young larval stage, spiders such as Oxyopes quadrifasciatus L. Koch. and O. elegans L. Koch. were considered as the principle enemies to the P. polytes. The other larval predators such as, Sycanus dichotomus Stal. (Heteroptera: Reduviidae), Calotes versicolor Fitzinger (Squamata: Agamidae), birds and Mantis attacked the older larvae. The population of parasitoids and predators increased with increasing number of eggs and larvae of P. polytes. There was a positive relationship between the egg densities of P. polytes and the number of predated eggs. Such relationship was observed in Papilio polyxenes Fabr. (Feeny et al. 1985), P. xuthus (Hirosie et al. 1980; Watanabe 1981), P. demoleus and P. polytes in Lower Sindh, Pakistan (Munir et al. 2007). Many egg predators (such as spiders and ants) attacked the eggs and each of

them ate many eggs during their attack. For all three seasons the egg mortality of P. polytes due to predation was 24.5%, higher than mortality due to parasitoid (6.6%). Predators were also found to be the most important mortality factor of the larval stages of P. polytes in all three seasons, because they killed more than 25% of each larval instar. The numbers of larvae P. polytes killed by predators were strongly correlated to population densities of each instar. The most important predator, spider was well adapted to living on the same host plant with P. polytes larvae. On grain crop in Australia, Pearce et al. (2004) reported that spiders were also among the most abundant predators. Since the spiders are voracious predators and combined with their high abundance, they played an important role in the reduction of the pest populations. Meanwhile another predator, S. dichotomus, was very active and it searched the larvae on every part of the host plant. This study also revealed that a member of the egg parasitized fluctuated closely with the total number of eggs of P. polytes. Obviously, the availability of eggs determined predatory success in the field. In a banana plantation Okolle (2007) reported a significant positive correlation between the density of egg of Erionota thrax L. (Lepidoptera: Hesperiidae) and its parasitoid, Ooencyrtus erionotae FerriĂ¨re (Hymenoptera: Encyrtidae) population. This finding was similar to previous studies on P. xuthus by Watanabe (1981), P. polyxenes by Feeny et al. (1985), and citrus leafminer, Phyllocnistis citrella Stainton (Lepidoptera: Gracillariidae) by Legaspi et al. (1999, 2001). Meanwhile, Fleming et al. (2005) reported that the egg parasitism affect the population dynamic of Heliconius charithonia L. (Lepidoptera: Nymphalidae). The effect of predation on the fifth instar was higher compared to other larval stage due to the presence of many predator species such as S. dichotomus, Mantis sp., birds and garden lizard during this stage. These predators were very active and they consumed large number of larvae. Hirosie et al. (1980) and Watanabe (1981) found that S. dichotomus and Mantis were very important predators of P. xuthus, while predation by vertebrate, particularly bird and garden lizard accounted for a low proportion of the mortality of the fourth and fifth instar larvae. Higher mortality rate in the fifth instar larvae resulted in a low pupal population. The relationship between the egg and its parasitoid in the secondary and primary wet seasons were higher than in the dry season. Higher rainfall and humidity in these seasons produced suitable host and the presence of food for adult. Legaspi et al. (1999) found that the higher parasitism on the leafminer, P. citrella in August compared to February was due to more rainfall in August which increased the humidity level. Furthermore they found that the presence of suitable hosts may also contribute to the increased incidence of the leaf miner parasites. The availability of young leaves in secondary and primary wet seasons affected on abundance of P. polytes and subsequently correlates with their natural enemies density. RiihimĂ¤ki et al. (2006) stated that the plant structure may have direct effects on herbivores as well as indirect effects mediated by natural enemies. Predator-prey

SUWARNO – Population dynamic of Papilio polytes

and parasitoid-host relationships more frequently occur in simple vegetation structures than in complex ones (Cudington and Yodzis 2002). CONCLUSION As a conclusion of this study, the abundance of P. polytes in the Firdauce orchard varied in different seasons. The biotic factor, mainly the attack of natural enemies was of considerable importance. Number of parasitized eggs and predated larvae were strongly influenced by egg and larval densities, respectively. Predators were the key mortality factor of immature stages in P. polytes in all seasons. ACKNOWLEDGEMENTS I acknowledge Assoc. Prof. Dr. Che Salmah Md Rawi and Prof. Dr. Abu Hassan Ahmad (School of Biological Sciences, Universiti Sains Malaysia) and Prof. Dr. Arsyad Ali (University of Florida) for editing this manuscript. Special thanks are extended to Dr. Zainal Abidin for the permission and accommodation in the Taman Firdauce orchard. REFERENCES Ba NM, Dakouo D, Nacro S, Karamage F (2008) Seasonal abundance of lepidopteran stemborers and diopsid flies in irrigated fields of cultivated (Oryza sativa) and wild rice (Oryza longistaminata) in western Burkina Faso. Int J Trop Insect Sci 28: 30-36. Baguette M, Schtickzelle N (2003) Local population are important to the conservation of metapopulation in highly fragmented landscapes. J Appl Ecol 40: 404-412. Barone JA (2000) Comparison of herbivores and herbivory in the canopy and under story for two tropical species. Biotropica 32: 307-317. Bryant RB, Thomas CD, Bale JS (2000) Thermal ecology of gregarious and solitary nettle-feeding nymphalid butterfly larvae. Oecologia 122: 1-10. Corbet AS, Pendleburry HM (1992) The butterflies of the Malay Peninsula. 4th ed. Malayan Nature Society, Kuala Lumpur. Cudington K, Yodzis P (2002) Predator-prey dynamics and movement in fractal environments. Amer Nat 160: 119-134. Davies ZG, Wilson RJ, Coles S, Thomas CD (2006) Changing habitat associations of a thermally constrained species, the silver-spotted skipper butterfly, in response to climate warming. J Anim Ecol 75: 247-256. Diez PA, Pena JE, Fidalgo P (2006). Population dynamic of Phyllocnistis citrella (Lepidoptera: Gracillariidae) and its parasitoids in Tafi Viejo, Tucuman, Argentina. Florida Entomol 89: 328-335. Dytham C (2003) Choosing and using statistics. A biologist’s guide. 2nd ed. Blackwell, Oxford. Ehrlich PR, Gilbert LE (1973) Population structure and dynamics of the tropical butterfly Heliconius ethila. Biotropica 19: 69-83. Feeny P, Blau WS, Kareiva PM (1985) Larval growth and survivorship of the black swallowtail butterfly in central New York. Ecol Monograph 55: 167-187. Fleming TH, Serrano D, Nassar J (2005) Dynamic of subtropical population of the zebra longwing butterfly Heliconius charithonia (Nymphalidae). Florida Entomol 88: 169-179. Freitas FAD, Zanuncio TV, Zanuncio JC, Conceicao PMD, Fialho MCQ, Bernardino AS (2005) Effect of plant age, temperature and rainfall on Lepidoptera insect pests collected with light traps in a Eucalyptus grandis plantation in Brazil. Ann Forest Sci 62: 85-90. Gomes-Filho A (2003). Seasonal fluctuation and mortality schedule for immatures of Hypna clytemnestra (Butler), an uncommon neotropical butterfly (Nymphalidae: Charaxinae). J Res Lepid 37: 37-45. Hayes JL (1981) The population ecology of a natural population of the pierid butterfly Colias alexandra. Oecologia 49: 188-200. Hirosie Y, Suzuki Y, Takagi M, Hiehata K, Yamasaki M, Kimoto H, Yamanaka M, Iga M, Yamaguchi K (1980) Population dynamic of the

citrus swallowtail, Papilio xuthus Linneaus (Lepidoptera: Papilionidae): Mechanism stabilizing its number. Res Pop Ecol 21: 260-285. Koh LP, Sodhi NS, Brook BW (2004) Ecological correlates of extinction proneness in tropical butterflies. Conserv Biol 18: 1571-1578 Legaspi JC, French JV, Schauff ME, Woolley JB (1999) The citrus leafminer, Phyllocnistis citrella (Lepidoptera: Gracillariidae) in South Texas: Incidence and parasitism. Florida Entomol 82: 305-316. Legaspi JC, French JV, Zuniga AG, Legaspi BC (2001) Population dynamic of the citrus leafminer, Phyllocnistis citrella (Lepidoptera: Gracillariidae) and its natural enemiesin Texas and Mexico. Biol Control 21: 84-90. Munir A, Siddiqui NY, Rafi MA, Pavulaan H, Wright D (2007) Bionomic studies of Papilio demoleus Linnaeus, the citrus butterfly, (Lepidoptera: Papilionidae) from Lower Sindh, Pakistan. Tax Rep Int Lepid Surv 8: 1-11. Nakayama T, Honda K (2004) Chemical basis for differential acceptance of two sympatric rutaceous plants by ovipositing females of a swallotail butterfly, Papilio polytes (Lepidoptera: Papilionidae). Chemoecology 14: 199-205. Nava DE, Parra JRP, Costa VA, Guerra TM, Consoli FL (2005) Population dynamic of Stenoma catenifer (Lepidoptera: Elachistidae) and related larval parasitoids in Minas Gerais, Brazil. Florida Entomol 88: 441-446. Okolle NJ (2007) Population dynamics, within-field and within-plant distribution of the banana skipper (Erionota thrax L.) (Lepidoptera: Hesperiidae) and its parasitoids, in Penang, Malaysia. [PhD Thesis]. Universiti Sains Malaysia, Penang. [Malaysia] Ono H, Nishida R, Kuwahara Y (2000) A dihydroxy--lactone as an oviposition stimulant the swallowtail butterfly, Papilio bianor, from the rutaceous plant, Orixa japonica. Biosci, Biotech, Biochem 64: 1970-1973. Pallant J (2005) SPSS Survival Manual. A steps by step guide to data analysing using SPSS for windows (version 12). 2nd ed. Allen & Unwin, New South Wales. Pearce S, Hebron WM, Raven RJ, Zalucki MP, Hassan E (2004) Spider fauna of soybean crops in south-east Queensland and their potential as predators of Helicoverpa spp. (Lepidoptera: Noctuidae). Aust J Entomol 43: 57-65. Perdikis DC, Fantinou AA, Lykouressis DP (2003) Constant rate allocation in nymphal development in species of Hemiptera. Physiol Entomol 28: 331-339. Pickens BA (2007) Understanding the population dynamic of a rare, polyvoltine butterfly. J Zool 273: 229-236. Rafi MA, Matin MA, Khan MR (1999a) Biology of eggs of citrus butterfly, Papilio demoleus L. (Lepidoptera: Papilionidae). Pak J Sci 51: 95-99. Rafi MA, Matin MA, Khan MR (1999b) Host preference of lemon butterfly Papilio demoleus L. in the Northern Barani areas of Pakistan. Pak J Sci 51: 93-94. Riihimäki J, Vehviläinen H, Kaitaniemi P, Koricheva J (2006) Host tree architecture mediates the effect of predator on herbivore survival. Ecol Entomol 31: 227-235. Saljoqi AUR, Aslam N, Rafi MA (2006) Biology and host preference of lemon butterfly, (Papilio demoleus L.). Environ Mon 6: 40-43. Stefanescu C (2000) Bird predation on cryptic larvae and pupae of a swallowtail butterfly. Bull GCA 17: 39-49. Stefanescu C, Jubany J, Dantart J (2006). Egg-laying by butterfly Iphiclides podalirius (Lepidoptera, Papilionidae) on alien plants: a broadening of host range or oviposition mistakes? Anim Biodiv Conserv 29: 83-90. Tiple A, Agashe D, Khurad AM, Kunte K (2009) Population dynamic and seasonal polyphenism of Chilades pandava butterfly (Lycaenidae) in central India. Curr Sci 19: 1774-1779. van Nouhuys S, Lei G (2004) Parasitoid-host metapopulation dynamic: the causes and consequences of phonological asynchrony. J Anim Ecol 73: 526-535. Warren MS. Hill JK, Thomas JA, Asher J. Fox R, Huntley B, Roy DB, Tefler MG, Jaffcoate S, Harding P, Jaffcoate P, Willis SG, GreatorexDavies JN, Moss D, Thomas CD (2001) Rapid responses of British butterflies to opposing forces of climate and habitat change. Nature 414: 65-69. Watanabe M (1981) Population dynamic of the swallowtail butterfly, Papilio xuthus L., in a deforested area. Res Pop Ecol 23: 74-93. Woods JN, Wilson J, Runkle JR (2008) Influence of climate on butterfly community and population dynamic in western Ohio. Environ Entomol 37: 696-706. Zalucki MP, Clarke AR, Malcolm SB (2002) Ecology and behaviour of first instar larval Lepidoptera. Ann Rev Entomol 47: 361-393.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 24-28

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110106

Butterfly diversity as a data base for the development plan of Butterfly Garden at Bosscha Observatory, Lembang, West Java TATI SURYATI SYAMSUDIN SUBAHARâ&#x2122;Ľ, ANNISA YULIANA Ecology and Biosystematics Research Group, School of Life Sciences and Technology, Bandung Institute of Technology, Jl. Ganesa No 10, Bandung 40132, West Java, Indonesia, Tel./fax.: +62-22-2534107, +62-22-2511575, ď&#x201A;Šemail: tati@sith.itb.ac.id, tatibio76@yahoo.com Manuscript received: 6 July 2009. Revision accepted: 12 November 2009.

ABSTRACT Subahar TSS, Yuliana A (2010) Butterfly diversity as a data base for the development plan of Butterfly Garden at Bosscha Observatory, Lembang, West Java. Biodiversitas 11: 24-28. Change of land use and the increasing number of visitors to Bosscha area was one factor for the development plan of butterfly garden in the area. The objectives of this research were to examine butterfly diversity and its potential for development plan of butterfly garden. Butterfly diversity and its richness conducted by standard walk methods. Host plant and larval food plant was recorded during butterfly survey. Public perception on the development plan of butterfly garden was examined by questionnaire. The results showed that 26 species of butterfly was found in Bosscha area and Delias belisama belisama was the most dominant species. Public perceptions consider that the development plan of butterfly garden will give benefit to the community; not only providing new insight (40.41%), additional tourism object (23.97%) and will gave aesthetical value (17.12%). Twelve local species should be considered for development plan of butterfly garden: Papilio agamemnon, P. demoleus, P. memnon, P. sarpedon, Delias belisama, Eurema hecabe, Danaus chrysippus, Argynnis hyperbius, Cethosia penthesilea, Hypolimnas misippus, Melanitis phedima and Euthalia adonia. Host plant: Bougainvillea spectabilis, Citrus aurantium, Lantana camara, Macaranga tanarius and food plants: Citrus aurantium, Cosmos caudatus, Eupatorium inulifolium, Gomphrena globosa, Hibiscus rosa-sinensis, Lantana camara, and Tithonia diversifolia. Key words: host plant, butterfly diversity, Bosscha Observatory, standard walk.

INTRODUCTION Urbanization and agriculture are some of the biggest threats to the world biodiversity that led to local species extinction due to habitat fragmentation and decreasing of open space (Connor et al. 2003; Hedblom 2007). Land use change and reducing open space for human settlement and agriculture are the general issue in Northern area of Bandung, West Java, including Bosscha Observatory area. So far, Bosscha has a unique role; as the only major Observatory in Indonesia, even in South East Asia (Anonymous 2007) has made Bosscha as the only one astronomical tourism objects in Indonesia. Bosscha Observatory visited by various groups of people: students, researchers etc and the number of visitor always increase every year. The number of tourist visits on the year 2006 was 55,419 people with an average of 4618 persons per month. In 2007, the number of visitors increased dramatically that was 72,037 people with an average per month of 4735 people (Anonymous 2007) and in the year of 2008, the number of visitors was 63,480 people (not published). Increasing number of visitors from time to time has led Bosscha Observatory as an educational heritage. The landscape of Bosscha was an agricultural area, however with the increasing number of local residents and visitors to the area has made the area as an urban resident with its all supporting activities. As consequences, open

space reduced and in the night time, light intensity from the residential area became one of the factors that disturb the activity of observatory which required minimal light condition (Ernanto 2006). Environmental conditions at Bosscha area with light pollution, increase number of visitors (who are interested to understand natural phenomena through star observation) and the limited capacity of the observatory require diversification of function of the area. Environmental changes can affect the biological diversity. Sundufu and Dumbuya (2008) study the habitat preferences of butterflies in the Bumbuna forest, Northern Sierra Leone showed that accumulated species richness and diversity indices in the disturbed habitats were lower than the forest. Learning from the San Francisco Bay, the impact of urbanization has caused 43% loss of butterfly species as a result of habitat fragmentation and loss of habitat for insects (Connor et al. 2003). Butterflies are good indicator species to monitor ecological changes in a habitat (Sreekumar and Balakrishnan 2001). Information on butterfly diversity in the region is very scarce. Tati-Subahar et al (2007) described butterfly stratification in Tangkuban Parahu (a mountainous area, Northern part of Bandung) from 1400-2000m above sea level (asl). Up till now there has been no report on butterfly diversity in Bosscha area and its vicinity. Therefore the objective of this research was to examine butterfly diversity and its potential for

SUBAHAR &YULIANA – Butterfly of Bosscha Observatory, Lembang

development plan of butterfly garden. Hope this result can contribute to the biodiversity database on development plan of butterfly garden as tourist object and also as natural laboratory for environmental education and conservation.

MATERIALS AND METHODS This research was conducted in Bosscha and Cihideung which was a hilly area about 15 km to the north of Bandung City, West Java, Indonesia. Bosscha observatory (107°36 'BT and 6°49' LS) was located at 1310 m asl (above sea level). The study site in Bosscha was an area of 6 ha consisted of 3 plots: The first plot was front park of the observatory: the park was covered by grass. In the middle of the area found Bougainvillea and Lantana camara. The second plot was a center park of the observatory which was covered by grass. During observations found some flowering plants at the road edge. The third plot was a back yard; there were shrubs, grasses and bamboo. The second location was Cihideung, known as a center for cultivation and trading of ornamental plants and fruit trees. This area lies along the road from south to the north of Bandung City. The study site (1032-1203 m asl) was 3 km in long covered by different type of flowering plants which consist of 4 plots: The first plot was the southern part, the flowering plants found only in one side of the road. At second plot, the flowering plant was located on both sides of the road. At plot 3, flowering plants found only in one side of the road and plot 4 was the middle of the flower garden which was closed to Bosscha. Survey on butterfly diversity conducted by standard walk methods (Pollard and Yates 1995; Tati-Subahar et al. 2007), i.e. walking along the plot while counting and recording the number of butterflies seen or encountered. The observation width was limited to about 5m. The presence and the number of known butterflies in each plot were directly recorded. After being examined and recorded morphologically by photograph, the butterfly released into natural habitat. No individual marking was conducted. Unidentified butterfly were collected using an insect net for later identification. The specific locations in which butterfly were observed, e.g. on flowers, other plant parts were also recorded. Surveyed was conducted in two periods, first period was on 8-24 May 2008 and the second was on 24-30 June 2008. Both periods of surveyed were the end of the rainy season and early dry season. Observation started from 07.00 am and finished around 12.00 am every day. Microclimate condition during the surveyed varied, temperature ranges from 21oC-26oC and relative humidity ranges from 72%-83 %. Species identification was conducted at Ecology and Biosystematic Laboratory of SITH-ITB Bandung follows Roepke (1932), Smart (1975), Preston and Mafham (2000), and Noerdjito and Aswari (2003). Identification of host and food plants conducted in Herbarium Bandungense-SITH ITB Bandung. Information on host and food plants for each species of butterfly was completed based on observations and

information from different locations (Nurcahya 2003; Soekardi 2002; Tati-Subahar et al. 2007). Host plants are the female butterfly lays their eggs on. The newly emerged (caterpillars) only eat the plant that they are hatched on. Food plants are the plants that adult butterfly forage on. Sorensen Index of Similarity (Odum 1971; MuellerDumbois and Ellenberg 1974) was used to compare butterfly community between Bosscha and Cihideung area. Potential utilization of butterfly as tourism object for environmental education and conservation were evaluated by distributing questioners to 86 students of Lembang district (about one km from Bosscha area). Evaluation on ecosystem services was analyzed following methods of Boyer and Polasky (2004). Community perception on the benefit of development plan of butterfly garden was evaluated by distributing 75 questioners randomly to Bosscha Observatory visitors every day during one week.

RESULTS AND DISCUSSION Diversity and abundance of butterfly During this research, 848 individuals of butterfly were recorded in Bosscha (26 species of 5 families), which were consisted of 4 species of Papilionidae, 4 species of Pieridae, 16 species of Nymphalidae, one species of Lycaenidae and one species of Hesperiidae. Butterfly survey in Cihideung found 41 species from 5 families which consisted 7 species of Papilionidae, 5 species of Pieridae, 26 species of Nymphalidae, two species of Hesperiidae and one species of Lycaenidae. These species were a new record of butterfly species from Bosscha and Cihideung, since there was not any previous documentation. Butterfly communities in Bosscha and Cihideung were almost similar (Sorensen Index of Similarity = 60.87%). Although Cihideung as a central of ornament plants with highest diversity of flowering plants, but individual number of butterfly observed during observation in Cihideung was lower than Bosscha (Table 1). As consequence, the total number of host plants and food plant recorded during periods of observation was lower. The highest relative abundance of butterfly species in Bosscha was Delias belisama belisama (32.19%) followed by Ypthima philomela philomela (20.64%), Papilio sarpedon (11.08%) and the rest of 14.50% were distributed to 19 species of butterfly that have a relative abundance about 0.76% per species (Figure 1a). In Cihideung, D. belisama belisama was also dominant with its relative abundance was 53, 21%, followed by Cethosia penthesilea (10.35%), and the rest of 34 species have a relative abundance about 0.3% per species (Figure 1b). Domination of D. belisama belisama in the two study site related to its characteristic as a polyfagous insect which has a wide range of host and food plants and could found in different habitats (Soekardi 2002). In determining the pattern of butterfly community, relative abundance of butterfly and plants resources was an important aspect that characterizes butterfly community (Yamamoto et al. 2007).

B I O D I V E R S IT A S 11 (1): 24-28, January 2010

Lycaenidae Hesperiidae

Some butterfly species were common in both locations, i.e. D. belisama, P. agamemnon, P. memnon, P. sarpedon, Eurema hecabe, Neptis hylas matuta, Argynnis hyperbius and Elymnias hypermnestra. The other species found only in one location, such as Hypolimnas bolina, Lethe manthara, Melanitis phedima, and Euthalia adonia which only found in Bosscha. During the observation, some species found only one individual, for example seven species (P. demoleus, Cethosia penthesilea, Hypolimnas bolina, Elymnias nesaea, Lethe manthara, Melanitis phedima and Pareba vesta vestoides) found only one

53.03

40 30

2.55

1.75

Papilio sarpedon

Species

1.75

1.11

0.30

34 Species

3.98

Neptis hylas matuta

7.48

Junonia atlites

7.80

Eurema hecabe

10.35

Papilio agamemnon

Elymnias hypermnestra

A 35

32.19

30 25 20.64

20 15

11.08

7.67

5.78

4.25

3.89 Mycalesis sudra

Papilio memnon

Papilio agamemnon

19 Species

0.22

0 Junonia iphita horsfieldi

2 0 2 4 0 1 2 1 1 0 2 1 2 1 49 2 1

Anthene lycaenina

1 1 11 0 1 0 33 175 1 2 0 0 0 0 4 3 0

Cethosia penthesia

16 1 1 2 5 1 25 1 333 1 0 11 2 1 7 1 1 2 3 5 6 65 0 3 2 11 1 1 47

Papilio sarpedon

30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46

49 1 0 0 36 0 94 0 273 7 2 21 0 0 14 0 0 10 0 0 25 1 1 6 0 0 65 0 11

Ypthima philomela

Nymphalidae

Papilio agamemnon agamemnon P. demoleus sthenelinus P. demolion P. helenus enganius P. memnon memnon P. polytes javanus P. sarpedon sarpedon Appias lyncida Delias belisama belisama D. hyparete Eurema alitha E. hecabe Leptosia nina Faunis canens Neptis hylas matuta N. nandina nandina Pantoporia nefte nefte Danaus chrysippus Danaida eryx furius Euploea mulciber Argynnis hyperbius javanica Cethosia penthesilea penthesilea Hypolimnas bolina iphigenia H. misippus misippus Junonia almana javana Junonia atlites atlites J. iphita horsfieldi J. orithya minagara Elymnias hypermnestra hypermnestra E. nesaea Lethe manthara L. rohria godana Melanitis leda simesa M. phedima phedima Mycalesis mineus M. sudra sudra Ypthima philomela philomela Pareba vesta vestoides Euthalia adonia adonia Pyrameis cardui cardui P. dejeani Cyrestis lutea lutea C. nivea nivea Anthene lycaenina Notocrypta paralysos Discophora celinde celinde

Delias belisama

Pieridae

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29

Delias belisama

Papilionidae

Species

Cihideung

Family

Bosscha

Occurrence

individual in Bosscha. While in Cihideung, 16 species (Papilio demoleus, P. demolion, P. polytes, Appias lyncida, Delias hyparete, Faunis canens, Neptis nandina, Pantoporia nefte, Junonia iphita, Junonia orithya, Mycalesis mineus, Ypthima philomela, Pareba vesta vestoides, Pyrameis dejeani, Cyrestis nivea and Discophora celinde) found only one individual during the observations. Those species were in a high risk condition when the disturbance occurred in the habitat as consequences the species would be very vulnerable or even extinct.

Relative abundance (%)

Table 1. Butterfly occurrence at Bosscha and Cihideung in MayJune 2008.

Relative Relativeabundance abundance (%) (%)

Species

B Figure 1. The relative abundance of butterfly in Bosscha (a) and in Cihideung (b).

Host plants and food plants of butterfly Studies on plant species in the same locations found at least seven species of flowering plants as food sources for butterfly, i.e. Citrus aurantium, Cosmos caudatus, Eupatorium inulifolium, Gomphrena globosa, Hibiscus rosa-sinensis, Lantana camara and Tithonia diversifolia (Table 2). As food plant, L camara visited by several species of butterfly such as D. belisama belisama, Elymnias hypermnestra, and Papilio sarpedon. In Cihideung, Gomphrena globosa the purple flower was not only as food plants for Cethosia penthesilea, but also as host plants for Argynnis hyperbius javanica and Junonia iphita. This phenomenon was also observed by Nurcahya (2003), where the butterfly choose plants and flowers with an attractive light color, such as red, orange, yellow, and purple flowers as their host plants.

SUBAHAR &YULIANA â&#x20AC;&#x201C; Butterfly of Bosscha Observatory, Lembang

Respondent (%)

During observations, Bougainvillea spectabilis, Citrus aurantium and Macaranga tanarius found as host plants for several butterflies (Table 2). For example Citrus sp. was a host plant of Papilionidae, such as Papilio memnon, P. helenus, and P. demoleus. There was evidence that related groups of butterfly species associated with particular plants (as larva host plants or imago food plants) and that phenomenon was also found by Queiroz (2002).

40.41

40 30 23.97

17.12

15.07

10 3.42

0 a

Option

Figure 2. Respondent perception (n=86) on the advantage of butterfly garden (a. to embellish the observatory landscape; b. media to get information out of astronomy; c. to add another perspective field of science; d. media to study human interaction with the environment; e. media to learn more about the butterfly). 50

Respondent (%)

Potential utilization of butterfly As an ecosystem, Bosscha area provide many benefits to humans not only as tourism object but also as butterfly habitat, which was also provide a number of important services. The open-access to Bosscha ecosystem and the characteristic of ecosystem itself as public good often neglected for its value of the ecosystem services. The presence of an open space with flowering plant, as habitat and food for butterfly, will give an additional value for environmental education. Students perception (40.41%) consider that the presence of butterfly garden will increase their insights on the butterfly science; 23.97% of the students thought butterfly garden can be an additional object besides the observatory (stars observation); 17.12% of the students thought butterfly garden will embellish the observatory landscape (Figure 2). Most of the students (40.91%) was curious on butterfly life cycle (Figure 3), they were interested to observed (i) how butterfly eggs attached to host plant; (ii) how developmental phase or metamorphoses of butterfly happened, from egg to the caterpillar or larvae; and (iii) how the pupae emergence and become a young butterfly with its beautiful color. From economic aspect, about 58% of observatory visitors or domestic tourists are willing to pay the additional cost Rp. 3000 or more to visit butterfly garden in Bosscha (Figure 4). Based on these results, the potential of butterfly diversity in the area can be managed as a tourist object for conservation and environmental education.

40.91

40 30

22.73

20.78

20 13.64

10 1.95

0 a

Option

Figure 3. Public expectation (n=86) on the availability of information in butterfly garden (a. color and pattern of butterfly; b. butterfly life cycle; c. interaction among butterfly population; d. butterflies in their habitat; e. the origin of butterfly species).

D C

11%

33%

Table 2. Host and food plants use by several butterfly in Bosscha and Cihideung Plants species Family Species Amaranthaceae Gomphrena globosa

Butterfly species Family Species Nymphalidae Argynnis hyperbius Cethosia penthesilea Junonia iphita Asteraceae Cosmos caudatus Nymphalidae Eupatorium inulifolium Nymphalidae Ypthima philomela Lycaenidae Anthene lycaenina Tithonia diversifolia Pieridae Delias belisama Nymphalidae Ypthima philomela Lycaenidae Anthene lycaenina Euphorbiaceae Macaranga tanarius Pieridae Delias belisama Malvaceae Hibiscus rosa-sinensis Nymphalidae Papilionidae Nyctaginaceae Bougainvillea spectabilis Nymphalidae Pareba vesta vestoides Rutaceae Citrus aurantium Papilionidae Papilio demoleus Papilio memnon Papilio polytes Verbenaceae Lantana camara Pieridae Delias belisama Nymphalidae Ypthima philomela Papilionidae Note: HP = Host plants; FP = Food plants

45%

Role HP HP HP HP HP HP HP HP HP FP HP HP FP HP & FP HP & HP HP & FP HP HP HP

< Rp. 5000

Rp 5000

Rp 10000

> Rp 10.000

Figure 4. Respondent willingness (n=75) to pay for future butterfly garden facilities in Bosscha. A < Rp. 5000, B = Rp. 5000, C Rp. 10000, D > Rp. 10000.

Development plan of butterfly garden Learning from the experience of several places such as "The San Francisco Bay Areaâ&#x20AC;? whose use the garden as a habitat for insect conservation area, one of the insect groups is butterfly, it seems that recreational activities could harmonize with conservation of beneficial insects (Connor et al.

B I O D I V E R S IT A S 11 (1): 24-28, January 2010

2003). In Indonesia, the experience of "Gita Persada" butterfly garden through ecological engineering by habitat modification has been successful in bringing back the butterfly which was lost from the forest area of Mount Betung, Lampungâ&#x20AC;&#x201C;Sumatra Island (Soekardi 2002). Another experience in Bantimurung-Bulusaraung National Park (South Sulawesi), a sustainable community-based on butterfly rearing had been successfully maintaining butterfly diversity in natural habitat (Harlina 2006). Experiences and success stories in some places where habitat rehabilitation and management of open space through butterfly garden can give an added-value of the area as educational tourism object. The implementation of development plan conducted in several activities such as creating a semi-natural rearing facility for butterfly by using native species. Based on the relative abundance and butterfly characteristics, there were 12 species to be considered, i.e., P agamemnon, P. demoleus, P. memnon, P. sarpedon, Delias belisama, Eurema hecabe, Danaus chrysippus, Argynnis hyperbius, Cethosia penthesilea, Hypolimnas misippus, Melanitis phedima and Euthalia adonia. Habitat modification will be the most important steps in order to prepare host and food plant for butterfly. These activities consist of planting and preparing for rearing facilities. At least seven species could be prepared as host and food plants such as Citrus aurantium, Cosmos caudatus, Eupatorium inulifolium, Gomphrena globosa, Hibiscus rosa-sinensis, Lantana camara and Tithonia diversifolia (Detail technical implementation will be described in separate article). The development plan of butterfly garden in Bosscha area is one of the optimization of ecosystem services. As an ecosystem, Bosscha area and its vicinity has a higher value than astronomical tourist object. Referring to Daily et al. (2002), a variety of ecosystem services provided by Bosscha was: the existence value of the area not only as a habitat for butterfly and other biotic components but can be extended to the whole open space as water catchments area and flooding controller for Bandung area (as an indirect value). Butterfly diversity could be used as a media to provide knowledge for the local and regional communities on the importance of maintaining biodiversity and improving the quality of environment. CONCLUSIONS Twenty six species of butterfly has been found in Bosscha area and 41 species in Cihideung. These results indicated that Cihideung area could be considered as a source for butterfly diversity in Bosscha area. Delias belisama belisama was the dominant species, but for development plan of butterfly garden there were 12 species to be considered i.e. Papilio agamemnon, P. demoleus, P. memnon, P. sarpedon, Delias belisama, Eurema hecabe, Danaus chrysippus, Argynnis hyperbius, Cethosia penthesilea, Hypolimnas misippus, Melanitis phedima and Euthalia adonia. Bougainvillea spectabilis, Citrus aurantium, Lantana camara, Macaranga tanarius, Cosmos caudatus, Eupatorium inulifolium, Gomphrena globosa, Hibiscus rosa-sinensis and Tithonia diversifolia could be

considered as plants that can guarantee the presence of butterfly in Bosscha. The advantages of the presence of butterfly garden in Bosscha area not only giving additional insight for the visitors of Bosscha Observatory but also educating the community on preserving biodiversity and the environment. ACKNOWLEDGEMENT We thank to Bosscha Observatory Management (Dr. Taufik Hidayat) for his permission in using the facility. To DGHE (Directorate General for Higher Education), National Education Department for its support to SITHITB Bandung.

REFERENCES Anonymous (2007) Bul Observ Bosscha 4: 1-30. Boyer T, Polasky S (2004) Valuing urban wetland: a review of nonmarket valuation studies. Wetland 24: 744-755. Connor EF, Hafernik J, Levy J, Moore VL, Rickman JK (2003) Insect conservation in an urban biodiversity hotspot: the San Francisco Bay area. J Insect Cons 6: 247-259. Daily C, Alexander S, Ehrlich PR, Goulder L, Lubchenco J, Matson PA, Mooney HA, Postel S, Schneider SH, Tilman D, Woodwell GM (2002) Ecosystem services: benefit supplied to human societies by natural ecosystems. Issues Ecol 6: 1-18. Ernanto D (2006) Sinyal SOS bagi Observatorium Bosscha. Harian Umum Sore Sinar Harapan. 26 April 2006. Harlina (2006) Butterfly management as tourist object in BantimurungBulusaraung National Park (TNB), Kabupaten Maros, Sulawesi Selatan [Thesis]. Bandung Institute of Technology, Bandung. [Indonesian] Hedblom M (2007) Birds and butterflies in Swedish urban and peri-urban habitats: a landscape perspective [Dissertation]. Swedish University of Agricultural Sciences, Uppsala. Mueller-Dombois DM, Ellenberg H (1974) Aims and method of vegetation ecology. John Willey and Sons, New York. Noerdjito WA, Aswari P (2003) Survey method and fauna monitoring: 4th seri butterfly Papilionidae. Division of Zoology, Research Centre for Biology, Indonesian Institute of Science, Bogor. [Indonesia] Nurcahya A (2003) Plant communiy and its potential in supporting butterfly survival (sub-ordo Rhopalocera) at Tanjung Putus and Pulau Tegal, Teluk Lampung [Undergraduate Thesis]. Bandung Institute of Technology, Bandung. [Indonesian] Odum EP (1971) Fundamental of ecology. 3rd ed. W.B. Saunders, London. Preston K, Mafham (2007) 500 butterflies: butterflies from arround the world. Grange Books, Rochester. Queiroz JM (2002) Host plant use among closely related Anaea butterfly species (Lepidoptera, Nymphalidae, Charaxinae). Braz J Biol 62: 657-663. Roepke W (1932) De vlinders van Java. E. Dunlop & Co, Batavia. Smart P (1975) The illustrated encyclopedia of the butterfly world. The Hamlyn Publishing Group, London. Soekardi H (2002) Diversity of Papilionidae in Gunung Betung forest, Lampung, Sumatra: rearing and habitat engineering as baseline for conservation [Dissertation]. Bandung Institute of Technology, Bandung. [Bahasa Indonesia] Sreekumar PG, Balakrishnan M (2001) Habitat and altitude preferences of butterflies in Aralam Wildlife Sanctuary, Kerala. Trop Ecol 42: 277-281. Sundufu A, Dumbuya R (2008) Habitat preferences of butterflies in the Bumbuna forest, Northern Sierra Leone. J Insect Sci 8: 64. Tati-Subahar SS, Amasya AF, Choesin DN (2007) Butterfly (Lepidoptera: Rhopalocera) distribution along an altitudinal gradient on Mount Tangkuban Parahu, West Java, Indonesia. Raff Bull Zool 55: 175-178 Yamamoto N, Yokoyama J, Kawata M (2007) Relative resource abundance explains butterfly biodiversity in island communities. Proc Natl Acad Sci USA 104: 10524-10529.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 29-33

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110107

Diversity and community structure of dung beetles (Coleoptera: Scarabaeidae) across a habitat disturbance gradient in Lore Lindu National Park, Central Sulawesi SHAHABUDDINâ&#x2122;Ľ Department of Agrotechnology, Faculty of Agriculture, Tadulako University, Tondo, Palu 94118, Central Sulawesi, Indonesia, Tel./fax.: +62-451429738, email: shahabsaleh@gmail.com Manuscript received: 12 November 2009. Revision accepted: 23 December 2009.

ABSTRACT Shahabuddin (2010) Diversity and community structure of dung beetles (Coleoptera: Scarabaeidae) across habitat disturbance gradient in Lore Lindu National Park, Central Sulawesi. Biodiversitas 11: 29-33. Dung beetles are important component of most terrestrial ecosystems and used to assess the effects of habitat disturbance and deforestation. This study aimed at comparing dung beetle assemblages among several habitat types ranging from natural tropical forest and agroforestry systems to open cultivated areas at the margin of Lore Lindu National Park (LLNP), Central Sulawesi (one of Indonesiaâ&#x20AC;&#x2122;s biodiversity hotspots). Therefore, 10 pitfall traps baited with cattle dung were exposed at each habitat type (n = 4 replicate sites per habitat type) to collect the dung beetles. The results showed that species richness of dung beetles declined significantly from natural forest to open area. However cacao agroforestry systems seemed to be capable of maintaining a high portion of dung beetle species inhabiting at forest sites. The closer relationship between dung beetle assemblages recorded at forest and agroforestry sites reflects the high similarity of some measured habitat parameters (e.g. vegetation structure and microclimate) between both habitat types, while species assemblages at open areas differed significantly from both other habitat groups. These results indicated that habitat type has importance effect on determining the species richness and community structure of dung beetles at the margin of LLNP. Key words: forest conversion, diversity, habitat selection, dung beetles marker.

INTRODUCTION The deforestation of tropical forests and their subsequent conversion to human-dominated land-use systems, such as agricultural land, is one of the most significant causes of biodiversity loss. This also occurs in Sulawesi where the deforestation rate has reached nearly 190,000 ha/year (Holmes 2000), and almost all lowland forests below 400 m asl. have been lost (FWI and GWF 2002). Dung beetles (Coleoptera: Scarabaeidae) are ecologically important since their dung burial activity could maintain soil fertility (Omaliko 1984), increase plant yields (Miranda et al. 2001) and dung-seed dispersal activity promotes plant regeneration (Andresen 2002, 2003). However, they have proved to be sensitive to habitat perturbation, such as canopy forest loss (Davis and Sutton 1998) and human habitat modification (Shahabuddin et al. 2005). Therefore, they widely used as biological indicators for evaluating the effects of habitat disturbance driven by human activities. Lore Lindu National Park (LLNP) in Central Sulawesi is one of the core areas for the protection of the biodiversity of Wallacea (Myers et al. 2000). Generally, forest habitats in the interior of LLNP are still relatively undisturbed while the margins of the park are characterized by a mosaic of near-primary forests, degraded forests, forest gardens and plantations of cacao, coffee, maize and

paddy rice as the most important crops. Although the forest conversion has strong negative effects on biodiversity (Lawton et al. 1998), some agroecosystems including traditional agroforestry systems, such as cacao and coffee cultivations established under the diverse layers of forest trees, can maintain a relatively high portion of tropical biodiversity (Harvey et al. 2006). Therefore, there is an increasing recognition that biodiversity conservation should also include efforts to a sustainable management of human-modified ecosystems outside of the protected areas, which are capable to maintaining at least a certain fraction of tropical biodiversity (Mendoza et al. 2005; Tscharntke et al. 2005; Bos et al. 2007). Studies on dung beetles have been conducted in some islands of Indonesia, such as in North Sulawesi (Hanski and Niemela 1990; Hanski and Krikken 1991), Sumatra (Gillison et al. 1996), and West Java (Noerdjito 2003), and Shahabuddin et al. (2005, 2007). More attention has to be paid to Sulawesi Island since with its unique geographical history (Whitten et al. 2002), about 75% of dung beetles species inhabited in Sulawesi are most likely endemic species (Hanski and Krikken 1991). However, most of those studies only focused on forest sites and did not take into account the relative contribution of land-use activities such as agroforestry on dung beetles diversity. This study aims to address the following research questions: (i) How forest conversion to land-use systems at

B I O D I V E R S IT A S 11 (1): 29-33, January 2010

the margin of LLNP affects the species richness and community structure of dung beetle and whether the species richness of dung beetles inhabiting natural forest is significantly higher than those in land-use system (agroforestry systems or annual cultures). (ii) How significant the contribution of agroecosystems to the total (regional) species richness of dung beetles is. (iii) What habitat variables will be the best predictors for explaining differences in dung beetle communities between land-use type.

(2005). On each site 10 traps were set up along a 50 m transect. Traps were baited and exposed synchronously to dung beetles three times during (May-July 2009). Traps were baited with ca. 20 g of fresh cattle (Bos taurus) dung collected from pasture land around the study sites. Trapped specimens were removed from the traps after two days and preserved in Scheerpelz solution as recommended by Krell (2007). Later on, samples were identified in the laboratory with available identification keys (e.g. Balthasar 1963). The reference collection of the Center for Biodiversity Research Tadulako University was used. Taxa, which could not be identified, were sorted to morphospecies.

MATERIALS AND METHODS Study area The study area is located on the northern margin of the Lore Lindu National Park in Central Sulawesi (LLNP)Indonesia. The Lore Lindu National Park, a local biodiversity hotspot is covering an area of 229,000 ha and located southeast of Palu, the province capital of Central Sulawesi. All study sites were selected in Palolo Valley in the vicinity of the villages of Bobo (01o07'10.2" S119o59'40.2" E) and situated at an altitude between 800 and 1000 m asl. Dung beetle communities were studied in four habitat types: - natural forest (NF), selectively logged forest (SF), agroforestry systems (cacao plantations with Gliricidia as shadow trees; AF) and annual cultures (maize fields, AC). For each habitat type three site replications were selected. Detailed description of each habitat type was shown in Table 1.

Table 1. Description of each habitat type studied. Habitat type Natural Forest

Habitat description

Lower montane forest; big emergent trees and numerous medium-sized trees form a multi-layered canopy; height of upper canopy layer 20-30m with single big emergent trees up to 40m; welldeveloped understorey layer of small trees/scrubs, ginger and rattan up to 4-8m high; herb layer dominated by Rubiaceae species and ferns. Selectively Single emergent trees up to 30 m; closed logged canopy layer 15-20 m high; herb layer forest 0.5-2m high and dominated by ferns and Rubiaceae sp. Some selective logging activities took place in all sites, however, the plots are so far just slightly affected. Agroforestry Ca. 5 yrs old cacao plantations (ca. 1 ha) system with Gliricidia sepium (Leguminosae) and Musa sp. as shaded trees; cacao trees up to 2-3 m high; Gliricidia sepium trees 7-9 high; some sites has herb layer with 20-30 cm high Annual Maize fields (0.5-1 ha). culture

Site codes NF1-3

SF1-3

CP1-3

Characterization of habitat type Several habitat parameters (vegetation structure and microclimate) were measured and tested for their potential to predict changes in the diversity and community structure of dung beetles (Shahabuddin et al. 2005).: 1). air temperature and relative humidity at about 1 m above ground at start and end of the exposing period were measured by using a digital thermo-hygrometer (CoronaR Model: GL 99), 2). Canopy cover visually estimated at four locations per site for a corridor of ca. 10 m beside the transect lines, 3). number of trees (dbh â&#x2030;Ľ 10 cm) measured at four of 5x5 m2 plots beside each transect, one plot every 10 m changing between both sides of the transect, and 4). herb layer coverage was estimated at four plots of 2x2 m2 randomly placed at ca. 5 m beside the transect line. Data analysis Species richness of dung beetles was estimated using the second-order jackknife extrapolation method (Colwell 2004), one of the best species richness predictor with respect to accuracy (e.g. Brose et al. 2003). As units for estimating the total species richness of individual sites, samples pooled for individual sample times (n = 4) were used). Effects of habitat type on species richness were tested using one-way ANOVA. Abundance data were log (n+1) transformed before analysis (Zar 1999). The BrayCurtis similarity index was used to quantify between-site (beta) diversity of dung beetles (Krebs 1999). The similarity matrices resulted was used to compare the similarity of species composition of dung beetles between habitat type and to construct a two-dimensional ordination of all samples using multidimensional scaling (Clarke 1993). The program Statistica 6.0 (StatSoft ,2001) were used to perform all statistical analyses. Second-order jackknife calculations were computed with the program EstimateS Version 7.00 (Colwell 2004) by randomizing the ranking of samples 100 times.

RESULTS AND DISCUSSION AC1-3

Specimens collection Dung beetles were sampled in 50m2 plots at 12 sites all using baited pitfall traps as described in Shahabuddin, et al.

Diversity of dung beetles A total of 1696 dung beetles were collected during study period. They belonged to 3 genera (dominated by Onthophagus) and 24 species. The five most predominant species were Onthophagus wallacei, O. fuscostriatus, O. limbatus, Copris macacus, and O. ribbei (Table 2).

SHAHABUDDIN â&#x20AC;&#x201C; Forest conversion, effects on dung beetles

habitat type. Three of those species were significantly highest in the natural forest (Figure 2 a-c), while the rest was highest in annual culture (Figure 2 d-f).

Species richness

Changes natural forest to other land-use type has significantly affect on species richness (ANOVA: F(3,8) = 7.46, p < 0.05, Figure 1a) but not on the abundance of dung beetles (Figure 1b). Although species richness was decreased from natural forest to open cultivated area, this study only detected a significant reduction of species richness on open cultivated area. Species richness at the natural forest, disturb forest and cacao agroforestry were nearly similar (Figure 1b). 16 14 12 10 8 6 4 2 0

Habitat type

Figure 1. Species richness (left) and abundance (right) of dung beetles in relation to habitat type. (NF = natural forest, SF = selectively logged forest, AF = agroforestry system, AC = annual culture). Arithmetic means Âą SD are given. Different letters on top of the bars indicate significant differences between habitats

Further analysis to show species level response to habitat changes revealed that six of the seven most abundant dung beetles species (total of â&#x2030;Ľ70 collected specimens) recorded in all habitats significantly respond to

Number of specimens

(a)

70 60 50 40 30 20 10

180 160 140 120 100 80 60 40 20 0

(d)

b b b NF

(b) a

20 a

15 10 b

0 NF

Number of specimens

40 30 20 10

(c)

20.0 ab ab

5.0 b

0.0 NF

Habitat type

10.0

(e)

-10

25.0

15.0

Habitat type 30.0

Habitat type

Number of specimens

Onthophagus wallacei Harold* O. fuscostriatus Boucomont* O. limbatus (Herbst) Copris macacus Lansberge* O. ribbei Boucomont Onthophagus. sp. 2 O. trituber Wiedemann Onthophagus sp. 6 Onthophagus sp. 5 Aphodius sp. 1 O. rudis Sharp Camponotus saundersi Harold O. holosericeus Harold* C. punctulatus Wiedemann Onthophagus sp. 3 Aphodius sp. 2 O. rectecormutus Lansberge O. forsteni Lansberge O. aureopilosus Boucomont* Onthophagus sp. 9 Onthophagus sp. 8 Onthophagus sp. 7 Onthophagus sp. 4 Onthophagus sp. 2

Number of specimens

Species

90 80

Number of specimens

100

Habitat Number of NF SF CP AC specimens 70 23 62 433 588 223 198 4 0 425 0 3 1 144 148 64 18 24 0 106 53 33 8 1 95 28 25 14 5 72 0 0 5 65 70 26 3 4 8 41 0 1 1 31 33 2 0 0 26 28 1 3 11 10 25 21 3 0 0 24 1 1 11 0 13 2 0 5 4 11 0 0 4 0 4 1 0 0 2 3 0 0 0 2 2 0 0 2 0 2 0 1 0 0 1 1 0 0 0 1 1 0 0 0 1 0 1 0 0 1 1 0 0 0 1 1 0 0 0 1

Habitat type

Number of specimens

Table 2. Number of collected specimens of individual dung beetle species in the four different habitat types. Species are ranked according to their total number of collected specimens. Species most likely endemic to Sulawesi according to Balthasar (1963) are indicated with an asterisk

30.0

(f) a

25.0 20.0 15.0 10.0 5.0

0.0 CP

Habitat type

Figure 2. Mean number of dung beetles in each of the four habitat types, given for the six most abundant species. a: Onthophagus fuscostriatus (ANOVA F3,8=9.47, p<0.05), b: Onthophagus ribbei (F3,8=22.31, p<0.05), c: Copris macacus (F3,8=5.99, p<0.05), d: O. wallacei (F3,8=8.70, p<0.05), e: Onthophagus limbatus (F3,8=1.04, p<0.01), f: Onthophagus trituber (F3,8=59.31, p<0.01). Different letters on top of the bars indicate significant differences between habitats.

This study documented that the conversion of natural forest to secondary forests and land use systems had a negative effect on dung beetle diversity. A decline of insect diversity related to forest disturbance and conversion of forests or forest plantations to agricultural habitats has been also shown for other taxonomic groups (e.g. Schulze 2004; Shahabuddin et al. 2007; Susilo et al. 2009). Although the structure of dung beetle communities changed between natural forest, selectively logged forests and the two landuse systems. Each of the tree habitat types following destruction of natural forest supported about 70% of the species richness found in the natural forest, thereby indicating a surprisingly little reduced diversity despite the great human-induced habitat transformation. Davis et al. (2001) reported a significant decrease of dung beetle diversity across a gradient of forest disturbance ranging from primary to logged and plantation forest in Northern Borneo. Recent analysis shows a strong and negative response of by tropical forest dwelling dung beetle communities to increasing modification of tropical forest and declining fragment size (Nichols et al. 2007). However, in general, the damage caused by logging and forest conversion seems to depend on the intensity of the logging and land-use. High intensity logging is known to have a negative effect on coprophagous beetles and other insect groups (Hill 1995; Davis et al. 2001) but a reducedimpact logging system has better preserved the coprophagous beetles assemblage of primary forest than

B I O D I V E R S IT A S 11 (1): 29-33, January 2010

conventional logging techniques (Davis 2000). This result could be explained by using intermediate disturbance hypothesis (reviewed by Collins and Glenns 1997) predicts that diversity will be highest in communities with intermediate levels of disturbance. If disturbances are too rare, the competitive dominants will eliminate other species and reduce diversity as equilibrium conditions develop. If disturbances are too frequent, most species will go locally extinct, which lowers diversity, because they can not tolerate repeated disturbances. Under intermediate levels of disturbance, diversity is maximized because disturbancetolerant species and competitively dominant species coexist. Species composition of dung beetles Change of habitat types has a tremendous effect on species composition. An ordination technique aimed to compare dung beetles community at habitat studied showed a higher distance between open area and natural forest, agroforestry cacao and annual culture compared to the distance among those three latter habitats (Figure 3). This result indicated that the structure of dung beetle communities differed between forest habitats and land-use systems. However, the dung beetle species did not appear to be restricted to only one habitat type causing a high overlap of the species composition between all sites (similar to the results from the community analysis using Sørensen indices). Of the 20 species recorded at forest sites, 60% (= 12 species) were also found at the cacao plantations. Although four of Onthophagus sp. and one of Aphodius sp. were only recorded at forest site they all could not be categorized as “forest-dependent species” since their abundance were very low (1 specimen per species) and those species could be collected in other landuse type if the sampling period was longer. Moreover, if the singleton was excludes about 90% of the species recorded at forest sites were also occurs at the cacao plantation. This species distribution indicating the high potential of agroforestry system for supporting the dung beetles diversity at the landscape level. Nevertheless, the contribution of agroforestry to the dung beetles (and overall) diversity depend on some factors such us management of shaded tree and the distance to the nearby forest (Bos et al. 2007; Clough et al. 2009). 1.2 CP2

1.0

Stress = 0.04

CP1

0.8 CP3

Dimension 2

0.6 0.4 SF1

0.2

Habitat parameters Canopy cover (%) Number of tree Herb coverage (%) Temperature (oC) Humidity (%)

NF 85.0 (5.0) 7.7 (1.2) 56.7 (5.8) 25.4 (1.3) 76.3 (5.5)

Habitat type SF AF AC 63.3 (2.9) 45.0 (5.0) 0.0 (0) 6.0 (1.0) 5.7 (0.6) 0.0 (0) 66.7 (5.8) 63.3 (2.9) 73.3 (2.9) 25.2 (1.2) 27.1 (2.3) 27.2 (1.7) 70.7 (1.2) 69.3 (0.6) 68.3 (3.1)

Besides effects of an uneven distribution of food among hab it at s a nd t he i n f l ue n ce o f so i l t yp e ( Ha n s ki and Cambefort 1991), microclimate conditions have a critical effect on habitat selection by coprophagous beetles (Barbero et al. 1999). Therefore, changes in microclimate following the alterations of vegetation structure at four studied land-use type may also affect coprophagous beetle populations. It is known that thermoregulation strategies and the ability to maintaining an optimal body temperature may restrict coprophagous beetles to a certain range of microclimatic conditions (Verdu et al. 2004, 2006). Closed canopy cover, lower temperature most likely suitable for O. ribbei, O. fuscostriatus, and Copris macacus because they more abundance in forest sites and cacao agroforestry system (see Figure 2 a-c). On the contrary O. wallacei, O. limbatus, and O. trituber were prefer to inhabit in drier habitat or sites with low canopy cover such as open cultivated area (see Figure 2 d-f). This is partly in line with previous study in Napu situated at southern part of Lore Lindu National Park. In that area O. ribbei, O. fuscostriatus, and Copris macacus were more abundance in cooler and shaded habitat (e.g. natural forest and cacao agroforestry) (Shahabuddin et al. 2005). In South Africa, dung beetles fauna were also showed a habitat preferences and grouped by Davis et al. (2002) as ‘shaded species” and “unshaded species”.

AC3

CONCLUSION

-0.2

-0.6

Table 3. Habitat characteristic at four habitat type studied in Lore Lindu National Park (NF=natural forest, SF= selectively logged forest, AF= agroforestry system, AC= annual culture)

NF3

0.0

-0.4

Relation with habitat parameter Changes of dung beetles diversity and composition may relate to some habitat parameters measured. Air temperature and herbs coverage increased from natural forest to annual culture, while humidity, canopy cover and number of tree showed a reverse pattern (Table 3). This result was concordance with previous study showed that some species of dung beetles response differently to changes of land-use type in Central Sulawesi (Shahabuddin et al. 2005).

AC1

SF3 SF2

AC2

NF2

NF1

-0.8 -1.0 -0.8 -0.6 -0.4 -0.2 0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

1.6

Dimension 1

Figure 3. Two-dimensional scaling plot based on Bray-Curtis indices quantifying similarity of dung beetle communities sampled at 12 sites belonging to four different habitat types. Lines connect sampling sites belonging to the same habitat.

Species richness and species composition of dung beetles changed following the conversion of natural forest to others land-use types at the margin of Lore Lindu National Park. Species richness of dung beetles tended to decrease with increasing of land-use intensity indicating a pronounced negative effect of land-use changes on the dung beetle fauna. However, contrasting to the open areas,

SHAHABUDDIN – Forest conversion, effects on dung beetles

cacao agroforestry systems with vegetation structure more resembling natural forest proved inhabited by a dung beetle fauna which was similar to the one found at forest sites. Although species richness of dung beetles at natural forest was higher compared to all others land-use types, agroforestry sites contributed a high level (at least 60%) to the diversity of dung beetle communities recorded at Palolo region. Although few species showed a high association with certain habitat type most of them were not restricted to certain habitat but seems to occupy a wide range of habitats indicating that majority of dung beetles in Central Sulawesi are generalist and do not have strongly pronounced habitat requirements. ACKNOWLEDGEMENTS This paper was a part of Fundamental Research funded by Directorate of Higher Education Ministry of Education Indonesia (Contract No. 155.20/H38.2/PL/2009). The author thank the head of Balai Taman Nasional Lore Lindu, who kindly provided permissions to conduct this study. REFERENCES Andresen E. (2002) Dung beetles in a Central Amazonian rainforest and their ecological role as secondary seed dispersers. Ecol Entomol 27: 257-270. Andresen E. (2003) Effect of forest fragmentation on dung beetle communities and functional consequences for plant regeneration Ecography 26 (1): 87-97. Balthasar V (1963) Monographie der Scarabaeidae und Aphodiidae der palaearktischen und orientalischen Region. Band 1. Verlag der Tschechoslawakischen Akademie der Wissenschaften. Prague. Barbero E, Palestrini C, Rolando A. (1999) Dung beetle conservation: effects of habitat and resource selection (Coleoptera: Scarabaeoidea). J Insect Cons 3: 75-84. Bos M, Höhn P, Shahabuddin S, Buchori D, Steffan-Dewenter I, Tscharntke T (2007) Insect responses to forest conversion and agroforestry management. In: Tscharntke T, Leuschner C, Guhardja E, Zeller M (eds). The stability of tropical rainforest margins: linking ecological, economic and social constraints of land-use and conservation. Springer, Berlin. Brose U, Martinez ND, Williams RJ (2003) Estimating species richness: sensitivity to sample coverage and insensitivity to spatial patterns. Ecology 84 (9): 2364-2377. Clarke KR (1993) Non-parametric multivariate analyses of changes in community structure. Aust J Ecol 18: 117-143. Clough Y, Dwi-Putra D, Pitopang R, Tscharntke T (2009) Local and landscape factors determine functional bird diversity in Indonesian cacao agroforestry. Biol Cons 142: 1032-1041 Collins SL, Glenn SM (1997) Intermediate disturbance and its relationship to within- and between-patch dynamics. N Z J Ecol 21 (1): 103-110. Colwell RK (2004) EstimateS: Statistical estimation of species richness and shared species from samples. Version 7. User's Guide and application. http://purl.oclc.org/estimates. Davis AJ, Holloway JD, Huijbregts H, Krikken J, Sutton SL. (2001). Dung beetles as indicators of change in the forests of Northern Borneo. Journal of Applied Ecology 38: 593−616. Davis AJ, Sutton SL (1998) The effects of rainforest canopy loss on arboreal dung beetles in Borneo: implications for the measurement of biodiversity in derived tropical ecosystems. Div Distrib 4: 167-173. Davis AJ. (2000). Does reduced-impact logging help preserve biodiversity in tropical rainforest? A case study from Borneo using dung beetles (Coleoptera: Scarabaeoidae) as indicators. Enviromental Entomology 29 (3): 467-475. Davis ALV, Van Arde RJ, Scholtz CH, Delport H (2002) Increasing representation of localized dung beetles across a chronosequence of regenerating vegetation and natural dune forest in South Africa. Global Ecol Biogeograph 11: 191-209. FWI, GFW [Forest Watch Indonesia, Global Forest Watch] (2002) The

State of the Forest: Indonesia. Forest Watch Indonesia, Bogor, Indonesia and Global Forest Watch, Washington, DC. Gillison AN, Liswanti N, Arief Rachman I (1996) Rapid ecological assessment Kerinci Seblat National Park buffer zone preliminary report on plant ecology and overview of biodiversity assessment. Working Paper No. 14. Jambi: Kerinci Seblat NP. Hanski I, Cambefort Y (ed) (1991) Dung beetle ecology. Princeton University Press, Princeton Hanski I, Krikken J (1991) Dung beetles in tropical forests in South-East Asia. In Hanski I, Cambefort Y (eds.). Dung beetle ecology. Princeton University Press, Princeton Hanski I, Niemela J (1990) Elevational distributions of dung and carrion beetles in Northern Sulawesi. In: Knight WJ, Holloway JD (eds). Insects and the rain forests of South East Asia (Wallacea). Royal Entomological Society of London, London, UK. Harvey CA, Gonzalez J, Somarriba E (2006) Dung beetle and terrestrial mammal diversity in forests, indigenous agroforestry systems and plantain monocultures in Talamanca, Costa Rica. Biodiv Cons 15: 555-585. Hill JK, Hamer KC, Lace LA, Banham WMT (1995) Effects of selective logging on tropical forest butterflies on Buru, Indonesia. J Appl Ecol 32: 754-760. Holmes D (2000) Deforestation in Indonesia: a review of the situation in Sumatra, Kalimantan and Sulawesi. World Bank, Jakarta, Indonesia. Krebs CJ (1999) Ecological methodology. 2nd ed. Addison-Welsey, California. Krell FT (2007). Dung beetle sampling protocols. Denver Museum of Nature & Science (DMNS)Technical Report, Denver, CO. Lawton JH, Bignell DE, Bolton B, Bloemers G,F, Eggleton P, Hammond PM, Hodda M, Holt RD, Larsen TB, Mawdsley NA, Stork NE, Srivastava DS, Watt AD (1998) Biodiversity inventories, indicator taxa and effects of habitat modification in tropical rain forest. Nature 391: 72-76. Mendoza CAO, Rios AM, Cano EB, Corte’s JLN (2005) Dung beetle community (Coleoptera: Scarabaeidae: Scarabaeinae) in a tropical landscape at the Lachua Region, Guatemala. Biodiv Cons 14: 801-822. Miranda CHB, dos Santos JC, Bianchin (2001) The role of Digitonthophagus gazella in pas-ture cleaning and production as a result of burial of cattle dung. Pasturas Tropicales 22 (1):14-18. Myers N, Mittelmeier RA, Mittelmeier CG, da Fonseca GAB, Kent J (2000) Biodiversity hotspots for conservation priorities. Nature 493: 853-858. Nichols E, Larsen T, Spector S, Davis AL, Escobar F, Favila M, Vulinec K (2007) Global dung beetle response to tropical forest modification and fragmentation: a quantitative literature review and meta-analysis. Biol Cons 137: 1-19. Noerdjito WA (2003) Keragaman Kumbang (Coleoptera). In: Amir M, Kahono S (eds.). Serangga Taman Nasional Gunung Halimun Jawa Bagian Barat. JICA Biodiversity Conservation Project, Bogor (Indonesian). Omaliko CPE (1984) Dung decomposition and its effects on the soil component of a tropical grassland ecosystem. Trop Ecol 25: 214-220. Schulze CH, Waltert M, Kessler PJA, Pitopang R, Shahabuddin, Veddeler D, Steffan-Dewenter I, Mühlenberg M, Gradstein SR, Tscharntke T (2004) Biodiversity indicator groups of tropical land-use systems: comparing plants, birds, and insects. Ecol Appl 14 (5): 1321-1333 Shahabuddin, Hidayat P, Noerdjito WA, Manuwoto S (2007) Response of coprophagus beetles (Coleoptera: Scarabaeidae) on changes of vegetation structure in various habitat types at Lore Lindu National Park, Central Sulawesi. Biodiversitas 8 (2): 1-6 (Indonesian). Shahabuddin, Schulze CH, Tscharntke T (2005) Changes of dung beetle communities from rainforests towards agroforestry systems and annual cultures. Biodiv Cons 14: 863-877. Susilo FX, Indriyati, Hardiwinoto S (2009) Diversity and abundance of beetles (Coleoptera) functional groups in a range of land use systems in Jambi, Sumatra. Biodiversitas 10 (2): 195-200. Tscharntke T, Klein AM, Kruess A, Steffan-Dewenter I, Thies C (2005) Landscape perspectives on agricultural intensification and biodiversity-ecosystem service management. Ecol Lett 8: 857-874. Verdu JR, Arellano L, Numa C (2006) Thermoregulation in endothermic dung beetles (Coleoptera: Scarabaeidae): effect of body size and ecophysiological constraints in flight. J Insect Physiol 52 (8): 854-860 Verdu JR, Diaz A, Galante E (2004) Thermoregulatory strategies in two closely related sympatric Scarabaeus species (Coleoptera: Scarabaeinae). Physiol Entomol 29: 32-38. Whitten AJ, Mustafa M, Henderson G S (2002) The ecology of Sulawesi. Periplus, Hong Kong. Zar JH (1999) Biostatistical analysis. 4th ed. Prentice-Hall Inc. New Jersey.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 34-39

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110108

Potentialities of line planting technique in rehabilitation of logged over area referred to species diversity, growth and soil quality PRIJANTO PAMOENGKASâ&#x2122;Ľ Department of Silviculture, Faculty of Forestry, Bogor Agricultural University, Kampus IPB Darmaga, P.O. Box 168, Bogor 16680, West Java, Indonesia, Tel. +62-251-8626806, Fax. +62-251-8626886, ď&#x201A;Šemail: prijantop@yahoo.com Manuscript received: 16 September 2009. Revision accepted: 31 December 2009.

ABSTRACT Pamoengkas P (2010) Potentialities of line planting technique in rehabilitation of logged over area referred to species diversity, growth and soil quality. Biodiversitas 11: 34-39. Human interventions in the utilization of tropical forest resources are experiencing unanticipated consequences. The selective logging practices generally cause considerable damage to vegetation and the soil surface. It is supposed that soil condition and vegetation growth rate is deteriorated and reduced. Therefore, scientist strongly argue that the only way to achieve sustainability of Indonesian natural forest will require that the production natural forest is managed through methods that are acceptable from the perspective of environment as well as timber production. This means that there will be a strong need and incentive for methods and innovative technology. For more than two decades, tropical rainforest in Indonesia have been managed intensively under the Indonesian selective cutting (TPI) and later on by the Indonesian selective cutting and replanting (TPTI) and then, selective cutting and line planting (TPTJ) system. TPTJ, as one example of selective cutting, recently become a proper alternative should be taken into consideration in the management of production natural forest in Indonesia by planting dipterocarp species in line. In this system, planting line (width 3 m) and intermediate line (width 17 m) are made alternately. The initial width of line is 3 m and to be expanded until 10 m within 5 years to introduce more light. The objective of this research was to assess growth and soil quality of TPTJ system. The object of research was TPTJ plot of various ages from 1 year to 7 years. For achieving the objective, 14 sample plots measuring 200 m x 200 m each, were laid out at research plots. The result showed that growth respond of Shorea leprosula toward the width of planting line was better comparing to Shorea parvifolia, but generally from this growth assessment it seems that individual tree has a successfully performance. In terms of soil quality, it was seen that planting line establishment of 3 m and more, does not cause soil properties decrease in the whole plots. Key words: selective logging, production forest, selective cutting and line planting, dipterocarp species, growth, soil quality.

INTRODUCTION Tropical rainforest is drawing much attention nowadays, primarily due to threats to its future sustainability is getting worse. Tropical forests generally have faster growth due to considerable amount of solar energy absorption, as well as important role in maintaining the balance of global carbon. In Indonesia the principal causes of natural forest destruction is commercial timber harvesting, especially dipterocarp species. The decrease and degradation of tropical forests affect not only the production of timber but also environment. The loss of biological diversity threaten the sustainable and development of forests ecosystem in the future. Maintaining biodiversity has become a global concern and requires the implementation of sustainable management practices at a range of spatial scale (production forests). The ecological importance of a species is very different and can have a direct effect on the community structure, and thus on overall biological diversity. For example, a dipterocarp species which support an endemic invertebrate fauna of hundred species evidently make a greater contribution to the maintenance of global biological diversity (Schulte 1996). In relation to this, rehabilitation of

degraded dipterocarps forests is most urgent matter from the viewpoints of both compensation or enrichment of ecosystems (for maintaining and the preservation of species population) and sustainable forests management. It is estimated that the growth and regeneration rate of dipterocarp species has decreased in the logged-over forest. Many of logged-over forests still contain enough residual dipterocarp trees, to give rise to a sufficient supply of ephemeral seedling stock. As a consequence of repeated selective cutting, residual dipterocarp trees are scarce or of lack high-valued timber. An additional source of valuable timber could be obtained by planting dipterocarps in line. Enrichment planting is applied to add natural regeneration where this is insufficient. The best known enrichment planting is line planting which has a number of variants throughout the tropics (Lamprecht 1989). The application in Indonesia, line enrichment planting means planting under the forest canopy. The common practice of high valued timber species, such as meranti, is planted in irregularly pattern as in TPTI system. This kind of traditional methods resulted in low growth performance. It is indicated by mean of annual increment diameter in permanent sample plot of less than 1 cm year-1. One of the problems in enrichment planting with

PAMOENGKAS – Line planting technique in rehabilitation of logged area

included in The study was conducted in PT. Sari Bumi Kusuma forest concession. The elevation is approximately 175-276 m above sea level. The area is originally covered with typical Mixed Dipterocarp Forests in which dominated by dipterocarpaceae species. The mean annual temperature is 25-28oC, while the mean rainfall is 3000 mm year -1. The soils in study site are dominated by Ultisol (Yellow-red Podsolic). Experimental design Vegetation collection was done in seven different ages of plantation, namely 1 to 7 years old. Method for data collection used combination of transects line and plot establishment (Moore and Chapman 1986). Several plots, i.e. size 20 m x 20 m for observation at tree level, size 10 m x 10 m for pole level, size 5 m x 5 m for sapling, and 2 m x 2 m for seedling, were established along the transect. Each transect line has 20 m width and 200 m length. Within one plot, there were two transects line. To ease the placement of observation plots, the sample units (plots) should possess similarity in terms of soil properties and slope (08%). Plots were installed at seven different ages, i.e. 1, 2, 3, 4, 5, 6, and 7 years old of plantation by using purposive sampling methods. The total sample plot was 7 plots x 2 replications are 14 plots. The each sample plot unit measuring of 200 m x 200 m or 4 ha, thus the totally area was 56 ha (Figure 1). The data collected in this research were primary data on diameter and height growth of Shorea leprosula and Shorea parvifolia at the ages of 1, 2, 3, 4, 5, 6, and 7 years, and soil data taken from depth of 010 cm and 10-20 cm, in the planting and intermediate line. The planting distance is 2.5 m x 20 m (200 plants ha-1). Within one sample plot, there were 3 planting line which serve as observation plots for diameter and height of S. leprosula and S. parvifolia, whereas soil sample was taken as composite sample. Through this false time series study (pseudo plots), there would be obtained some ideas concerning several indications of growth recovery. 1.5 m

1.5 m

17 m Intermediate line Planting distance

dipterocarps is that most species are adapted to narrow ecological niches, which prohibits successful establishment of most dipterocarps on planting. The most important conditions for success of line planting are light control, species choice and tending intensively (Weidelt 1996). The most critical treatment is improvement of light conditions of the seedlings and saplings by release cutting or canopy opening. The growth of South East Asian dipterocarp forests varies from 8 m3 ha-1 year-1 (Appanah and Weinland 1993) to 17 m3 ha-1year-1 (Evans 1982). Based on the findings, the new planting technique (line planting) is implemented to accelerate growth of dipterocarps species. In the management of production forest, its productivity becomes a criterion which should be considered. Productivity itself depends on site quality and the developed silvicultural technique. The main challenge in rehabilitation of logged over production forest is creating growth site condition which is suitable for growth of dipterocarp species. For the purpose of increasing productivity of logged over natural production forest, the application of selective cutting and line planting System (TPTJ) which afterwards change into the so called ”intensive TPTI” (intensive Indonesian selective cutting and planting) or ”SILIN” (intensive silviculture) is one alternative which should be considered in managing production natural forest by planting dipterocarp species in line. In this system, after selective cutting is practiced, alternate line of 3 m wide for planting and 17 m wide for conservation as intermediate line are made. The initial 3 m width of planting line will be expanded to 10 m within 5 years, to allow more lights to penetrate. It seems that this system give impression that it will probably create considerable effect on vegetation, soil quality, and finally on plant growth. Soil quality constitutes a complete description of specific condition of the soil for carrying out its function (Karlen et al. 1997). For this purpose, study on the change of soil quality, which up to now is rarely discussed, is very important in the practice of intensive TPTI. Information on soil quality which constitutes a sensitive indicator can be used to answer several questions related to probable decrease in forest productivity and amount of carbon stored in forest soil. Besides that, information on soil quality will help in investigating several key components of forest ecosystem, such as productivity and sustainability of forest management system, soil and water conservation, and contribution of forest area on global carbon cycle (O’Neill and Arnacher 2004). Therefore, soil quality is an important aspect in relation with issue of sustainability and environment (Lal 1998). The objective of this research is to analyze vegetation aspects, plant growth at several ages of one to seven years, and condition of soil quality at age of one to five years.

2.5 m

20 m Inter-line distance Planting line

Figure 1. Lay-out of TPTI intensive (TPTJ) implementation

MATERIALS AND METHODS Study area description The research was conducted from June through August 2007 in Seruyan Forest Cluster, Central Kalimantan which

Data analysis Statistical analysis. Vegetation data will be analyzed to find out species density and species composition in a certain plantation plot. From the above vegetation data,

B I O D I V E R S IT A S 11 (1): 34-39, January 2010

average of the stand height and diameter were calculated. On the other hand, data of soil physical, soil chemical and soil biological are analyzed by using Mann-Whitney test to compare median value of planting line and intermediate line. Analytical network process. The soil quality evaluation was obtained by clustering the result of weight count of every soil quality indicator. Some of soil parameters used was bulk density, soil aggregate stability, C–organic, N– total and C–mic. The mentioned weight value is found by using Analytical Network Process (Saaty 1996). The final value of soil quality is the results of multiplication between weighted value and score value.

RESULTS AND DISCUSSION Species composition The species composition was clustered into dipterocarpaceae and non-dipterocarpaceae. The mentioned group species can be seen in Table 1.

Table 1. Species composition and density in each study plot Stand (age)

Species density (N/ha)

Tree Pole Sapling 1 108 (13%) 90 (11%) 1040 (11%) 2 103 (25%) 115 (26%) 1160 (29%) 3 115 (10%) 175 (3 %) 940 (9%) 4 160 (24%) 64 (19%) 336 (19%) 5 154 (19%) 80 (15%) 464 (45%) 6 141 (28%) 100 (26%) 384 (58%) 7 115 (13%) 144 (39%) 400 (44%) Note: Figures in the brackets showed percentage species.

Seedling 5250 (12%) 12000 (11%) 4875 (15%) 3300 (94%) 4700 (15%) 6800 (60%) 3400 (64%) of dipterocarp

In general, it can be said that the number of species per ha in all level is fluctuating, there is no increase tendency or consistent decrease starting from plot of 1 year to 7 years. As regards with contribution of dipterocarp species towards the total number of species in all level, it showed that non dipterocarp species is the largest except for seedling in plot of 4, 6, and 7 years old, and sapling in plot of 6 years. Some tree species of dipterocarpaceae family which is classified commercial found in the plot among others are S. leprosula, Shorea laevis, Shorea johorensis, etc. In general sense, the total of regeneration and tree level in all research plots classified over average if it is referred to the regulation of Indonesian selective cutting and replanting system (TPTI). In our study, as resulting from selective cutting of the overstorey is expected to create numerous canopy openings, and increased light availability and changed the species composition. This finding in line with the research conducted by Griffis et al. (2001), and Angers et al. (2005) who stated that enhancing light availability induces advance regeneration and change coverage in the understorey, even the species richness of e layer.

The results showed that the proportion of dipterocarps species in all plots study is smaller compared to non dipetrocarp species. Referring to the contribution tree level of dipterocarp species in primary forests which reaches 18.4% (not presented), it appears that the value of dipterocarp species in plot study were still accepted. In the plots of primary forest are found as many as 38 species per ha for tree level. The same finding is also reported by Mauricio (1991) in Philippines who recorded 31 species per ha. An average, the total number of species found in the plot study were 35 species per ha. In average, the total seedling from dipterocarp species was relatively balanced compared to non dipterocarp. This was caused that dipterocarps species had trouble for competition with other species to dense crown condition. This species is very sensitive towards light so that have preference towards certain micro condition. Based on the regeneration observation in seedling level, selective cutting and line planting system apparently gave stimulus as indicated by density in plantation plots. In the case of sapling regeneration, it appears that the recruited seedling which already existed before, has not reached the sapling stage may be due to increasing competition. It is recommended Potentialities of line planting that in all plots should be subjected to some silvicultural treatment, such as liberation to give better growing space for dipterocarps species by reducing competition. In relation to biodiversity, Matthews et al. (2000) stated that forest ecosystems, which can be defined by presence of tree canopies that cover more than 10% of a site have not escaped the loss of biodiversity. Thus, the application of line planting system on management of logged-over forest caused on acceptable level changes on vegetation aspect. Growth respond Data of diameter and height growth of S. leprosula and S. parvifolia at ages of 1 to 7 years can be seen in Table 2. The diameters of two dipterocarps species, i.e. S. parvifolia, and S. leprosula were measured in planting line, which have different width, as consequences of increasing width of planting line according to increasing age of plantations. Table 2. Growth performances for species planted in study plot Species Shorea leprosula

Shorea parvifolia

Stand age (yr) 1 2 3 4 5 6 7 1 2 3 4 5 6 7

MAI diameter (cm yr-1) 0.78 (n= 356) 1.57 (n=158) 2.09 (n=162) 2.07 (n= 55) 2.08 (n= 15) 2.08 (n=12) 2.19 (n= 84) 0.70 (n= 15) 1.17 (n= 6) 2.30 (n= 17) 1.85 (n= 10) 1.35 (n=9) 1.48 (n=9) 1.60 (n=12)

MAI height (cm yr-1) 249.73 (n=356) 154.53 (n=158) 175.10 (n=162) 166.35 (n= 55) 168.89 (n= 15) 173.13 (n= 12) 175.11 (n= 84) 209.00 (n=15) 125.00 (n=6) 165.49 (n= 17) 136.25 (n = 10) 148.00 (n=9) 137.50 (n=9 ) 133.81(n=12)

PAMOENGKAS – Line planting technique in rehabilitation of logged area

Table 2 showed the growth performance of line planting. In general, it can be said that the growth of S. leprosula was better than S. parvifolia, though some trees failed to survive due to natural causes. At seven year of age, S. leprosula showed the greatest of Mean Annual Increment (MAI) in diameter of 2.19 cm year-1, followed by age 3, 6, 5, 4, 2, 1 year at 2.09 cm, 2.08 cm, 2.08 cm, 2.07 cm, 1.57 cm, and 0.78 cm respectively. One can say that there was a consistent increase in diameter growth, with increasing age of plants. S. leprosula seems to be benefited slightly from more open canopy (Weidelt and Banaag 1982). This finding was confirmed in this study. The results showed the importance of a sufficient amount of overhead light when planting S. leprosula. Mean Annual Increment in diameter of S. parvifolia was 0.70 cm to 2.30 cm per year. Generally, from this growth, it seems that individual tree has a successfully growth performances in terms of diameter and height. We recommend opening up the canopy along the line before planting. Lamprecht (1989) stated that the greater the width the better the light conditions. It seems that generalization when planting dipterocarps are dangerous. Dipterocarp species have various requirements for light in the planting line. At the present, line planting method is applied for enrichment planting under the TPTJ system in Indonesia. This system has been successfully implemented in 6 forest concessions model and now is being expanded to more than 20 concessions managing mostly large proportion of logged-over dipterocarp forests. The success of line planting depends on light penetration. Therefore, the most critical treatment is improvement of light conditions by release cutting or canopy opening. Light deficiency is the main factor causing death of plants. Light conditions are related to the line width and stand height. The line width changes depending on the light requirements of the species planted, for example, relatively light demanding species such as S. leprosula and S. parvifolia. Consequently, the planting line is expanded to introduce more light (Weidelt, 1996). Hence, direction of line, width, and maintenance method of line planting are important for its success. Appanah and Weinland (1993) emphasized the importance of removal of the overstorey in line planting. Dipterocarp seedlings are naturally adapted to germinate and establish in the forest understorey where it is dark and moist. Complete removal of the canopy may lead to high mortality of the dipterocarp seedlings, while removing too little may suppress growth. Using the line planting has advantage, among others, a wider range of dipterocarps species can be used and consequently, mortality due to heat and water stress will be reduced. In relation to our results, Dupuy and Chazdon (2008) suggested that the choice of management system for these secondary forests can strongly influence the direction and rate of succession. Management systems that mimic natural canopy gaps could enhance tree species diversity and favor the regeneration of shade tolerant species, potentially accelerating convergence of secondary stand to old-growth forest composition.

The difference of soil properties between planting line and intermediate line To find out whether there is a difference soil physical, chemical and biological property between planting line and intermediate line in every age is done with. Statistical test Mann-Whitney (Table 3) was used to asses soil physical, chemical and biological properties between planting line. Table 3. Median of soil physical, chemical and biological properties in all plot studied Soil 4 years 3 years 2 years 1 year properties PL IL PL IL PL IL PL IL Agregat stb 70.50a) 73.13a 73.98a 74.70a 75.61a 69.33a 67.13a 69.82a C-org 1.02a 0.85a 1.37a 0.86a 1.13a 1.25a 0.92a 1.59a N-total 0.09a 0.08a 0.12a 0.08a 0.10a 0.11a 0.09a 0.12a C-mic 917.40a973.30a880.20a541.70a933.40a867.40a746.00a721.50a Note: Number followed by the same letter at the same row showed that there were no significant differences at α level of 0.5%, PL = Planting Line, IL = Intermediate Line

C organic in all research plots including low category, namely in the range between 0.85% to 1.59% and included low for N – total namely between 0.08 to 0.12% with variability between plots is relative small. The changing responses of soil C – organic and N – tot is extremely influenced by biotic factor vegetation, abiotic (temperature and moisture) and management (Parton et al. 1987 in Handayani and Prawito 1998). Further was added by Gregorich et al. (1997) that soil organic matter level caused by two reasons first decomposition process increased due to environmental changes (soil moisture, aeration and soil temperature). Both C and N input is low due to the decrease of organic residue input in an ecosystem. Other possible cause is composition species as important actor for C – organic formation in which each species has difference in carbon of land C – organic input. Handayani (2004) stated that the lower level of total C and N may have resulted from a combination of lower C inputs because of less biomass C returned and greater C losses because of aggregate disruption. The trends toward lower of total C and N in disturbed land is probably caused by the breakdown of aggregates (Gupta and Germida 1988; Blair et al. 1995), and greater organic matter oxidation following deforestation (Handayani et al. 2001) The above opinions are met with field finding. As observed, in general sense soil C – organic in the whole research plots can be classified low. The low of C – organic and N – total in the related primary forest were due to increasing of decomposition processes in which implicates to C and N pool decrease. Other possibility low C – organic and N – tot is indigenous value from the primary forest ecosystem or in other words soil organic matter in the primary forest governed by environmental condition. The actual condition of land closure which began to dense was assumed to affect microorganism activities result in C – mic content was high in plots of 3 years old and 4 years old. The behavior of C – mic is in complex which is affected by soil temperature, soil moisture, pH, texture and microbial activities The results showed no difference in soil physical properties tested namely

B I O D I V E R S IT A S 11 (1): 34-39, January 2010

aggregate stability in the planting line and intermediate line,. and oil chemical properties, namely C- organic and N-total, . The same also observed in C-mic. Based on statistical test assessment, soil physical, chemical or land biological properties in 1 year plants (3 m width) did not differ with condition in intermediate line. The same conclusion was obtained in 2 years old plant (4 m width planting line), 3 years old plant (6 m width), 4 years old plant (10 m width). In other words that establishment of planting line did not affect soil properties decrease in the whole plots studied. Assumption or perception that planting line establishment in silviculture system will change soil properties was not proved. Nutrient leaching had not occurred in the planting line due to the fact that planting line was still shaded by meranti plants and there was still accumulation of fresh organic matter which resists the leaching process. Soil quality value Soil quality value in each plot evaluated through threshold soil quality is depicted in Table 4. The soil quality indicator used to decide soil quality value is bulk density, aggregate stability, C-organic, N-total and C-mic. Due to there is a dependent influence inter soil quality indicator mentioned so determination of soil quality value in the research plots were done by analytic network process (ANP). Table 4. Category of soil quality value Value of soil quality 8-10 6-7.9 4-5.9 2-3.9 0-1.9

Category Very good Good Moderate Low Very low

Final value of soil quality was obtained by multiplication between scoring value (0-10) as a result of laboratory analysis with weighted value. The final weighted value of C-mic is bigger compared with other Indicators. C-mic gives biggest contribution (0.302) towards soil quality as compared to other indicators, such as bulk density (0.075), aggregate stability (0.121), Corganic (0.268), and N-total (0.234). This indicates that Cmic plays an important role in deciding soil quality. The soil quality value which obtained from multiplication between scoring value and weighted value, can be seen in Table 5.

Table 5. The final soil quality value in each study plot Plot Primary forest 4 years 3 years 2 years 1 year

Soil quality value 4.707 3.937 4.205 4.130 4.055

Category Moderate Low Moderate Moderate Moderate

Table 5 showed that soil quality in the whole plots is in category low to medium. The soil quality conditions in the area of TPTJ of 3, 2 and 1 year old exist in medium class, and followed by 4 years old at 4.205, 4.130, 4.055, and 3.937 respectively. The value of soil quality in plantation of 4 years old were lower compared to the others plot because of less N-total and C-organic level at 0.08% and 0.97% respectively. These proved that lower level of total N and C-organic in the ecosystem of 3 years old plantation may have resulted from the combination of lower C inputs because of less biomass returned or faster decomposition processes. In other words, silviculture practices should be directed into site improvement such as creating micro climate in planting line, to allow more lights to penetrate and to increase decomposition processes. Soil quality is a critical component of sustainable agriculture, while in term soil quality is relatively new, it is well known that soils vary in quality and that soil quality changes in response to their use and management (Handayani 2001). Type of land use is sustainable only when soil quality is maintained or improved. In this case, a quantitative assessment of the changes in soil quality provides a measure of sustainable management. Carter (2002) stated that a major component of sustainable land use is to sustain and improve the quality of the soil resource base. Monitoring is important, but the usefulness of the data will only be realized if it is used in management decisions to correct or improve the quality of the soil resource. Increasing forest productivity that maintaining soil quality has been a continuing and fundamental management issue for forest soil researchers. Line planting system so called intensive silviculture, allowing foresters to use intensive management practices that increase productivity above native levels while avoiding activities that degrade soil quality (Carter and Foster 2006). This is clearly demonstrated by this research shows growth and soil quality tends to recover after planting, implying that forest productivity increased both by shortening the rotation and increasing carrying capacity (Burger 2009). Logged-over forest invariably decrease soil organic matter content and open nutrient cycles (Olson et al. 2000). In turns, soil productivity decreases. The successes of line planting system using intensive silviculture in dipterocarps plantations have increased diameter growth by around 2 cm per year depending on the site. In relation to this, Fox et al. (2007) stated that practice on soil treatment, fertilization, and weed control have increased pine plantation productivity by 5-10 m3 ha-1 year-1. This increased productivity has been achieved by understanding forest response to each forest practices as well as the cumulative effects of intensive silviculture.

CONCLUSIONS In general, plant growth of S. leprosula was better than that of S. parvifolia. Diameter and height growth of S. leprosula showed positive respond, whereas for S. parvifolia this condition was not favorable. The overview

PAMOENGKAS â&#x20AC;&#x201C; Line planting technique in rehabilitation of logged area

of the line planting indicates the plantations following this technique have good growth performance that leads to an increase in resource base. The salient criteria of line planting under TPTJ system considers that to be followed by the forest managers to make it sustainable. Establishment of planting line does not cause soil properties decrease in the whole study plots so that perception that planting line establishment in TPTJ system will cause changes towards soil properties is not proved. Status of soil quality in primary forest and planting area was in the categories of ranging from low (3.937) to moderate (4.707). The soil quality improvement occurred in the area of TPTJ while soil quality status at 4 years old TPTJ included in the low category.

REFERENCES Appanah S, Weinland G (1993) Planting quality timber trees in Peninsular Malaysia: a review. Malayan For Rec 38: 221. Angers VA, Messier C, Beaudet M, Leduc A (2005) Comparison composition and structure in old-growth and harvested (selection and diameter limit cuts) northern hardwood stands in Quebec. Forest Ecol Manage 217:275-293. Blair GJ, Lefroy RDB, Lisle L (1995) Soil carbon fractions based on their degree of oxidation and the development of carbon management index for agricultural system. Aus J Agric Res 46: 1459-1466. Burger JA (2009) Management effects on growth, production and sustainability of managed forest ecosystems: past trends and future directions. Forest Ecol Manage 258: 2335-2346. Carter MR (2002) Soil quality for sustainable land management: organic matter and aggregation interactions that maintain soil functions. Agron J 94: 38-47. Carter MC, Foster CD (2006) Milestones and millstones: a retrospective on 50 years of research to improve productivity in loblolly pine plantations. Forest Ecol Manage 277:137-144. Dupuy JM, Chazdon RL (2008) Interacting affects of canopy gap, understory vegetation and leaf litter on tree seedling recruitment and composition in tropical secondary forests. Forest Ecol Management 255: 3716-3725. Evans J (1982) Plantation forestry in the tropics. Oxford University Press, Oxford. Fox TR, Allen HL, Albaugh TJ, Rubilar R, Carlson CA (2007) Tree nutrition and forest fertilization of pine plantations in the Southern United Stated. Southern Journal of Applied Forestry 31: 5-11. Gregorich EG, Carter MR, Doran JW, Pankhurst CE, Dwyer LM (1997) Biological attributes of soil quality. In: Gregorich EG, Carter MR

(eds). Soil quality for crop production and ecosystem health. Elsevier, Amsterdam. Griffis KL, Crawford JA, Wagner MR, Moir WH (2001) Understory response to management treatments in northern Arizona ponderosa pine forests. Forest Ecol Manage. 146:239-245. Gupta VVSR, Germida JJ (1988) Distribution of microbial biomass and its activities in different soil aggregate size classes as influenced by cultivation. Soil Biol Biochem 20: 777-789. Handayani IP, Prawito P (1998) Dinamika mineralisasi karbon dan nitrogen pada lahan alang-alang (Imperata cylindrica). J Tanah Tropika 7: 93-102. Handayani IP (2001) Comparison of soil quality in cultivated fields and grassland. J Tanah Tropika 12: 135-143. Handayani IP, Prawito P, Lestari P, Coyne MS (2001) Potential of carbon sequestration after reforestation and grass establishment on tropical degraded soils. In: Thielges BA, Satrapradja SD, Rimbawanto A (eds) In situ and ex situ conservation of commercial tropical trees. GMU and ITTO, Yogyakarta. Handayani IP (2004) Soil quality changes following forest clearance in Bengkulu, Sumatera. Biotropia 22: 1-16. Karlen DL, Mausbach MJ, Doran JW, Cline RG, Harris RF, Schuman GE (1997) Soil quality: a concept, definition, and framework for evaluation (a guest editorial). Soil Sci Soc Amer J 61: 4-10. Lal R (1998) Soil quality and sustainability. In: Lal R, Blum WH, Valentine C, Stewart BA (eds). Methods for assessment of soil degradation. CRC Press, Florida. Lamprecht H (1989) Silviculture in tropics. Tropical forest ecosystems and their tree species: possibilities and methods for their longterm utilization. GTZ Eschborn, Germany. Matthews E, Payne R, Rohweder M, Murray S (2000) Pilot analysis of global ecosystems: forest ecosystems. World Resources Institute. Washington DC. Moore PD, Chapman SB (1986) Methods in plant ecology. Blackwell, Oxford. Olson RK, Schnoeneberger MM, Aschmann SG (2000) An ecological foundation for temperate agroforestry. In: Garret HE, Rietveld WJ, Fisher RF (eds) North American Agroforestry : An Integrated Science and Practice. American Society of Agronomy. Madison. Oâ&#x20AC;&#x2122;Neill K, Arnacher M (2004) Forest inventory and analysis: soil quality indicator. FIA fact sheet series. USDA Forest Service, Washington. Saaty TL (1996) The analytic hierarchy process: planning, priority setting, resource allocation. RWS Publications, Pittsburgh. Schulte A (1996) Dipterocarp forest ecosystem theory based on matter balance and biodiversity. In: Schulte A, Schoene D (eds) Dipterocarp forest ecosystems: towards sustainable management. World Scientific, Singapore. Weidelt HJ (1996) Sustainable management of dipterocarp forests: opportunities and constraints. In: Andreas S, Dieter S (eds) Dipterocarp forest ecosystems. World Scientific, Singapore. Weidelt HJ, Banaag VS (1982) Aspect of management and silviculture of Philippine dipterocarp forest. GTZ, Eschborn, Germany.

B I O D I V E R S IT A S Volume 11, Number 1, January 2010 Pages: 40-45

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110109

Fuel characteristics and trace gases produced through biomass burning BAMBANG HERO SAHARJO1,♥, SHIGETO SUDO2, SEIICHIRO YONEMURA2, HARUO TSURUTA2 1

Forest Fire Laboratory, Faculty of Forestry, Bogor Agricultural University, PO. BOX 168, Bogor 16001, West Java, Indonesia, Tel.:+ 62-251-8421-929, Fax.: + 62-251-8626-886, email: bhsaharjo@gmail.com 2 Greenhouse Gas Emission Team, National Institute for Agro-Environmental Science. 3-1-3 Kannondai, Tsukuba, Ibaraki 305-8604, Japan Manuscript received: 10 November 2009. Revision accepted: 30 December 2009.

ABSTRACT Saharjo BH, Sudo S, Yonemura S, Tsuruta H (2010) Fuel characteristics and trace gases produced through biomass burning. Biodiversitas 11: 40-45. Indonesian 1997/1998 forest fires resulted in forest destruction totally 10 million ha with cost damaged about US$ 10 billion, where more than 1 Gt CO2 has been released during the fire episode and elevating Indonesia to one of the largest polluters of carbon in the world where 22% of world’s carbon dioxide produced. It has been found that 80-90% of the fire comes from estate crops and industrial forest plantation area belongs to the companies which using fire illegally for the land preparation. Because using fire is cheap, easy and quick and also support the companies purpose in achieving yearly planted area target. Forest management and land use practices in Sumatra and Kalimantan have evolved very rapidly over the past three decades. Poor logging practices resulted in large amounts of waste will left in the forest, greatly elevating fire hazard. Failure by the government and concessionaires to protect logged forests and close old logging roads led to and invasion of the forest by agricultural settlers whose land clearances practices increased the risk of fire. Several field experiments had been done in order to know the quality and the quantity of trace produced during biomass burning in peat grass, peat soil and alang-alang grassland located in South Sumatra, Indonesia. Result of research show that different characteristics of fuel burned will have the different level also in trace gasses produced. Peat grass with higher fuel load burned produce more trace gasses compared to alang-alang grassland and peat soil. Key words: peat soil, peat grass, alang-alang, biomass burning, trace gas.

INTRODUCTION In 1997/1998 forest fires at least 20 million of Indonesian peoples were directly and indirectly affected by fire. Black smokes contain many air pollutants: CO, CO2, SO2, NO(x) and NH4 and bacteria such as Streptococcus cause many diseases such as diarrhea, pneumonia, bronchitis and brain disturbance. Thousand of people in Riau, Jambi, South Sumatra, and Central, West and East Kalimantan were hospitalized for medical treatment. Black smoke that continued for at least two months in southern Sumatra at which time the lack of sunshine reduced food production and caused many peoples to seek emergency food sources. Hundreds of peoples also died in Papua (Irian Jaya) because the transportation of food and other supplies could not reach their areas due to smoke. Biomass burning is the burning of living and dead vegetation, including grasslands, forests and agricultural lands following the harvest for land clearing and land-use change. Biomass burning is not restricted to one geographical region, but is rather a truly global phenomenon (Levine 1996). Biomass burning is a significant global source of gaseous and particulate emissions to the atmosphere. Gases produced by biomass burning include: (i) GHG gases, CO2, CH4 and N2O that lead to global warming, (ii) chemically active gases, NO, CO, CH4 and NMHCs, which lead to the photochemical production of ozone (O3) in the troposphere, and (iii)

CH3Cl, and CH3Br which lead to the chemical destruction of ozone in the stratosphere (Levine 1985). Direct effects on atmospheric composition and chemistry and climate, biomass burning perturbs other components and processes in the earth’s system, including (i) the biogeochemical cycling of nitrogen (N2O and NO) and carbon (CO2, CO and CH4) gases from the biosphere to the atmosphere, (ii) water run-off and evaporation, and hence, impacts the hydrological cycle, (iii) the reflectivity and emissivity of the land, which in turn changes the radiative properties of the land and hence, impacts climate, and (iv) the stability of ecosystems which in turn impacts biological diversity. Fire is a significant source of gases and particulate to the atmosphere: environmentally important gases produce by fire includes carbon dioxide, carbon monoxide, methane, non-methane hydrocarbons and oxides of nitrogen. Fire also produces large amounts of small, solid particles or “particulate matter”, which absorb and scatter incoming solar radiation, and hence the impact of our planet as well as provoking a variety of human health problems (Levine 1996). Fire can therefore be considered one of the local points of the multiple relationships between humans and the environmental changes in fire patterns can be taken as indicator of change in land-use patterns and overall environmental conditions (Malingreau and Gregorie 1996). Forest fires destroy large forest areas that serve as habitat for biodiversity. They directly eliminate plants and

SAHARJO et al. – Greenhouse gases produced through biomass burning

animals and also result in forest degradation that leads to a decrease in the survival rate of the species. For example, the fires in 1997-1998 resulted in 33% population decline of the Orangutan (Pongo pygmaeus) in Borneo (Rijksen and Meijaard 1999). Moreover, fires in 1982-1983 in Kutai National Parks (East Kalimantan) resulted in widespread mortality of reptiles and amphibians (MacKinnon et al. 1996) and lost of fruit trees that caused the population of fruit-eating birds such hornbills declined dramatically (Nasi et al. 2002). The most severe fires in the last twenty years occurred in 1997, affecting approximately 11.7 million hectares, mostly lowland peat and swamp forests, timber plantations and agricultural areas (Tacconi 2003). This study consisted of two parts: field experiment and laboratory experiment (analysis). First, test biomass burning was conducted at three different land type namely alang-alang or cogon (Imperata cylindrica) grassland, peat soil and peat grass. These land types are suspected of being responsible for much of the smoke produced during the 1997/98 fire episode in Indonesia. More than 2 Mha of peat lands were burned during the 1997/98 fires (Tacconi et al. 2006). In addition, when these vulnerable ecosystems were accounted for, Murdiyarso and Adiningsih (2007) estimate that the total carbon emissions during 1997/98 fires was 5.3 Gt CO2.

MATERIALS AND METHODS Research Site Alang-alang grassland The research in alang-alang grassland was carried out in the area belonging to the PT. Musi Hutan Persada, Barito Pacific Group, South Sumatra. The mean annual rainfall is about 2,000 mm and monthly rainfall is about 209 mm ranging from 92 mm in July to 278 mm in February. According to the Schmidt and Fergusson (1951) system, the climate of this area belongs to rainfall type A. Mean maximum air temperature in this area is 32.6°C in August, mean minimum air temperature is 22.6°C in December and mean annual relative humidity is about 85%. The soil is red yellow podsolic and the USDA soil classes are: Haplaquox, Dystropepts, Kandiudults and Haplaquox. This area has the following characteristics: Slopes < 8%, 5-125 m altitude, drainage varies from well-drained to imperfectly drained, soil mineral depth is 101-150 cm, Cation Exchange Capacity (CEC) is 4.9 - 17.9 me gr-1, base saturation is 5.621.8%, available phosphorus is 0.4-3.2 ppm, total nitrogen is 0.11-0.20%, organic carbon is 1.09-3.51%, free salinity, and soil fertility based on Indonesian criteria is low (Hikmatullah et al. 1990). Understorey vegetation dominated by Imperata cylindrica, Eupatorium pubescens, Clidemia hirta, Tetracera sp, Artocarpus anisophyllus, Macaranga javanica, and Dillenia grandifolia. Peat soil and peat grass The test burn in the peat grass site was done in peat soil that (dominated by hemic peat) was located in Teluk Pulai the area which belongs to PT. SBA Wood Industry, South

Sumatra. The mean annual rainfall is about 2,800 mm and monthly rainfall ranges from 70 mm in August to 365 mm in March. According to the Schmidt and Fergusson (1951) system, the climate of this area belongs to rainfall type B. Mean maximum air temperature in this area is 38.8°C in June, mean minimum air temperature is 21.2°C in January and mean annual relative humidity is about 89.9%. The peat type is hemic dominant. The area has the following characteristics: Slopes < 8%, 0-5 m altitude, drainage varies from well-drained to imperfectly drained, high salinity, peat depth is 0.5-2.5 m and soil fertility based on Indonesian criteria is low or poor (Suwarso 1997). Measurement and Analysis Data presented in this paper were derived from field experiments both in mineral soil (alang-alang grassland) and peat land (peat soil and peat grass). Alang-alang grassland Three of 3 m x 3 m plot were established in the alangalang grassland, where 1.5 m of firebreaks established surrounded the plot. Fuel available consisted of grasses and litter. Smoke samples from the burning site were taken by using evacuated canisters. Smoke samples were taken at different phases of combustion, flaming, smoldering and glowing. Samples taken were analyzed in the Greenhouse Gas Laboratory of National Institute for Agro-Environmental Science, Tsukuba, Japan for analysis. Gasses analyzed are CO, CO2, N2O, CH4, CH3Br, CH3Cl, and CH3 I. Peat soil Three different size plots were established in the hemic site. First and second plots, 1 m x 2 m, were surrounded by a 30 cm wide and 50 cm deep canal. The third plots were 1 m x 1.5 m size. Peat moisture content was measured directly using digital moisture content thermocouple. Flame temperature was measured using data logger which the sensors (thermocouples) put in the peat surface in the plot. Logs and branches were put in the canal and fires were started using gasoline. Flame temperature was monitored for a half days following ignition. Smoke samples were taken by using portable pump which connected to the vacuum plastic. Smoke samples accumulated in vacuum plastic then transferred into the vacuum bottles. Smoke samples were taken at combustion during flaming, smoldering and glowing phases of the burn. Smoke samples that storage in the vacuum bottles were then sent to the Greenhouse Gas Laboratory of National Institute for Agro-Environmental Science, Tsukuba, Japan for analysis. Gasses analyzed are CO, CO2, N2O, CH4, CH3Br, CH3Cl, and CH3I. Peat grass Three plot of 1 m x 1 m size was established in the peat grass dominated by ferns (Gleichenia linearis, Lygodium scandens, Nephrolepis fascigera and Stenochlaena palustris). Before burning was conducted, 1 m firebreaks were established around each plot in order to prevent fire

B I O D I V E R S IT A S

11 (1): 40-45, January 2010

from jumping to non target areas. Burning was conducted by igniting the dead fuels within live grass community. Smoke samples were taken by using portable pump which connected to the vacuum plastic. Smoke was taken at combustion during flaming, smoldering and glowing phases of the burn. Smoke samples in the vacuum plastic then transferred to the vacuum bottle which finally sent to the Greenhouse Gas Laboratory of National Institute for Agro-Environmental Science, Tsukuba, Japan for analysis. Gasses analyzed are CO, CO2, N2O, CH4, CH3Br, CH3Cl, and CH3 I. Trace gas measurement Four kinds of gas chromatograph (GC) systems were used for whole analysis on trace gases measurement. Methane (CH4) gas was measured by using Shimadzu GC9A Flame Ionization detector (FID) which attached to GC with back flush system. Carbon Dioxide (CO2) gas was measured by using Thermal Conductivity Detector (TCD) of the same column system with methane gas measurement. Carbon Monoxide (CO) gas was measured by using Trace Analytical Systems Inc. RGA3 of Reduced Gas Detector with 5 m Molecular Sieve 13X packed column system. 1 ml STP gas was injected for analysis of each above 3 gas species. For halogenated species for example: CFCs, CH3Br, and CH3I, Shimadzu GC-14B ECD (Electron Capture Detector) system with clyofocusing method used. CP PoraBOND Q capillary column (0.32 mm i.d. x 25 m) was used for separation. ECD was also used for N2O analysis with three dimensional separation systems with using 3 Poapak Q (3.0 mm i.d. x 1 m) stainless steel columns. For CH4, CO2, CO, N2O, standard gas were made by Nippon Sanso Industry with concentration of 2.0 ppm and 10.0 ppm for CH4, 350 ppm and 500 ppm for CO2, 150 ppb and 20 ppm for CO, and 300 ppb and 1 ppm for N2O respectively. For halogenated species, standard gas measurement were made by Taiyo Toyo Sanso Industry, Japan with concentration of 100 ppt, 1.00 ppb, and 10.0 ppb for all gases containing CFC12, CFC-11, CH3Cl, CH3Br, CH3I. Precision of whole measurement were within 10 percent. Statistical analysis A completely random design analysis of variance was used to test for differences among subplots, based on the following model (Steel and Torrie 1981): Ymn = U + Tm + Emn Ymn = fuel and fire behavior parameter at m subplot in n replication U = mean of the treatment population sampled Tm = treatment (slashing, drying, burning) Emn = random component

To detect significance difference among fuel and fire behavior parameter among subplot (p  0.05), the Duncan test was used (Steel and Torrie 1981).

RESULTS AND DISCUSSION Weather condition and fire behavior Data in Table 1 show weather conditions when burnings were conducted. The peat grass site air temperature was 28-32C which was the lowest. Relative humidity at peat soil was 60-65% which was the lowest of the site. Fuel load varied from 10 to 20 ton ha-1. Fuel moisture content varied from 10% to a maximum of 70%. Flame temperature during burning was 500-600C at the alang-alang site and at the other two sites, 700-800C. Burning time for alang-alang grasslands was the fastest compared to other type of fuel from different land type where the 9 m2 plot of alang-alang required less than 3 minutes for each plot. Table 1. Weather condition and fire behavior parameter Parameter

Alang-alang 33-35 Air temperature (C) Relative humidity (%) 70-80 Wind speed (m sec -1.) 2-3 Fuel potency (ton ha-1) 10-20 Moisture content (%) 10-70 500-600 Flame temperature (C)

Land type Peat soil 32-33 60-65 2-3 15-20 30-60 > 830

Peat grass 28-32 75-85 1-2 10-15 20-60 700-800

Green house gases emission (CO2, CO, CH4 and N2O) Most tropical fires are set or spread accidentally or intentionally by humans and are related to several causative agents; some of them limited to subsistence livelihood, others to commercial activities (Qadri 2001). Emissions from burning the cleared vegetation depend on the degree of combustion that is achieved, i.e. the proportion of biomass consumed by fire (Brinkman 2009). If fire is continued to be used for land preparation in peat areas the status of peat become critically endangered (Saharjo 2007). Forest fires release toxic gases like carbon monoxide (CO), ozone (O3), nitrogen dioxide (NO2), hydrocarbons, aldehydes, particles and polycyclic aromatic hydrocarbons (PAHs) (Ostermann and Brauer 2001). It is known that exposure to these gases can cause acute respiratory infections. In 2000, Southeast Asia contributed 12% of global GHG emissions, amounting to 5,187.2 MtCO2-eq, including emissions from LUCF (ADB 2009) CO2 emission Peat soil, with higher moisture content compared to alang-alang and peat grass, had the highest CO2 emission (Table 2). This is understandable due to smoldering combustion was dominant. In alang-alang grassland, because the fuel was relatively dry, flaming combustion dominated. Flaming combustion accounts for most of the fuel consumption in grass fires. However, the emissions from smoldering combustion in grass fires also play an important role in tropical dry season tropospheric chemistry (Granier et al. 1996) and in tropical and temperate forests consume a much larger percentage of the fuel by smoldering combustions (Ward et al. 1992). Grassland management has the potential to sequester carbon by 0.11- 1.50 tCO2 ha -1 per year (ADB 2009).

SAHARJO et al. – Greenhouse gases produced through biomass burning Table 2. CO2 , CO, CH4 and N2O emission (ppm) during burning in alang-alang, peat soil and grass Land type Alang-alang Peat soil Peat grass CO2 (1494.31246.1)b (21466.77731.5)c (9906.75794.0)a CO (400.7136.5)a (1331.7409.0)b (6717.34584.8)c CH4 (7.15.8)a (475.3250.4)b (507.7149.1)c N2O (1201.0185.8)a (1828.0347.8)b Note: Means are significantly different when standard errors are followed by different letters (p0.05) Parameter

Peat grass combustion emitted the second largest CO2 (Table 2). This can be understood because grass that lives in the peat has quite high moisture content (an average of more than 50%) which then slows the combustion processes. These high moisture contents of peat grass required more energy and time to reach the ignition temperature. This situation can be seen clearly through flame temperature of peat grass burning that vary from 700-800C. Carbon dioxide can account for 99% of the C emissions in efficient burns that is burns consuming most, if not all, of the available fuels, but account for only 50% in low-intensity smoldering fire (Debano et al. 1998). CO emission Carbon monoxide emission in peat grass was the highest among the three sites (Table 2). High moisture content of peat grass resulted in incomplete burning and lowintensity fire. Low combustion resulted in high CO emission during combustion compared to alang-alang grassland and peat soil. Field fire experiments show that fires in the peat grass was slow and required much time to reach the ignition temperature of 360C needed for spreading compared to alang-alang and peat grass. The resulting low-intensity fire, produced CO, which is commonly produced with incomplete combustion of moist fuels. High emissions of CO have been measured on the fire line of relatively low-intensity fire (Sandberg et al. 1975) and the amount of CO emitted by fire is a function of combustion efficiency, increasing as efficiencies drop (Debano et al. 1998). The emission factor (amount of the compound released per amount of fuel consumed) of CO for peat grass (0.636 mol CO/mol CO2) was the highest compared to peat soil (0.0509 mol CO/mol CO2) and alang-alang (0.0082 mol CO/mol CO2). The emission ratio of CO/CO2 from peat grass was higher than that reported by Ward (1986) and Greenberg et al. (1984), but less than reported by Lacaux et al. (1993). CH4 emission The first step in the combustion of vegetation is the pyrolitic decomposition of plant matter immediately ahead of the

flames, this releases a wide array of reduced and partially oxidized organic compound with CH4 being the most abundant species (Andreae et al. 1996). Peat grass combustion emitted more CH4 than alang-alang grassland and peat soil. High CH4 emission resulting from peat grass combustion is believed to be due to the fuel bed characteristics of grass, especially on compaction and pattern (Table 2). Physically the performance of peat grass seems to be the same as alang-alang grassland but then combustion is actually different. The peat grass grows in bunches which are rather thick at the bottom compared to alang-alang grassland. This condition affects the combustion processes. During the early, flaming stage, when the temperature is high and the combustion zone is well oxygenated, the emissions of partial combustion products are low. The proportion of the total combustion process which is made up by each stage is mainly controlled by the physical attributes of the fuel and to a lesser degree by the weather during the fire (Scholes et al. 1996). This emission ratio of CH4 observed in our study from peat grass (Table 3) was higher than that reported by Andreae (1991) and Akeredolu and Isichei (1991) but less than reported by Lacaux et al. (1993), both from savannas and forests.

Table 3. Emission factors reported for vegetation fires Emission factor (Original units) Carbon monoxide (CO) 0.055-0.145 g CO2/g CO2 0.05-0.3 mol CO/mol CO2 6.1 mol CO/mol CO2

Emission factor (g kg-1)

230 (forest) 63-73 (savannas) 59-97 (grasslands) 0.636 mol CO/mol CO2 (peat grass) 0.0509 mol CO/mol CO2 (peat soil) 0.0082 mol CO/mol CO2 (alangalang) Methane (CH4) 0.0062-0.016 mol/mol 0.012 mol/mol

0.3130.89 mol CH4/mol CO2 0.3130.89 mol CH4/mol CO2 0.0018-0.016 mol/mol 0.0354 mol/mol (peat grass) 0.0093 mol/mol (peat soil) 0.000028 mol/mol (alang-lang) Nitrogen Monoxide plus Dioxide 0.061-0.417 g N-NOx/g N-fuel 0.3-3.5 x 10-3 mol NOx/mol CO2

1.01 x 10-4 mol/mol(peat grass) 6.10 x 10-5 mol/mol(peat soil)

1.0-1.8 1.8-4.3 4.5 9.47 (forest) 1.8-2.3 (savannas) 1.7-3.4 (grassland) 1.3-2 (savannas) 6-8 (forests)

Reference Ward (1986) Greenberg et al. (1984) Lacaux et al. (1993) Scholes et al. (1996) Scholes et al. (1996) Scholes et al. (1996) This study This study This study

Andreae (1991) Akreedolu and Isichei (1991) Ward et al. (1992) Bonsang et al. (1991) Greenberg et al. (1984) Scholes et al. (1996) Scholes et al. (1996) Scholes et al. (1996) Lacaux et al. (1993) Lacaux et al. (1993) Hao et al. (1990) This study This study This study

Ward (1990) Lobert et al. (1991) 16.1 (forest) Scholes et al. (1996) 4.4-5.1 (savannas) Scholes et al. (1996) 4.2-6.8 (grasslands) Scholes et al. (1996) This study This study

B I O D I V E R S IT A S

11 (1): 40-45, January 2010

N2O emission Peat grass combustion emitted more N2O than the peat soil (Table 2). Peat grass is an N-rich a fuel compared to alang-alang grassland which is usually used as a poor land indicator. N2O emission ratio from peat grass was higher than peat soil, but les than reported by Lobert et al. (1991) and Scholes (1996). Production of NO(x) by burning is largely dependent on the N content of the fuel consumed. Both NO and NO2 are reactive gases that are emitted from combustion (Lobart and Warnatz 1993). NO is a thermally stable product of combustion. Although NO2 is less stable than NO, its abundance increases in smoldering fire (Clements and McMahon 1980). CH3Br, CH3Cl and CH3I emission during burning in the alang-alang, peat soil and peat grass Peat grass produces more CH3Br, CH3Cl and CH3I emission (Table 4) during burning compared to alang-alang grassland and peat soil. The amount of chlorine and bromine released to the atmosphere during a fire depend on chlorine and bromine concentration in the fuel burned, the contribution of the fuel to the aboveground biomass and the percentage of fuel burned (McKenzie et al. 1996) and also depend on fire intensity and combustion efficiency (Reinhard and Ward 1995). CH3Cl, CH3Br together with CH4 play a significant role in stratospheric ozone chemistry (Mano and Andreae 1994). CH3Br (methyl bromide) releases atomic bromine, which leads to the catalytic chemical destruction of stratospheric ozone, which is very similar to the catalytic destruction of stratospheric ozone by chlorine (Levine 1996) and the discovery that methyl bromide is an important combustion product of biomass burning identified a previously unknown and very important connection between biomass burning and the chemical destruction of stratospheric ozone. Table 4. CH3Br, CH3Cl and CH3I emission during burning in the alang-alang, peat soil and peat grass Land type Alang-alang Peat soil Peat grass CH3Br (543.0508.1)a (25.63.4)b (41.19.8)c CH3Cl (1420.7416.1)a (280.083.3)b (694.7340.3)c CH3I (38.611.2)a (1.00.6)b (1.50.8)c Note: Means are significantly different when standard errors are followed by different letters (p0.05) Parameter

Biomass burning releases a large amount of aerosol into the atmosphere, which in turn reduces the solar radiation absorbed at the Earth’s surface and it can reduce the rainfall over a large area. For urban and industrial areas, there is evidence that air pollution can suppress rain and snow (Rosenfeld 2000; Givati and Rosenfeld 2004). The aerosol (smoke) entrained in a cloud may modify its development by increasing the number of, and depressing the size of, ice crystals within them (Sherwood 2002; Sherwood et al. 2006; Jiang et al. 2008). This will suppress precipitation in the cloud and also change the electrical properties of the cloud. It is known that the highest measured lightning currents with the largest charge

transfers are associated with positive lightning. These positive cloud-to-ground (+CG) lightning strikes, even though a small percentage (10%-12%) of the total, are the most 15 efficient for fire ignition. Some recent examples are given in McGuiney et al. (2004); Wotton and Martell (2005).

CONCLUSIONS Peat grass, where smoldering combustion was dominant due to the high moisture content, produced more trace gasses (CO, CH4, N2O, CH3Cl, CH3Br and CH3I) than alang-alang grassland and peat soil. Dry fuel combustion was dominated by flaming combustion in the alang-alang grassland resulted in lower trace gas production. Fuel characteristics (moisture content, type, pattern and potency) and environmental condition (relative humidity, wind speed and air temperature) determine trace gasses production during burning.

REFERENCES ADB (2009) The economic of climate change in Southeast Asia: regional review. ADB, Manila, The Philippines. Akeredolu F, Isichei AO (1991). Emissions of carbon, nitrogen and sulfur from biomass burning in Nigeria. In: Levine JS (ed.) Global biomass burning: atmospheric, climatic and biospheric implications. MIT Press, Cambridge, Mass. Andreae MO (1991) Biomass burning: Its history, use and distribution and its impact on environmental quality and global climate. In: Levine JS (ed.) Global biomass burning: atmospheric, climatic and biospheric implications. MIT Press, Cambridge, Mass. Andreae MO, Atlas E, Cachier H, Coffer WR, Harris GW, Helas G, Koppmann R, Lacaux JP, Ward DE (1996) Trace gas and arosol emission from savanna fires. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Brinkman A (2009) GHG emissions from palm oil production. Brinkman Consultancy, The Netherlands. Clements HB, McMahon CK. 1980. Nitrogen oxides from burning forest fuels examined by thermogravimetry and evolved gas analysis. Thermochimica Acta 35 (2): 133-265. Debano L, Neary DG, Ffolliott PF (1998) Fire’s effects on ecosystems. John Wiley and Sons, Inc. Givati A, Rosenfeld D (2004) Quantifying precipitation suppression due to air pollution. J Appl Meteorol 43: 1038-1056 Granier C, Hao WM, Brasseur G, Muller JF (1996) Land use practices and biomass burning: Impact on the chemical composition of the atmosphere. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Greenberg JP, Zimmerman PR, Heidt L, Pollock W (1984) Hydrocarbon and carbon monoxide emissions from biomass burning in Brazil. J Geophys Res 89: 1350-1354. Hao WM, Liu MH, Crutzen PJ (1990) Estimates of annual and regional releases of CO2 and other trace gasses to the atmosphere from fires in the tropics based on the FAO statistics fro the period 1975-1980. In: Crutzen PJ, Goldammer JG (eds) Fire in the tropical biota: ecosystem processes and global challenges, Ecol Stud vol. 84. Springer, New York. Hikmatullah A, Hidayat U, Affandi E, Suparma Y, Chendy TF, Buurman P (1990) Handbook on land and soil map units, sheet Lahat (1012), Sumatra. Research Center for Soil and Agro-climate. Bogor. [Indonesian] Jiang JH, Su H, Schoeberl RM, Massie TS, Colarco P, Platnick S, Livesey J N (2008) Clean and polluted clouds: Relationships among pollution, ice clouds, and precipitation in South America. Geophys Res Lett 35. L14804, doi: 10.1029/2008 GL034631.

SAHARJO et al. â&#x20AC;&#x201C; Greenhouse gases produced through biomass burning Lacaux JP, Cachier H, Delmas R (1993) Biomass burning in Africa: An overview of its impact on atmospheric chemistry. In: Crutzen PJ, Goldammer JG (editors). Fire in the environments: the ecological, atmospheric, and climatic importance of vegetation fires. John Wiley and Sons, New York. Levine JS (1996) Introduction. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Levine JS (ed) (1985) The photochemistry of atmospheres: Earth, the other planets and comets. Academic Press, San Diego, CA. Lobart JM, Warnatz J (1993) Emissions from the combustion processes in vegetation. In: Crutzen PJ, Goldammer JG (editors). Fire in the environments: The ecological, atmospheric, and climatic importance of vegetation fires. John Wiley and Sons, New York. Lobert JM., Scharffe DH, Hao WM, Kuhlbusch TA, Seuwen R, Warneck P, Crutzen PJ (1991) Experimental evaluations of biomass burning emissions: Nitrogen and carbon containing compounds. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Malingreau JP, Gregorie JM (1996) Developing a global vegetation fire monitoring system for global change studies; A frame work. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Mano S, Andreae MO (1994) Emission of methyl bromide from biomass burning. Science 263: 1255-1257 Rijksen HD, Meijaard E (1999) Our vanishing relative: The status of wild orang-utans at the close of the twentieth century. Kluwer, Amsterdam. McGuiney E, Shulski M, Wendler G (2004) Alaska lightning climatology and application to wildfire science. Proceedings of the 85th AMS Annual Meeting, San Diego, CA, 9-13 Jan. 2004 MacKinnon K, Hatta G, Halini H, Mangalik A (1996) The Ecology of Kalimantan. Periplus, Singapore. McKenzie LM, Ward DE, Hao WM (1996) Chlorine and Bromine in the biomass of tropical and temperate ecosystem. In: Levine JS (ed.). Biomass burning and global change. Vol. 1. MIT Press, Cambridge, Mass. Murdiyarso D, Adiningsih ES (2007) Climate anomalies, Indonesian vegetation fires and terrestrial carbon emissions. J Mitig Adapt Strat Glob Change 12: 101-112. Nasi R, Meijaard E, Applegate G, Moore P (2002) Forest fire and biological diversity. Unasylva 53 (209): 36-40 Ostermann K, Brauer M (2001) Air quality during haze episodes and its impact on health. In: Peter E, Radojevic M (eds.) Forest fires and regional haze in Southeast Asia. Nova Science Publishers, Huntington, New York.

Qadri ST (2001) Fire, smoke and haze. ASEAN-ADB, Manila. Reinhard TE, Ward DE (1995) Factor affecting methyl chloride emission from forest biomass combustion. Env Sci Tech 29: 825-832 Rosenfeld D (2000) Suppression of rain and snow by urban and industrial air pollution. Science 287: 1793-1796 Saharjo BH (2007) Shifting cultivation in peatlands. J Mitig Adapt Strat Glob Change 12: 135-146. Sandberg DV, Pickford SG, Dfarley EF (1975) Emissions from slash burning and the influence of flame retardant chemicals. J Air Poll Contr Assoc 25: 278-281 Schmidt FH, Ferguson JHA (1951) Rainfall type based on wet and dry period ratios of Indonesia with western New Guniea. Verhandelinegn No. 42, Kementerian Perhubungan, Djawatan Meteorologi dan Geofisika, Djakarta. Scholes RJ, Ward DE, Justice CO (1996) Emissions of trace gases and aerosol particles due to vegetation burning in southern hemisphere Africa. J Geophys Res 101 (D19): 23,677-23,682 Sherwood SC (2002) A microphysical connection among biomass burning, cumulus clouds and stratospheric moisture. Science 295 (5558): 1272-1275 Sherwood SC, Phyllips VTJ, Wettlaufer JS (2006) Small ice crystals and the climatology of lightning. Geophys Res Lett 33: L05804, doi:10.1029/2005GL025242 Steel RG, Torrie JH (1981) Principles and pocedures of statistics: a biometrical approach. 2nd ed. McGraw-Hill, New York. Suwarso (1997) Swamp forest vegetation damage caused by illegal logging in the concession area; case studies on forest group of Tanjung Koyan-Sungai Lumpur, Simpang Tiga Village, Sub-District Tulung Selapan, District OKI, Province South Sumatra. [M.ScThesis]. Pasca Sarjana IPB, Bogor. [Indonesia] Tacconi L (2003) Fires in Indonesia: causes, costs and policy implications. CIFOR Occasional Paper No. 38. CIFOR, Bogor. Tacconi L, Rios MTS, Wells A (2006) Justice in the forest: rural livelihoods and forest law enforcement. CIFOR. Bogor Ward DE, Susott RA, Kaufmann JB, Babitt RE, Cummings DL, Dias B, Holben BN, Kaufmann YJ, Rasmussen RA, Setzer AW (1992) Smoke and fire characteristics for cerrado and deforestation burns in Brazil: BASE-B experiment. J Geophys Res 97 (14): 14,601-14,619 Ward DE (1986) Chracteristics emissions of smoke from prescribed fires for source appointment. In: Proceeding of the annual meeting of the Pacific Northwest in section air pollution control association. Wotton BM, Martell DL (2005) A lightning fire occurrence model for Ontario. Can J Forest Res 35: 1389-1401.

BIODIVERSITAS Volume 11, Number 1, January 2010 Pages: 46-54

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic) DOI: 10.13057/biodiv/d110110

Characterizing forest reduction in Ketapang district, West Kalimantan, Indonesia ASEP SUNJAYA ADHIKERANA1, JITO SUGARDJITO2,♥ 1

Fauna and Flora International – Indonesia Programme, Jl. Harsono, R.M. No.1, Ragunan, Jakarta 12550, Indonesia 2 Research Centre for Biology, The Indonesia Institute for Sciences, Jl. Raya Jakarta-Bogor Km. 46, Cibinong-Bogor 16911, West Java, Indonesia, Tel.: +62-21-8765056 Fax.: +62-21-8765068, ♥e-mail: jitos@cbn.net.id Manuscript received: 16 November 2009. Revision accepted: 29 December 2009.

ABSTRACT Adhikerana AS, Sugardjito J (2010) Characterizing forest reduction in Ketapang district, West Kalimantan, Indonesia. Biodiversitas 11: 46-54. We have characterized deforestation in the Ketapang district forests when we implemented the Orangutan (Pongo pygmaeus wurmbii) Conservation in Trans-boundary Landscape between Central and West Kalimantan provinces. For the purpose of evaluating the changes in land use and land cover in the study areas, a series of Landsat imageries have been analyzed. Each of the Landsat imagery data set for all study areas was initially classified using unsupervised classification into 13 different land-cover types. Ground truth checks were undertaken for Ketapang district forests and Sungai Puteri peat swamp forest, from which the results were used for the supervised land use classification of these two study areas. Between 1992 and 2000 there was only small conversion of primary forest into secondary forests. During this period barren land remained extensive about 30.17% of the total area of Ketapang district. Both agriculture and plantation areas substantially increased 56% and 55% respectively during 2003, while at the same time the extent of both primary and peat swamp forests were considerably reduced up to 15% and 28% respectively. The most striking conversion was from secondary forest to agricultural land and from peat swamp forest to swamp areas. A fraction of lowland forest was also converted into oil-palm plantation which was extended with considerable size into agricultural land. The patterns of land use changes detected in this study indicated a number of possible causes that trigger deforestation in this district include, the local government policy and market demand. Key words: deforestation, habitat fragmentation, Landsat, imagery, land use.

INTRODUCTION For more than four decades Indonesia has been relied its economic development on forest resources. In the 1980s, the forestry sector became the second highest contributor to foreign exchange in Indonesian economy after oil and gas sector (FWI/GFW 2002). In this decade, Indonesia became the biggest producer of plywood in the world. It was able to manage 75% of the world market demands. Despite the success of controlling plywood market in the world, the rainforest which is the pool of biodiversity has been reduced drastically and has changed the land use and land cover in the region. Up to 1950 Indonesia was still densely forested, but 40% of the forests existing in that year were cleared in the following 50 years. In round numbers, forest cover fell from 162 million hectares to 98 million hectares during 50 years period (FWI/GFW 2002). This phenomenon makes Indonesia belong to one of the countries with the highest tropical forest loss rate in the world. The annual rate of deforestation was at least 1.7 million hectares between 1985 and 1997, and it has been even higher at about 2 million hectares lost annually since 1997 (Scotland 2000). Conversion, degradation, and fragmentation threaten the integrity of forested ecosystem. Holmes (2002) suggested that without any conservation measures, tropical lowland evergreen forest would be diminished by 2005 in

Sumatra and after 2010 in Kalimantan due to deforestation. He estimated the rate of deforestation in Kalimantan during the period of 1985 until 1997 reaching 8.5 million hectares, with a loss of 21%, or 706,000 ha per year. Curran et al. (2004) suggested that deforestation in Kalimantan was not primarily due to local human population density, smallholder agricultural clearing, or paved roads. Kalimantan has relatively low human population density and growth rates (MacKinnon et al. 1996). However, the spontaneous interior migration which caused slash and burn cultivation can not be neglected. Kalimantan is also distinctive because of the dominance of the timber industry and the commercial value of stock of Dipterocarpaceae forests. Over the past two decades, the volume of dipterocarp timber exports from Borneo (Kalimantan, Sarawak, and Sabah) exceeded all tropical wood exports from tropical Africa and Latin America combined (ITTO 1996). There has been no simple explanation for deforestation in Indonesia. What so ever the causes of deforestation, the reduction of forest habitats is still continuing and it is therefore, sound regional land use planning is critical to protecting lowland forest habitats from increasingly human pressures. During the implementation of “Landscape-based Conservation of Orangutans in Trans-boundary Landscape between Central and West Kalimantan Provinces” we were able to characterize the patterns of deforestation in the

ADHIKERANA & SUGARDJITO – Forest reduction in Ketapang, West Kalimantan

region in order to understand at local to regional scales how changes in land use and land cover related to forest habitats reduction in the district of Ketapang, West Kalimantan. MATERIALS AND METHODS Digital data For the purpose of evaluating the changes in land use and land cover in the study areas, namely: Ketapang district, West Kalimantan, Indonesia and Sungai Puteri peat swamp forest (within the district) over 18 years (from 1989 to 2007), a series of Landsat imageries were analyzed. However, only six series of Landsat 7 ETM Path 121 Row 61 were available for the areas of Ketapang District (i.e. years of 1992, 2000, 2002, 2003, 2004 and 2005), and seven series of Landsat 5 ETM Path 121 Row 61 were available for analyzing Sungai Puteri peat swamp forest, namely those of 1989, 1990, 1999, 2000, 2002, 2005, and 2007 images. The area of Ketapang district analyzed lies within the coordinates: 0°45’00”-3°00’0” S and 108°30’0”111°30’0”E. and that of Sungai Puteri peat swamp forest lies within the following coordinates: 1o15’00”-1o55’00”S and 109o58’00”-110o25’00”E, where rivers are the main natural barrier for this area (namely Pawan river – ca. 75 m width – in the south, and Rangkung river – ca. 5 m width – in the north) in addition to the main road from Ketapang to Sukadana. The source of digital data for Ketapang district was the Ministry of Forestry, while those of Sungai Puteri peat swamp forest was analyzed from satellite imageries. Image pre-processing The subset extracted from each Landsat-TM image was geometrically corrected and geo-referenced using 1:50,000 topographic maps of the study area obtained from the National Agency for Survey and Mapping of Indonesia, where image-to-image rectification was undertaken. Radiometric correction was undertaken using Envi 4.1 and ERDAS 9.1 for combining and manipulating band-widths. Later, image enhancement was carried out in order to obtain images with good quality of visual and spectral data. Vegetation classification Each of the Landsat imagery data set for all study areas was initially classified using Unsupervised Classification into 13 different land-cover types: Primary forest, Secondary forest, Peat swamp forest, Mangrove forest, Coastal fishery, Swamp, Savannah, Agriculture, Bushes, Plantation, Mining, Settlement, and Barren land. Two other classes were also identified, namely undetermined area and no data due to cloud coverage. Ground truth checks were undertaken for Ketapang district and more details for Sungai Puteri peat swamp forest, from which the results were used for the Supervised Landuse Classification of these two study areas. The final land use classification for Sungai Puteri peat swamp forest consists of mangrove forest, peat swamp forest, riparian forest, oil-palm plantation, settlement, agriculture and settlement, swamp area, bushes, and barren land. The extent of each land cover area was calculated directly using ArcGIS 9.2.

Patterns of landuse changes and deforestation rate In order to establish the land cover changes that occurred within the study areas, a post classification change detection analysis of the available imageries was performed. For this purpose, two sets of information classes are available: (i) West Kalimantan province and Ketapang district comprising the data derived from 1992 to 2005.Landsat images, which are further grouped into two series of changes, namely “from 1992 to 2000 land use change” and “from 2000 to 2005 land use change”. This was done to account for the impacts of forest fire occurring in 1996-1997 on the calculation of deforestation rate analyzed in this study; (ii) Sungai Puteri peat swamp forest comprising the data derived from satellite images recorded during the 1989-2007 period, which are grouped into two series of changes, namely “from 1989 to 1999 land use change” and “from 1999 to 2007 land use change” for the same reason as before. Each “from to land use change” was detected with overlapping the relevant imageries, with which the extent of any significant change from one class to another could be estimated. Deforestation is defined as the conversion of forested areas to non-forest land use such as arable land, urban use, logged area or wasteland. Tree height and percent crown cover are the quantitative forest parameters used widely in classifying forest from non-forest areas, as well as different categories of forest classes. Deforestation rates were estimated for the whole West Kalimantan region, the area of Ketapang district, and that of Sungai Puteri peat swamp forest. The deforestation rate for West Kalimantan province and Ketapang district was estimated for the following periods: 1992 to 2000, 2000 to 2002, 2002 to 2003, 2003 to 2004, 2004 to 2005, and 2000 to 2005. Sungai Puteri peat swamp forest was estimated for the following intervals: 1989 to 1990, 1990 to 1999, 1999 to 2000, 2000 to 2002, 2002 to 2005, 2005 to 2007, 1989 to 1999, and 1999 to 2007. Two models have been applied for calculating deforestation as follows (Puyravaud 2003): q = {(A2/A1)1/(t2 – t1)} – 1 r = [1/(t2 – t1)]*ln(A2/A1)

(1) (2)

where A1 and A2 are the forest cover at time t1 and t2, respectively. In these models, the rate of deforestation can be expressed as the percentage of remaining forest that is cleared per year. Equation (1) is proposed by FAO (1995) as to derive the rate of change from the Compound Interest Law, while equation (2) denotes the rate of the Compound Interest Law itself as proposed by Puyravaud (2003). Basically there are no significant differences in the calculation results between the two models, but here they are presented for the comparison purpose only. RESULTS AND DISCUSSION Land-cover changes Ketapang District Unfortunately the available imageries data were stained by cloud coverage and poor mosaic that made some areas

BIODIVERSITAS

11 (1): 46-54, January 2010

were undetermined, and yet these areas are presumed to be the spots of forest fire occurring in 1997 (Table 1). This constraint has made it impossible to predict changes in these areas. There seems to be virtual changes from “forest” to “non-forest” during the period of 1992 until 2000, but substantial changes from “forest” to “non-forest” occurred during the period of 2000 to 2005, when about 27.11% of forested areas converted into non-forested areas (Figure 1; Figure 3). During the period of 1992 to 2000 there was only very small conversion of primary forest into secondary forest; there was a slight succession from secondary forest to primary forest as well as from swampy, agriculture, plantation, settlement, and barren areas into secondary forest. During this period barren land remained extensive about 30.17% of the total area of Ketapang district (Table 2).

Figure 1. Changes in the extent of forest and non forest areas in Ketapang district during the period of 1992 to 2005.

Table 1. Land use of Ketapang district (ha) during the period of 1992 to 2005. 1992

2000

2002

2003

Primary forest 439,573.00 439,563.00 348,264.55 294,236.73 Secondary forest 1,282,921.55 1,280,955.12 606,809.64 758,412.54 Peat swamp forest 617,189.01 617,184.52 717,390.51 513,910.97 Mangrove forest 13,405.21 12,428.40 37,555.84 32,870.24 Coastal fishery 1,537.25 Swamp 65,420.08 65,420.12 158,077.42 359,438.55 Savannah 2,291.48 Agriculture 224,527.01 224,527.27 486,051.59 752,872.63 Bushes 893,639.32 333,077.43 Plantation 87,346.12 87,346.23 73,774.43 114,706.91 Mining 508.80 2,706.88 Settlement 3,607.07 3,607.14 2,073.26 18,193.09 Barren land 1,108,143.36 1,108,143.10 6,214.76 237,146.94 Undetermined 139,302.40 140,803.35 130,505.01 124,431.10 No data (cloud) 130,310.86 131,757.43 211,747.10 126,779.51 Total 4,111,745.67 4,111,735.68 3,672,612.25 3,672,612.25 Source: Available GIS data for Ketapang district acquired by FFI and analysed for this study.

2004

2005

290,553.96 761,265.07 510,349.10 26,282.87 1,594.24 692,711.70 2,291.51 753,599.83 114,737.29 2,706.93 17,730.75 26,835.45 455,670.23 16,283.32 3,672,612.25

292,018.00 756,141.05 511,813.03 32,870.93 1,594.24 359,850.62 2,291.48 753,599.80 332,861.04 114,737.26 2,706.90 17,730.71 26,835.41 347,770.74 119,791.04 3,672,612.25

Table 2. Details of landuse changes (ha) in Ketapang district during the period of 1992 to 2000.

From (in 1992)

Peat Mangrove swamp Swamp Agriculture Plantation Settlement Barren land forest forest 439,573.00 1,280,953.88 617,184.14 13,005.21 65,420.08 224,527.01 87,346.12 3,607.07 1,107,132.78 Primary forest

Secondary forest

To (in 2000) Primary forest Secondary forest Peat swamp forest Mangrove forest Swamp Agriculture Plantation Settlement Barren Land Total

439,563.00 0.01 1.00 1,280,950.41 0.29 0.11 0.02 0.57 0.09 0.25 617,181.62 0.16 0.43 0.44 0.04 0.01 0.14 13,004.51 0.02 0.08 0.01 0.01 0.49 0.03 65,419.15 0.12 0.02 0.72 0.50 0.02 0.16 224,525.07 0.12 0.08 0.14 0.23 0.04 0.13 87,345.67 0.01 2.39 0.96 0.16 0.27 0.58 0.18 439,564.00 1,280,953.88 617,184.14 13,005.21 65,420.08 224,527.01 87,346.12

3,607.02 0.03 3,607.07

To (in 2000) Remaining (ha) Converted (ha)

439,563.00 1,280,950.41 617,181.62 13,004.51 65,419.15 224,525.07 87,345.67 1 3.47 2.52 0.71 0.94 1.93 0.46

3,607.02 1,107,127.99 0.04 4.80

0.0002

0.0003

0.0004

0.0055

0.0014

0.0009

0.0005

0.01

0.0011

2.47 1.30 0.02 0.27 0.51 0.21 0.02 1,107,127.99 1,107,132.78

0.0004

ADHIKERANA & SUGARDJITO – Forest reduction in Ketapang, West Kalimantan

Both agriculture and plantation areas substantially increased in year 2003, and at the same time the extent of both primary and peat swamp forests reduced considerably. Further, a ground truth survey has also confirmed such a conversion. Mining activities started in 2002 has been identified in a relatively small area, but it increased substantially in the year 2003. This mining however, has taken place in secondary forests. Such mining seemed to create “savannah-like” landscape with barren land more exposed in extent. During the period of 2000 to 2005 the primary forest remained secured, but substantial changes occurred in other land use classes (Table 3). As it has been shown here the abandoned areas might have been the new settlement areas (‘transmigration” sites) which were deserted by the migrants. This is a normal case for Kalimantan, where migrants returned to their original site after the government supports no longer provide their life in a few years. The conversion of plantation area into bushes could be due to the desertion of “transmigration” areas. Substantial conversion of agricultural land to barren land could also be similar to the above mentioned case. The most striking conversion was from secondary forest to agricultural land suggesting the impact of economic development during this period, and from peat swamp forest to swamp areas might be due to peat subsidence caused by canal construction. The barren land remains extensive in 2005 suggesting that this type of land has not been managed for economic development purposes. Deforestation has always been the subject of concerns since the early 60s (SAF 1983), and it is an ongoing disturbance within human-dominated landscapes. The type, intensity, and frequency of deforestation have long been known to be attributable to the extent of human activities on the landscape (Curtis

1956; Burgess and Sharpe 1981; Zipperer et al. 1990). Settlement area changed substantially leaving only 5.80% of the original area in 2000, where it extensively changed into bushes. However, significant succession occurred from secondary forest into primary forest and from barren land into secondary forest, and additionally though in more less extensive area from other land use into secondary forest. Sungai Puteri peat swamp forest The main land use being concerned in this forest block is due to the fact that it is an extensive peat land with an average depth of 7 m (range: 2-15 m) with a reduction of about 35% of its size in 1989 (Table 4). The forest areas in this forest block including peat, mangrove, and riparian forests have reduced more in the period 1989 to 1999 as compared to the 1999-2007 period, whereas at the same time, non-forest areas increased substantially at the first period (1989-1999) and gradually at the second period (Figure 2, Figure 4).

Figure 2. Changes in the extent of forest and non forest areas in the Sungai Puteri peat swamp forest blocks during the period of 1989 to 2007.

Table 3. Details of land use changes (ha) in Ketapang district during the period of 2000 to 2005.

From (in 2000)

Peat Mangrove swamp Swamp Agriculture Plantation Settlement Barren land forest forest 439,563.00 1,280,955.12 617,910.82 12,428.40 65,420.03 224,527.18 87,346.14 3,607.05 1,108,143.01 Primary forest

Secondary forest

To (in 2005): Primary forest Secondary forest Peat swamp forest Mangrove forest Coastal fishery Swamp Savanna Agriculture Bushes Plantation Mining Settlement Water Body Barren Land Total

22,730.15 78.42 146,191.31 57,371.74 20,522.77 830.43 2,082.53 1,719.37 18,361.85 439,563.00 1,280,955.12

To (in 2005): Remaining (ha) % Converted (ha) %

150,205.36 100 0 0

439,563.00

287,368.95 637,868.69 3,886.63 79,367.71 358,532.54 6,461.21 10,447.30

71.36 1,167.18 6,148.47

116,360.79 2,521.16 1,454.54 29,826.61 586.73 44,770.65 4,630.95 592.78 374.28 1,679.97 416.97 3,373.04 510.64 42,573.53 413.12 617,910.82 12,428.40

637,868.69 358,532.54 49.80 58.02 643,086.43 259,378.28 50.20 41.98

432.96 3.21 213.51 6,894.29 7,584.68 1,106.17 2,416.05 29.41 214.57 28,876.69 23,076.07 7,925.02 39.74 2,836.51 88,683.13 6,985.86 6,529.48 40,324.10 591.66 24,533.32 23,852.61 179.76 90.09 7,081.67 183.25 4,038.87 211.27 12,709.12 73,135.35 6,514.95 65,420.03 224,527.18 87,346.14

6,148.47 28,876.69 88,683.13 23,852.61 49.47 44.14 39.50 27.31 6,279.93 36,543.34 135,844.05 63,493.54 50.53 55.86 60.50 72.69

76.97 28.17 614.74

1,556.76 104,652.42 52,814.86 307.49

209.06 285.69 107.61 3,607.05

153,155.52 544.68 466,058.95 144,457.66 39,176.05 249.33 5,049.77 6,678.35 133,441.17 1,108,143.01

209.06 5.80 3,397.99 94.20

133,441.17 12.04 974,701.84 87.96

364.86 1,919.94

BIODIVERSITAS

11 (1): 46-54, January 2010

Table 4. Land use of Sungai Puteri peat swamp forest blocks (ha) during the period of 1989 to 2007. Landuse Mangrove forest Peat swamp forest Riparian forest Oil-palm plantation Settlement Agriculture and Settlement Swamp Bushes Barren land Total

1989 221.01 125,760.12 317.11 256.10 16,582.01 1,445.14 2,568.16 147,149.65

1990

1999

244.12 123,725.10 317.13 219.01 17,817.13 2,249.09 2,578.07 147,149.65

2000

131.50 91,928.74 345.32 383.83 50,756.18 501.32 263.45 2,839.32 147,149.65

174.46 88,850.65 297.36 195.61 50,410.78 491.96 4,329.09 2,399.74 147,149.65

2002

2005

145.62 86,553.60 298.66 342.31 53,279.77 2,264.52 4,265.19 147,149.66

2007

132.09 84,301.97 283.49 394.03 244.25 55,734.58 58.25 947.43 5,053.56 147,149.65

132.09 81,512.68 251.15 5,077.35 244.07 53,169.67 246.36 1,072.24 5,444.05 147,149.65

Table 5. Details of land use changes (ha) in Sungai Puteri peat swamp forest blocks during the period of 1989 to 1999.

From (in 1989) To (in 1999): Peat swamp forest Mangrove forest Riparian forest Swamp Agriculture Agriculture and Settlement Bushes Settlement Barren land Total To (in 2007): Remaining (ha) % Converted (ha) %

Peat swamp Mangrove Riparian Agriculture and Agriculture Bushes Settlement forest forest forest Settlement 128,262.35 221.48 315.65 16,305.08 210.84 1,639.63 249.09 90,606.41 54.22 414.53 36,171.60 102.19 263.46

Barren land 2,568.13

649.95 128,262.35

128.35 93.12 221.48

291.11 24.54 315.65

825.55 15,220.62 5.42 253.48 16,305.08

13.14 11.88 185.82 210.84

364.36 1,275.27 1,639.63

2.30 128.08 118.72 249.09

117.16 43.28 106.66 2,301.02 2,568.13

90,606.41 70.64 37,655.94 29.36

128.354 57.95 93.12 42.05

291.105 92.23 24.54 7.77

15,220.624 93.35 1,084.45 6.65

185.823 88.14 25.01 11.86

0.000 0.00 1,639.63 100.00

118.716 47.66 130.37 52.34

2,301.022 89.60 267.10 10.40

Table 6. Details of land use changes (ha) in Sungai Puteri peat swamp forest blocks during the period of 1999 to 2007.

From (in 1999): To (in 2007): Peat swamp forest Mangrove forest Riparian forest Swamp Agriculture Agriculture and Settlement Bushes Oil-palm plantation Settlement Barren land Total To (in 2007): Remaining (ha) % Converted (ha) %

Peat Mangrove Riparian swamp forest forest forest 91,928.85 130.50 345.32 81,093.58 0.02 7.06 8,182.30

89.99 -

253.11

40.51

33.93

Swamp

Agriculture Barren and Bushes Settlement land Settlement 52,909.43 293.19 263.46 383.84 2,939.32

Agriculture

501.32

50.65

70.98 158.90 67.65

651.47 437.42 9.00 1,514.06 91,928.85

130.50

41.57 345.32

81,093.58 88.21 10,835.27 11.79

89.985 68.96 40.51 31.04

253.105 73.30 92.22 26.70

760.44 41.87 0.62 45,427.42

12.98 0.01

248.33

20.90 131.81

47.07 81.28 104.93

1.78

280.21

202.07 1.72 501.32

132.43 4,628.41 50.22 1,866.24 52,909.43

293.19

15.13 263.46

40.94 190.19 383.84

2,706.04 2,939.32

158.896 45,427.423 31.70 85.86 342.43 7,482.01 68.30 14.14

280.207 95.57 12.98 4.43

15.127 5.74 248.33 94.26

190.194 2,706.043 49.55 92.06 193.65 233.28 50.45 7.94

ADHIKERANA & SUGARDJITO â&#x20AC;&#x201C; Forest reduction in Ketapang, West Kalimantan

1989

1999

2002

ADHIKERANA & SUGARDJITO â&#x20AC;&#x201C; Forest reduction in Ketapang, West Kalimantan

1992 2000 2003 2005 Figure 3. Cumulative of forest reduction and land use changes in Ketapang district excluding Sungai Puteri forest block during the period of 1992 and 2005. Forest and nonforest classifications are based on a Landsat Thematic Mapper time series 1992, 2000, 2002, 2003, 2004, and 2005.

2007 51

Figure 4. Cumulative of forest reduction and land use changes in Sungai Puteri peat swamp forest block during the period of 1989 and 2007. Forest and nonforest classifications are based on a Landsat Thematic Mapper time series 1989, 1990, 1999, 2000, 2002, 2005, and 2007. Classifications are shown for 1989, 1999, and 2007.

BIODIVERSITAS

11 (1): 46-54, January 2010

In details, peat swamp forests seemed to be subject for conversion to agriculture, and subsistence agriculture. These conversions have been accompanied by small settlement which are identified as “agriculture and settlement” in the table. Further, it was followed by settlement and later to oil-palm plantation. During the period of 1989 to 1999, the conversion of peat swamp forest into agriculture area was considerably extensive, almost 28% of its original size in 1989. However, there were more than 1,000 ha of non-forest areas, especially agriculture, bushes, settlement and barren land, reestablished into peat swamp forests (Table 5). During the period of 1999 to 2007, conversion of peat swamp forest into agricultural land remained substantial although it was much lower compared to previous period but, a fraction of this forest was also converted into oilpalm plantation. The oil-palm plantation was even extended with considerable size came from agricultural land and peat swamp forest (Table 6). Barren land was also increased and it is formed from a more varied land use. The intensity of peat swamp forest converted into barren land was quite significant during this period, and the field survey confirmed that most of which were abandoned agriculture. At the same time, a number of land use types also re-established peat swamp forest, namely swamp, agriculture, agriculture and settlement, settlement, and barren land, with a total of about 900 ha, of which were abandoned land. The striking changes of settlement, agriculture, and plantation areas into bushes and barren land in Ketapang district could have been due to migrant desertion. Sunderlin and Resosudarmo (1996) described two types of migrants, namely “regular” who receive full government assistance, whereas the “spontaneous” one who receive partial or did not receive at all of government assistance. They suggested that forest cover removal for the migrant settlement site has been the direct effect of regular migrants. Dick (1991) has been assessed that spontaneous migrants could be the single most important cause of forests degradation, although, Sunderlin and Resosudarmo (1996) suggested that it might not be necessarily the case. The desertion of migrant resettlement sites in Ketapang district could always happen due to insufficient incomes that force the migrants move to other sites which is posing another land pressure on other forested land. Further, Cernea and Schmidt-Soltau (2003) showed that forcing resettlement, i.e. “transmigration”, could push people to face impoverishment risks such as landlessness, joblessness, homelessness, marginalization, food insecurity, increased morbidity and mortality, loss of access to common property, and social disarticulation. The migrant people in Ketapang district might have been exposed to such risks that made them move away from their designated sites. On the other hand, barren land both in Ketapang district and Sungai Puteri peat swamp forest blocks remains extensive without any assessment on economic benefits. Such barren land is certainly prone to wild fire during the dry season and it can be attributable to people perception or skill to utilize marginal land. As Zube (1987) pointed out that land use patterns could be perceived in many perspectives,

where they could indicate landscape function, economic opportunity, and environmental amenities, and each with different value orientation. Estate plantation companies could perceive such barren land as less economic opportunity since its conversion into plantation might need considerable investment to recover the land. At the same time, the local people could have the same perception with different reasoning, for example they do not know how to recover the barren into arable land. When such underskilled local people are not assisted to gain more capacity in technological skill, the barren land would always extent in the future. This indicates that capacity development for people living in surrounding forest habitat is urgently needed. O’Connor et al. (2003) indicated that socioeconomic and political conditions influence the effectiveness of conservation interventions, and therefore, improvement of local people capacity will certainly need appropriate socio-economic and political will in sustaining the environmental conservation. Mikkelson et al. (2007) also suggest that inequality of economic opportunity could induce biodiversity loss through a number of devastating human activities. In Kalimantan, concession-based timber extraction, plantation establishment, and weak institutions have resulted in highly fragmented and degraded forests (Ross 2000) and one of the main impacts is habitat fragmentation. Forest habitat fragmentation is considered by many to be the most important threat to biological diversity, and is the primary cause of the present biodiversity extinction crisis (Wilcox and Murphy 1985; Laurance et al. 2003; Wilcove et al. 1986). Rate of deforestation Tables 7a and 7b have shown the rate of deforestation both in the Ketapang district excluding Sungai Puteri and in the Sungai Puteri block of forest alone respectively. Despite forest fire disaster in 1997, there was virtually lack of deforestation in Ketapang district during the period of 1992 to 2000. Deforestation was significantly increased from 2000 to 2003, then it slowed down from 2003 to 2005 (Table 7). In average, the deforestation rate in Ketapang district during the period of 2000 to 2005 was 3.93% per year. When we compared to the whole area of Ketapang district, deforestation in Sungai Puteri peat swamp forest blocks was low. In 2000-2002 it was only 1.31% contrasting to 6.14% for Ketapang district. In average the annual deforestation rate in Sungai Puteri during the period of 1989 to 2007 was 2.41%. There has been no simple explanation for ongoing deforestation in Indonesia. It was suggested that unsynchronized government policies, institutional arrangement and market demands are being the main triggers of deforestation (Adhikerana 2002). The rate of deforestation has been enhanced by the change of government policy towards decentralization implemented in 2001 which allows local district to issue small logging parcel leases of 1 km2. This has resulted in the virtually uncontrolled harvest of remaining accessible lowland forests. Further, widespread oil palm plantation establishment is also converting lowland forest. In this study, a substantial development activities including

ADHIKERANA & SUGARDJITO â&#x20AC;&#x201C; Forest reduction in Ketapang, West Kalimantan

agriculture, plantation, and mining in Ketapang district during the period of 2000 to 2005 were majority took place in both secondary and peat swamp forests. A similar case was also found in Sungai Puteri peat swamp forest blocks, where agriculture, settlement, and oil-palm plantation took place mostly in the peat swamp forests although, this peat swamp forest is not allowed to be converted in accordance to the Presidential Decree No 32/1990. Despite the average depth of this peat swamp forest is 7 meters there has been a significant conversion in the extent of peat swamp forest in this area. However,, to some extent such a significant changes of land cover might have been attributed to forest fire occurring in 1997 and this was confirmed during the ground truth where forest fire spots are now become barren land and swamp areas. This might be due to the fact that Kalimantanâ&#x20AC;&#x2122;s rainforests are driven by El Nino Southern Oscillation events. When forest fragmentation and deforestation increased it could transforms El Nino from regenerative to a high destructive phenomenon, one that triggers droughts and wild fires with increasing frequency and intensity, disrupts fruting of dipterocarp trees, interrupts wildlife reproductive cycles, and erodes the basis for rural livelihoods (Curran et al. 1999). Further, forest habitat fragmentation has been shown to induce changing in the primate behavior (Stickler 2004). In Kalimantan recently, orangutan has been found to raid the timber industrial plantation area in order to feed the cambrium of Acacia mangium, the tree species planted for pulp and paper industries. This phenomenon has never been seen when the forests habitat still intact and unlogged. This suggests resource and habitat limitation have constrained the species well-being.

Unfortunately, people are commonly view peat swamp forest as naturally unproductive. That is why the majority of peat swamp forests suffer from land use conversion. The ecological functions of peat swamp forest as carbon store and geological source of organic rock are mainly neglected, and especially deforestation towards peat swamp forest is confirmed to have a severe impact on the global atmosphere. The tropical deforestation would releases 1.5 billion tonnes of carbon each year into the atmosphere (CSIRO 2007). A report suggested that tropical deforestation, including both the permanent conversion of forests to croplands and pastures, and the temporary or partial removal of forests for shifting cultivation and selective logging, is estimated to have released on the order of 1-2 PgC/year or 15% to 35% of annual fossil fuel emissions during the 1990s (Moutinho and Schwartzman 2005). Therefore, the avoided deforestation scheme is now gaining more concerns from the global communities, which actually presents an alternative economic development for the local people through carbon trade mechanism, such as the Reducing Emission from Deforestation and Forest Degradation (REDD). While protecting the peat swamp forest habitat for orangutan population in Sungai Puteri, it could also gain an invaluable benefit from the REDD initiative. It was predicted that Long Term Carbon Accumulation Rate (LORCA) in this peat swamp forest may probably fall between 0.4 and 0.8 t C/ha per year. (Anshari et al. 2009). However, it is not sure whether this peat swamp forest would survive or not under the present disturbances related to global climate change and economic development program.

Table 7. Deforestation rate in Ketapang District (in Ketapang district forests excluding Sungai Puteri, and in Sungai Puteri Peatforest blocks).

CONCLUSIONS

From

Interval (years)

q (%)

r (%)

1992 2000 2002 2003 2004 2000

2000 2002 2003 2004 2005 2005

Ketapang district 8 2 1 1 1 5

0.01 6.14 6.33 0.28 0.14 3.85

0.01 6.34 6.54 0.28 0.14 3.93

1989 1990 1999 2000 2002 2005 1989 1999 1989

Sungai Puteri peat swamp forest 1990 1 1.59 1999 9 3.24 2000 1 3.34 2002 2 1.31 2005 3 0.88 2007 2 1.68 1999 10 3.08 2007 7 1.50 2007 18 2.38

1.61 3.29 3.39 1.32 0.89 1.69 3.12 1.51 2.41

The Ketapang district forests are well known for being the home of orangutan, the endangered great ape species. A newly discovered orangutan population in Sungai Puteri peat swamp forest blocks could be used as a tool to protect this peat swamp forest as the habitat of orangutan.

This study shows that the rate of deforestation in Ketapang district of West Kalimantan remarkably increased during the period of 2000 to 2004 reaching to 6.54%. Although it was slowing down during 2004 to 2005, the average rate from 2000 to 2005 was 3.39% per year. Despite the high rate of deforestation in Indonesia as a whole which reached 1.08 million hectares during 20002005, or even that in Kalimantan with 1.23 million hectares, the extent of deforestation in Ketapang district was merely 74,590 ha per year. The patterns of land use changes detected in this study indicated a number of possible causes of deforestation in this district. The local government policy and market demand could be those of so many triggers of deforestation, in addition to impoverishment risks experienced by the local people, especially the migrants. Deforestation patterns in the study areas have created forest fragmentation, from which the structural change of landscape can increase the probability of natural disturbance and of exposure of the species diversity and wildlife population to deterioration. Appropriate forest conservation measures combined with pro-poor development, through for example REDD scheme, will certainly help lessen the human pressures on the lowland forests in this district as well as the peat swamp forest blocks of Sungai Puteri.

BIODIVERSITAS

11 (1): 46-54, January 2010

ACKNOWLEDGEMENT The HCVF team of FFI-Ketapang program has helped collected data and undertaken ground truth check in the field. Their supports are deeply appreciated. We also acknowledge the technical assistance of Darkono for preparation of digital maps of this paper. The local people of Sungai Puteri, Sungai Tolak and other villages have also been helpful in the field. Their supports will be unforgettable. The government of Ketapang district has also been very supportive to the study, about which great appreciation is presented. We are grateful to Orangutan Conservation Support Program for granting fund to this fieldwork. REFERENCES Adhikerana AS (2002) The triggers of deforestation in Indonesia. Jambi Express 18 Maret 2002. (Bahasa Indonesia) Anshari ZG, Momberg F, Adhikerana A, Darkono, Nuriman M, Sahir M (2009) Horizontal and vertical distribution of carbon, nitrogen and sulfur concentrations in Sungai Putri Peat Forest, Ketapang District of West Kalimantan Province, Indonesia: variations in fluvial-deltaic and coastal reservoirs deposited in tropical environments. AAPG Hedberg Conference, Jakarta, Indonesia, April 29-May 2, 2009. Burgess RL, Sharpe DM (eds) (1981) Forest island dynamics in mandominated landscapes. Springer, New York. Cernea MM, Schmidt-Soltau K (2003) Biodiversity conservation versus population resettlement: risks to nature and risks to people. The International Conference on Rural Livelihoods, Forests and Biodiversity, Bonn, Germany, 19-23 May 2003. CSIRO (2007) Confirmed: deforestation plays critical climate change role. Science Daily, May 11, 2007. Retrieved October 22, 2008. Curran LM, Chaniago I, Paoli GD, Astani D, Kusneti M, Leighton M, Neraista CE, Haeruman H (1999) Impact of El Nino and logging on canopy tree recruitment in Borneo. Science 286: 2184-2188. Curran LM, Trigg SN, McDonald AK, Astiani D, Hardiono, YM, Siregar P, Caniago I, Kasischke E (2004) Lowland forest loss in protected areas of Indonesian Borneo. Science 303: 1000-1003. Curtis JT (1956) The modification of mid-latitude grasslands and forests by man. In: Thomas WL (ed). Man’s role in changing the face of the earth. University of Chicago Press, Chicago. Dick J (1991) Forest land use, forest use zonation, and deforestation in Indonesia: a summary and interpretation of existing information.

Background paper to UNCED for the State Ministry for Population and Environment (KLH) and the Environmental Impact Management Agency. BAPEDAL, Jakarta. FAO (1995) Forest resource assessment 1990. Global Synthesis. FAO, Rome. FWI/GFW (2002) The state of the forest: Indonesia. Global Forest Watch, Washington DC. Holmes D (2002). Deforestation in Indonesia: a review of the situation in Sumatra, Kalimantan and Sulawesi. World Bank, Jakarta. ITTO (International Tropical Timber Organization) (1996). Annual review and assessment of world tropical timber situation 1995. ITTO Japan, Yokohama. Laurance WF, Rankin de Merona JM, Andrade A, Laurance SG, D’Angelo S, Lovejoy TE, Vasconcelos HL (2003) Rain-forest fragmentation and the phenology of Amazonian tree communities. J Trop Ecol 19: 343-347. MacKinnon KS, Hatta G, Halim H, Mangalik A (1996) The ecology of Kalimantan. Periplus, Singapore. Mikkelson GM, Gonzalez A, and Peterson GD (2007) Economic inequality predicts biodiversity loss. Plos One 2 (5): 444. doi:10.1371/journal.pone.0000444 O’Connor C, Marvier M, Kareiva P (2003) Biological versus sociopolitical priority-setting in conservation. Ecol Lett 6: 706-711 Puyravaud, JP (2003) Standardizing the calculation of the annual rate of deforestation. Forest Ecol Manag 177: 593-596. Ross ML (2000) Timber booms and institutional breakdown in Southeast Asia. Cambridge University Press, Cambridge. SAF (1983) Terminology of forest science technology, practice and products. Society of American Foresters, Bethesda, Maryland. Scotland N (2000) Indonesia country paper on illegal logging. The World Bank-WWF Workshop on Control of Illegal Logging in East Asia. Jakarta, 28 August 2000. Stickler CM (2004) The effects of logging on primate-habitat interactions: a case study of redtail monkeys (Cercopithecus ascanius) in Kibale National Park, Uganda. (Thesis). Graduate School of the University of Florida, Florida. Sunderlin W, Resosudarmo IAP (1996) Rates and causes of deforestation in Indonesia: towards a resolution of the ambiguities. CIFOR Occasional Paper No. 9. CIFOR, Bogor. Wilcove DS, McLellan CH, Dobson AP (1986) Habitat fragmentation in the temperate zone. In: Soule ME (ed). Conservation biology: the science of scarcity and diversity Sinauer, Sunderland. Wilcox BA, Murphy DD (1985) Conservation strategy: the effects of fragmentation on extinction. Amer Nat 125: 879-887. Zipperer WC, Burgess RL, Nyland RD (1990) Patterns of deforestation and reforestation in different landscape types in central New York. Forest Ecol Manag 36: 451-471. Zube EH (1987) Perceived land use patterns and landscape values. Landscape Ecol 1: 37-45.

GUIDANCE FOR AUTHORS BIODIVERSITAS, the Journal of Biological Diversity publishes scientific articles, i.e. original research and review in all biodiversity aspects of plants, animals and microbes at the level of gene, species, and ecosystem. Scientific feedback (short communication) is only received for manuscript, which criticize published article before. Manuscripts will be reviewed by managing editor and invited peer review according to their disciplines. The only articles written in English (U.S. English) are accepted for publication. This journal periodically publishes in January, April, July, and October. In order to support reduction of global warming as a consequence of transportation vehicles emission and forest degradation for paper manufacturing, management of the journal prefer receiving manuscripts via e-mail rather than in hard copy. Manuscript and its communications can only be addressed to the managing editor; better to “CC” to one of the communicating editor for accelerating evaluation. A letter of statement expressing that the author (s) is responsible for the original content of manuscript, the result of author(s)’s research and never been published must be attached. Manuscript is typed at one side of white paper of A4 (210x297 mm2) size, in a single column, double space, 12-point Times New Roman font, with 2 cm distance step aside in all side. Smaller letter size and space can be applied in presenting table. Word processing program or additional software can be used, however, it must be PC compatible and Microsoft Word based. Names of sub-species until phylum should be written in italic, except for italic sentence. Scientific name (genera, species, author), and cultivar or strain should be mentioned completely at the first time mentioning it, especially for taxonomic manuscripts. Name of genera can be shortened after first mentioning, except generating confusion. Name of author can be eliminated after first mentioning. For example, Rhizopus oryzae L. UICC 524, hereinafter can be written as R. oryzae UICC 524. Using trivial name should be avoided, otherwise generating confusion. Mentioning of scientific name completely can be repeated at Materials and Methods. Biochemical and chemical nomenclature should follow the order of IUPAC-IUB, while its translation to Indonesian-English refers to Glossarium Istilah AsingIndonesia (2006). For DNA sequence, it is better used Courier New font. Symbols of standard chemical and abbreviation of chemistry name can be applied for common and clear used, for example, completely written butilic hydroxytoluene to be BHT hereinafter. Metric measurement use IS denomination, usage other system should follow the value of equivalent with the denomination of IS first mentioning. Abbreviation set of, like g, mg, mL, etc. do not follow by dot. Minus index (m-2, L-1, h-1) suggested to be used, except in things like “per-plant” or “per-plot”. Equation of mathematics does not always can be written down in one column with text, for that case can be written separately. Number one to ten are expressed with words, except if it relates to measurement, while values above them written in number, except in early sentence. Fraction should be expressed in decimal. In text, it should be used “%” rather than “gratuity”. Avoid expressing idea with complicated sentence and verbiage, and used efficient and effective sentence. Manuscript of original research should be written in no more than 25 pages (including tables and picture), each page contain 700-800 word, or proportional with article in this publication number. Invited review articles will be accommodated. Title of article should be written in compact, clear, and informative sentence preferably not more than 20 words. Name of author(s) should be completely written. Running title is about five words. Name and institution address should be also completely written with street name and number (location), zip code, telephone number, facsimile number, and e-mail address. Manuscript written by a group, author for correspondence along with address is required. First page of the manuscript is used for writing above information. Abstract should not be more than 200 words, written in English. Keywords is about five words, covering scientific and local name (if any), research theme, and special methods which used. Introduction is about 400600 words, covering background and aims of the research. Materials and Methods should emphasize on the procedures and data analysis. Results and Discussion should be written as a series of connecting sentences, however, for manuscript with long discussion should be divided into sub titles. Thorough discussion represents the causal effect mainly explains for why and how the results of the research were taken place, and do not only reexpress the mentioned results in the form of sentences. Concluding sentence should preferably be given at the end of the discussion. Acknowledgments are expressed in a brief. Figures and Tables of maximum of three pages should be clearly presented. Title of a picture is written down below the picture, while title of a table is written in the above the table. Colored picture and photo can be accepted if information in manuscript can lose without those images. Photos

and pictures are preferably presented in a digital file. JPEG format should be sent in the final (accepted) article. Author could consign any picture or photo for front cover, although it does not print in the manuscript. There is no appendix, all data or data analysis are incorporated into Results and Discussions. For broad data, it can be displayed in website as Supplement. Citation in manuscript is written in “name and year” system; and is arranged from oldest to newest and from A to Z. The sentence sourced from many authors, should be structured based on the year of recently. In citing an article written by two authors, both of them should be mentioned, however, for three and more authors only the family (last) name of the first author is mentioned followed by et al., for example: Saharjo and Nurhayati (2006) or (Boonkerd 2003a, b, c; Sugiyarto 2004; El-Bana and Nijs 2005; Balagadde et al. 2008; Webb et al. 2008). Extent citation as shown with word “cit” should be avoided, and suggested to refer an original reference. APA style in double space is used in the journal reference as follow: Journal: Carranza S, Arnold EN (2006) Systematics, biogeography and evolution of Hemidactylus geckos (Reptilia: Gekkonidae) elucidated using mitochondrial DNA sequences. Mol Phylogenet Evol 38: 531-545.

Saharjo BH, Nurhayati AD (2006) Domination and composition structure change at hemic peat natural regeneration following burning; a case study in Pelalawan, Riau Province. Biodiversitas 7: 154-158. Book: Rai MK, Carpinella C (2006) Naturally occurring bioactive compounds. Elsevier, Amsterdam. Chapter in book: Webb CO, Cannon CH, Davies SJ (2008) Ecological organization, biogeography, and the phylogenetic structure of rainforest tree communities. In: Carson W, Schnitzer S (eds) Tropical forest community ecology. Wiley-Blackwell, New York. Abstract: Assaeed AM (2007) Seed production and dispersal of Rhazya stricta. 50th annual symposium of the International Association for Vegetation Science, Swansea, UK, 23-27 July 2007. Proceeding: Alikodra HS (2000) Biodiversity for development of local autonomous government. In: Setyawan AD, Sutarno (eds) Toward mount Lawu national park; proceeding of national seminary and workshop on biodiversity conservation to protect and save germplasm in Java island. Sebelas Maret University, Surakarta, 17-20 July 2000. [Indonesian] Thesis, Dissertation: Sugiyarto (2004) Soil macro-invertebrates diversity and inter-cropping plants productivity in agroforestry system based on sengon. [Dissertation]. Brawijaya University, Malang. [Indonesian] Information from internet: Balagadde FK, Song H, Ozaki J, Collins CH, Barnet M, Arnold FH, Quake SR, You L (2008) A synthetic Escherichia coli predator-prey ecosystem. Mol Syst Biol 4: 187. www.molecularsystemsbiology.com Publication manuscript “in press” can be cited and mentioned in reference (bibliography); “personal communications” can be cited, but cannot be mentioned in reference. Research which not be published or “submitted” cannot be cited. Some annotation. Manuscript typed without sign link (-) (except repeated word in Indonesian). Usage of letter “l” (el) to “1” (one) or “O” (oh) to “0” (null) should be avoided. Symbols of α, β, χ, etc. included through facility of insert, non altering letter type. No space between words and punctuation mark. Progress of manuscript. Notification of manuscript whether it is accepted or refused will be notified in about three months since the manuscript received. Manuscript is refused if the content does not in line with the journal mission, low quality, inappropriate format, complicated language style, dishonesty of research authenticity, or no answer of correspondence in a certain period. Author or first authors at a group manuscript will get one original copy of journal containing manuscript submitted not more than a month after publication. Offprint or reprint is only available with special request. NOTE: Author(s) agree to transfer copy right of published paper to BIODIVERSITAS, Journal of Biological Diversity. Authors shall no longer be allowed to publish manuscript completely without publisher permission. Authors or others allowed multiplying article in this journal as long as not for commercial purposes. For the new invention, authors suggested to manage its patent before publishing in this journal.

NOTIFICATION: All communications are strongly recommended to be undertaken through email.

ISSN: 1412-033X (printed edition) ISSN: 2085-4722 (electronic)

GENETIC DIVERSTY

Genetic variations among Indonesian native cattle breeds based on polymorphisms analysis in the growth hormone loci and mitochondrial DNA SUTARNO The genetics of glutamate oxaloacetate transaminase (GOT) in Pinus merkusii Jungh. et de Vriese ISKANDAR ZULKARNAEN SIREGAR, TEDI YUNANTO

1-4

5-8

SPECIES DIVERSTY

Systematic studies of Australian stipoid grasses (Austrostipa) based on micromorphological and molecular characteristics BETTY MAULIYA BUSTAM

9-14

Begonia “Tuti Siregar” (Begonia listada Smith &Wasshausen x Begonia acetosa Vellozo): a new cultivar from Bali Botanic Garden, Indonesia

15-18

HARTUTININGSIH-M. SIREGAR, I MADE ARDAKA, MUSTAID SIREGAR ECOSYSTEM DIVERSTY

Population dynamic of the swallowtail butterfly, Papilio polytes (Lepidoptera: Papilionidae) in dry and wet seasons SUWARNO Butterfly diversity as a data base for the development plan of Butterfly Garden at Bosscha Observatory, Lembang, West Java TATI SURYATI SYAMSUDIN SUBAHAR, ANNISA YULIANA Diversity and community structure of dung beetles (Coleoptera: Scarabaeidae) across a habitat disturbance gradient in Lore Lindu National Park, Central Sulawesi SHAHABUDDIN Potentialities of line planting technique in rehabilitation of logged over area referred to species diversity, growth and soil quality PRIJANTO PAMOENGKAS Fuel Characteristics and Trace Gases Produced through Biomass Burning BAMBANG HERO SAHARJO, SHIGETO SUDO, SEIICHIRO YONEMURA, HARUO TSURUTA Characterizing forest reduction in Ketapang District, West Kalimantan, Indonesia ASEP SUNJAYA ADHIKERANA, JITO SUGARDJITO

19-23

24-28

29-33

34-39

40-45 46-54

Front cover: Begonia “Tuti Siregar” (PHOTO: HARTUTININGSIH-M. SIREGAR)

Published four times in one year

PRINTED IN INDONESIA ISSN: 1412-033X (printed)

ISSN: 2085-4722 (electronic)