Biodiversitas vol. 13, no. 3, July 2012

ISSN: 1412-033X E-ISSN: 2085-4722

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B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 107-113

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130301

A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers ASHWANI KUMAR1,♥, NEELAM VERMA2 1

Department of Biotechnology, Shri Ram Group of Colleges, Muzaffarnagar 251001, Uttar Pradesh, India. Tel. +91-7417076417, ♥ email: ashwani_biotech@yahoo.co.in 2 Department of Biotechnology, Punjabi University, Patiala 147002, Punjab, India.

Manuscript received: 6 June 2012. Revision accepted: 31 August 2012.

ABSTRACT Kumar A, Verma N. 2012. A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers. Biodiversitas 13: 107-113. Several DNA marker systems and associated techniques are available today for fingerprinting of plant varieties. A total of 5 RAPD and 8 ISSR primers were used. Amplification of genomic DNA of the 6 genotypes, using RAPD analysis, yielded 164 fragments that could be scored, of which 47 were polymorphic, with an average of 9.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 6 (AKR-1) to 10 (AKR-4) and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16% (AKR-4) to a maximum of 41% (AKR-4), with an average of 29.6%. The 8 ISSR primers used in the study produced 327 bands across 6 genotypes, of which 114 were polymorphic. The number of amplified bands varied from 7 (ISSR 7) to 12 (ISSR 1&3), with a size range of 250-2,800 bp. The average numbers of bands per primer and polymorphic bands per primer were 40.87 and 14.25, respectively. Percentage polymorphism ranged from 24% (ISSR 4) to 53.84% (ISSR 2), with an average percentage polymorphism of 35.59% across all the genotypes. The 3′-anchored primers based on poly (AC) and poly (AT) motifs produced high average polymorphisms of 53.84% and 40.81%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 35.59% polymorphic DNA markers in Tribulus terrestris as compared to 29.6% for RAPD markers. Clustering of genotypes within groups was not similar when RAPD and ISSR derived dendrogram were compared, whereas the pattern of clustering of the genotypes remained more or less the same in ISSR and combined data of RAPD and ISSR. Key words: Genetic diversity, Phylogenetic analysis, Tribulus terrestris, RAPD, ISSR Abbreviations: CTAB: Cetyltrimethylammonium bromide, ISSR: Inter simple sequence repeat, RAPD: random amplified polymorphic DNA, PCR: Polymerase chain reaction, PCA: principal component analysis, TT: Tribulus terrestris, UPGMA: unweighted pair group method

INTRODUCTION Puncture vine (Tribulus terrestris) is an important medicinal weed found wildly in India, most parts of central and east Africa and in other Asian countries including Sri Lanka, China and Japan. Tribulus terrestris belonging to the Zygophyllaceae family grows naturally in moist places in woods, low mountains and hills. Its geographical distributions are wide in East Asia, particularly in China.

This plant species, commonly called puncture vine, is a perennial herb highly valued in Chinese traditional medicine. In addition, the claimed therapeutic values of T. terrestris include treatment for dyspepsia, poor appetite, fatigue and psychoneurosis. Recent years have seen a highly accelerated demand for T. terrestris, fruits which has inevitably led to destructive over-harvesting and depletion of its natural resources (Van Valkenburg and Bunyapraphatsara 2001).

Molecular markers are highly heritable and exhibit enough polymorphism to discriminate genotypes. They can be very useful in identifying varieties at early stages of growth and characterize the genotype comprehensively since they are available in very high numbers and are distributed throughout the genome. Application of

molecular markers as complementary approach for genetic characterization has been reported in many crops (Karp et al. 1998). ISSR (Gupta et al. 1994; Zietkiewicz et al. 1994; Bornet and Branchard 2001) markers are considered superior to RAPD (Qian et al. 2001). RAPD analysis provides high resolution and can be carried out on small amount of DNA (Carter and Sytsma 2001; Jolner et al.

2004). However, it is well known that RAPD markers can be sensitive to changes in reaction conditions, resulting in low reproducibility and inconsistencies in amplification efficiencies among samples (Weising et al. 1995). ISSR markers have been used to characterize gene bank accessions (Blair et al. 1999) as well as to identify closely related cultivars (Fang and Roose 1997). The potential supply of ISSR marker depends on the variety and frequency of microsatellites, which changes with species and the SSR motifs that are targeted (Depeiges et al. 1995).

ISSR primers with a given microsatellite repeat should reflect the relative frequency of that motif in a given genome and would provide an estimate of the motif’s abundance. A large number of polymorphic markers are required to measure genetic relationships and genetic diversity in a reliable manner (Santalla et al. 1998). This limits the use of morphological characters and isozymes,

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which are few or lack adequate levels in Tribulus terrestris.

Molecular genetic markers have developed into a powerful tool to analyze genetic relationships and genetic diversity.

Both RAPD and ISSR remain attractive options despite availability of sophisticated techniques because they are easy, quick, simple and economical. Present study reports molecular characterization of 6 cytotypes of T. terrestris, from Northwest India employing RAPD and ISSR techniques. Our aims also were to: (i) provide a better understanding of the phylogenetic relationships between Tribulus population, and (ii) determine the degree to which PCR based markers such us as RAPDs or ISSRs are able to assess the variation and genetic relationship within Tribulus terrestris population.

MATERIALS AND METHODS Several experiments were carried out, however, only the optimized protocol is described here.

Plant material and DNA preparation Here, we used the set of 6 cytotypes of Tribulus terrestris (Table 1). The leaves of Tribulus terrestris were collected from naturally grown population of the six different districts (Patiala, Delhi, Meerut, Muzaffarnagar, Baghpat and Haridwar) of Northwest India in 2006, and their identity was confirmed and voucher specimens were deposit in the herbarium, Department of Botany, Punjabi University, Patiala, India.

Young and healthy leaves from single trees of each accession were used for DNA isolation by CTAB method with minor modifications (Doyle and Doyle 1990; SaghaiMaroof et al. 1984). Essentially, the extraction buffer composition was 4% w/v CTAB, 1.4MNaCl, 100 mM TrisHCl (pH 8), 20 mM EDTA, 2% PVP w/v, and 0.2% 2mercapto ethanol v/v. DNA was treated with bovine pancreatic RNAse and extracted once with phenol: chloroform (1:1) and twice with chloroform: iso-amyl alcohol (24:1). After precipitation with iso-propanol, a 70% ethanol wash was given. DNA was dissolved to appropriate dilution in TE buffer and quantified in a spectrophotometer.

Primer screening A preliminary experiment on 6 randomly selected Tribulus accessions was carried out to select most suitable

primers for identification. 6 random primers and 8 ISSR primers were screened for repeatability, scorability, and their ability to distinguish within varieties. Random primers AKR-1, AKR-2, AKR-3, AKR-4, AKR-5 and AKR-6 procured from (Imperial LifeSci Ltd. India); and primers ISSR1-8 from (custom made from Bangalore Genei) exhibited maximum efficiency of discrimination in terms of resolving power (Prevost and Wilkinson 1999). These primers were employed for varietal identification.

Amplification conditions for RAPD and ISSR-PCR and gel electrophoresis In order to select optimal primers for effective use in RAPD-PCR analysis, 6 random primers (10 bp) of the Imperial biotech company were screened using TT population genomic DNA; 5 primers that generated clear bands and reproducible fragments were selected for further investigation (Table 2).

RAPD assay was carried out in 25 μl reaction mixture containing 2.5 μl 10x Taq DNA-polymerase PCR amplification buffer, 10 mM dNTPs, 1.0 U/μl of Taq DNA polymerase (Genei, India), 15 pmoles of 10-mer primer (Operon Technologies Inc, USA) and 50 ŋg of genomic DNA. Amplification was performed in PCR (Techne, India).

The sequential PCR steps involved: 1 cycle of 2 min at 93°C, 2 min at 35°C and 2 min at 72°C followed by 44 cycles of 1 min at 93°C, 1 min at 36°C and 2 min at 72°C. The last cycle was followed by 10 min extension at 72°C. The amplified products were resolved in 1.8% agarose gel (1xTBE) followed by Ethidium Bromide staining and the bands detected then photographed using a gel documentation system (BioRad, USA) (Figure 1).

A total of 8- primers were tested to amplify DNA banding patterns using the total genomic DNA (Table 2).

These were: ISSR-1 (GAGA)4 GAT, ISSR-2 (GAGA)4GAAC, ISSR-3 (GAGA)4 GAAT, ISSR-4 (ACC)4Y, ISSR-5 (GACA)4, ISSR-6 (GATA)4, ISSR7(GA)9 C and ISSR-8 (GA)9 A. The PCR reaction mixture (25 µL) was composed of 50 ng of total cellular DNA (2 µL), 10 µM of primer (2.5 µL), 2.5 µL of Taq DNA polymerase reaction buffer, 0.35 µL of Taq DNA polymerase and 10 mM of each dNTPs (DNA polymerization mix). Each reaction mixture was overlaid with 25 µL of mineral oil to avoid evaporation during PCR cycling. Amplifications were performed in a DNA

Table 1. Localities and their geographical coordinates from which Tribulus terrestris samples were collected for the morphological and chromosomal characterization.

Coll. no.

01 02 03 04 05 06

Locality New Delhi; Railway Line, Near Old Delhi railway station Uttaranchal, Haridwar; PAC Play ground, Jwalapur Uttar Pradesh, Baghpat; Railway Line, Near Baraut railway station Punjab, Patiala; Road side, Punjabi Univ. Patiala Campus Uttar Pradesh, Meerut; Play ground, CCS Univ. Meerut Campus Uttar Pradesh, Muzaffarnagar; Waste Land of Titawi village

Geographical coordinates 28º36” N, 77º12” E 29º48” N, 78º36” E 28º00” N, 77º00” E 30º09” N, 76º17” E 29º00” N, 77º00” E 29º09” N, 77º43” E

Chromosome no.

(2n) 12 48 36 24 24 24

Ploidy Diploid Octaploid Hexaploid Tetraploid Tetraploid Tetraploid

Accession No. 49318 49316 49320 49321 49319 49317

KUMAR & VERMA – Phylogenetic analysis of Tribulus terrestris based on RAPD and ISSR

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Table 2. Analysis of banding patterns generated by RAPD and ISSR assay for the six Tribulus terrestris cultivars

Primer name and sequence RAPD AKR-1 (3’TGTGTGCCAC5’) AKR-3 (3’AGGCTGTGCT5’) AKR-4 (3’GTGCCGTTCA5’) AKR-5 (3’GGGTGGGTAA5’) AKR-6 (3’CCGACAAACC5’) ISSR ISSR-1 (GAGA)4 GAT ISSR-2 (GAGA)4GAAC ISSR-3 (GAGA)4 GAAT ISSR-4 (ACC)4Y ISSR-5 (GACA)4 ISSR-6 (GATA)4 ISSR-7 (GA)9C ISSR-8 (GA)9A

Total no. of amplified products

Expected PCR Annealing Polymorphic % Polymorphism product size temperature bands (bp) (0C)

23

6

26

25 50 27 39 Total = 164

7 8 10 16 47

28 16 37 41 Average = 29.6

49 26 51 50 42 45 35 29 Total = 327

20 14 18 12 15 17 8 10 114

40.81 53.84 35.29 24 35.71 37.77 22.85 34.48 Average = 35.59

amplification thermo cycler (Techne India). The apparatus was programmed to execute the following conditions: a denaturation step of 7 min at 94°C followed by 30 cycles each composed of 30 s at 94°C, 45 s at the primer’s specific melting temperature (Tm) at 52-60°C and 2 min at 72°C. A final extension step of 7 min also at 72°C was run at the end of the last PCR cycle.

PCR reactions were electrophorezed on 1.4% agarose gels in 10xTBE buffer by loading 25 µL of the reaction mixture into prepared wells. Gels were run for 4-5 h at 90 V, stained with ethidium bromide (10 µg ml−1) according to Sambrook et al. (1989), and ISSR banding patterns were visualized using a UV transilluminator (Figure 2).

Statistical data analysis For RAPD and ISSR analysis, the banding patterns were recorded using a gel documentation system (Bio-Red Gel Doc 1000), and the image profiles and molecular weight of each band were determined by Molecular Analyst/pc (Version 1.2) software. The fragment size scored ranged from 250 to 1000 bp. Both weak bands with negligible intensity and smearing bands were excluded from final data analysis. The bands with the same molecular weight and mobility were treated as identical fragments. In the data matrices, Amplicons were scored as discrete variables, using 1/0 as presence/absence of homologous bands for all samples. The data matrices were analyzed by the SIMQUAL program of NTSYS-pc (Version 1.8), and similarities between accessions were estimated using the Jaccard coefficient. For phylogenetic analysis, Dendrograms were produced from the distance matrices using the unweighted pair group method with arithmetic averages (UPGMA). In order to compare the levels of genetic diversity between the Tribulus species (Sneath and Sokal 1973). Finally, a principal component analysis (PCA) was performed in order to highlight the resolving power of the ordination.

200 to 2,500 360C for all 5 for all products primers

300-1500 250-2500 250-1500 250-2800 250-2000 250-2000 250-2000 300-2000

52 52 52 60 52 42 55 55

RESULT AND DISCUSSION Marker profile and discrimination of genotypes Molecular markers generated by RAPD and ISSR target different regions of the genome, though in a random manner. Theoretically, a combination of RAPD and ISSR markers would give a better coverage of genome. Primer wise and technique wise results are given in Table 2 Amplification of genomic DNA of the 6 TT genotypes, using RAPD analysis, yielded 164 fragments that could be scored, of which 47 were polymorphic, with an average of 9.4 polymorphic fragments per primer. Number of amplified fragments with random primers ranged from 6 (AKR-1) to 10 (AKR-4) and varied in size from 200 bp to 2,500 bp. Percentage polymorphism ranged from 16% (AKR-4) to a maximum of 41% (AKR-4), with an average of 29.6%. Polymorphism was high enough to enable discrimination of all the varieties, though none of the primers discriminated all the accessions independently. The 8 ISSR primers used in the study produced 327 bands across 6 genotypes, of which 114 were polymorphic. The number of amplified bands varied from 7 (ISSR 7) to 12 (ISSR 1&3), with a size range of 250-2,800 bp. The average numbers of bands per primer and polymorphic bands per primer were 40.87 and 14.25, respectively.

Percentage polymorphism ranged from 24% (ISSR 4) to 53.84% (ISSR 2), with an average percentage polymorphism of 35.59% across all the genotypes. The 3′anchored primers based on poly (AC) and poly (AT) motifs produced high average polymorphisms of 53.84% and 40.81%, respectively. ISSR markers were more efficient than the RAPD assay, as they detected 35.59% polymorphic DNA markers in Tribulus terrestris as compared to 29.6% for RAPD markers. This deduced that these primers employed in the study returned a high degree of confidence in identification.

B I O D I V E R S IT A S 13 (3): 107-113, July 2012

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A

B

D

E

C

Figure 1. A. Screening of 6 RAPD primers for Tribulus terrestris genotypes DNA fingerprinting; and RAPD-PCR products of 6 different Tribulus terrestris DNA samples with random primers: B. AKR-1, C. AKR-2, D. AKR-3, E. AKR-4, F. AKR-5. From left to right: Lane M. 100bps DNA ladder, lane 1. Haridwar, 2. Meerut, 3. Delhi, 4. Baghpat, 5. Muzaffarnagar, and 6. Patiala

A

B

C

D

E

F

G

H

I

Figure 2. A. Screening of ISSR primers; and ISSR profile of 6 Tribulus terrestris genotypes with primer: B. ISSR-1, C. ISSR-2, D. ISSR-3, E. ISSR-4, F. ISSR-5, G. ISSR-6, H. ISSR-7, I. ISSR-8. From left to right: Lane M. 100bps DNA ladder, lane 1. Haridwar, 2.

Meerut, 3. Delhi, 4. Baghpat, 5. Muzaffarnagar, and 6. Patiala

KUMAR & VERMA – Phylogenetic analysis of Tribulus terrestris based on RAPD and ISSR

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5

5

5

2

3

3

3

6

6

6

2

1

1

4

2

4

1

4

A

B

C

Figure 3. Dendrograms generated using UPGMA analysis, showing relationships between Tribulus terrestris genotypes, using: A. RAPD, B. ISSR, C. combining both RAPD and ISSR data. Note: 1. Haridwar, 2. Meerut, 3. Delhi, 4. Baghpat, 5. Muzaffarnagar, and 6.

Patiala

Genetic relations and utilization of diversity Although major emphasis of this work was to generate DNA profiles of the varieties, the marker data was also used to study genetic relations among varieties. Cluster analysis was used to generate three dendrograms based on the Jaccard coefficient of RAPD, ISSR and combined of both for all 6 plant samples (Figure 3.A, 3.B, 3.C respectively). The resultant dendrograms from RAPD and ISSR showed that the populations were divided into two groups but some differences were observed in the allocation of populations to these groups in the RAPD and ISSR data sets, while combined data represent three groups of Tribulus population. In the dendrograms derived from the RAPD data set the Jaccard coefficient ranged from 0.61 to 1.00. Populations from Muzaffarnagar, Meerut, Delhi, Patiala and Haridwar all formed one main group, and another second group was composed of Baghpat population. Within the latter group, populations of Meerut and Delhi constituted one sub-group and populations from Patiala and Haridwar clustered into the other subgroup.

Populations from Patiala and Haridwar appeared to be closer to each other than clusters of the other populations.

Dendrograms derived from the ISSR data set the Jaccard coefficient ranged from 0.525 to 1.00. Populations from Muzaffarnagar and Delhi formed one group, and another second main group was composed of Meerut, Patiala and Haridwar and Baghpat population. Within the latter group, populations of Meerut and Baghpat constituted one subgroup. Populations from Meerut and Baghpat also appeared to be closer to each other than clusters of the other populations In the combined dendrogram derived from RAPD and ISSR analysis, the Jaccard coefficient ranged from 0.582 to 1.00. Populations from Muzaffarnagar and Delhi formed one group, Patiala and Haridwar formed the second group and remaining samples those from Meerut and Baghpat,

formed the third group. Populations of Meerut and Baghpat also appeared to be closer to each other than another clustered populations of Tribulus terrestris. The dendrogram based on RAPD showed some variation with in the clustering of genotypes. The ISSR and combined dendrogram indicated a similarity that Meerut and Baghpat populations were closer to each other than other populations. All the Tribulus genotypes could be discriminated based on these total 13 RAPD and ISSR primers.

PCA output produced by NTSYS using the simple matching matrix for RAPD and ISSR combined data In order to visualize the data, you may wish to present the PCA graph, which gives a good three-dimensional picture of the variation. Principal Component Analysis associated with the Minimum Spanning Tree, based on the six genotypes of Tribulus and provided complementary information to cluster analysis, as it allows a graphical presentation of the distribution of the cultivars in a threedimensional plot (Figure 4.A, 4.B, 4.C). In this representation Populations from Meerut and Baghpat are the most similar cultivars similar like ISSR-PCA analysis.

The Eigenvalues indicate that three components provide a very good description of the data, as account for 69.68% of the standardized variance. The analysis of Eigenvectors provides information about the traits responsible for the separations along the first three Principal Components. PC1 had 29.2124% of the total variation. This Principal Component is responsible for the individualization of Delhi and Muzaffarnagar populations. PC2 exhibited 21.6469% of the total variability, and is responsible for the separation of Meerut and Baghpat from Haridwar and Patiala populations. A part from the other cultivars PC3 had 18.9246% of the total variation. PC3 is responsible for the separation of Haridwar and Patiala population.

B I O D I V E R S IT A S 13 (3): 107-113, July 2012

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BAGHPAT

BAGHPAT M.NAGAR

BAGHPAT

A

B

C

Figure 4. Three-dimensional plot of principal component analysis of using Tribulus terrestris genotypes using: A. RAPD, B. ISSR, C. combining both RAPD and ISSR analyses.

The evolution of varieties in distinct agro-climatic zones demonstrates significant levels of variation in response to the selection pressure in the zones (Singh et al.

1998). It is, therefore, not surprising to find significant levels of polymorphism among the 6 genotypes of puncturevine in RAPD (29.6%) and ISSR (35.59%) markers. The RAPD technique has been applied to assess molecular polymorphism in Vigna (Kaga et al. 1996), mung bean (Santalla et al. 1998; Lakhanpaul 2000). The success of our study in identifying polymorphism is due to the use of a number of randomly selected prescreened highly informative primers.

study of a big collection should provide a better knowledge about genetic diversity and its relationship with geographical origin. This information could be very valuable in the management and cultivation of plant genetic resources for the herbal drug formulation research.

CONCLUSION

REFERENCES

With the increasing use of DNA fingerprinting in plant and its potential use in herbal drug industry, the preparation of good quality and quantity DNA has become a major concern. The extraction from tissue needs to be simple, rapid, efficient and inexpensive when many samples are used, such as in population studies, molecular breeding and screening of raw herbal drug materials.

Here we report a study on the genetic variation of Tribulus terrestris populations of Northwest India by means of random amplified polymorphic DNA (RAPD) and ISSR fingerprinting. Our aim was to investigate the genetic variation within population of Tribulus terrestris.

Accessions with the most distinct DNA profiles are likely to contain the greatest number of novel alleles. It is these accessions that are likely to uncover the largest number of unique and potentially agronomic useful alleles. This strategy has resulted in a high proportion (50%) of new and useful quantitative trait loci alleles in rice and tomato (Tanksley and McCouch 1997). This information may assist in the development of an effective cultivation strategy on the Tribulus terrestris. Further studies are envisaged to quantify the genetic gain in populations derived from genotypes with distinct DNA profiles. The

Blair MW, Panaud O, McCouch SR. 1999. Inter simple sequence repeat (ISSR) amplification for analysis of microsatellite motif frequency and fingerprinting in rice. Theor Appl Genet 98: 780-792.

Bornet B, Branchard M. 2001. Nonanchored inter simple sequence repeat (ISSR) markers: Reproducible and specific tools for genome fingerprinting. Plant Mol Biol Rep 19: 209-215.

Carter MC, Sytsma MD. 2001. Comparison of the genetic structure of North and South American populations of a clonal aquatic plant. Biol Invasions 3: 113-118.

Depeiges A, Goubely C, Lenoir A, Cocherel S, Picard G, Raynal M, Grellet F, Delseny M. 1995. Identification of the most represented repeat motifs in Arabidopsis thaliana microsatellite loci. Theor Appl Genet 91:160-168 Fang DQ, Roose MZ. 1997. Identification of closely related citrus cultivars with inter simple sequence repeat markers. Theor Appl Genet 95: 408-417.

Gupta M, Chyi YS, Romero-Severson J, Owen JL. 1994. Amplification of DNA markers from evolutionarily diverse genomes using single primers of simple-sequence repeats. Theor Appl Genet 89: 998-1006.

Jolner SK, Sastad SM, Taberlet P, Brochmann C. 2004. Amplified fragment length polymorphism versus random amplified polymorphic DNA markers: clonal diversity in Saxifraga cernua. Mol Ecol 13: 8186.

Kaga AN, Tomooka N, Egawa Y, Hosaka K, Kamijima O. 1996. Species relationships in subgenus Ceratotropis (genus Vigna) as revealed by RAPD analysis. Euphytica 88: 17-24 Karp A, Issar PG, Ingram DS. 1998. Molecular tools for screening biodiversity. Chapman & Hall, London, UK.

ACKNOWLEDGMENTS This work was supported by the Department of Biotechnology and Department of Botany, Punjabi University, Patiala 147 002, Punjab, India.

KUMAR & VERMA – Phylogenetic analysis of Tribulus terrestris based on RAPD and ISSR Lakhanpaul S, Chadha S, Bhat KV. 2000. Random amplified polymorphic DNA (RAPD) analysis in Indian mungbean [Vigna radiata (L.) Wilczek] cultivars. Genetica 109: 227-234.

Prevost A, Wilkinson MJ. 1999. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor Appl Genet 98 (1): 107-112.

Qian W, Ge S, Hong DY. 2001. Genetic variation within and among populations of a wild rice Oryza granulata from China detected by RAPD and ISSR markers. Theor Appl Genet 102: 440-449.

Saghai-Maroof MA, Soliman KM, Jorgensen RA, Allard RW. 1984.

Ribosomal DNAsepacer-length polymorphism in barley: mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81: 8014-8019.

Sambrook J, Fritschi EF, Maniatis T. 1989. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, New York.

Santalla M, Power JB, Davey MR. 1998. Genetic diversity in mungbean germplasm revealed by RAPD markers. Plant Breed 117: 473-478.

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Singh AK, Smart J, Simpson CE, Raina SN. 1998. Genetic variation vis-avis molecular polymorphism in groundnut, Arachis hypogaea L. Genet Resour Crop Evol 45: 119-126.

Sneath PHA, Sokal RR. 1973. Numerical Taxonomy. Freeman, San Francisco.

Tanksley SD, McCouch SR. 1997. Seed banks and molecular maps: unlocking genetic potential from the wild. Science 277: 1063-1066.

Van Valkenburg JLCH, Bunyapraphatsara N, (eds). 2001. PROSEA. Plant resources of South-east Asia, No. 12(2): Medicinal and poisonous 332 plants – 2., Backhuys, Leiden.

Weising K, Nybom H, Wolff K, Meyer W. 1995. DNA Fingerprinting in Plants and Fungi. CRC Press, Boca Raton, FL, USA. Zietkiewicz E, Rafalski A, Labuda D. 1994. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20: 176-183.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 114-117

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130302

In Silico chloroplast SSRs mining of Olea species ERTUGRUL FILIZ1,♼, IBRAHIM KOC2 1

Department of Crop and Animal Production, Cilimli Vocational School, Duzce University, Duzce, Turkey. Tel. +90 380 681 7312, ext.7406, Fax: +90 380 681 73 13, ď‚Šemail: ertugrulfiliz@gmail.com 2 Department of Molecular Biology and Genetics, Gebze Institute of Technology, Kocaeli, Turkey Manuscript received: 4 June 2012. Revision accepted: 21 June 2012.

ABSTRACT Filiz E, Koc E. 2012. In Silico chloroplast SSRs mining of Olea species. Biodiversitas 13: 114-117. Simple sequence repeat (SSR) markers are highly informative and have been widely applied as molecular markers in genetic studies. The purpose of present study is to analyze the occurrence and distribution of chloroplast SSRs in genic and intergenic regions from Olea species viz., Olea europaea, Olea europaea subsp. cuspidate, Olea europaea subsp. europaea, Olea europaea subsp. maroccana, Olea woodiana subsp. woodiana by using bioinformatics tools. We identified 1149 chloroplast SSRs (cpSSRs) in all genome and a total of 340 (29.6%) was located in genic regions. It was observed that the most abundant repeat types were found mononucleotide SSR (66.7 %) followed by trinucleotide SSR (28.3 %), dinucleotide (2.7%), tetranucleotide (1.5%) and pentanucleotide (0.8%). cpSSRs located in genic regions were identified only mono- and trinucleotide motifs, the most abundant of which was trinucleotide (16.2%) followed by mononucleotide (14.3%). All types of repeat motif (mono-, di-, tri-, tetra- and pentanucleotide) were detected except hexanucleotide motifs. According to SSRs analysis, the most abundant observed motifs were identified for mono-, di-, tri-, tetra- and pentanucleotide cpSSRs A/T, AT/TA, AAG/CTT, AAAG/AGTTT, and AATCC/ATTGG respectively. This study results provided scientific base for phylogenetics, evolutionary genetics and diversity studies on different Olea species in future.

Key words: Olea, olive, chloroplast SSRs, SSR mining, in silico analysis

INTRODUCTION Olea is a genus of about 40 species in the family Oleaceae and it is distributed warm temperate and tropical regions of southern Europe, Africa, southern Asia and Australasia. Olea known as olive (Olea europaea L.), is the most important oil crop in the Mediterranean region and has been used for pharmaceutical, industrial and consumer purposes (Hannachi et al. 2010). Olive is also the second most important oil fruit crop cultivated worldwide after oil palm (Baldoni and Belaj 2009). Olea species were evaluated by using various genetic markers such as SSRs genotyping (Taamalli et al. 2008; Ercisli et al. 2011, Gomes et al. 2009, Carriero et al. 2002); RAPDs genotyping (Bogani et al. 1994; Belaj et al. 2004); AFLPs genotyping (Owen et al. 2005; Baldoni et al. 2006), SNPs (Reale et al. 2006), Ribosomal DNA polymorphisms (Hess et al. 2000, Besnard et al. 2007), and organelle DNA polymorphisms (Besnard et al. 2002; Intrieri et al. 2007).

Simple sequence repeats (SSRs) or microsatellites are tandem repeated motifs which are 1-6 nucleotide length and located in all prokaryotic and eukaryotic genomes (Zane et al. 2002). Microsatellites are being used for mapping and tagging of genes, marker assisted selection (MAS), genome mapping and functional genomics (Kalia et al. 2011). SSRs are associated with coding and noncoding regions and can be found nuclear, chloroplast and mitochondrial genome (Provan et al. 2001; Rajendrakumar et al. 2007). The chloroplast genome

includes about 120-130 genes and usually ranges in size from 120-200 kb (Sugiura 1992) and is characterized by haploidy and a lack of recombination and uniparental inheritance. Therefore, chloroplast markers (cpSSRs) are more effective indicator of population genetic structure than nuclear SSRs markers (Birky 1995; Petit et al. 1995).

In the last decade, chloroplast SSRs (cpSSRs) have been widely used for plant taxonomic and phylogenetic studies, diversity analysis and population genetics (Provan et al.

2001). The main objective of this study was to analysis SSRs in chloroplast genome (cpDNA) of Olea species for their occurrence and distribution in both coding and noncoding regions.

MATERIAL AND METHODS All the chloroplast genome sequences of olive (Olea europaea, Olea europaea subsp. cuspidate, Olea europaea subsp. europaea, Olea europaea subsp. maroccana, Olea woodiana subsp. woodiana) were downloaded in FASTA format from GenBank (ftp://ncbi.nlm.nih.gov/genbank/ genomes/) (Table 1.). The identification of chloroplast microsatellites was carried out by MISA perl script (http://pgrc.ipk-gatersleben.de/misa/). The minimum motif repeat size were set to 8 for mononucleotide, 5 for dinucleotide, 3 for trinucleotide, tetranucleotide, pentanucleotide and hexanucleotide in locating the microsatellites with maximum differences two SSRs was

FILIZ & KOC – Chloroplast SSRs of Olea

100. SSRs were searched in full chloroplast genome as well as separate coding and non-coding regions for each species.

Also, GC content was calculated.

Table 1. Details of chloroplast genomes in Olea species

Plant species

Plastome Accession size (bp) Number

Olea europaea O. europaea subsp. cuspidata O. europaea subsp. europaea O. europaea subsp. maroccana O. woodiana subsp. woodiana

155888 155862 155875 155896 155942

NC_013707 NC_015604 NC_015401 NC_015623 NC_015608

G+C content (%) 37.80% 37.81% 37.81% 37.81% 37.79%

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frequency of SSRs in Olea species in present study (1.47 SSR per kb) is lower than found in loblolly pine EST-SSRs (42.9 SSR per kb), some cereal species EST-SSRs (6 SSR per kb) (Varshney et al. 2002), palm EST-SSRs (4.4 SSR per kb) (Palliyarakkal et al. 2011).

Distribution of cpSSRs The investigation of different types of SSR repeats showed that the percentage of occurrence of mononucleotide SSR (66.7 %) was the highest followed by trinucleotide SSR (28.3%), dinucleotide (2.7%), tetranucleotide (1.5%) and pentanucleotide (0.8%) (Figure 1.).

Table 2. The number of the genic and intergenic cpSSRs based on motif size for each species Mono G I 33 122 33 119

Di GI 0 6 0 6

Tri G I 35 31 35 30

Tetra G I 0 4 0 3

Olea europaea Olea europaea subsp. cuspidata Olea europaea 33 120 0 6 35 30 0 subsp. europaea Olea europaea 33 120 0 6 35 30 0 subsp. maroccana Olea woodiana 32 121 0 7 36 28 0 subsp. woodiana Total Note: G: genic, I: intergenic

Penta Hexa G I G I Total 0 2 0 0 233 0 2 0 0 228

Hexanucleotide

Pentanucleotide

Tetranucleotide

Trinucleotide

Dinucleotide

Abundance of SSRs A total of five Olea species chloroplast complete genome sequences were evaluated for chloroplast SSRs and we found 1149 SSRs, of which 340 (29.6%) were localized in genic regions and the 809 (70.4%) cpSSRs were localized in intergenic regions (Table 2.). G+C content of the species are closely similar frequency ranging from 37.79 to 37.81% (Table 1.)

Mononucleotide

RESULTS AND DISCUSSION

Figure 1. Frequencies (%) of different repeats in genic and intergenic regions.

2 0 0 230

AATCC/ATTGG

4 0

AAACT/AGTTT

2 0 0 229

AAAG/CTTT

3 0

AAG/CTT

Figure 2. The most abundant cpSSRs motifs in all chloroplast genomes.

2 0 0 229

AT/TA

Total number of cpSSRs in the chloroplast genomes ranged from 228 to 233 and the density of microsatellites ranged from 1.46-1.49 cpSSR per kb. Olea europaea subsp. cuspidate chloroplast has the least number of cpSSRs (228) while Olea europaea chloroplast had the most abundant cpSSRs (233). The density of microsatellites were found for Olea europaea, Olea europaea subsp. cuspidate, Olea europaea subsp. europaea, Olea europaea subsp. maroccana and Olea woodiana subsp. woodiana 1.49, 1.46, 1.47, 1.47 and 1.47 cpSSR per kb respectively. An average frequency was 1.47 cpSSR per kb which was higher than some cereal species cpSSRs (1.36 cpSSR per kb) (Melotto-passarin et al. 2011), Solanaceae species cpSSRs (1.26 cpSSR per kb) (Tambarussi et al. 2009), Solanum lycopersicum EST-SSRs (1.3 SSR per kb) (Gupta et al. 2010). However, an average

3 0

AT/T

1149

The most abundant genic cpSSRs type was trinucleotide (16.2%) followed by mononucleotide (14.3%). This finding supports that triplet SSR repeats can be located easily within coding regions (Hancock and Simon 2005). Our results revealed that A/T repeats (98%) were found to be more abundant than the G/C (2%) motifs. These results were consistent with SSRs analysis of major cereal organelle genome data (Rajendrakumar et al. 2008) (Figure 2.).

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For dinucleotide repeats, AT/TA motif was the most common dinucleotide repeat with a frequency of 83.9%.

These results are in agreement with previous findings (Gandhi et al. 2010; Rajendrakumar et al. 2007; Kuntal et al. 2012). The higher AT/TA frequencies in chloroplast genomes can be explained to be the conclusion of the high A/T content of the genomes. Among the trinucleotide repeats, AAG/CTT motif was the most common (28.9%) followed by AAT/TTA (24.9%) and AAC/GTT (16%) and this finding was not consistent with earlier studies results (Rajendrakumar et al. 2007; Melotto-passarin et al. 2011; Kuntal et al. 2012). In tetranucleotide SSR motifs, the maximum frequency of 47.1% was showed by AAAG/CTTT followed by AAAT/ATTT (29.4%). In pentanucleotide SSR motifs, frequency of AAACT/ AGTTT and AATCC/ATTGG motifs were found to be equal (50%). Interestingly, there are no any hexanucleotide repeats in all chloroplast genomes. A total of 340 cpSSRs (30.5%) were found to be in genic regions while a total of 809 cpSSRs (69.5%) were found to be in intergenic regions and genic cpSSRs were identified only mono and trinucleotide motifs. In general, intergenic cpSSR (69.5%) in Olea species were more abundant than genic cpSSR (29.6%) and this result is consistent with earlier studies Asteraceae (Timme et al. 2007), Fabaceae (Saski et al.

2005), and Solanaceae (Daniell et al. 2006), Saccharum (Melotto-passarin et al. 2011). According to the analysis, the most abundant genic cpSSRs were found for trinucleotide motifs (57.2%). The SSRs in genes show a higher mutation rate (instability) than nonrepetitive regions in genome. Also, SSRs variations affect gene expression, inactivation of gene activity, and/or a change of function (Li et al. 2004). Thus, our results corroborate this hypothesis that the most number of genic trinucleotide cpSSRs may cause dynamic evolution and mutational forces in exons and exhibit genetic and phenotypic variations. The number of plastome genes with cpSSR were identified (Table 3.) and we found Olea europaea, Olea europaea subsp. cuspidate, Olea europaea subsp. europaea, Olea europaea subsp. maroccana with 49 genes while Olea woodiana subsp. woodiana has 46 genes.

Table 3. Frequency (%) of plastome genes with cpSSRs

Species Olea europaea Olea europaea subsp. cuspidata Olea europaea subsp. europaea Olea europaea subsp. maroccana Olea woodiana subsp. woodiana

Number Genes of with plastome cpSSR genes 130 49 130 49 130 49 130 49 130 46

Genes with cpSSR % 37.7% 37.7% 37.7% 37.7% 35.4%

Many studies revealed that large numbers of SSRs are distributed in genic regions of genomes, containing proteincoding genes and expressed sequence tags (ESTs). SSRs distributions play key role for history of genome evolution and mutational processes (Morgante et al. 2002). In this study, we investigated the distribution of different types of

SSR in coding and non-coding regions of five different species of Olea genus by using bioinformatic tools. The location of SSR in the genome determines its functional role like gene regulation, development and evolution (Kalia et al. 2011). According to analysis, intergenic cpSSRs are predominant over genic cpSSRs and as expected, trinucleotide repeats are more common in coding regions.

Except mononucleotide and trinucleotide repeats, other classes of repeats were low in number in the chloroplast genomes.

CONCLUSION SSR markers are very informative because they are codominant and highly polymorphic. In addition, SSRs markers are highly mutable loci and they can be used for characterization of genome and a particular region can be identified in the genome. This study results can be targeted towards the diversity and evolutionary studies of Olea species.

REFERENCES Baldoni L, Belaj A. 2009. Olive. In: Vollmann J, Rajean I (eds) Oil crops.

Handbook of plant breeding. Vol 4. Springer, New York.

Baldoni L, Tosti N, Ricciolini C, Belaj A, Arcioni S, Pannelli G, Germana MA, Mulas M, Porceddu A. 2006. Genetic structure of wild and cultivated olives in the central Mediterranean basin. Ann Bot 98: 935942.

Belaj A, Rallo L, Trujillo I, Baldoni L. 2004. Using RAPD and AFLP markers to distinguish individuals obtained by clonal selection of ‘Arbequina’ and ‘Manzanilla de Sevilla’ olive. HortScience 39: 15661570.

Besnard G, Henry P, Wille L, Cooke D, Chapuis E . 2007. On the origin of the invasive olives (Olea europaea L., Oleaceae). Heredity 99: 608-619.

Besnard G, Khadari B, Baradat P, Berville A. 2002. Combination of chloroplast and mitochondrial DNA polymorphism to study cytoplasm genetic differentiation in the olive complex (Olea europaea L.). Theor Appl Genet 105: 139-144.

Birky CW Jr. 1995. Uniparental inheritance of mitochondrial and chloroplast genes: Mechanisms and evolution. Proc Natl Acad Sci USA 92: 11331-11338.

Bogani P, Cavalieri D, Petruccelli R, Roselli G. 1994. Identification of olive tree cultivars by using random amplified polymorphic DNA. Acta Hortic 356: 98-101.

Carriero F, Fontanazza G, Cellini F, Giorio G. 2002. Identification of simple sequence repeats (SSRs) in olive ( Olea europaea L.). Theor Appl Genet 104 (2-3): 301-307.

Daniell H, Lee SB, Grevich J, Saski C et al.. 2006. Complete chloroplast genome sequences of Solanum bulbocastanum, Solanum lycopersicum and comparative analyses with other Solanaceae genomes. Theor Appl Genet 112: 1503-1518.

Ercisli S, Ipek A, Barut E. 2011. SSR marker-based DNA fingerprinting and cultivar identification of olives (Olea europaea). Biochem Genet.

49 (9-10): 555-61.

Gandhi SG, Awasthi P, Bedi YS. 2010. Analysis of SSR dynamics in chloroplast genomes of Brassicaceae family. Bioinformation 5 (1): 16-20.

Gomes S, Martins-Lopes P, Lopes J, Guedes-Pinto H. 2009. Assessing Genetic Diversity in Olea europaea L. using ISSR and SSR Markers.

Plant Mol Biol Rep 27: 365-373.

Gupta S, Tripathi KP, Roy S, Sharma A. 2010. Analysis of unigene derived microsatellite markers in family Solanaceae. Bioinformation 5 (3): 113-121.

Hancock JM, Simon M. 2005. Simple sequence repeats in proteins and their significance for network evolution. Gene 345: 113-118.

FILIZ & KOC – Chloroplast SSRs of Olea Hannachi H, Breton C, Msallem M, Hadj SB, Gazzah M, Bervillé A. 2010. Genetic relationships between cultivated and wild olive trees (Olea europaea L. var. europaea and var. sylvestris) based on nuclear and chloroplast SSR markers. Nat Res 1: 95-103.

Hess J, Kadereit W, Vargas P. 2000. The colonization history of Olea europaea L. in Macaronesia based on internal transcribed spacer 1 (ITS-1) sequences, randomly amplified polymorphic DNAs (RAPD) and intersimple sequence repeats (ISSR). Mol Ecol 9: 857-868.

Intrieri MC, Muleo R, Buiatti M. 2007. Chloroplast DNA polymorphisms as molecular markers to identify cultivars of Olea europaea L. J Hort Sci Biotechnol 82: 109-113.

Kalia RK, Rai MK, Kalia S, Singh R, Dhawan AK. 2011. Microsatellite markers: an overview of the recent progress in plants. Euphytica 177: 309-334.

Kuntal H, Sharma V, Daniell H. 2012. Microsatellite analysis in organelle genomes of Chlorophyta. Bioinformation 8 (6): 255-259.

Li YC, Korol AB, Fahima T, Nevo E. 2004. Microsatellites within genes: Structure, function, and evolution. Mol Biol Evol 21 (6): 991-1007.

Melotto-passarin DM, Tambarussi EV, Dressano K, De Martin VF, Carrer H. 2011. Characterization of chloroplast DNA microsatellites from Saccharum spp. and related species. Genet Mol Res 10 (3): 20242033.

Morgante M, Hanafey M, Powell W. 2002. Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes.

Nat Genet 30: 194-200.

Owen CA, Bita EC, Banilas G, Hajjar SE, Sellinakis V, Aksoy U, Hepaksoy S, Chamoun R, Talhook SN, Metzidakis I, Hatzopoulos P, Kalaitzis P. 2005. AFLP reveals structural details of genetic deiversity within cultivated olive germoplasm from the Eastern Mediterranean. Theor Appl Genet 110: 1169-1176.

Palliyarakkal MK, Ramaswamy M, Vadivel A. 2011. Microsatellites in palm (Arecaceae) sequences. Bioinformation 7 (7): 347- 351.

Petit RJ, Bahrman N, Baradat P. 1995. Comparison of genetic differentiation in maritime pine (Pinus pinaster Ait.) estimated using isozyme, total protein and terpenic loci. Heredity 75: 382-389.

117

Provan J, Powell W, Hollingsworth PM. 2001. Chloroplast microsatellites: new tools for studies in plant ecology and evolution.

Trends Eco Evol 16: 142-147.

Rajendrakumar P, Biswal AK, Balachandran SM, Srinivasarao K, Sundaram RM. 2007. Simple sequence repeats in organellar genomes of rice: frequency and distribution in genic and intergenic regions.

Bioinformatics 23: 1-4.

Rajendrakumar P, Biswal AK, Balachandran SM, Sundaram RM. 2008. In silico analysis of microsatellites in organellar genomes of major cereals for understanding the phylogenetic relationships. In Silico Biology 8: 9-18.

Reale S, Doveri S, Dı ´az A, Angiolillo A, Lucentini L, Pilla F, Martı ´n A, Donini P, Lee D. 2006. SNP-based markers for discriminating olive (Olea europaea L.) cultivars. Genome 49: 1193-1205.

Saski C, Lee SB, Daniell H, Wood TC, Tomkins J, Kim HG, Jansen RK. 2005. Complete chloroplast genome sequence of Glycine max and comparative analyses with other legume genomes. Plant Mol Biol 59: 309-322.

Sugiura M. 1992. The chloroplast genome. Plant Mol Biol 19: 149-68.

Taamalli W, Guena F, Bassi D, Daoud D, Zarrouk M. 2008. SSR marker based fingerprinting of Tunisian olive (Olea europaea L.) varieties. J Agron 7:176-181.

Tambarussi EV, Melotto-passarin DM, Barbosa AL, Brigati JB et al..

2009. In silico analysis of simple sequence repeats from chloroplast genomes of Solanaceae species. Crop Breed Appl Biotech 9: 344352.

Timme R, Kuehl EJ, Boore JL, Jansen RK. 2007. A comparative analysis of the Lactuca and Helianthus (Asteraceae) plastid genomes: identification of divergent regions and categorization of shared repeats. Am J Bot 94: 302-312.

Varshney RK, Thiel T, Stein N, Langridge P, Graner A. 2002. In silico analysis on frequency and distribution of microsatellites in ESTs of some cereal species. Cell Mol Biol Lett 7: 537 - 546.

Zane L, Bargelloni L, Patarnello T. 2002. Strategies for microsatellite isolation: a review. Mol Ecol 11: 1-16.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 118-123

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130303

Taxonomy of Indonesian giant clams (Cardiidae, Tridacninae) UDHI EKO HERNAWAN♥ Biotic Conservation Area of Tual Sea, Research Center for Oceanography, Indonesian Institute of Sciences. Jl. Merdeka, Katdek Tual, Southeast Maluku 97611. Tel. +92-916-23839, Fax. +62-916-23873, ♥email: udhi_e_hernawan@yahoo.com Manuscript received: 20 December 2010. Revision accepted: 20 June 2011.

ABSTRACT Hernawan E. 2012. Taxonomy of Indonesian giant clams (Cardiidae, Tridacninae). Biodiversitas 13: 118-123. A taxonomic study was conducted on the giant clam’s specimens deposited in Museum Zoologicum Bogoriense (MZB), Cibinong Indonesia. Taxonomic overviews of the examined specimens are given with diagnostic characters, remarks, habitat and distribution. Discussion is focused on specific characters distinguishing each species. From seven species known to distribute in Indonesian waters, there are six species, Tridacna squamosa Lamarck, 1819; T. gigas Linnaeus, 1758; T. derasa Roding, 1798; T. crocea Lamarck, 1819; T. maxima Roding,1798; and Hippopus hippopus Linnaeus, 1758. This study suggests the need for collecting specimen of H. porcellanus Rosewater, 1982. Important characters to distinguish species among Tridacninae are interlocking teeth on byssal orifice, life habits, presence of scales and inhalant siphon tentacles.

Key words: Tridacninae, taxonomy, Museum Zoologicum Bogoriense

INTRODUCTION Giant clams, the largest bivalve in the world, occur naturally in association with coral reefs throughout the tropical and subtropical waters of the Indo-Pacific region.

From the southeast Pacific westwards to East Africa, its distribution extends up north to the Red Sea (bin Othman et al. 2010). They can generally be found in marine shallow water habitats (1-20 m) and are restricted in only clear waters due to their phototrophic characteristic (Jantzen et al. 2008). Their strong requirement of photosynthetic light is a consequence of their symbiotic relationship with zooxanthellae of the genus Symbiodinium (Hirose et al.

2006). Scientifically, there has been an increasing interest to the clams for more than the last four decades because their high commercial value has led the natural population to extinction. Vulnerable status, or even local extinction, for some species has been reported in Indonesia (Raymakers et al. 2003), Malaysia (Shau-Hwai and Yasin 2003) and several regions in Pacific (UNEP-WCMC 2010).

Recently, there are ten described species of the living giant clams, in only two genera, Tridacna and Hippopus (bin Othman et al. 2010). Three sub-genera are within Tridacna, Tridacna sensu stricto, consisting only T. gigas (Linnaeus, 1758); Persikima consisting T. derasa (Roding, 1798) and T. tevoroa (Lucas, Ledua and Braley 1990); and Chametrachea comprising T. squamosa (Lamarck, 1819), T. crocea (Lamarck, 1819), T. maxima (Roding, 1798), T. rosewateri (Sirenko and Scarlato 1991) and T. costata (Richter, Roa-Quiaoit, Jantzen, Al-Zibdah, and M. Kochzius 2008). The genera Hippopus comprises of two species, H. hippopus (Linnaeus, 1758) and H. porcellanus (Rosewater 1982). There has been much discussion as to whether the giant clams should be placed still on their own

family (Tridacnidae) or revised to be subfamily Tridacninae, included in family Cardiidae. Recently, based on sperm ultrastructure and molecular phylogenetic studies, the clams are belonging to family Cardiidae, subfamily Tridacninae (Schneider and Foighil 1999; Keys and Healey 2000).

Globally, various studies focusing on many aspects of the clams have been done, from biological experiments to mariculture and conservation strategy, for example Keys and Healy (2000), Buck et al. (2002), Kinch (2002), Harzhauser et al. (2008), and Teitelbaum and Friedman (2008). In context of Indonesia, taxonomic notes of giant clams are rare although Indonesian waters are the habitat for most of giant clams species in the world (bin Othman et al. 2010). This paper reports a taxonomic study of giant clams specimens deposited in Museum Zoologicum Bogoriense, Cibinong, Bogor, Indonesia.

MATERIALS AND METHODS The study was conducted in June 2009 as a part of Workshop on Marine Taxonomy 2009, organized by Research Center for Oceanography. The giant clams specimens were observed from the dry collection of Museum Zoologicum Bogoriense (MZB), Cibinong Indonesia. All specimens were personally described, reidentified and determined based on Braley and Healy (1998), Newman and Gomez (2002), Dharma (2005), and ter Porten (2007). An overview of the examined specimens was given with diagnostic characters, remarks, habitat and distribution. Discussion is focused on specific characters distinguishing each species.

HERNAWAN – Giant clams of Indonesia

RESULTS AND DISCUSSION Results From the ten extant giant clams, seven species are known to inhabit Indonesian waters, i.e. Tridacna squamosa Lamarck, 1819; T. gigas Linnaeus, 1758; T. derasa Roding, 1798; T. crocea Lamarck, 1819; T. maxima Roding,1798; Hippopus hippopus Linnaeus, 1758 and H. porcellanus Rosewater, 1982 (Newman and Gomez 2002; bin Othman et al. 2010). However, the specimen collection of giant clams in MZB is obviously not complete since only six species are deposited (Figure 1). None of the specimens is H. porcellanus Rosewater, 1982. Here is the taxonomic overview.

Class Bivalvia Linnaeus, 1758 Subclass Heterodonta Neumayr, 1884 Order Veneroida H. and A. Adams, 1856 Superfamily Cardioidea Lamarck, 1809 Family Cardiidae Lamarck, 1809 Subfamily Tridacninae Goldfuss, 1820

Genus Hippopus Hippopus hippopus (Linnaeus, 1758) (Figure 1A) Syn. Chama hippopus Linnaeus, 1758: 691; Hippopus maculatus Lamarck, 1801: 117.

Material examined. No. Lam 1327; Figure 1 (1); 7 specimens, paired valves (height : length; 6,05 : 8,10 cm; 6,62 : 9,25 cm; 5,85 : 8,12 cm; 5,95 : 7,05 cm; 5,00 : 8,10 cm; 5,72 : 8,18 cm; 5,27 : 7,35 cm; 5,82 : 8,05 cm); Loc. Laratuka Strait, Flores; Date 1953; Coll. Fr. L. Viannay, Det. U. E. Hernawan (22 June 2009) Diagnostic characters. Solid shell, thick and heavy; equivalve, inequilateral, strongly inflated and longer than high (maximum length 40 cm, commonly 20 cm). Umbo position is in midline. Outline of shell fan-shape; posterior and ventral margin meet at an angle less than 900; anterior and ventral margin meet an angle less than 900; posterioventral and anterioventral margin meet at an angle more than 900. Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve.

Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculptured with 9 to 14 large radial fold with 2 to 3 small rib-like at each interstices; anteriodorsal margin with interlocking crenulations, byssal orifice presence on anteriodorsal area but without byssal gape.

Inner margin with irregular crenulations correspond with the sculpture of outer surface. Coloration on outer surface whitish, irregular reddish blotches arranged in irregular concentric bands; inner surface porcelaneous white.

Inhalent siphon without tentacles.

Remarks. This species is easily distinguished to the other giant clams by the presence of irregular semi-tubular spines and numerous riblets on the principal ribs and interstices. Irregular reddish blotches arranged in irregular concentric bands are the other specific character.

Habitat. Found on sandy areas in coral reefs, sometime on seagrass beds adjoining reefs; not attached by a byssus to the substrate.

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Distribution. Tropical eastern Indian Ocean to western Pacific, from Andaman Islands to eastern Melanesia; north to southern Japan and south to Queensland Genus Tridacna Tridacna (Persikima) derasa (Roding, 1798) (Figure 1B) Syn. Tridacna serrifera Lamarck, 1819; Persikima whitleyi Iredale, 1937 Material examined. No. Lam 899; Figure 1(2); 1 specimen, paired valves (11 cm in height and 16,5 cm in length); Loc. Maluku; Date (?); Coll. Rykschroeff, Det. U. E. Hernawan (22 June 2009) Diagnostic characters. Shell solid, thick and heavy; equivalve, inequilateral, inflated and longer than high (maximum length 60 cm, commonly 50 cm). Umbo position posterior. Outline of shell fan-shaped; rounded margin. Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve. Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculptured with 7 to 12 large radial fold with 7 to 12 small rib-like at each interstices; no scales and spines on the fold; anteriodorsal margin with non-interlocking crenulations, byssal orifice presence on anteriodorsal area with small byssal-gape (less than a half of anteriodorsal margin length). Inner margin with distinctively crenulations, correspond to the small rib-like sculpture at the interstices. Coloration on outer surface white, inner surface porcelaneous white. The inhalant siphon with tentacles.

Remarks. This species has outer surface without scales or spines, smoother than the others.

Habitat. Found in coral reefs, shallow water to a depth of 20 m.

Distribution. Tropical western Pacific, from western Indonesia to eastern Melanesia; north to the Philippines and south to New South Wales.

Tridacna (Tridacna) gigas (Linnaeus, 1758) (Figure 1C) Syn. Chama gigas Linnaeus, 1758.

Material examined. No. Lam 899; Figure 1(3); 1 specimen, paired valves (13.44 cm in height and 22,5 cm in length); Loc. Maluku; Date (?); Coll. Rykschroeff, Det. U. E. Hernawan. (22 June 2009).

Diagnostic characters. Shell solid, thick and heavy; equivalve, equilateral, inflated and longer than high (maximum length 137 cm, commonly 80 cm). Umbo position midline. Outline of shell fan-shaped; rounded margin. Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve. Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculptured with 4 to 6 deep radial fold; forming distinctively elongate-triangular projections on ventral free margin (V-shape) with 7 to 12 small rib-like at each interstices; no scales and spines on the fold; anteriodorsal margin with non-interlocking crenulations, byssal orifice presence on anteriodorsal area with small byssal-gape (less than a half of anteriodorsal margin length). Inner margin with indistinctively crenulations. Coloration on outer surface white, inner surface porcelaneous white. The

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inhalant siphon without tentacles.

Remarks. The specific characters are deep radial folds forming V-shape projection in ventral view. Outer surface relatively smooth and extremely large shell.

Habitat. Found in sand, coral reefs, shallow water to a depth of 20 m.

Distribution. Eastern Indian Ocean and tropical western Pacific, from southwestern Myanmar and western Indonesia to Micronesia and eastern Melanesia; north to southern Japan and south to Queensland and New Caledonia.

Tridacna (Chametrachea) crocea (Lamarck, 1819) (Figure 1D) Syn. Tridacna crocea Lamarck, 1819: 106; Tridacna cumingii Reeve, 1862: pl. 7, fig. 7a (part); Tridacna ferruginea Reeve, 1862: pl. 8, fig. 8a-b.

Material examined. No. Lam. 443; Figure 1(4); 2 specimens, paired valves (6,65 cm in height and 8,05 cm in length,); Loc. (?); Date (?); Coll. Duwens, Det. U. E. Hernawan. (22 June 2009).

Diagnostic characters. Shell solid, thick; equivalve, inequilateral, inflated and longer than high (not exceeding 15 cm in length, commonly 11 cm). Umbo position posterior. Outline of shell fan-shaped; rounded margin.

Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve. Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculpture with 6 to 8 low radial fold with scales, closely spaced, near free ventral margin; no spines; no small rib-like at each interstices; anteriodorsal margin with non-interlocking crenulation; byssal orifice presence on anteriodorsal area with wide byssal-gape (more than a half of anteriodorsal margin length). Inner margin with indistinctively crenulations. Coloration on outer surface creamy white; inner surface: porcelaneous, white; often pinkish. The inhalant siphon with tentacles.

Remarks. The smaller species with wide byssal gape.

Outer surface with fingernail-shape scales, closely spaced, undulate, arranged regularly on radial fold, but only near free ventral margin.

Habitat. Found in coral reefs, burrower, totally embedded, burrowed into a coral boulder on the reef top, only shell margin and mantle are visible to the diver.

Distribution. Tropical eastern Indian Ocean to western Pacific, from Andaman Islands to Fiji Islands; north to Japan and south to New Caledonia and Queensland.

Tridacna (Chametrachea) maxima (Roding, 1798) (Figure 1E) Syn. Tridachnes maxima RĂśding, 1798: 171, no. 184; Tridacna elongata Lamarck, 1819: 106; Tridacna rudis Reeve, 1862: pl. 5, fig. 4a-c; Tridacna compressa Reeve, 1862: pl. 6, fig. 5; pl. 7, fig 5b; Tridacna cumingii Reeve, 1862: pl. 7, fig. 7b (part); Tridacna acuticostata Sowerby, 1912: 30-31, fig.

Material examined. No. Lam 1301; Figure 1(5); 1 specimen, right valve (6,79 cm in height and 13,22 cm in length); Loc. (?); Date (?); Coll. (?), Det. U. E. Hernawan.

(22 June 2009).

Diagnostic characters. Shell solid, thick; equivalve, inequilateral, inflated and longer than high (not exceeding 35 cm in length, commonly 25 cm). Outline of shell fanshaped; rounded margin; anterior about twice in length than posterior. Umbo position posterior. Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve. Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculpture with 6 to 8 low radial fold with fingernail-shape scales, closely spaced, arranged regularly on radial fold, near free ventral margin; anteriodorsal margin with non-interlocking crenulations; byssal orifice presence on anteriodorsal area with medium byssal-gape (about a half of anteriodorsal margin length).

Inner margin with distinctively crenulations. Coloration of outer surface, white; inner surface, porcelaneous, white, pale yellowish tinge near the inner margin. The inhalant siphon with tentacles.

Remarks. The elongate giant clam. Shell growth widely elongated in anterior position. The scales presence only near the upper margin.

Habitat. Found in coral reefs, coral burrower partially embedded into a coral boulder on the reef top, sandy bottoms, firmly attached to coral head, in shallow and clean waters with majority living at less than 7m.

Distribution. Widespread in the Indo-West Pacific, from East Africa, including Madagascar, the Red Sea and the Persian Gulf to eastern Polynesia; north to Japan and south to New South Wales and Lord Howe Island.

Tridacna (Chametrachea) squamosa (Lamarck, 1819) (Figure 1F) Syn. Tridacna squamosa Lamarck, 1819: 106; Chama squammata Rumphius 1705; female Littoralis’, pl. 42, fig. A. Material examined. No. Lam. 890; Figure 1(6); 3 specimen, 2 paired valves, 1 right valve (9,15 cm in height and 12,7 cm in length); Loc. (?); Date (?); Coll.

Rykschroeff; Det. U. E. Hernawan. (22 June 2009) Diagnostic characters. Shell solid, thick; equivalve, inequilateral, inflated and longer than high (not exceeding 40 cm in length, commonly 30 cm). Outline of shell fanshaped; rounded margin. Umbo position midline. Hinge with 1 ridge-like cardinal tooth, 2 lateral teeth on right valve, 1 lateral tooth on left valve. Pallial line presence but no pallial sinus; 1 adductor muscle scar. Outer surface sculptured with 5 to 6 low radial fold with more than 6 small rib-like at each interstices. Scales presence; no spines. Anteriodorsal margin with non-interlocking crenulations; byssal orifice presence on anteriodorsal area with medium byssal-gape (about a half of anteriodorsal margin length). Inner margin with distinctively crenulations. Coloration of outer surface, white; inner surface, porcelaneous, white. The inhalant siphon with tentacles.

Remarks. This species have specific character, viz. blade-like, spoon-shape scales, arranged regularly on radial fold. The scales still presence near the umbo, but being smaller.

Habitat. Found in coral reefs, littoral and shallow water to a depth of 20 m, live not embedded into a coral boulder, attached by the byssus to the substrate.

HERNAWAN – Giant clams of Indonesia

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10 cm

4 cm

A

6 cm

B

4 cm

D

C

6 cm 6 cm

E

F

Figure 1. Giant clams specimens deposited in MZB; outer surface view; L (left valve), R (right valve). A. Hippopus hippopus (Linnaeus, 1758), L; B. Tridacna (Persikima) derasa (Roding, 1798), R; C. Tridacna (Tridacna) gigas (Linnaeus, 1758), L; D. Tridacna (Chametrachea) crocea (Lamarck, 1819), R; E. Tridacna (Chametrachea) maxima (Roding, 1798), R; F. Tridacna (Chametrachea) squamosa (Lamarck, 1819), R

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Distribution. Widespread in the Indo-West Pacific, from East Africa, including Madagascar, the Red Sea, but not the Persian Gulf, to eastern Melanesia; north to southern Japan and south to Queensland and New Caledonia.

Discussion The fact that specimens of only six species of the giant clams are deposited in the MZB suggests the need for collecting specimens of H. porcellanus Rosewater, 1982.

The distribution of H. porcellanus is the smallest among distribution of six other species. It is distributed in eastern part of Indonesian waters. In a larger scale, it can be found also in Philippine (Braley and Healy 1998; Newman and Gomes 2002).

Tridacninae can be easily distinguished from other bivalves based on its large shell with a strong radial fold in a few number and brightly colored mantel. Each shell has only one adductor muscle scar where a pedal retractor muscle attached, but no pallial sinus. The shell ligament is external with hinge teeth. One character easily separating Tridacna and Hippopus is the teeth on byssal orifice of opposed valves. Hippopus has interlocking teeth, while Tridacna does not. In turn, Tridacna bears a byssal gape which is not present in Hippopus. Additionally to the shell character, mantle character can also be used to differentiate living Hippopus and Tridacna. When it fully opens, Tridacna’s mantle expands laterally beyond the ventral margin shell. On the contrary, Hippopus’s mantle expands without passing through the ventral margin shell.

Phylogenetically, T. squamosa, T. maxima, and T. crocea are grouped in a single clade taxonomically known as subgenus Chametrachea (Benzie and Williams 1998) because of the character of their life habit attaching and boring coral substrate. T. squamosa is unique for its spoonlike scales. T. crocea embeds totally its shell into coral substrates, whilst only half part of T. maxima shell is embedded into coral substrate. Subgenus Tridacna sensu stricto and Persikima do not attach to their substrate. They are separated each other based on the presence of tentacles in the inhalant siphon. T. derasa has low and weak radial folds on its shells. In contrast, T. gigas is specific for its remarkable large, smooth shell and strong U-shape radial folds.

CONCLUSION Despite many field observations reporting that seven species of the giant clams inhabit Indonesian waters, the MZB deposits specimens of six species (Tridacna squamosa Lamarck, 1819; T. gigas Linnaeus, 1758; T. derasa Roding, 1798; T. crocea Lamarck, 1819; T. maxima Roding, 1798 and Hippopus hippopus Linnaeus, 1758), suggesting the need for collecting specimen of H. porcellanus Rosewater, 1982. Important characters to distinguish species among Tridacninae are interlocking teeth on byssal orifice, life habits, presence of scales and inhalant siphon tentacles.

ACKNOWLEDGEMENTS The author thanks to the staffs of Malacological Laboratory, Museum Zoologicum Bogoriense for their assistance in examining specimens; Pitra Widianwari, Dr. Rosichon Ubaidillah, Dr. Teguh Triyono, Dr. Hari Sutrisno and Dr. Yayuk R. Suhardjono for the fruitful discussion.

REFERENCES Adams H, Adams A. 1856. The genera of recent Mollusca arranged according to their organization, volume 2. John Van Voorst, London.

Benzie JAH, Williams ST. 1998. Phylogenetic relationships among giant clam species (Mollusca: Tridacnidae) determined by protein electrophoresis. Mar Biol 132: 123-133.

Buck BH, Rosenthal H, Saint-Pauli U. 2002. Effect of increased irradiance and thermal stress on the symbiosis of Symbiodinium microadriaticum and Tridacna gigas. Aquat Liv Res 15: 107-117.

Braley R, Healy JM. 1998. Superfamily Tridacnoidea. 332-336. In Beesley PL, Ross GJ B, Wells A. (eds). Mollusca: the southern synthesis. Fauna of Australia 5(A). CSIRO Publishing, Melbourne, 563 pp.

Dharma B. 2005. Recent and fossil Indonesian shells. Hackenheim, Germany.

Goldfuss SGA. 1820. Handbuch der zoologie. Johann Leonard, Schrag, Nurnberg.

Harzhauser M, Mandic O, Piller WE, Reuter M, Kroh A. 2008. Tracing back the origin of the Indo-Pacific mollusc fauna: Basal Tridacninae from the Oligocene and Miocene of the Sultanate of Oman.

Paleontology. 51 (1): 199-213.

Hirose E, Iwai K, Maruyama T. 2006. Establishment of the photosymbiosis in the early ontogeny of three giant clams. Mar Biol 148: 551-558.

Iredale T. 1937. Mollusca. In: Whitley GP (ed) The Middleton and Elizabeth Reefs, South Pacific Ocean. Aust Zool 8: 232-261.

Jantzen C, Wild C, El-Zibdah M, Roa-Quiaoit HA, Haacke C, Richter C. 2008. Photosynthetic performance of giant clams, Tridacna maxima and T. squamosa, Red Sea. Mar Biol 155: 211-221.

Keys JL, Healy JM. 2000. Relevance of sperm ultrastructure to the classification of giant clams (Mollusk, Cardioidea, Cardiidae, Tridacninae). Geological Society London Special Publications 177: 191-205.

Kinch J. 2002. Giant clams: their status and trade in Milne Bay Province, Papua New Guinea. Traffic Bulletin 19(2): 1-9.

Lamarck JB. 1809. Philosophie zoologique, Tome 1 Dentu, Paris.

Lamarck JB. 1819. Histoire naturelle des animaux sans vertèbres. Paris.

Lamarck JB. 1801. Système des animaux sans vertèbres. Deterville, Paris.

Linnaeus C. 1758. Systema naturae per regna tria naturae, secundum classes, ordines, genera, species, cum characteribus, differentiis, synonymis, locis. Tomus I, Holmiae, Laur Salvius.

Lucas JS, Ledua E, Braley RD. 1990. A new species of giant clam (Tridacnidae) from Fiji and Tonga. Australian Centre for International Agricultural Research, Working Paper 33: 1-8.

Newman WA, Gomez ED. 2002. On the status of giant clams, relics of Tethys (Mollusk: Bivalvia: Tridacnidae). In: Moosa MK, Soemodihardjo S, Sugiarto A, Romimohtarto K, Nontji A, Soekarno, Suharsono (eds.). Proceeding of the 9th International Coral Reef Symposium. Bali, 23-27 October 2000. 2: 927-936.

Neumayr M. 1884. Zur morphologie des bivalvenschlosses. KaiserlichKo¨nigliche Akademie der Wissenschaften, Wien. MathematischNaturwissenschaftliche Klasse 88: 385-418. bin Othman AS, Goh GHS, Todd PA. 2010. The distribution and status of giant clams (Family Tridacnidae), a short review. Raffles Bull Zool 58 (1): 103-111. ter Poorten JJ. 2007. Results of the Rumphius Biohistorical Expedition to Ambon (1990). Part 13. Mollusca, Bivalvia, Cardiidae. Zoological Mededelingen 81 (15): 259-301.

HERNAWAN – Giant clams of Indonesia Raymakers C, Ringuet S, Phoon N, Sant G. 2003. Review of the Exploitation of Tridacnidae in the South Pacific, Indonesia and Vietnam. TRAFFIC Europe, Brussels.

Reeve LA. 1862. Monograph of the genus Tridacna. Conchologia Iconica 14, London.

Richter C, Roa-Quiaoit H, Jantzen C, Al-Zibdah M, Kochzius M. 2008.

Collapse of a new living species of giant clam in the Red Sea. Curr Biol 18 (17): 1349-1354.

Röding PF. 1798. Museum Boltenianum, Part 2.— Hamburg.

Rosewater J. 1982. A new species of Hippopus (Bivalvia: Tridacnidae).

The Nautilus 96: 3-6.

Rumphius GE. 1705. D’Amboinsche Rariteitkamer, Behelzende eene Beschryvinge van allerhande zooweeke als harde Schaalvisschen, te weeten raare Krabben, Kreeften, en diergelyke Zeedieren, alsmede allerhande Hoorntjes en Schulpen, die men in d’Amboinsche Zee vindt: Daar beneven zommige Mineraalen, Gesteenten, en soorten van

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Aarde, die in d’Amboinsche, en zommige omleggende Eilanden gevonden worden, 28.— Amsterdam.

Schneider JA, Foighil DO. 1999. Phylogeny of giant clams (Cardiidae: Tridacninae) based on partial mitochondrial 16S rDNA gene sequences. Mol Phylogenet Evol 13(1): 59-66.

Shau-Hwai AT, Yasin Z. 2003. Status of giant clams in Malaysia. SPC Trochus Inform Bull 10: 9-10.

Sirenko BI, Scarlato OA. 1991. Tridacna rosewateri sp. n. A new species of giant clam from the Indian Ocean. La Conchiglia 22 (261): 4-9.

Sowerby GB. 1912. Notes on the shells of Tridacna and descriptions of a new species. Proc Malacol Soc London 10: 29-31.

Teitelbaum A, Friedman, K. 2008. Successes and failures in reintroducing giant clams in the Indo-Pacific region. SPC Trochus Inform Bull 14: 19-26.

UNEP-WCMC. 2010. CITES Species Database, UNEP World Conservation Monitoring Centre, Cambridge, UK. http://www.cites.org/eng/resources/ species.html.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 124-129

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130304

The exploration and diversity of red fruit (Pandanus conoideus L.) from Papua based on its physical characteristics and chemical composition MURTININGRUM1,♼, ZITA L. SARUNGALLO1, NOUKE L. MAWIKERE2 1

Department of Agriculture Technology, Papua State University. Manokwari 98314, West Papua, Indonesia. Tel. +62-986-214991, Fax: +62-986-214991. ď‚Šemail: ningrum_mrt@yahoo.com 2 Department of Agriculture, Papua State University, Manokwari, West Papua, Indonesia Manuscript received: 20 December 2010. Revision accepted: 20 June 2011.

ABSTRACT Murtiningrum, Sarungallo ZL, Mawikere NL. 2012. The exploration and diversity of red fruit (Pandanus conoideus L.) from Papua based on its physical characteristics and chemical composition. Biodiversitas 13: 124-129. The aim of this study was to determine the diversity of red fruit based on its physical characteristics and chemical composition. Exploratory survey method and laboratory research (pure experiment) were used to assess the physical character and chemical composition of crude red fruit. Physical character of each accession showed a variation on fruit color (dark red and red); the fruit and single fruit (drupa) length ranged from 21-71 cm and 1.2-1.8 cm, respectively. The dried red fruit contains 2.03- 3.50% ash, 3.12-6.48% protein, 11.21-30.72% fat, 43.86-79.66% carbohydrate, 3.7821.88 mg/100g vitamin C, 2.00-3.14 mg/100g vitamin B1, 0.53-1.11% Ca, 8.32-123.03 ppm Fe, and 0.01-0,33% P, with total carotenoids and total tocopherol ranging from 332.65-3309.42 ppm and 964.52-11917.81 ppm. The clustering analysis result of red fruit based on its physical characteristic and chemical composition showed that the related accessions was U Saem and Tawi Magari, having a 25% similarity. The accession U Saem and Tawi Magari had the highest level of similarity in total carotenoids and total tocopherol.

Furthermore, they also perform similar physical characteristics by having triangular cylinder shape, red flesh color, and fruit length category.

Key words: red fruit, Pandanus conoideus, accession, physical, chemical, cluster

INTRODUCTION The genus Pandanus is a complex plant with highest species diversity. It is believed that there are approximately 600-700 species of this genus around the world. These plants are prevalent in tropical areas, especially in the Pacific islands, Malaysian islands and Australia (Wagner et al. 1990; Jong and Chau 1998). In Indonesia, a total of 100 species, 60 species and 20 species are found in Papua, Borneo and Maluku, respectively (Purwanto 2007). Some species of the genus Pandanus are very important for people whom live in the highlands of the Papua and West Papua Provinces, one of which is Pandanus conoideus L. P. conoideus is known with different local names in Indonesia, such as pandan seran (Maluku), saun (Seram), sihu (Halmahera), while Papuan call this plan as buah merah which literally means red fruit. This plant is also used by people of Papua New Guinea and it is commonly known as marita (Pidgin) (Stone 1997).

Red fruit grows at wider altitudes ranging from the coast up to 1700 m above sea level (Wiriadinata 1995).

These plants spread almost all over of Papua and West Papua territory. However, the tree is predominantly in Jayawijaya Mountains, Jayapura, Manokwari, Nabire, Timika, and Sorong (sub-district of Ayamaru) (Budi and Paimin 2004). Besides the difference of the deployment, red fruit also composed of different accessions, which is used by local people for various purposes. Papua and West

Papua society, especially who lives in the surrounding mountains of Arfak and Wamena, utilize red fruit as food.

They may consume it directly, or used it as a sauce for sago and sweet potato, or consumed directly (Sadsoeitoeboen 1999).

The diversity of red fruit accession which spread in Papua and West Papua is not yet known. It is probably because the public have not intensively cultivated these plants, and only a few people who has tried to cultivate this crops. The diversity of red fruit accessions can be identified based on their physical characteristics and chemical composition. Chepalium of red fruit consists of a tubular (cylindrical) triangle-shaped, bright yellow to dark red with a length of 42-70 cm (100-110 cm), and 9.6-11 cm in diameter (circumference 30-34.5 cm), which is the center of the pedicel chepalium white; and composed by many single fruit (drupa). Drupa or single fruit has triangularshape with pericarp (layer of single fruit) and contains fat (pulp) yellow or red that is surrounding seed (Walujo et al.

2007). Physical character of a fairly prominent red fruit varies in form of fruit (pericarp), drupa size, and color, while its chemical composition varies mainly on the content of carotene, vitamins, and minerals.

The aim of this study was to determine the diversity of red fruit in some areas of Papua and West Papua Provinces based on their physical traits and chemical composition. It can be assumed that by identifying the chemical components in red fruit flesh, there is an opportunity of red

MURTININGRUM et al. – Diversity of Papuan Pandanus conoideus

fruit to be utilized as a good source of natural antioxidants for food and non food products.

MATERIALS AND METHODS The selected areas for exploration were the central areas of red fruit (P. conoideus) distribution. Those areas were (i) in the Province of West Papua, Indonesia consisted of Manokwari, Teluk Bintuni, and South Sorong Districts, and (ii) in the Province of Papua, there were Nabire and Jayawijaya Districts (Figure 1). In Manokwari District, sampling was conducted in the Sub-district of Masni representing lowland and Sub-district of Minyambou representing highland. In Teluk Bintuni District, sampling was carried out in the Sub-district of Merdey representing the lowland area. In South Sorong District, sampling was taken in Kampung Susai, Sub-district of Aifat, belonging to middle latitudes. In Nabire District, sampling was performed in Kampung Rawawudo and Kalisemen, Subdistricts of Nabire representing the lowlands; while in the Jayawijaya District sampling conducted in the Sub-district of Kelila which classified as highland.

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This study used exploratory survey which was included: (i) inventory of red fruit accessions that are known by the name of their local community, and (ii) observation in the community to examine the use of red fruit based on the society knowledge. After red fruit accessions exploration in every area of research, as much as 1-3 accessions of red fruit which is related to the criteria of most widely grown and consumed by local people was taken as a sample.

Moreover, further observations of physical character and chemical composition were performed on selected accessions.

Laboratory analysis was conducted to study the physical characteristics and chemical composition of drupa. Observation of the physical characteristics which are consists of the cross-sectional shape of fruit, fruit color, fruit length, and drupa length. Analysis of chemical composition on the drupa include water content (oven method), ash (furnace method), fat (Soxhlet extraction), protein (micro Kjeldahl), levels of calcium (Ca), iron (Fe), phosphorus (P), vitamin B1 and vitamin C (oxidometry method), and total carotene (spectrophotometry) and total tocopherol (spectrophotometry) (AOAC 1999).

1 2 3 4

5

Figure 1. Study area to explore the diversity of red fruit (Pandanus conoideus) in Indonesian Papua. Note: 1. Manokwari, 2. South Sorong, 3. Teluk Bintuni, (West Papua Province), 4. Nabire, 5. Jayawijaya (Papua Province).

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Data of physical character and chemical composition was analyzed using a Cluster Analysis. Dendogram character diversity among accessions was constructed by Unweighted Pair-Group Method with Arithmetic (UPGMA) using NTSYS-pc computer program version 2.0 (Rohlf 1993).

RESULTS AND DISCUSSION Exploration of the red fruit accessions The exploration result of some target area showed that there was a variety on physical and chemical characteristics of each accession of red fruit. In Manokwari, there were 18 accessions of red fruit. It is included two accessions in the Sub-district of Masni and 16 accessions in the Sub-district of Minyambouw. In Teluk Bintuni (Sub-district of Merdey) they were 32 accessions and in South Sorong (Sub-district of Aifat) and Jayawijaya (Sub-district of Kelila) there were 12 accessions, moreover in Nabire, there were only 11 accessions. Total red fruit accessions which found in Papua and West Papua were 85 accessions.

The naming for these red fruit accessions based on the traditional classification which was called ‘emic’ by the local peopled. The local people differentiated each accession based on the characteristics of its growth, fruit size, fruit color, drupa size, and utilization. The naming for the red fruit accessions in every region started by the name of a specific area. For example in Jayawijaya it was named as Tawi, the letter U in South Sorong District. In Subdistrict of Minyambou, it was called Hib/Him/Hit in and while in District Teluk Bintuni, it was called Mongk.

The interview results showed that not all accessions of red fruit had a potential to be utilized by the local community. Trees that had been cultivated, rich in oil, highly productive, and resistance to disease are accessions that had the potential to be developed. Based on this evaluation, it was found that the accessions which have a potential to be developed were 7 accessions of Manokwari (1 accession in Masni, 3 accessions in Minyambouw and 3 accessions of Teluk Bintuni (Sub-district of Merdey), there were also 3 accessions each from Nabire, Jayawijaya and South Sorong, so overall there were 16 accessions in total (Table 1).

It is believed that this study might reveal the potential natural resources of Papua based on community local knowledge. Red fruit accessions have not highly-cultivated, therefore commonly planted accession was a primitive native race, and even some of them are taken directly from the nature.

Physical characteristics of red fruit The identification of accessions of red fruit in some areas of Papua and West Papua shows a diversity of physical characteristics (Table 1). Generally, the crosssectional units (cores) of red fruit had triangular shaped and yellow white color. Outer shape of red fruit is not always influenced by the shape of cross-section. For example, there are accessions of red a fruit that form a triangular cross-section, but has round fruit shape extending from the base to the tip of fruit. The color of the set pieces of red

fruit is red to dark red, which is influenced by the content of carotenoid and environmental conditions. In addition, there are some set of pieces of red fruit color such as orange and yellow color, but this accessions are rarely found in society.

The size and length of red fruit can be classified into long pieces (size > 50 cm), medium (size 49-53 cm), and short (size < 35 cm). The highland generally have medium fruit size (42 cm) to long size (80.2 cm), while the lowland areas tend to have a length ranging from 59 to 66 cm. In lowland areas, the size of the fruits is more varied, that is in short (25-29 cm) to long (70 cm).

The variation of length of the red fruit drupa ranging from 1.2 to 1.8 cm. Accession of MMS-M has a shortest grain size length (1.2 cm), while accessions MHY-M, and MHB-M and MTM-N have the longest drupa length (1.8 cm). Long fruit size does not corresponded to the length of drupa. For instance, the accessions MID-M has long pieces (62 cm) but has a shorter length of drupa (1.3 cm). MMWM accessions have short pieces (21 cm), and a longer length of drupa (1.7 cm) (Table 1). Regarding to the height of growth, red fruit in the highland have relatively long size (1.5-2.0 cm), in the lowland varies from short (1.2 cm) to long (1.7 cm), whereas in the middle land have moderate to long grain length (1.4-1.6 cm).

Diversity of red fruit accessions physical character populations in one area may differ in other populations. It is assumed that the growth of red fruit is dependent on the type of its ecogeography. This phenomenon was similar to Jatropha curcas L. which was growing scattered in some areas of Mexico in differences altitude, average temperature, and type of climate. It has a different protein content (19-33%) and fat (46-64%), and also has different physical characteristics, especially shape and size of seeds (Makkar et al. 1998; Herrera et al. 2010). Diversity of physical characteristics, especially those controlled genetically are very useful as a source for red fruit breeding program. In agricultural technology, physical characteristics are required to accomplish the equipment design for handling, processing, and storage (Asoegwu et al. 2006).

Chemical composition of red fruit The chemicals compositions of selected red fruit accessions were varied among others (Table 2 and Table 3). The average value was ash 2.03-3.50%, protein 3.126.48%, fat 11.21-30.72%, carbohydrate 43.86-79.66%, vitamin C 3.78- 21.88 mg/100g, vitamin B1 0.97-3.14 mg/100g, calcium (Ca) 0.53-1.11%, iron (Fe) 8.32123.03%, phosphorus (P) 0.01-0.33%, total carotenoids 333-3309 ppm and total tocopherol 964-11918 ppm. Table 2 shows that the drupa containing the highest fat compared to other proximate components. It can be said that the red fruit is a good source of oil. Drupa of accessions from lowlands are higher in fat than accessions from medium and highland. Red fruit accession with the highest fat content was MMS-M (30.72%).

A variety of fat content can be affected by a variety of plants, genetic, climate conditions, level of maturity, harvest time, and method of extraction (Idouraine et al.

1996; Egbekun and Ehieze 1997). Younis et al. (2000)

MURTININGRUM et al. – Diversity of Papuan Pandanus conoideus

states that the yield of Cucurbita pepo L. African plant oils that grows in the highlands to the low temperatures have a higher fat content than those grown in the lowlands to high temperatures. Although accessions of MMS-M were found in the lowland, the growth has average air temperature which is sufficient for their growth. As a result, it contains the highest fat content (30.72%). Merdey sub-district where MMS-M accession found has an average air temperature of 27.4oC and humidity of 82.83%. Required air temperature to grow crops of red fruit is 23-33oC and in moderate humidity (Budi and Paimin 2004).

Drupa of the accessions which were found in the lowlands also contain higher vitamin C, iron (Fe), phosphorus (P), total carotenoids, and total tocopherol than accessions originating from medium and highland. The highest content of vitamin C is produced by MMW-M accession (21.88 mg/100g), the highest of vitamin B1 is produced by MUSW-S accessions (3.14 mg/100g), the highest calcium levels produced by MUSW-S and MTMW accessions (0.90%), the highest iron levels resulting in MID-M accessions (123.03 ppm), the highest phosphorus

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levels resulting in MUA-S accessions (0.33%), the highest total carotenoids is produced by MTM-N accessions (3309.42 ppm), and the highest total tocopherol is produced

by MID-M accessions (11917.81 ppm) (Table 3).

Cluster analysis Of the 16 accessions of red fruit used in this study, each accession showed different characteristics. The differences are due to the red fruit habitat. The habitat of plants is influenced by sunlight, weather or climatic conditions, temperature, humidity, and the availability of nutrients which can be absorbed by plant. It is also known that habitat of plants be affected the physical characteristics and chemical composition of the plant.

Even though there were different characteristics on each accession, there were also similar characteristics of the 16 red fruit-accessions as shown in Table 1, 2 and 3. The similarities in some of red fruit plant were evaluated to determine the genetic relationship by Cluster Analysis.

Pattern of each red fruit accession similarity depicted in the dendogram of physical and chemical characters (Figure 2).

Table 1. Physical character red fruit (Pandanus conoideus) from Papua, Indonesia No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Origin of accession (district/sub-district) Manokwari/Masni Manokwari/ Minyambow Manokwari/ Minyambow Manokwari/ Minyambow Teluk Bintuni /Merdey Teluk Bintuni /Merdey Teluk Bintuni /Merdey South Sorong /Aifat South Sorong /Aifat South Sorong /Aifat Nabire/Nabire Nabire/Nabire Nabire/Nabire Jayawijaya/Kelila Jayawijaya/Kelila Jayawijaya/Kelila

Locally name Idebebcs Hityom Himbiak Hibnggok Monsmir Memyer Memiwuk U Saem U Sauw U Aupat Tawi Bilim Tawi Muni Tawi Kubu Tawi Ugi Tawi Magari Tawi Kenen

Red fruit accession MID-M MHY-M MHB-M MHG-M MMS-M MMY-M MMW-M MUSM-S MUSW-S MUA-S MTB-N MTM-N MTK-N MTU-W MTM-W MTK-W

Core Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder Triangular cylinder

Fruit flesh color Dark red Red Dark red Red Red Red Red Red Red Red Red Red Dark Red Red Red Red

Fruit length (cm) 62/long 76/ long 71/ long 42/ long 68/ long 70/ long 21/short 66/ long 61/ long 59/ long 52/ long 53/ long 54/ long 75/ long 60/ long 60,1/long

Length of Drupa (cm) 1.3 1.8 1.8 1.7 1.2 1.3 1.7 1.6 1.4 1.6 1.5 1.8 1.6 1.6 1.6 1.5

Table 2. Proximate composition of 16 accessions red fruit (Pandanus conoideus) Red fruit accessions MID-M MHY-M MHB-M MHG-M MMS-M MMY-M MMW-M MUSM-S MUSW-S MUA-S MTB-N MTM-N MTK-N MTU-W MTM-W MTK-W

Water (%,bb)

Ash (%,bk)

Protein (%,bk)

Carbohydrate (%, bk)

Fat (%,bk)

40.82±0.08 52.70±1.04 51.18±0.06 46.95±0.72 40.26±0.40 43.96±0.01 51.93±0.91 51.53±0.29 47.10±0.13 45.18±0.39 41.57±1.29 44.07±2.61 41.42±1.01 51.26±0.38 42.99±0.03 49.02±0.64

2.62±0.04 2.77±0.09 2.99±0.00 3.50±0.28 2.10±0.08 2.09±0.07 2.64±0.11 2.45±0.31 2.78±0.07 2.03±0.01 2.31±0.07 3.15±0.28 2.95±0.00 2.76±0.16 3.03±0.02 2.65±0.87

4.01±0.04 5.53±0.31 5.78±0.02 6.48±0.09 5.54±0.26 5.30±0.15 4.33±0.15 4.77±0.35 5.37±0.65 5.20±0.18 6.22±0.07 5.69±0.25 5.45±0.37 5.50±0.18 3.12±0.10 5.15±0.02

71.15±0.19 71.19±0.78 74.67±0.36 78.81±0.59 61.64±0.14 71.43±0.44 68.33±0.65 77.77±0.17 64.96±0.41 71.66±1.11 79.66±0.45 68.01±1.82 79.33±0.52 75.66±0.36 66.46±0.21 74.24±0.26

22.23±0.20 20.50±0.56 16.55±0.34 11.21±0.22 30.72±0.19 21.18±0.36 24.70±0.39 15.00±0.13 26.88±0.18 21.10±0.92 11.81±0.32 23.15±1.86 12.27±0.15 16.07±0.37 27.39±0.09 17.96±0.36

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Table 3. Vitamins, minerals, total of carotene and tocopherol composition of 16 accessions red fruit (Pandanus conoideus) Red fruit accessions MID-M MHY-M MHB-M MHG-M MMS-M MMY-M MMW-M MUSM-S MUSW-S MUA-S MTB-N MTM-N MTK-N MTU-W MTM-W MTK-W

Vit. C (mg/100 g) 20.61±0.93 16.18±0.59 8.02±0.16 9.97±1.20 12.53±0.11 18.90±0.00 21.88±1.27 9.42±0.61 8.45±0.28 10.30±0.13 3.78±0.08 5.64±0.82 7.33±1.25 4.39±2.30 7.40±1.07 15.41±1.57

Vit. B1 (mg/100 g) 1.88±0.02 2.60±0.12 2.30±0.04 2.39±0.15 0.97±0.03 1.09±0.01 2.47±0.00 2.11±0.00 3.13±0.02 2.22±0.00 2.00±0.10 2.97±0.13 3.09±0.16 2.54±0.11 2.21±0.11 2.04±0.06

Ca (%, bk)

Fe (ppm)

0.60±0.00 0.77±0.02 0.83±0.01 0.74±0.00 0.68±0.00 0.54±0.00 0.58±0.00 0.90±0.05 1.11±0.05 0.59±0.01 0.55±0.00 0.79±0.04 0.71±0.01 0.57±0.04 0.90±0.04 0.53±0.02

123.03±2.09 20.86±0.12 14.65±1.73 16.27±0.86 22.52±0.91 11.83±1.58 17.18±1.20 39.37±0.36 21.88±1.01 22.29±0.51 29.07±0.90 26.69±3.72 26.6±4.76 8.32±0.99 24.54±0.02 12.34±0.79

P (%, bk) 0.11±0.00 0.01±0.00 0.01±0.00 0.02±0.00 0.32±0.00 0.25±0.00 0.31±0.00 0.29±0.00 0.31±0.00 0.33±0.02 0.08±0.00 0.07±0.00 0.07±0.00 0.02±0.00 0.01±0.00 0.01±0.00

Total carotenoids (ppm) 2584.82±224.78 748.86±18.39 332.58±92.36 704.04±37.72 1264.28±38.96 1137.98±37.24 593.89±27.94 547.96±51.29 857.90±15.62 603.16±4.63 759.12±16.72 3330.51±902.91 1185.80±198.52 388.75±11.95 545.80±63.46 730.63±106.65

Total tocopherol (ppm) 11917.81±72.32 5927.11±512.26 2988.76±26.57 6778.49±293.79 2294.12±211.48 1180.54±46.37 2424.49±101.38 2853.23±7.15 1043.04±49.21 964.52±39.39 4529.84±1178.36 6736.36±1625.27 6419.41±723.01 1848.96±150.63 2599.00±297.95 3665.85±521.64

MID-M MMY-M MHY-M MHB-M MTM-N MHG-M MMW-M MUSW-S MUSM-S MTM-W MTU-W MUA-S MTB-N MTK-W MMS-M MTK-N 0.09

0.13

0.17

0.21

0.25

Coefficient

Figure 2. Dendogram of similarity of physic characters and chemical composition in 16 accessions red fruit (Pandanus conoideus) from Papua and West Papua Provinces. Note: Abbreviation of the red fruit accession follows Table 1.

The results of clustering analysis by UPGMA method (Unweighted Pair Group Method with Arithmetic) shows that the patterns of accessions was not based on region of origin, but grouped randomly according to character similarity. These results provide evidence that the

accessions of red fruit in Papua and West Papua were very heterogeneous. Several factors that can cause a high diversity of plant characters were (i) the occurrence of hybridization between accessions, (ii) gene mutation, (iii) migration, (iv) introduction, and (v) the difference of

MURTININGRUM et al. â&#x20AC;&#x201C; Diversity of Papuan Pandanus conoideus

ecogeographic. Natural hybridization can lead to high diversity in populations derived when the different characters are passed down from elders (Grant 1971).

Migration or movement of an individual or population of plants from one place to another followed by the occurrence of geographical isolation and hybridization can lead to gene flow, which ultimately leads to increase the diversity of plant characters (Nagy 1997; Mawikere 2007).

MTK-N accession was grouping in a single group and differ from 15 other accessions, with the similarity just as much as 9%. It is indicated that the MTK-N accessions from Nabire have a fairly distant genetic relationship with other accessions, both accessions from the same region or from other areas. Characters that distinguish the MTK-N with other accessions were their chemical characters.

Accessions that have the closest genetic relationship were accession MUSM and MTM-W, with 25% similarity of character. Although comes from different regions of origin, accessions MUSM-S (South Sorong) and MTM-W (Jayawijaya) have the same chemical and physical components i.e. the highest content of total carotenoids, total tocopherol, the fruit-sectional shape (triangle), color of flesh (red), and fruit length (length). This phenomenon indicates that the red fruit that comes from a region not necessarily have a closer genetic relationship compared to other regions.

CONCLUSION It can be concluded that in order to have more accurate data of genetic relationships among red fruit accessions, the identification cannot be measured only by the physical characteristics and chemical composition, but also on other characters such as molecular traits. It might be due to the facts that physical characteristics and chemical composition were still more influenced by ecogeographic conditions of plants.

The dried red fruit contains 2.03-3.50% ash, 3.126.48% protein, 11.21-30.72% fat, 43.86-79.66% carbohydrate, 3.78-21.88 mg/100g vitamin C, 2.00-3.14 mg/100g vitamin B1, 0.53-1.11% Ca, 8.32-123.03 ppm Fe, and 0.01-0,33% P, with total carotenoids and total tocopherol ranging from 332.65-3309.42 ppm and 964.5211917.81 ppm. The results of clustering analysis based on physical characteristics and chemical composition of 16 accessions of red fruit showed that the accessions MTK-N into a single cluster was different from 15 other accessions, with only 9% similarity of character. Accessions that have the closest genetic relationship were accession MUSM-S and MTM-W with of 25% similarity of character. Accession MUSM-S and MTM-W were similar on their chemical components of the highest content of total carotenoids and especially total tocopherol and also common features of the physical character of which is fruit sectional shape (triangle), fruit flesh color (red) and fruit length.

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ACKNOWLEDGEMENTS The authors would like to thank to the Directorate for Research and Community Service, Directorate General for Higher Education for funding through Competition Research Grant XIV No. 037/SP3/PP/DP2M/II/2006.

Thanks to Sergius Wamafma and Sefnad Asmorom for their assist in the study.

REFERENCES Asoegwu SN, Ohanyere SO, Kanu OP, Iwueke CN. 2006. Physical properties of African oil bean seed (Pentaclethra macrophylla).

Agricultural International: the CIGR Ejournal. Manuscript FP 05 006.

Vol. 8. August, 2006.

AOAC [Association of Analytical Chemist]. 1999. Official methods of analysis of the Association of Official Analytical Chemist. 16th ed.

AOAC, Inc. Arlington, V.A. Budi IM, Paimin FR. 2004. Red fruit. Penebar Swadaya, Jakarta.

[Indonesia] Egbekun MK, Ehieze MU. 1997. Proximate composition and functional properties of fullfat and defatted beniseed (Sesamum indicum L.).

Flour Plant Foods Human Nutr 51: 35-41.

Grant V. 1971. Plant speciation. Columbia Univ Pr, New York.

Herrera JM, Ayala ALM, Makkar H, Francis G, Becker K. 2010.

Agroclimatic conditions, chemical and nutritional characterization of different provenances of Jatropha curcas L from Mexico. Eur J Sci Res 39 (3): 396-407.

Idouraine A, Kohlhepp EA, Weber CW. 1996. Nutrient constituents from eight lines of naked seed squash (Cucurbita pepo L.). J Agric Food Chem 44: 721-724.

Jong TT, Chau SH. 1998. Antioxidative activities of constituents isolated from Pandanus odoratissimus. Phytochemistry 49 (7): 2145-2148.

Makkar HPS, Becker K. 1998. Jatropa curcas toxicity: Identification of toxic principle(s). In: Garland T, Barr AC (eds) Toxic plants and other natural toxicants. CAB International, New York, N.Y. Mawikere NL, Hartana A, Guhardja E, Suharsono, Aswidinnoor H. 2007.

Genetic diversity and relationships of coconut germplasm in East Malesia region based on RAPD markers. Zuriat 18 (1): 81-92.

[Indonesia] Nagy ES. 1997. Frequency-dependent seed production and hybridization rates: Implication for gene flow between locally adapted plant populations. Evolution 51 (3): 703-714.

PurwantoY. 2007. The long hunt pandan. Trubus 38 (Oct 2007): 100-104.

[Indonesia] Rohlf FJ. 1993. NTSYS-pc. Numerical taxonomy and multivariate analysis system. Exeter Software, New York.

Sadsoeitoeboen MJ. 1999. Pandanaceae: Aspects of botany and ethnobotany in the Life Arfak Tribe in Irian Jaya. [Thesis]. Graduate Programme, Bogor Agricultural University, Bogor. [Indonesia] Stone BC. 1997. Pandanus Parkinson. In: Verheij EWM, Coronel RE (eds). Prosea Vegetable Resources of Southeast Asia: Fruits Edible.

PT Gramedia Pustaka Utama, Jakarta. [Indonesian] Wagner WL, Herbst DR, Sohmer SH. 1990. Manual of the flowering plants of Hawaii, Vol. 2. University of Hawaii Press/Bishop Museum Press, Honolulu, Hawaii.

Walujo EB, Keim AP, Sadsoeitoeboen MJ. 2007. Study of ethnotaxonomy Pandanus conoideus Lamarck to bridging the local and scientific knowledge (in Indonesian). Berita Biologi 8 (5): 391-404.

Wiriadinata H. 1995. Plant domestication red fruit (Pandanus conoideus Lam) in Jayawijaya, Irian Jaya. In: Research and Development Project of Biological Resources. Proceedings of the Seminar on Biological Resources Research and Development 1994/1995; Bogor, January 11, 1995. Center for Biological Research and Development (LIPI). Bogor. [Indonesia] Younis YMH, Ghirmay S, Al-Shihry SS. 2000. African Cucurbita pepo L.: Properties of seed and variability in fatty acid composition of seed oil. Phytochemistry 54 (1): 71-75.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 130-134

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130305

Assessment of biodiversities and spatial structure of Zarivar Wetland in Kurdistan Province, Iran MAHDI REYAHI-KHORAM♥, KAMAL HOSHMAND♥♥ Department of Environment, Hamadan Branch, Islamic Azad University, Hamadan, Iran, P.O.BOX: 734, Professor Mosivand st. Tel: +988114494170, Fax: +988114494170, Email: ♥ phdmrk@gmail.com; ♥♥ kamal_hoshmand@yahoo.com Manuscript received: 3 July 2010. Revision accepted: 22 December 2011.

ABSTRACT Reyahi-Khoram M, Hoshmand K. 2012. Assessment of biodiversities and spatial structure of Zarivar Wetland in Kurdistan Province, Iran. Biodiversitas 13: 130-134. Wetlands are valuable ecosystems that occupy about 6% of the world’s land surface. Iran has over 250 wetlands measuring about 2.5 million hectares. Zarivar wetland (ZW) is the only natural aquatic ecosystem in Kurdistan province in Iran. The present research was carried out during 2009 through 2010 with the aim of recognizing the capabilities and limitations of ZW through documentary, extensive field visits and also direct field observations during the years of study. Geographic Information System (GIS) has been used to evaluate the land as a main tool. The results of this research showed that ZW has a great talent regarding diversity of bird species and the ecological status of wetland has caused the said wetland welcome numerous species of birds. The results of this research showed that industrial pollutions are not considered as threats to the wetland but evacuation of agricultural runoff and development of Marivan city toward the wetland and the resulting pollution load could be introduced as an important part of the wetland threats. It is recommended to make necessary studies in the field of various physical and biological parameters of the wetland, and also the facing threats and opportunities.

Key words: aquatic, biodiversity, environment, wetland, Zarivar

INTRODUCTION The environment is a giant and sophisticated set of different processes that have emerged due to gradual evolution of living beings and their interference with nonliving parts on earth. Wetlands are ecosystems that provide numerous goods and services that have an economic value, not only to the local population living in its periphery but also to communities living outside the wetland area (Reyahi Khoram et al. 2011, 2012). Wetlands are valuable ecosystems that occupy about 6% of the world’s land surface. They comprise both land ecosystems that are strongly influenced by water, and aquatic ecosystems with special characteristics due to shallowness and proximity to land. The Convention on Wetlands is an intergovernmental treaty, adopted on 2nd February 1971, in Ramsar, a northern coastal city in Iran; its aim is to promote the conservation and wise use of wetlands, acknowledging that these are extremely important ecosystems for the conservation of biological diversity and welfare of human communities. At present the Ramsar list includes 1933 wetlands of international importance, summing up 189 million of hectares protected in the 160 member states (Ramsar Convention of Wetlands 2011).

Iran has over 250 wetlands measuring about 2.5 million hectares. 22 out of these wetlands measuring 1.8 million hectares have been registered as international wetland in Ramsar Convention (Ramsar Convention of Wetlands 2011). Although the ecological value of wetlands is 10

times as forests and 200 times as farmlands, but unfortunately, 6 international wetland sites of 22 registered wetland sites of Ramsar conversion with the area of 583000 hectares (over 30% of the area of the wetlands of the country registered with Ramsar Convention) are exposed to the threat and acute ecological changes, so that their name is in Montreux Record (Ramsar Convention of Wetlands 2009). Although various different classifications of wetlands exist, a useful approach is one provided by the Ramsar Convention on Wetlands. It divides wetlands in to three main categories of wetland habitats: marine (coastal) wetlands; man-made wetlands and inland wetlands. Inland wetlands refer to such areas as lakes, rivers, streams and creeks, waterfalls, marshes, peat lands and flooded meadows (Schuyt 2004).

Zarivar wetland (ZW) is inland wetland according to mentioned classification. ZW is the only natural aquatic ecosystem in Kurdistan province in Iran. This wetland has formed due to sever erosion of geological formations of the region. This important ecological zone is located in the northwest of Marivan city and situated in the north of Zagros fold belt. According to the classification of wetland habitats approved by Ramsar convention, ZW is in the sweet water section of permanent reservoirs of permanent sweet water wetlands (more than 8 hectares). From a tectonic point of view, Marivan region is an active region and its fold belongs to the middle of the third geological period. ZW is located between 46○, 06', 11" to 46○, 9', 16" eastern longitudes and between 35○, 30', 53" to 35○, 35',

KHORAM & HOSMAND â&#x20AC;&#x201C; Recognition of Zarivar Wetland (ZW), Iran

12" northern latitude with altitude of 1285 meters from sea level, and 2 Kilometers far from Marivan city in Kurdistan province (Figure 1) The aim of this research is to determine characteristics of ZW and providing management strategies which tourists could visit the attractions without damaging the area.

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used to evaluate the land as a main tool. The software used was Arc View (version 3.2a) with the Universal Transverse Mercator (UTM) projection and scale was 1/50,000 (Demers 2009).

RESULTS AND DISCUSSION MATERIALS AND METHODS The present research was carried out during 2009 through 2010 to recognizing the capabilities and limitations of ZW in Kurdistan province in Iran through documentary, extensive field visits and also direct field observations during the years of study. Through the period, using the map, Global Positioning System (GPS) and in some cases through afoot surveying or using car, the geographical location of aquatic species of the region were identified. In this research, valid academic resources were used for identification of Birds, Mammals, Reptiles and Amphibians (Latifi 2000; Mansoori 2008; Ziaie 2008). To identify and define ecological resources of the region, digital maps were used and on this basis the topology situations as well as ground cover of studied area have been accomplished. In addition, Geographic Information System (GIS) has been

Physical and hydrological status ZW covering 3292 hectares was officially declared as a wildlife refuge in 2009 by Department of Environment (DoE) of Iran. The semi humid to humid climatic conditions of the area surrounding the ZW caused formation of a unique forest covering in the mountains of this region, and despite the numerous devastating consequences, it has still beautiful landscapes. The pastures of ZW were one of the first grade and appropriate pastures of Iran. But today, due to irregular use, its ecological balance has changed. ZW was more extensive with a spherical zone shape was made in the past due to function of some faults with northwestern-southeastern alignment and falling of its middle part. Other natural functions of ZW are related to sweet water wetland that creating an appropriate medium for growth of plants, fish and also living of migrating and native birds and animals.

Zarivar Wetland

Marivan City

Figure 1. Location of the study area, Zarivar Wetland, Marivan City, Kurdistan Province in Iran

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B I O D I V E R S IT A S 13 (3): 130-134, July 2012

The surroundings of wetland are Forest Mountains related with Zagros Mountains. The water of sweet water wetland is supplied from a number of springs on the bottom of wetland and also climatic precipitations and numerous springs surrounding the wetland. Due to the changes in the volume of wetland water in the seasons of the year, its water level changes. Its minimum depth is 6 meters and maximum 12 meters. The Zarivar basin with a population of 70,445 (over 85% city dwellers) has two sub basins; Marivan sub basin with the area of 5000 hectares and Zarivar sub basin with the area of 10,827 hectares. The volume between the minimal and maximal figures of wetland water height reaches 19 million cubic meters, and the average annual water evacuation of springs at the bottom of wetland reaches 13 million cubic meters. The average annual water entering the wetland is about 54 million cubic meters, of which 41 million cubic meters is supplied through surface runoff and the remainder through the springs at the bottom of wetland. The average annual precipitation of the region is 786 millimeters. The area of wetland varies due to the changes of water volume during various seasons. The wetland medium perimeter is about 22 Km, relative humidity of 58% and the average annual evaporation of approximately 1900 mm. The highest point in the studied area is 1895 meters above sea level, on northwest of wetland.

The soils around the wetland and shallow lands are deep with brownish grey color. Most of these soils are of silt clay or loam silt clay. The soil textile is light in the water surface parts and is heavy in depth. The underground water table in these types of soil has been estimated 1 to 2 meters. ZW acts in the center of the basin as water flow regulator. The status of springs at the bottom of wetland is not precisely known. Some deem it likely that the water of springs is supplied through carset resources. On this basis, the said springs are related with the confined aquifer from geohydrological point of view. An aquifer may be defined as a formation that contains sufficient saturated permeable material to yield significant quantities of water to springs or wells. Confined aquifers, also known as pressure aquifers, occur where groundwater is confined under pressure grater than atmospheric by overlying relatively impermeable strata. Therefore, if the wetland water level is manipulated by measures such as dam construction, the pressure of water due to increasing water level of wetland will prevent from water flow of wetland bottom springs.

Field studies showed that the industrial plants are not located around the wetland. Hence ZW is not exposed to industrial wastewater pollution and the source of wetland pollution is related to surface runoff and compared to the flow rate of springs, it is more important as regards quality and quantity.

Based on the existing reports, the amount of five-day Biochemical Oxygen Demand (BOD5) and Chemical Oxygen Demand (COD) of wetland water is very small so that BOD5 of wetland water has been reported between 1 to 2 milligrams per liter and the amount of Dissolved Oxygen (DO) was acceptable (Rahnamai 1996). Ghaderi and Ghafouri (2006) showed in their research that from the pollutants transferred to ZW and regarding the intensity of

pollution production, the non point source pollution related to agricultural activities was first rank among other pollutant as community wastewater, solid waste grassland pollution and forest. These pollutions are transferred directly to wetland and threaten the biological systems of ZW. In this situation, the results obtained from the experiments made on the wetland water show that the amount of DO of wetland water is in favorable limit and the amount of the measured BOD and COD is acceptable.

It is obvious that the low amount of pollution indices including BOD and COD indicate that the amount of selfpurification capacity of wetland is favorable and the reedy surrounding the wetland has an effective role in diminishing the pollution.

The results of this research showed that industrial pollutions are not considered as threats to the wetland but evacuation of agricultural runoff and development of Marivan city toward the wetland and the resulting pollution load could be introduced as an important part of the wetland threats. Also evacuation of urban and rural wastewaters inside the wetland will create sever problems for the ecological status of the wetland.

Biological status ZW is considered among the important habitats of the province, and its surrounding regions are appropriate habitat for various animals. Reedy that is connected to the wetland coasts and the reedy that have been formed the islands inside the wetland, are among the main habitats for the birds and amphibious of wetland. Each year numerous species of migrating birds including waterfowl and wader birds spend part of their winter times and birth giving season in this wetland. The biological statuses of wetland include the following sections.

Animal phone The said wetland is important as concerns biological conditions. First its surrounding reedy's is ecologically appropriate for egg laying of the birds and amphibious.

Also the wetland water ecosystem provides appropriate conditions for the life of various amphibious and aquatics (Rahnamai 1996). The most important known biological elements existing within ZW adopted by its environment is as follows: Phytoplanktons: Blue-green algae Brown algae Diatom Diatom Diatom Green algae Green algae Green algae Zooplanktons: Cyclops Daphnia Diaphanosoma Phyllodiaptomus Rotifer

(Microcystis sp.)

(Macrocystis sp.)

(Cymbella sp.)

(Navicula sp.)

(Synedra sp.)

(Chalmydomonas sp.)

(Chlorella sp.)

(Scenedesmus sp.)

(Cyclops sp.)

(Daphnia sp.)

(Diaphanosoma sp.)

(Phyllodiaptomus sp.)

(Rotifera sp.)

KHORAM & HOSMAND â&#x20AC;&#x201C; Recognition of Zarivar Wetland (ZW), Iran Fishes: Caspian shemaya, Blea Common carp Crucian carp Eel Grass carp & White amur Mosquito fish Parma levantskĂĄ Silver carp Stone morocco Birds: Common moorhen Eurasian Coot Grate crested grebe Great cormorant Grey heron Lesser white-fronted goose Little grebe Mallard Purple heron Squacco heron Water rail Mammals: Otter, european oteter Water vole Wetland cat Wild boar Reptiles: Caspian pond turtle Common grass snake Tesselated snake Amphibians: Green toad Green tree frog Wetland frog

(Chalcalburnus sp.)

(Cyprinus carpio) (Carassius auratus) (Mastacembelus mastacembelus) (Ctenopharyngodon idella) (Gambusia affinis) (Capoeta damascina) (Hypophthalmichthys molitrix) (Pseudorasbora parva) (Gallinula chloropus) (Fulica atra) (Podiceps cristatus) (Phalacrocorax carbo) (Ardea cinerea) (Anser erythropus) (Tachybaptus ruficollis) (Anas platyrhynchos) (Ardea purpurea) (Ardeola ralloides) (Rallus aquaticus) (Lutra lutra) (Arvicola terrestris) (Felis catus) (Sus scrota) (Mauremys caspica) (Natrix natrix) (Natrix tessellate) (Bufo viridis) (Hyla cinerea) (Rana ridibanda)

It is to be noted among the said fish species, Stone morocco (Pseudorasbora parva), Crucian carp (Carassius auratus) and Mosquito fish (Gambusia affinis (species are introduced to the said ecosystem and the remainders are native.

Plant flora The flora of ZW includes the flora of the regions surrounding the wetland and the parts inside it. The ecological status around the lake, namely penetration of dry regions inside the water or more movement of water toward dry regions makes an inseparable tie between hydrophilic and xerophytes units. The natural changes of the water level of wetland during various seasons are accompanied by stress on the plant coverage, but its role in biodiversity of the coverage around the wetland is positive and causes formation of different units around it.

Aquatic plants of ZW are divided into four categories of emergent plants, submerged plants, floating-leaved plants and floating plants. Emergent plants grow in the humid lands of the margins of ZW and may penetrate into wetland from the humid lands. The emergent plants in ZW are Common reed (Phragmites australis), cats tail (Typha sp.), ruch (Juncus sp.), flowering rush (Butomus umbellatus), cypress grass (Cyperus rotundus), alkali bulrush (Scirpus maritimus) and sedge (Carex sp.).

Submerged plants spend their entire life cycle under the water unless the flowering stage. This group of aquatic

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plants is rooted in the water bed. Submerged plants could be observed from near shore to the deepest part of ZW. The most area covering these plants could be observed in the distance between water bed of wetland just in vicinity of farmlands in south of ZW and the eastern and southeastern coasts. Submerged plants in ZW are: common bladderwort (Utricularia neglecta), common hornwort (Ceratophyllum demersum), pondweed (Potamogeton Sp.), and longroot smartweed (Polygonum amphibium).

The third category of aquatic plants of ZW includes floating-leaved plants whose leaves are floating on the water surface and their roots are inside the bed. The most location of accumulation of this type of wetland plants is in the distance between the wide band of straw farms in the southern coasts of ZW and eastern coasts of wetland in contact with slope lands on which recreation installations have been constructed. The most species of floating-leaved plants on ZW is white lotus water lily (Nymphaea alba).

The fourth category of aquatic plants of this wetland is floating plants which are also known as free floating plants with leaves and stems floating on water surface. The root of this category of aquatic plants in the water column is free, with no contact with the bed. Although the root is free and there is no dependence on the bed, these plants in ZW are only observed in shallow waters (depth of 0.5 to 1 meter). The most extensive species of floating plants in ZW is Common duckweed (Lemna minor) species.

Based on the results, ZW has a grate capability regarding diversity of birds, Fishes, Mammals, Reptiles, Amphibians and flora species and the ecological status of wetland has caused the said wetland hosts numerous ecotourists every year including school and university study groups as local and international tourists.

CONCLUSION AND RECOMMENDATIONS The obtained results show that this wetland is very talented in the field of ecotourism, bird watching and recreation. It is obvious that investment of public and private sectors will help to realization of potential talents of the wetland and the native people will enjoy its graces. On this basis, they will show more interest in protection and maintenance of the wetland. Local people have good experiences and have inherited such experiences from their ancestors. In this situation, providing an initial trainings related to environmental conservation and ecotourism, they may take part in the related economic activities like local accommodation centers department, restaurants, visitors' guide production of handicraft, and other tourist services.

Certainly training to villagers about the values of ZW, guarantees sustainable ecotourism in the area and so implementing ecotourism programs must provide an economic base for preservation and restoration of ZW. It is obvious that programming must be made in such a way as to guarantee the preservation and durability of the areas, increasing income of local people and also increasing the fans of nature and environment.

Study and investigation of the area can be recommended to determine and identify the borders of

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wetland. Also, Comprehensive study and topographic surveying of the status of actual evaporation and potential evaporation of the wetland water is recommended. Since environmental management of ZW is very important, it is suggested that the authorities consider and Efforts for declaration of ZW as an International Wetland according to Ramsar Convention.

ACKNOWLEDGEMENTS This research was supported by Department of Environment (DoE) of Islamic Republic of Iran, to which the authorsâ&#x20AC;&#x2122; thanks are due. The authors also thank Khairollah Moradi, the head of Kurdistan Provincial Directorate of Environmental Protection, for their collaboration in this study.

REFERENCES Demers M. 2009. Fundamental of Geographic Information System. 4th ed.

Johne Wiley & Sons, New York.

Ramsar Convention of Wetlands. 2011. The Ramsar List of Wetlands of International Importance. The Ramsar Convention of Wetlands. http://www.ramsar.org/pdf/sitelist.pdf Ghaderi N, Ghafouri AM. 2006. Comparative assessment of natural (forest and range) versus manmade (agriculture and urbane) environment in lake Zarivar, Iranian J For Range Protect Res 4: 1927.

Latifi M. 2000. Snakes of Iran. DoE of Iran, Tehran.

Mansoori J. 2008. A guide to the birds of Iran. Farzan Book Publisher, Tehran.

Rahnamai MT. 1996. Sustainable protect and enjoying of Zarivar lake.

Kurdistan Provincial Directorate of Environmental Protection, Sanandaj.

Ramsar Convention of Wetlands. 2009. List of wetlands of international importance included in the Montreux record. http://www.ramsar.org/cda/en/ramsar-documents-montreuxmontreux-record/main/ramsar/1-31-118%5E20972_4000_0 #remove Reyahi-Khoram M, Norisharikabad V, Vafaei H. 2011. Review of seasonal wetlands, case study: AG-GOL Wetland in Hamadan Province, IRAN. Proceeding of 4th USM-JIRCAS Joint International Symposium, Universiti Sains Malaysia, Penang, Malaysia, 18-20 January 2011.

Reyahi-Khoram M, Norisharikabad V, Vafaei H. 2012. Study of biodiversity and limiting factors of Ag-gol Wetland in Hamadan Province, Iran. Biodiversitas 13: 135-139.

Schuyt K, Brander L. 2004. Living waters conserving the source of life (The Economic Values of the Worldâ&#x20AC;&#x2122;s Wetlands). Swiss Agency for the Environment, Forests and Landscape (SAEFL), Amsterdam.

Ziaie H. 2008. A filed guide to the mammals of Iran. DoE of Iran, Tehran.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 135-139

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130306

Study of biodiversity and limiting factors of Ag-gol Wetland in Hamadan Province, Iran MAHDI REYAHI-KHORAM1,♥, VAHID NORISHARIKABAD2, HOSHANG VAFAEI1 1

Department of Environment, Hamadan Branch, Islamic Azad University, Professor Mosivant st., P.O.Box 65138-734, Hamadan, Iran. Tel: +98-8114494170, Fax: +98811 4494170, ♥email: phdmrk@gmail.com 2 Graduate School of the Environment and Energy, Islamic Azad University-Science and Research Branch, Tehran, Iran.

Manuscript received: 24 June 2012. Revision accepted: 30 July 2012.

ABSTRACT Reyahi-Khoram M, Norisharikabad V, Vafaei H. 2012. Study of biodiversity and limiting factors of Ag-gol Wetland in Hamadan Province, Iran. Biodiversitas 13: 135-139. Ag-gol Wetland is one of the important and seasonal wetlands of Iran. This wetland is located on southeast of Hamadan Province and was declared as a prohibited hunting area by Department of Environment (DoE) of Iran.

Identifying the characteristics, capabilities and limiting factors of Ag-gol Wetland is the most important objective of this study. The present study was conducted during 2007 through 2010. Documentary and observation methods have been used to access to information. For general identification of the area, digital maps and Geographic Information System (GIS) were used. Identifying capabilities and limitation factors of the studied area was made through extensive field inspections and direct field observations. Bird species of the region were identified and statistics of the population of bird species were gathered. In this research, valid academic resources were used. According to field inspections and studies, 46 species of waterfowl birds and wader birds of Iran have been identified at this wetland. The results of this research showed that Ag-gol Wetland has a high potential regarding the variety of waterfowl and wader birds. Continuous study in order to determine the depth of wetland water in different time intervals to estimate the volume of wetland water during different seasons of the year is recommended.

Key words: biodiversity, environment, prohibited hunting area, seasonal wetland

INTRODUCTION Global warming and climate change could severely impact streams, wetlands, and other sensitive areas. The water levels and ecology of the wetlands are sensitive to atmospheric change. Wetlands all over the world have been lost or are threatened in spite of various international agreements and national policies. This is caused by: the public nature of many wetlands products and services; user externalities imposed on other stakeholders; and policy intervention failures that are due to a lack of consistency among government policies in different areas. All three causes are related to information failures which in turn can be linked to the complexity and invisibility of spatial relationships among groundwater, surface water and wetland vegetation (Conly and Van Der Kamp 2001).

Wetlands measure about 885 million hectares which 1888 wetlands measuring and 185 million hectares have been registered in Ramsar Convention. Iran has over 250 wetlands measuring about 2.5 million hectares. 22 out of these wetlands measuring 1.8 million hectares have been registered as international wetland in Ramsar Convention (Ramsar Convention of Wetlands 2010). In this situation, only a small part of the world's wetlands (0.3%) is located in Iran (Reyahi-Khoram and Hoshmand 2012). Ramsar Convention classified wetlands in 3 main types; marine (coastal) wetlands; man-made wetlands and inland

wetlands. Many of Inland wetlands are seasonal and particularly in the arid and semiarid area, may be wet only periodically. Seasonal wetland, known commonly as vernal ponds, are isolated from river and other water bodies and characterized by a seasonally fluctuating water level, often drying out completely for some part of the year. Seasonal wetlands are particularly important to amphibian populations and often are small. These habitats provide breeding sites for wetland wildlife that are not populated by predatory fish or other major predators.

Throughout the world, endemic flora and fauna are adapted to the physical and chemical environment found in seasonal wetlands, which are biodiversity hotspots for a broad array of organisms. Several species of small mammals apparently readily use seasonal pools and we do not have a clear understanding of how important seasonal wetlands are to these species (Peter 2005). Seasonal wetlands are beginning to achieve recognition as important habitats because of its richness in biodiversity and the habitat they provide for plants and animals during periods of high water levels. Seasonal wetland dry up in summer and only contain water during wetter months of the year.

As a result of this periodic drying, species requiring water year-round are not able to survive.

Ag-gol Wetland is one of the important and seasonal wetlands of the country; this wetland is located on southeast of Hamadan Province, 30 km northeast of

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Malayer city and is situated between 34ยบ, 32', 10'' and 34ยบ, 33', 2'' northern latitudes and between 49ยบ,1',23'' and 49ยบ,1',27'' eastern longitudes (Figure 1). Ag-gol Wetland was declared as a prohibited hunting area by Department of Environment (DoE) of Iran in second half of 2009.

Thereafter, it was protected as per the law.

Identifying the characteristics, capabilities and limiting factors of Ag-gol Wetland is the most important objective of this study. The said research may provide a means of identifying the threatened species and critically endangered species and also determine the effective causes of protecting and survival of the said species and may contribute to improvement of the programming and management overall study area.

MATERIALS AND METHODS The present study was conducted in Ag-gol Wetland, Hamadan Province, Iran, during 2007 through 2010.

Documentary and observation methods have been used to access to information. This means that identifying capabilities and limitation factors of the studied area was made during the research years through extensive field inspections and direct field observations.

During the period, using the map, camera, Global Positioning System (GPS) and in some cases through afoot surveying, the status of Ag-gol Wetland was identified.

Means, the status of Ag-gol Wetland was inspected in all

seasons of the year. Regarding that this wetland is one of the seasonal wetlands of Iran, it was not possible to easily inspect all parts of the wetland in all of seasons, but the hummocks islands of the wetland were also inspected and studied afoot over the swamp. Bird species of the region were identified and statistics of the population of bird and animal species were gathered. In this research, valid academic resources were used for identification of birds and animals (Mansoori 2008) and (Brati 2009). In order to determine the volume of annual precipitation, the figures and information gathered via the meteorological stations around the wetland were used. For general identification of the area, digital maps and Geographic Information System (GIS) were used and on this basis, the topological status of the area was identified. The software used was Arc View (version 3.2a) with the Universal Transverse Mercator (UTM) projection and scale was 1/50,000 (Demers 2009).

RESULTS AND DISCUSSION Physical and hydrologic status Ag-gol Wetland is a nearly plain wetland, which it's lowest part has a height of 1653.65 meters from free sea level. The villages surrounding this wetland are Eslamabad, Nasirabad, Majidabad, Ghasemabad and Tootal. The bed of Ag-gol Wetland is comprised- up to 70%- of hard formations, namely lime stones, sand stones and hard schist. About 30% are shale formations, which their final

Hamadan Province

Iran, Islamic Republic

A

Zarivar Wetland

Malayer City

B Figure 1. Location of the study area, A. Hamadan Province, Iran, B. Malayer City, C. Ag-gol Wetland

Marivan City C

REYAHI-KHORAM et al. â&#x20AC;&#x201C; Biodiversity of Ag-gol Wetland, Iran

destruction and decomposition leads to small granules such as silt and clay. The wetland's adjacent lands include alluvium plains, flood plains and closed basins. Alluvium plains include Kardkhord soil series (with mean texture), Ghahavand soil series (including heavy texture) and Ahmadabad soil series (with mean alluvium texture). Also flood plains include Ghasemabad soil series (with very heavy alkali texture) and Abdolrahim soil series (with heavy texture and high salinity and alkali). Closed basins are meant the soil around the wetland, which are comprised of heavy-texture alkali soils with low slope.

Its general slope from the focal point of wetland toward the north is 0.03% and from the center toward the south is 0.02%. The real area in closed curve of 1653.7 meters is 116 hectares, and the area in the curves of 1653.7 and 1654 is about 500 hectares, and the area in the curves of 1654 and 1654.5 is 313 hectares. In recent years, governmental organizations have constructed flood gate around the wetland in order to protect the wetland and also to prevent from water flood in the adjacent lands and villages. This flood gate measure 10 meters in width and 1.5 meters in height and has encompassed the surrounding area of wetland. On this basis, the area of the wetland restricted in this flood gate is estimated over 500 hectares. From the time of construction of this flood gate, during winter and spring seasons, water height has increased in the wetland (Figure 1C).

Due to irregular entry of water and evaporation, the surface of wetland is changing; but the wetland area during winter and spring seasons reaches to over 500 hectares and as the water advances into the middle parts, two hummocks islands will appear that their height reaches 6 meters. Also during these days, the average water height in Ag-gol Wetland reaches 1 meter and in the best conditions, the amount of water available in the wetland is about 5 million cubic meters.

The results showed that the average precipitation in the surface of Ag-gol Wetland watershed equals about 271 millimeters per year. Also the area of Ag-gol Wetland basin is equivalent to 125 Square Kilometers. Therefore, the amount of water from atmospheric precipitations in wetland basin is equivalent to 30 million cubic meters per year, which a part of it flows toward the farms and agricultural lands through the constructed channel. So, the sources of supplying water of this wetland are precipitations in the surface of the wetland's basin and also the seasonal channels and streams floodway which is connected to Ghare-Chay River somehow. The waters coming to the wetland (during watery seasons) accumulate on the surface of wetland and then flow as flood from north of wetland toward north of the region. Based on the approvals made in the 180th session of ministry of Energy's water allocation commission, 2 million cubic meters water right per year has been confirmed as right of environmental water need of Ag-gol Wetland per year. When the water current from Ghare-Chay reaches zero, farmers begin taking water from the wetland. It is obvious that in this case, Ag-gol Wetland act such as a reservoir for agricultural affairs of villagers of the region. Also under some conditions, farmers of the region use pumping

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systems to take the wetland's water. The studies showed that this wetland is dry during July, August, September, October and November months and hence, wetland impounding begins in December each year.

Biological and ecological status Ag-gol Wetland is considered as one of the important habitats of the province. Every winter, a large number of migratory birds, including waterfowl and wader birds spend a part of their winter times and birth giving season in this wetland. In Ag-gol Wetland, the wader bird species exceed than waterfowl birds and wader birds are usually was show in the margins of wetland, and these regions are regarded as wader birds. The trend of changes related to species population of index species such as waterfowl birds and wader birds of Ag-gol Wetland has been estimated by using the numerous statistics methods. Study of general changes show the reduction of the number of population from July to December and increase in the number of birds from March to May and further reduction in June. The species that were identified in this wetland in June are mostly reproductive species.

According to field inspections and studies, 46 species of waterfowl birds and wader birds of Iran have been identified at this wetland. The identified species include 14 families. From among these, Scolopacidae are a highly diverse family of birds related to Ag-gol Wetland and this Family is equals 27% of bird species of wetland. Also the most variety of species in this wetland is related to Order Charadriiformes. The diversity of Birds in Ag-gol Wetland is presented in Table 1.

Water quality Field studies showed that the industrial plants are not located around the wetland. Hence Ag-gol Wetland is not exposed to urban and industrial wastewater pollution.

According to the results of experiments, in spring (which the amount of wetland water is in its highest amount) the volume of Dissolved Oxygen (DO) was between 6 to 8.5 mg/L, the average of 5- day Biochemical Oxygen Demand (BOD5) and Chemical Oxygen Demand (COD) is respectively equal to 8 and 19 mg/L and the amount of Total Dissolved Solid (TDS) was between 164 to 576 mg/L. Also the amount of Total Suspended Solid (TSS) was between 80 to 624 mg/L. Discussion The results of this research showed that Ag-gol Wetland has a high potential regarding the variety of waterfowl and wader birds. The ecological status of wetland has made it welcome numerous species of waterfowl and wader birds. These species include winter migrating species and reproductive species, which are seen in this wetland in winter and spring. Regarding this characteristic, it seems that this wetland has a high potential of ecotourism attractions such as bird watching and similar recreations. It is obvious that investments by public and private sectors will promote the potentials of the wetland and the native peoples will benefit from it, and hence they will show more interest in protecting and

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Table 1: Diversity of birds in Ag-gol Wetland, Iran Family

Scientific name

Accipitridae Anatidae

Aquila clanga Anas acuta Anas crecca Anas platyrhynchos Anser anser Aythya ferina Tadorna ferruginea Tadorna tadorna Ardea cinerea Bubulcus ibis Casmerodius albus Egretta garzetta Charadrius alexandrinus Charadrius dubius Charadrius hiaticula Charadrius mongolus Vanellus vanellus Ciconia ciconia Ciconia nigra Glareola pratincola Arenaria interpres Larus genei Larus ridibundus Phalaropus fulicarius Phalaropus lobatus Phalacrocorax carbo Phoenicopterus ruber Fulica atra Himantopus himantopus Recurvirostra avosetta Actitis hypoleucos Calidris alpine Calidris minuta Gallinago gallinago Lymnocryptes minimus Numenius arquata Tringa nebularia Tringa ochropus Tringa tetanus Chlidonias leucopterus Sterna albifrons Sterna hirundo Sterna nilotica Sterna repressa Plegadis falcinellus Platalea leucorodia

Ardeidae

Charadriidae

Ciconiidae Glareolidae Laridae

Phalacrocoracidae Phoenicopteridae Rallidae Recurvirostridae Scolopacidae

Sternidae

Threskiornithidae

maintaining the wetland. The results of this research showed that industrial pollutions are not a threat to the Aggol Wetland. Regarding shortage of surface and underground water resources, the owners of industries show no interest in construction of industrial plant adjacent to the wetland. The results obtained from the experiments made on the wetland water also indicate that the amount of DO in the wetland water is favorable and the amount of BOD and COD is acceptable. It seems that the negligible pollution related to BOD and COD is due to the migration of birds and presence of aquatics in this wetland, and mentioned parameters are not as threat for the wetland. The

measurements related to the amount of TDS and also TSS in water show that these amounts are available in an extensive range and this could be attributed to different climatic conditions such as wind blow, storm, precipitation and flood.

Based on the results of this research, the issue of water and lack of its management is the most basic challenge and threat to the Ag-gol Wetland; such that the farmers of the region use pumping systems to take water from the wetland in aridity conditions, which is contrary to environmental rules, regulations and standards. Regarding that Ag-gol Wetland has been introduced as a prohibited hunting area

by DoE of Iran, it is necessary that the Hamadan Provincial Directorate of Environmental Protection and also Hamadan water resources management company enforce laws and prevent from wetland water taking and protect the wetland.

As mentioned above, in the 180th session of ministry of Energy's water allocation commission, 2 million cubic meters water right per year has been confirmed as right of environmental water need of Ag-gol Wetland per year, although this amount does not conform to the amount of evaporation (only during non-dry seasons) from the surface of wetland in December, January, February, March, April, May and June; namely, the amount of water evaporation during the said months reaches to 500 millimeters and the volume of evaporated water equals over 2 million cubic meters.

CONCLUSION AND RECOMMENDATION Seasonal wetlands are flooded in the winter and begin to dry out in the summer. With due regard to the limited water resources in the studied area and according to the results of this research, it can be concluded that, the water quantity management of the study area is very important and is the key strategy in the management of Ag-gol Wetland. In similar survey done in United State, it is found that regional stresses to the shallow groundwater system such as pumping or low Great Lake levels can be expected to affect even drier wetland types (Skalbeck et al. 2008).

Quinn and Hanna (2003) suggested the development of a comprehensive flow and salinity monitoring system and application of a decision support system (DSS) to improve management of seasonal wetlands in the San Joaquin Valley of California. In another study, it also suggested the use of a number of sensor technologies that have been deployed to obtain water and salinity mass balances for a 60,000 ha tract of seasonally managed wetlands in the San Joaquin River Basin of California (Quinn et al. 2010). In another study, the importance of two hummocks islands in the middle of Ag-gol Wetland was emphasized and promoting the level of protection of the region was recommended (Reyahi-Khoram et al. 2010a,b).

The authors sure that water quantity management of the study area can improve the environmental conservation and biodiversity of two hummocks islands and enhancement of stockholders. Continuous study in order to determine the depth of wetland water in different time intervals to estimate the volume of wetland water during

REYAHI-KHORAM et al. â&#x20AC;&#x201C; Biodiversity of Ag-gol Wetland, Iran

different seasons of the year is recommended. Also, it is suggested that future studies should focus on determine and identify the borders of wetland, topographic surveying of wetland in order to draw level curves with 10 cm distances, the status of actual evaporation and potential evaporation of the wetland water.

ACKNOWLEDGEMENT This research was supported by Department of Environment, Islamic Republic of Iran, to which the authorsâ&#x20AC;&#x2122; thanks are due. The authors also thank Mohammad Pour, the head of Hamadan Provincial Directorate of Environmental Protection, for their collaboration in this study.

REFERENCES Brati A. 2009. Physical and biological study of the Ag-gol Wetland, Hamadan Province. Hamadan Provincial Directorate of Environmental Protection, Hamadan.

Conly FM, Van Der Kamp GG. 2001. Monitoring the hydrology of Canadian Prairie Wetlands to detect the effects of climate change and land use changes. Environ Monit Assess 67 (1-2): 195-215.

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Demers M. 2009. Fundamental of Geographic Information System. John Wiley & Sons, New York.

Mansoori J. 2008. A guide to the birds of Iran. Farzan Book, Tehran.

Peter WCP. 2005. A review of vertebrate community composition in seasonal forest pools of the northeastern United States. Wetlands Ecol Manag 13: 235-246 Quinn NWT, Hanna WM. 2003. A decision support system for adaptive real-time management of seasonal wetlands in California. Environ Model Software 18 (6): 503-11.

Quinn NWT, Ortega R, Rahilly PJA, Royer CW. 2010. Use of environmental sensors and sensor networks to develop water and salinity budgets for seasonal wetland real-time water quality management. Environ Model Software e 25 (9): 1045-58.

Ramsar Convention of Wetlands. 2010. The Ramsar list of wetlands of international importance, the Ramsar convention of wetlands, http://www.ramsar.org/pdf/sitelist.pdf Reyahi-Khoram M, Hoshmand K. 2012. Assessment of biodiversities and spatial structure of Zarivar Wetland in Kurdistan Province, Iran.

Biodiversitas 13 (3): 130-134.

Reyahi-Khoram M, Norisharikabad V, Abdollahi R. 2010a. Survey on Khan Gormaz Protected Area (KGPA) and its ecotourism attractions; Proceeding of the 4th International Colloquium on Tourism & Leisure.

Bangkok, 6-9 July 2010.

Reyahi-Khoram M, Riazi B, Norisharikabad V. 2010b. Study of the nests of Sterna nilotica in Ag-gol Wetland of Hamadan Province. Wetland J 5: 37-42.

Skalbeck JD, Reed DM, Hunt RG, Lambert JD. 2008. Relating groundwater to seasonal wetlands in southeastern Wisconsin, USA. Hydrogeol J 17 (1): 215-28.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 140-144

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130307

Diversity of soil macrofauna on different pattern of sloping land agroforestry in Wonogiri, Central Java MARKANTIA ZARRA PERITIKA, SUGIYARTO♥, SUNARTO Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University Surakarta. JI. Ir. Sutami 36a Surakarta 57126, Central Java, Indonesia. Tel./fax: +62-271-663375. ♥email: sugiyarto_ys@yahoo.com Manuscript received: 27 December 2010. Revision accepted: 1 April 2011.

ABSTRACT Peritika MZ, Sugiyarto, Sunarto. 2012. Diversity of soil macrofauna on different pattern of sloping land agroforestry in Wonogiri, Central Java. Biodiversitas 13: 140-144. The purposes of this study were to determine the diversity level of soil macrofauna on different patterns of sloping land agroforestry, in Wonogiri District, Central Java, and to find out the relationship between environmental factors and the level of soil macrofauna diversity. The study was conducted by sampling at three different patterns of agroforestry, namely: pattern of mixed agroforestry (PAC), pattern of teak agroforestry (PAJ), and the pattern of sengon agroforestry (PAS). The field sampling used two methods, namely pit fall traps to obtain above ground macrofauna, and hand sorting methods to obtain underground macrofauna, on land slope of 39%, 35%, and 27%. The data were collected to determine the diversity index of soil macrofauna; and the environmental factors were also measured. The relationship between environmental factors and the diversity index of soil macrofauna was presented in Pearson's correlation analysis. The results showed that the pattern of sloping land agroforestry in Wonogiri District, Central Java had different diversity index of soil macrofauna. The average diversity index of surface macrofauna was the PAC (0.710), PAS (0661), and PAJ (0.417). The average diversity index of underground macrofauna was the PAC (0.887), PAS (0.860), and PAJ (0.843). The diversity index of soil macrofauna in various patterns of sloping land agroforestry showed that there was a correlation with environmental factors.

Key words: soil macrofauna, diversity, agroforestry, sloping land

INTRODUCTION Most areas in Indonesia are hilly or mountainous areas that create the sloping lands (Setyawan et al. 2006).

Sloping lands are scattered in the tropics. Around 500 million people use them for farming (Craswell et al. 1997).

Wonogiri is one of the regions having many mountains and hills with an area of 182,236.02 ha consisting of different types of land, among others: alluvial, litosol, regosol, andosol, grumusol, mediterranian and latosol. Wonogiri has a harsh topography. Most of the land is rocky and dry and is not good for agricultural purposes (BPS 2010).

One technology being assessed as appropriate with the conditions of sloping lands is the application of agroforestry, a land management system with the basis of sustainability, which increases the overall land production, simultaneously or sequentially combines the production of agricultural crops (including tree plants) and forest plants and/or animals on the same land unit, and implements new ways of managing appropriate to the local population culture (Kartasubrata 1991; Damanik 2003).

Agroforestry is appropriate for the management of watershed area (flood and landslide control) with a variety of considerations, namely the land rehabilitation which can improve the physical fertility (improving land structure and water content), chemical fertility (increasing levels of organic matter and nutrient availability) and land biology (increasing activity and diversity), land morphology (the

formation of solum); and it has an important role in rehabilitating degraded land (Wongso 2008).

The largest land reforestation program in the developing countries has been done in China under China's Sloping Land Conversion Program (SLCP), having the goal of converting 14.67 million hectares of cropland to forests by 2010 (4.4 million of which is on land with slopes greater than 25°) and an additional goal of afforesting a roughly equal area of wasteland by 2010 (Bennett 2008). This program is proven to improve soil quality and increase rural household incomes (Grosjean and Kontoleon 2009; Xu et al. 2010).

Agroforestry as a system of land use is more acceptable by the society because it is profitable for the socio-economic development, and as a venue for farmers’ community empowerment and conservation of natural resources and rural areas environment management. This pattern is considered very suitable to be developed in Solo’s upstream of watershed area that has many aslant areas (Soedjoko 2002). One of Solo’s upstream of watershed area is located in Wonogiri, Central Java.

Soil macrofauna with size of more than 2 mm consists of miliapoda, isopods, insects, mollusks and earthworms (Maftu'ah et al. 2005). Soil macrofauna has an important role in the decomposition of land organic matter in the supply of nutrients. They will eat dead vegetable substances, then the material will be extracted in the form of dirt (Rahmawaty 2004).

PERTIKA et al. â&#x20AC;&#x201C; Diversity of macrofauna on sloping land agroforestry

The relation between soil macrofauna diversity and ecosystem function is very complex and mostly unknown.

The concern to conserve of soil macrofauna biodiversity is very limited (Lavelle et al. 1994; Sugiyarto 2008).

Currently, there is no research about the diversity of soil macrofauna found in various patterns of sloping land agroforestry, in Wonogiri District, Central Java, Indonesia.

Given the importance of soil macrofauna role in the ecosystem and relatively limited information about the existence of soil macrofauna in various patterns of sloping land agroforestry, it is necessary to make inventory about the diversity of soil macrofauna on the area.

This research in soil macrofauna diversity on Sloping land agroforestry in Wonogiri District, Central Java was done by identifying and quantifying soil macrofauna diversity in various patterns of Sloping land agroforestry and explained the relationship between environmental factors and levels of soil macrofauna diversity.

MATERIALS AND METHODS Study area The research was conducted in sloping land area of Semagar Duwur Village, Girimarto Subdistrict, Wonogiri District, Central Java, Indonesia.

Procedure Sampling point determination There were three observation stations with the slope of 39%, 35%, and 27%, respectively. Three patterns of agroforestry were determined in each station namely: mixed agroforestry pattern (PAC), teak agroforestry pattern (PAJ) and sengon agroforestry pattern (PAS).

Then, with simple random sampling method, sampling points were randomly determined in each pattern.

Soil macrofauna sampling The method of pit fall traps was used to get surface macrofauna and the method of hand sorting was used to get underground macrofauna (Suin 1997; Maftuâ&#x20AC;&#x2122;ah et al. 2005).

Identification of soil macrofauna Identification of soil macrofauna was done with reference to some books, including Borror et al. (1989), and Suin (1997).

Measurement of environmental factors At each station, the biotic environmental factors were recorded,

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namely the amount of vegetation types. Then, at each sampling point, several abiotic environmental factors were measured, both the characters of physics and chemistry, namely: (i) physical characteristics (intensity of sunlight, air relative humidity, air temperature, soil temperature); (ii) chemical characteristics (soil pH, soil organic matter).

Data collection techniques The species of soil macrofauna were identified and counted normally. Environmental factors variables were taken using its own measuring instruments on sites directly or in the laboratory indirectly.

Data analysis The data were used for calculating the Diversity Index.

Then, Pearson correlation analysis is performed to determine the relationship of diversity indices with environmental factors.

Table 1. Environmental factors in a variety of agroforestry patterns on sloping land area of Semagar Duwur Village, Girimarto Subdistrict, Wonogiri District, Central Java.

Biotic

Abiotic Physical properties Chemical properties No. of Relative Air Soil Soil Site Sunlight vegetation air tempetempeSoil organic intensity types humidity rature rature pH matter (Lux) (%) (C) (C) (%) PAC I 10 4,040 48.3 28.0 25.7 5.75 3.10 PAC II 12 8,783 62.3 27.3 27.3 5.19 4.24 PAC III 9 9,303 55.7 30.7 26.3 5.62 3.85 Average 10 7,375 55.4 28.7 26.4 5.52 3.73 PAJ I 7 33,310 51.0 33.3 29.0 5.24 3.49 PAJ II 2 16,490 58.3 29.0 29.3 5.36 5.39 PAJ III 9 20,053 54.0 31.7 27.7 5.81 3.07 Average 6 23,284 54.4 31.3 28.7 5.47 3.98 PAS I 7 27,136 46.0 33.0 29.3 5.55 3.46 PAS II 5 9,430 51.3 30.7 28.0 5.29 4.62 PAS III 3 7,752 59.3 28.7 28.0 6.12 2.70 Average 5 14,773 52.2 30.8 28.4 5.65 3.59 Note for Table 1, 2 and 3: I, II, III: The name of the station (Station I, II, III); PAC: Patterns of Mixed Agroforestry, PAJ: Patterns of Teak Agroforestry, PAS: Patterns of sengon agroforestry

Table 2. The number of individuals, the number of species and diversity index of soil macrofauna at each research station Surface macrofauna Number Site No. of Diversity of individuals index species PAC I 19 7 0.787 PAC II 65 8 0.684 PAC III 41 8 0.660 Average 42 8 0710 PAJ I 13 5 0.710 PAJ II 24 5 0.524 PAJ III 1610 12 0.017 Average 549 7 0.417 PAS I 40 7 0.614 PAS II 11 5 0.711 PAS III 37 8 0.659 Average 29 7 0.661

Underground macrofauna Number Number of Diversity of individuals index species 17 11 0.858 44 22 0.903 31 14 0.899 31 16 0887 29 9 0.792 15 8 0.818 23 15 0.919 22 11 0.843 18 9 0.827 24 13 0.892 26 13 0.861 23 12 0.860

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RESULTS AND DISCUSSION

Underground macrofauna were found of 46 species which were divided into two phyla, namely annelids and arthropods. The phylum Annelida was only found in one class namely Chaetopoda. The phylum Arthropods was found consisting of five classes, namely, Insecta, Diplopoda, arachnids, Chilopoda and Malacostraca.

Wallwork (1970) explained that the Phylum Arthropoda was a group of soil animals, which generally showed the highest dominance among the organisms making up the community of soil animals. The species was 29 species of 46 species and were originated from the Insecta class. This was in accordance with the revelation of Borror et al.

(1989) that the Insecta class was the dominant animal on earth. Borror et al. (1989) stated that the ants were the most common group and were widespread in terrestrial habitats.

Ants were groups of animal which species and populations were abundant.

Dominant surface macrofauna which was found mostly come from the family Formicidae (ants). Leptomyrmex rufipes of the subfamily Dolichoderinae macrofauna was the dominant surface macrofauna, and was the most commonly found species. Dolichoderinae were predators of soft beetles, such as aphids (Shattuck 1999). Solenopsis invicta was a species of fire ants which were commonly found as pests of plants. This species was easily spread in suitable habitats (NPS 2010). Genus Ponera were distributed along Indo-Australia (Taylor 1967; Csosz and Seifert 2003).

The life activity of soil macrofauna could not be separated from the influence of environmental factors. The activity of soil organisms is generally influenced by various

Environmental factors intensely determined the structure of soil animal communities. Since, one of soil animals was soil macrofauna being part of the soil ecosystem, therefore, in studying soil animal ecology, physico-chemical soil factors were always measured (Suin 1997) (Table 1).

The intensity of sunlight received by the ecosystem was an important determinant of primary productivity, which henceforth would affect species diversity and nutrient cycling (Mokany et al. 2008). Air humidity could be affected by light intensity. Air humidity was higher when light intensity was lower (Sulandjari et al. 2005).

Temperature was influenced by the radiation of sunlight received by earth (Lakitan 2002). Sulandjari et al. (2005) stated that the lower the light intensity the lower the temperature. Fluctuation was also influenced by weather conditions, topography and soil conditions (Suin 1997).

Soil pH value could be related with soil organic matter content. Decomposition of organic matter tended to increase the acidity of the soil due to the production of organic acids (Killham 1994; Makalew 2001).

From Table 2, it can be concluded that the PAC had a positive influence on the diversity index of soil macrofauna, but the PAS and PAJ gave a different effect.

Different influences of PAS and PAJ to soil macrofauna diversity index could be due to differences in affecting environmental factors.

Supporting capacity of agroforestry patterns to the life of surface macrofauna could directly be associated with a number of vegetation types (Table 1).

PAC had highest number of vegetation types, so it could be Table 3. Dominant soil macrofauna in agroforestry of Wonogiri District, Central Java concluded that the supporting capacity of PAC to the life of surface Site Surface macrofauna Underground macrofauna macrofauna was high. More diverse PAC I Ant A (Subfamily Dolichoderinae) Phyllophaga sp. species of plant provided more food PAC II Leptomyrmex rufipes Camponotus nigriceps supply to the soil surface PAC III Leptomyrmex rufipes Byturus sp. macrofauna. On the other hand, PAJ PAJ I Solenopsis invicta Microtermes sp. had the lowest number of vegetation PAJ II Leptomyrmex rufipes Ponera sp. types, so the supporting capacity for PAJ III Leptomyrmex rufipes Oniscus sp. soil surface macrofauna life was also PAS I Solenopsis Invicta Phyllophaga sp. low. From the observation can be PAS II Ponera sp. and Allonemobius fasciatus Blatella sp. seen that the diversity index PAS III Leptomyrmex rufipes Blatella sp. macrofauna in the soil and the number of species found greater Table 4. Correlation analysis between the level diversity of soil macrofauna and value when compared with the soil environmental factors. surface macrofauna.

The study found surface Pearson correlation value macrofauna with the amount of 27 Surface Underground species from a single phylum namely Environmental factors macrofauna ID macrofauna ID Arthropods. This arthropod phylum PAC PAJ PAS PAC PAJ PAS was consisting of two classes namely The intensity of sunlight -0.996 0.551 -0.801 0.986 -0.493 -0.836 Insecta and Arachnids. Insecta class Air relative humidity -0.787 -0.159 0.359 0.916 0.092 0.415 was found consisting of six order of Temperatures -0.480 0.115 -0.500 0.248 -0.048 -0.552 Hymenoptera, Coleoptera, Lepidoptera, Soil temperature -0.667 0.905 -0.845 0.831 -0.932 -0.876 Soil pH 0.587 -0.588 -0.421 -0.770 0.641 -0.362 Hemiptera, Blatodea, and Orthoptera.

Soil organic matter -0.762 0.419 0.541 0.899 -0.479 0.488 Arachnids class was found consisting No. of vegetation types -0.017 -0.516 -0.465 0.264 0.573 -0.518 of only one order namely Araneae.

PERTIKA et al. – Diversity of macrofauna on sloping land agroforestry

factors, including climate (rainfall, temperature etc.), soil (acidity, moisture, temperature, nutrients etc.) and vegetation (forests, grasslands, shrubs and other) (Hakim et al. 1986).

The analysis showed that the Pearson correlation value between soil macrofauna diversity indices with abiotic environmental factors ranging from 0.017 to 0.996. Pearson correlation values were positive and some were negative.

Correlation coefficient (r) could be translated in several levels, namely: a) r = 0, no correlation; b) 0 <r ≤ 0.200, the correlation is very low / very weak; c) 0.200 <r ≤ 0.400, the correlation is low / weak but certain; d) 0.400 <r ≤ 0.700, significant correlation; e) 0.700 <r ≤ 0.900, the correlation is very high, robust; f) 0.900 <r ≤ 1, the correlation is very high, very robust, reliable (Hasan 2001 ).

The increase of light intensity could decrease the soil macrofauna diversity index and vice versa. The upswing of light intensity might result in some underground macrofauna to be dead due to the underground environmental conditions that was too hot. The intensity of sunlight was also affected by canopy closure. The thick canopy allowing sunlight to reach the ground floor reduced, and vice versa (Sanjaya 2009; Sitompul 2002).

Mokany et al. (2008) stated that the intensity of sunlight affect species diversity. Suhardjono (1998) stated that research in the Bogor Botanical Gardens show there is more animal on the forest floor with less sunlight than the one with much sunlight.

Air relative humidity will decrease the diversity of surface macrofauna index. It agrees with the statement of Purwanti (2003) that the increase in air humidity can interfere the oxygen uptake (respiration) of surface macrofauna. The disruption of the process led to the decrease of soil macrofauna diversity. It might be because of the unability of soil macrofauna to survive or to migrate to another location.

The increase of relative air humidity will increase diversity index of soil macrofauna and vice versa. It is in accordance with the results of research conducted by Sugiyarto (2000) regarding the diversity of soil macrofauna at various age of sengon stands at Forest Police Resort (RPH) Jatirejo, Kediri. It shows the same thing that there is a positive correlation between the relative air humidity with soil macrofauna. The correlation between two variables is 0.04 for surface macrofauna and 0.05 for underground macrofauna.

An increase of air temperatures will reduce soil macrofauna diversity index. Lakitan (2002) stated that air temperature was affected by radiation of sunlight received by the earth. The higher the light intensity is, the higher the air temperature (Sulandjari et al. 2005). Temperature which is too high would cause some physiological processes, such as reproductive activity, metabolism, and respiration, to be disrupted (Kevan 1962; Sugiyarto 2007). The disruption of physiological processes of soil macrofauna will then affect the diversity.

An increase in soil temperatures will lower soil macrofauna diversity index and vice versa. Soil temperatures which are too high would cause some physiological processes such as reproductive activity,

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metabolism, and respiration, to be disrupted (Kevan 1962; Sugiyarto 2007). Disruption of physiological processes of soil macrofauna would then affect diversity. It is in accordance with the research of Handayani (2008) regarding the inventory diversity of soil macrofauna in carrot crop (Daucus carota L.) which was fertilized with various organic and inorganic fertilizers. It showed soil temperature having negative correlation with the diversity of soil macrofauna particularly on Coleoptera order.

An increase in acidity would increase the diversity index of soil macrofauna and vice versa. High number of soil acidity means having a low pH (pH below 7). In tropical environments where some soil has been sour for a long period of time, soil fauna have evolved its tolerance to low pH. Most of the macrofauna including diggers species such as worms and termites tend to decline its abundance in large amounts in acidic soil conditions, with most activities are limited to layers of waste where the pH is significantly higher and usually alkaline (DPI 2010).

An increase of soil organic matter would increase the diversity index of soil macrofauna and vice versa. Soil macrofauna improves decomposition of organic residues, although its role depends on the nature of the material in it (Karanja et al. 2006). The more the organic material available the bigger the number of individuals of soil macrofauna, because it is able to protect against environmental stresses both the high temperature environment and the possible presence of predators (Sugiyarto 2007). TSK (2008) states that soil macrofauna take nutrients from the soil organic matter, so the availability of adequate soil organic matter will affect the survival of soil macrofauna.

CONCLUSION Based on research results, it can be concluded that various patterns of sloping land agroforestry had a different index of soil macrofauna diversity. There was a correlation between index diversity of soil macrofauna with environmental factors in various patterns of sloping land agroforestry.

REFERENCES Bennett MT. 2008. China's sloping land conversion program: Institutional innovation or business as usual? Ecol Econ 65 (4): 699-711 Borror DJ, Triplehorn CA, Johnson NF. 1989. An introduction to the study of insects. 6th ed. Saunders, Philadelphia.

BPS. 2010. Wonogiri in Figure 2010. Central Bureau of Statistics, Wonogiri. [Indonesia] Craswell E, Sajjapongse A, Howlett D, Dowling A. 1997. Agroforestry in the management of sloping lands in Asia and the Pacific. Agrofor Syst 38 (1-3): 121-137.

Csosz S, Seifert B. 2003. Ponera testacea Emery, 1895 Stat. N.-A sister species of P. coarctata (Latreille, 1802) (Hymenoptera, Formicidae).

Acta Zool Hung 49 (3): 201-214.

Damanik RIM. 2003. Technology of agroforestry in dryland (West Nusa Tenggara Province). Study Program of Forest Science, Faculty of Agriculture, University of North Sumatra, Medan. [Indonesia] DPI [Department of Primary Industries]. 2010. Major affects of soil acidification Chapter 4. Department of Primary Industries, Victoria, Australia. www.dpi.vic.gov.au

144

B I O D I V E R S IT A S 13 (3): 140-144, July 2012

Grosjean P, Kontoleon A. 2009. How sustainable are sustainable development programs? The case of the Sloping Land Conversion Program in China. World Dev 37 (1): 268-285 Hakim N, Nyakpa MY, Lubis AM, Nugroho SG, Dika MA, Go BH, Bailley HH. 1986. Principles of soil science. University of Lampung, Bandar Lampung.

Handayani P. 2008. Diversity inventory of soil macrofauna at carrots cultivation (Daucus carota L.) with different organic and inorganic fertilizers. [Hon. Thesis]. Study Program of Soil Science. Faculty of Agriculture, Sebelas Maret University, Surakarta. [Indonesia] Hasan I. 2001. Principles of statistics matter 2 (Inferensif Statistics). 2nd ed. Bumi Aksara. Jakarta. [Indonesia] Karanja NK, Ayuke FO, Swift MJ. 2006. Organic resources quality and soil fauna: Their role on the microbial biomass, decomposition and nutrient release patterns in Kenyan soils. Trop Subtrop Agroecosyst 6: 73-86.

Kartasubrata J. 1991. Planning and implementation aspects based on some successful agroforestry projects in Indonesia. In: Mellink W YS Rao, MacDicken KG (eds) Agroforestry in Asia and the Pacific. RAPA Publication 1991/5. Bangkok, FAO Regional Office for Asia and the Pacific and Winrock International Institute for Agricultural Development.

Kevan DKM. 1962. Soil animals. HE & G Witherby, London.

Killham K. 1994. Soil ecology. Cambridge University Press. Cambridge.

Lakitan B. 2002. Principles of climatology. Raja Grafindo Persada, Jakarta. [Indonesia] Lavelle P, Dangerfield MT, Fragoso C et al.. 1994. The relationship between soil macrofauna and tropical soil fertility. In Woomer PL, Swift MJ (eds). The biological management of tropical soil fertility.

John Wiley and Sons, Chichester.

Maftuâ&#x20AC;&#x2122;ah E, M Alwi, Willis M. 2005. Potential of soil macrofauna as bioindicator of peat land quality. Bioscientiae 2 (1):1-14.

Makalew ADN. 2001 Diversity of soil biota on non tillage agroecosystems. School of Graduates, Bogor Agricultural University, Bogor. [Indonesia] Mokany A, Wood JT, Cunningham SA. 2008. Effect of shade and shading history on species abundances and ecosystem processes in temporary ponds. Freshwater Biol 53 (10): 1917-1928.

NPS [National Park Service]. 2010. Integrated pest management manual (fireants). National Park Service, US Department of the Interior, Washington DC. www.nature.nps.gov/biology/ipm/manual/fireants.cfm.

Purwanti. 2003. Diversity of soil macrofauna in different types and combination of intercropping crop under sengon stand (Paraserianthes falcataria (L.) Nielson) in Forest Police Station (RPH) Jatirejo Kediri, East Java. [Honorary Thesis]. Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University Surakarta. [Indonesia] Rahmawaty. 2004. Study on soil mesofauna diversity in Forest Nature Recreation Sibolangit (Sibolangit Village, Sibolangit Subdistrict, Deli

Serdang District, North Sumatra Province). [Honorary Thesis]. Study Program of Forest Management, Department of Forestry, Faculty of Agriculture, University of North Sumatra, Medan. [Indonesia] Sanjaya A. 2009. Soil macrofauna diversity in different stands types at Alas Kethu, Wonogiri District, Central Java. [Honorary Thesis].

Department of Biology, Faculty of Mathematics and Natural Sciences, Sebelas Maret University Surakarta. [Indonesia] Setyawan A, Wilopo W Suparno, Supriyanto. 2006. Know your natural disaster of landslide and its mitigation. Inovasi Online 8 (Nov. 2006). http://io.ppi-jepang.org/article.php?id=196. [Indonesia] Shattuck S. 1999. Australian ants: Their biology and identification.

CSIRO Publishing. Melbourne.

Sitompul SM. 2002. Radiaion in agroforestry system. In: Hairiah K, Utami SR, Lusiana B (eds) WaNuLCAS, Simulation model for agroforestry systems. International Centre for Research in Agroforestry, Southeast Asian Regional Research Programme, Bogor, Indonesia. [Indonesia] Soedjoko SA. 2002.

Management of land resources. www.mayong.staff.ugm.ac.id. [Indonesia] Sugiyarto. 2000. Diversity of soil macrofauna in different sengon age stands at RPH Jatirejo, Kediri District. Biodiversitas 1 (2): 47-54.

[Indonesia].

Sugiyarto. 2008. Soil macrofauna conservation in agroforestry systems.

National Seminar on Education Biology: Quality Improvement of Biological Learning through Applied Biology Enrichment Materials.

Study Program and Alumni Association of Bioscience, Graduate Program, Sebelas Maret University Surakarta, 24 Mei 2008.

[Indonesia] Sugiyarto, Efendi M, Mahajoeno E, Sugito Y, Handayanto E, Agustina L. 2007. Preference of soil macrofauna to crops residue at different light intensity. Biodiversitas 7(4): 96-100. [Indonesia] Suhardjono YR. 1998. Litter insects: taxon diversity and their role in the Bogor Botanical Gardens. J Biota 3 (1): 16-24. [Indonesia] Suin N M. 1997 Soil animals ecology. Bumi Aksara. Jakarta. [Indonesia] Sulandjari, Pramono S, Wisnubroto S, Indradewa D. 2005. Microclimate relationship with the growth and yield of sarpagandha (Rauvolfia serpentina Benth.). Agrosains 7 (2): 71-76. [Indonesia] Taylor RW. 1967. A monographic revision of the ant genus Ponera Latreille (Hymenoptera: Formicidae). Pac Insect Monogr 13: 1-112.

TSK [Tim Sintesis Kebijakan]. 2008. Utilization of soil biota for sustainable agriculture productivity on acid dryland. Pengembangan Inovasi Pertanian 1 (2): 157-163. [Indonesia] Wallwork JB. 1970. Ecology of soil animals. Mc Graw-Hill. London.

Wongso SA. 2008. Agroforestry, anticipation of erosion and landslides. www.perumperhutani.com [Indonesia] Xu J, Tao R, Xu Z, Bennett MT. 2010. Chinaâ&#x20AC;&#x2122;s Sloping Land Conversion Program: Does expansion equal success? Land Econ 86 (2): 219-244

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 145-150

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130308

Fish biodiversity in coral reefs and lagoon at the Maratua Island, East Kalimantan HAWIS H. MADDUPPA1,♥, SYAMSUL B. AGUS1, AULIA R. FARHAN2, DEDE SUHENDRA3, BEGINER SUBHAN1 1

Department of Marine Science and Technology, Faculty of Fisheries and Marine Sciences, Bogor Agricultural University. Jl. Agatis No.1, Bogor 16680, West Java, Indonesia. Tel./Fax +62 251 8623644, email: hawis@ipb.ac.id 2 Agency for Marine and Fisheries Research and Development, Ministry of Marine Affairs and Fisheries, Republic of Indonesia. Kompleks Bina Samudera, Jl. Pasir Putih I, East Ancol, North Jakarta 14430, Indonesia 3 Fish Quarantine Centre Class II, Tanjung Priok. Jl. Enggano Raya No. 16 Tanjung Priok, Jakarta 14320, Indonesia Manuscript received: 10 June 2012. Revision accepted: 22 July 2012.

ABSTRACT Madduppa HH, Agus SB, Farhan AR, Suhendra D, Subhan B. 2012. Fish biodiversity in coral reefs and lagoon at the Maratua Island, East Kalimantan. Biodiversitas 13: 145-150. Fishes are one of the most important biotic components in the aquatic environment. They are filling different habitats, including coral reef and lagoon. This study aims to (i) assess biodiversity in coral reef and lagoon in Maratua Island, East Kalimantan, and (ii) compare the fish community indices (Shannon-Wiener diversity, Evenness, and Dominance) between the coral reef and lagoon. A total of 159 fish species of belonging to 30 families were observed during five visual census of the study period. The number of species on coral reefs is higher (121 species) than in the lagoons (47 species). Relative abundance (%) of each species also varied and did not form a specific pattern. However, a clear cluster between the coral reef and lagoon habitats from fish relative abundance based on multivariate analysis and dendogram Bray-Curtis Similarity was revealed. The Evenness index value (E) ranged from 0.814 to 0.874, the dominance index (C) ranged from 0.023 to 0.184, and the Shannon-Wiener diversity index (ln base, H') ranged from 1.890 to 4.133. Fish biodiversity in coral reefs was higher (H'= 3.290±0.301) than in the lagoon (H' = 2.495±0.578).

Key words: diversity index, Maratua Island, lagoon, fish visual census, reef fishes

INTRODUCTION Fishes are one of the most important biotic components in the aquatic environment. They fill a very specific habitat by meeting a variety of waters substratum. Several studies have mentioned the importance of glittering fish communities in ecosystem processes through trophic relationships with other biotic components (e.g. Carrasson and Cartes 2002). Coral reefs are used by fishes as a territorial (Robertson et al. 1976), a feeding ground (Reese 1981), as a place to hide (Hixon 1991), and as a reproduction and spawning ground (Wootton 1992). Fish reach their high biodiversity in coral reef ecosystem (Allen and Werner 2002), especially in Indonesia (Allen and Adrim 2003). Indonesian coral reefs harbour more than 2000 fish species (Allen and Adrim 2003). Consequently, Indonesia has been declared as the centre of marine biodiversity (Allen 2008; Allen and Werner 2002).

Lagoon ecosystem is one of shallow water ecosystems that are separated by barrier islands or coral from larger water systems, which is characterized by predominant sand substratum (Hutomo and Moosa 2005). Sedberry and Carter (1993) states that due to differences in characteristics of the substrate between coral reef and lagoon area, the fish community will be different in terms of composition, relative abundance and biomass. In addition, lagoon surrounded by a coral reef may have

similar to the biota of coral reef ecosystem, because corals may grow on hard substrate at its leeside with a variety of coral lifeforms (Hutomo and Moosa 2005).

Maratua Island, one of the islands in Derawan Islands, has a land area of 384.36 km2 and sea area of 3735.18 km2, which is fringed with coral reefs, lagoon and uplifted atoll (Tomascik et al. 1997). Geographically the island is situated on a peninsula north of Berau seas (02°15'12" N and 118°38'41" E). Maratua’s climatic conditions in the region are affected by the rainy season (October-May) and dry season (July to September). Oceanographic factors influenced the seasonal movement of currents and Indonesian Trough Flow (Arlindo) from the Pacific to the Indian Ocean through the Straits of Makassar (Wyrtki 1961). These conditions create an environment that supports a high biodiversity of marine organisms, especially in fish.

However, documentations of the biodiversity of marine organisms such as fish in Maratua Island are poorly known.

The island’s coral reefs are threatened by many anthropogenic pressures like other Indonesian reef system that potentially lead many organisms to extinction.

Therefore, this study aims to (i) assess fish biodiversity at the coral reefs and lagoons in Maratua Island, East Kalimantan, and (ii) compare the fish community indices between the reef and lagoon.

B I O D I V E R S IT A S 13 (3): 145-150, July 2012

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MATERIAL AND METHODS Study sites and periods The reef fish communities on the island of Maratua were observed at the two stations on the reef slope (Maratua 1, and Maratua 2) and three stations for the lagoon (Tanjung Duwata, Karang Bentukan, and Buar) (Figure 1). The study was conducted on 27-28 November 2005 and November 13 to 14, 2006 (Table 1).

Table 1. Information on data collection for each site Sites Coral reefs: Maratua 1 Maratua 2 Lagoons: Tanjung Duwata Karang Bentukan Buar

Geographical position N E

Study period

2° 16' 51.806" 118° 33' 45.003" 13-11-2006 2° 15' 42.705" 118° 33' 26.999" 14-11-2006 2° 9' 50.809" 118° 38' 45.014" 27-11-2005 2° 7' 8.309" 118° 42' 23.888" 27-11-2005 2° 12' 14.387" 118° 37' 12.515" 28-11-2005

Data collection The location of the study sites were determined initially by snorkeling and observing the coral reef conditions and representative areas (3-7 meters depth). Data obtained by the method of fish visual census along 50 m transect line (English et al. 1997). A total of two transects were laid on coral reefs, and three transects on lagoon. The fish transect lines are straight and follow the depth contour. The basic unit of data collection for the fish visual census was 50 m x 5 m (250 m2). The procedure was to wait for at least 10 minutes before surveying reef fish species (Halford and Thompson 1994), and the approximate time of fish census was up to 60 minutes for each transect. The SCUBA observer swims slowly along transect and recording fish encountered within 2.5 m on both side and 5 m above transect. After data collection, reef fish identification was confirmed by using several fish identification books, i.e.

Allen (2000) and Lieske and Myers (2001).

Figure 1. Map of study sites: the reef slope (station 1 and 2) and the lagoon (stations 3, 4 and 5) on the island Maratua within Derawan Islands, East Kalimantan. Insert: Maratua Island in relation to Indonesia indicated by a red box.

MADDUPPA et al. – Fish biodiversity of Maratua Island, East Kalimantan

Data analysis Shannon-Wiener Diversity Index (H’; ln basis), Evenness Index (E), and Simpson Dominance Index (C) (Odum 1971; Magurran 1988; Ludwig and Reynolds 1988; Smith 2002), as well as species richness and their relative abundance were compared among sites. In addition, functional categories (target, major and indicator) were analyzed. The target fish is individual fish that has economic value and food resources for fishermen. This group is also known as the economically important fish or fish consumption. The indicator fish is group of fish that determines the health of coral reefs. This is caused by the strong relationship between the fish and coral, such as Family Chaetodontidae. Member of chaetodonthids used coral reefs as a source of food. The major fish is fish groups that are not included in the targets and indicators.

This group is generally found in large quantities and many of them are traded as marine ornamental fish. Non-metric Multidimensional Scaling (MDS) and similarity dendogram were used to visualize the differences in fish communities of the two different habitats (coral reefs and lagoons) (Shepard 1962; Kruskal 1964) by using PRIMER5 software (Clarke and Gorley 2001). MDS was based on Bray-Curtis similarities. The quality of the MDS plot is indicated by the stress value. Values <0.2 give a potentially useful 2-dimensional picture, stress <0.1 corresponds to a good ordination and stress <0.05 gives an excellent representation (Kruskal 1964; Field et al. 1982; Clarke 1993). This approach has been widely applied to multivariate analysis of the various communities (Field et al. 1982; Clarke and Green 1988).

RESULTS AND DISCUSSION Fish community structure A total of 2145 individuals from 160 species of fish belonging into 30 families were observed in this study (Table 2). The number of species on coral reefs was higher (121 species) than in the lagoons (47 species). List of families that have three top species composition in coral reef areas were Pomacentridae, Labridae and Acanthuridae, while in the lagoon were Pomacentridae, Chaetodontidae, and Nemipteridae (Figure 2). These families are also mainly observed on the other coral reefs around Indonesia (Estradivari et al. 2007; Ferse 2008).

Fish abundance in the coral reef was higher (690±163 individual/250 m2) than in the lagoon (255±92 individual/250 m2), and relative abundance (%) of each species also varied over study sites (Table 2). A clear difference between the reef and lagoon habitats based on MDS and dendogram was revealed (Figure 3). This is similar with other study that showed differences in fish community based on the abundance of species from the results of the multivariate analysis (Madduppa et al. 2012).

The presence of reef fish in the waters depends on coral health indicated by the percentage of live coral cover. It is very possible because of the live reef fish associated with the shape and type of coral as a shelter, protection and

147

places to look for food (Madduppa 2006). Besides the health of coral, reef structure complexities have enriched reef fishes (Nybakken 1992).

Reef fish communities observed in this study were grouped into three functional categories (Table 2). The target fish families were observed in the study sites as follows: Haemulidae, Nemipteridae, Serranidae and Pomacanthidae. The target fish in the coral reef stations (40±8 species/250 m2) were greater than in the lagoons (7±2 species/250 m2). The presence of the target fish in the coral reef ecosystem are due to searching for food (feeding ground) or spawning and nursery. In the Philippines, many coral reef fishes are caught for the small-scale fisheries including surgeonfish, groupers, and snappers (Amar et al.

1996).

A total of 16 species of fish were found in all indicators of the study sites. Groups of fish indicators also showed a similar pattern in the area where the coral reef was higher (7±1 species/250 m2) than in the lagoon (3±2 species/250 m2). Due to their greatest biodiversity in coral reef ecosystems (Allen and Werner 2002) and their interdependence to the health of coral reef ecosystem (Hourigan et al. 1988), chaetodontid has been considered as a reliable way to indirectly assess the changes of a coral reef and monitor it through time (Tanner et al. 1994; Markert et al. 2003). Many members of Chaetodontidae are highly dependent on live coral polyps, and multiple studies have proven that they are corallivorous (e.g. HarmelinVivien and Bouchon-Navaro 1983; Pratchett 2005; Reese 1981; Birkeland and Neudecker 1981; Alwany et al. 2003; Madduppa 2006).

The major groups of fish on the coral reefs stations were higher (39±7 species/250 m2) than in the lagoon (13±4 species/250 m2). Function and role of the fish were not yet clear but could be as one link in the ecological system and food webs in coral reef ecosystems. Major fish species are found mainly from the family Pomacentridae.

Fish biodiversity The Evenness index (E) value ranging from 0814 to 0874, the dominance index (C) 0023-0184 range, and the value of diversity index (H ') ranged from 1.890-4.133.

Fish communities in reef areas were more diverse (H '= 3290±0301) than in the lagoon (H' = 2495±0578) (Figure 4). The value of diversity can indicate the level of stress or pressure received by the species from the environment (Lardicci et al. 1997). In the lagoon, reef fish communities in the lagoon which is located outside near reef edge (Station 3, see Figure 1) was higher in comparison with two other lagoon stations which are located in the inside (Station 4 and 5). This is similar to Hutomo and Moosa (2005) that fish communities in the lagoon near coral reefs is highly influenced by the benthic community (including corals).

Diversity of species of reef fish have a close relationship with the characteristics of the substrate in the area, such as the existence of the herbivorous fishes of the family Scaridae, because of dead coral covered with macroalgae (Madduppa et al. 2012). The fish will tend to

B I O D I V E R S IT A S 13 (3): 145-150, July 2012

148

Number of Species

30

Table 2. List of taxa, number of individual (±SE individual/250 m2), species richness (±SE species/250 m2), fish categorical function (±SE species/250 m2, T=Target, M=Major, I=Indicator), and relative abundance (%) for each species at each site (1 and 2 = reef slope, 3-5 = lagoon).

Lagoon

25

Coral reefs

20 15 10 5

Acanthuridae Apogonidae Balistidae Belonidae Blenniidae Caesionidae Carangidae Chaetodontidae Cirrhitidae Diploprionini Ephippidae Gobidae Haemulidae Holocentridae Labridae Lethrinidae Lutjanidae Microdesmidae Mullidae Nemipteridae Platycephalidae Plesiopidae Pomacanthidae Pomacentridae Scaridae Serranidae Siganidae Tetraodontidae Theraponidae Zanclidae

0

Family

Figure 2. Comparison of the number of species of fish from each family were found among the coral reefs and lagoons

Figure 3. Multivariate analysis: (left) Bray-Curtis Similarity and (right) non-metric MDS (multidimensional scaling) of the fish community based on the relative abundance of each species per station on coral reefs (1 and 2) and in the lagoon (3, 4 and 5)

4,5 Lagoon

Community index

4

Coral reef

3,5 3 2,5 2 1,5 1 0,5 0 H'

E

C

Figure 4. Comparison of reef fish community index values between the lagoon and reef slope are seen from the value of Shannon-Wiener diversity (H '), Evenness (E), and Dominance (C)

cluster in certain forms of corals and generally have limited movement compared to other invertebrates that have the same size cite. Because the reef fish communities have a close relationship with the coral reefs as a habitat, so that if a high percentage of dead coral will cause a significant decrease in the number of fish species and individuals associated with coral reefs cite. The current study observed that some fishes found in a specific habitat, but many species were found in more than one habitat. Generally each species has specific habitat preferences (Hutomo 1986), and they have two different interaction mode in coral reefs: direct interaction (e.g. as a refuge from predators or prey, interactions in search of food the relationship between corals and fish that live on the reef biota, including algae) and indirect interaction due to reef structure and hydrology and sediment (Choat and Bellwood 1991).

Taxa (family/species) Acanthuridae Acanthurus auranticavus Acanthurus gahhm Acanthurus leucocheilus Acanthurus leucosternon Acanthurus lineatus Acanthurus thompsoni Naso annulatus Naso brevirostris Naso lituratus Naso sp.

Paracanthurus hepatus Zebrasoma scopas Apogonidae Apogon cavitienis Apogon cookie Apogon doederleini Apogon kauderni Sphaeramia nematoptera Balistidae Balistapus undulatus Balistoides viridescens Odonus niger Pseudobalistes flavimarginatus Pseudobalistes fuscus Belonidae Strongylura incisa Meiacanthus vittatus Caesionidae Caesio cuning Caesio lunaris Caesio teres Caesio xanthonota Carangidae Gnathanodon speciosus Chaetodontidae Chaetodon auriga Chaetodon collare Chaetodon collare Chaetodon decussatus Chaetodon decussatus Chaetodon ephippium Chaetodon guttatissimus Chaetodon melapterus Chaetodon meyeri Chaetodon rafflesi Chaetodon trifascialis Chaetodon trifasciatus Chaetodon vagabundus Chelmon rostratus Hemitaurichthys polylepis Parachaetodon ocellatus Cirrhitidae Paracirrhites arcatus Paracirrhites forsteri Paracirrhites nisus Diploprionidae Diploprion bifasciatum Platax orbicularis

Cate- Relative abundance (%) gory 1 2 3 4 5 T T T T T T T T T T T T

1.3 0.8 0.8 0.1 0.8 0.2 0.4 2.2

2.3 1.1 0.4 2.1 0.2 0.2 - 1.9 6.4 - 11.3 2.1 -

-

M M M M M

- 21.3 - 5.8 2.7 4.0 - 1.3 0.5 0.8 -

-

M M M M M

- 0.2 0.1 - 3.0 - 5.1 1.6 0.2 -

-

-

M M

1.1 0.1

0.3

-

-

T T T T

0.1 0.2 0.1 0.2 0.1 - 0.3

-

-

-

-

-

T

-

0.2

I I I I I I I I I I I I I I I I

0.4 0.2 0.6 0.1 0.2 0.1 1.3 -

0.2 0.2 0.6 0.2 2.3 0.4 -

M M M

-

-

M M

- 0.6 4.1 -

-

0.6 0.8 0.3 0.9 - 0.6 0.8 2.2 0.6 0.3 12.9 -

-

- 9.7 6.3 2.7 31.5 - 2.4 -

-

-

MADDUPPA et al. – Fish biodiversity of Maratua Island, East Kalimantan Platax pinnatus Platax teira Gobiidae Gobi sp.

Istigobius nigroocellatus Haemulidae Plectorhinchus sordidus Plectorhinchus flavomaculatus Plectorhinchus chaetodonoides Holocentridae Myripristis amaena Sargocentron caudimaculatus Labridae Bodianus mesothorax Cheilinus fasciatus Cheilinus oxycephalus Cheilinus trilobatus Choerodon zamboanga Choerodon anchorago Gomphosus caeruleus Gomphosus varius Halichoeres hortulanus Halichoeres melanurus Halichoeres ornatissimus Halichoeres purpurascens Labroides dimidiatus Thalassoma grammaticum Thalassoma lunare Lethrinidae Gymnocranius griseus Monotaxis grandoculis Lutjanidae Lutjanus decussatus Lutjanus ehrenbergii Lutjanus fulviflamma Lutjanus kasmira Lutjanus quinquelineatus Lutjanus semicinctus Lutjanus vitta Microdesmidae Ptereleotris evides Mullidae Parupeneus bifasciatus Upeneus sp.

Upeneus vittatus Nemipteridae Parascolopsis eriomma Pentapodus caninus Pentapodus trivittatus Scolopsis bilineatus Scolopsis ghanam Scolopsis lineatus Scolopsis margaritifer Scolopsis trilineatus Platycephalidae Thysanophrys arenicola Plesiopidae Calloplesiops altivelis Pomacanthidae Centropyge eibli Centropyge multispinis Centropyge nox Pygoplites diacanthus Pomacentridae Abudefduf sexfasciatus Abudefduf vaigiensis Amblyglyphidodon curacao

M M

0.4 0.1

-

-

-

-

M M

1.1 -

-

1.4

-

2.7

T T T

0.2 0.7 0.2 7.8

-

-

-

T T

1.5 0.4

-

3.3 -

-

T T T T T T T T T T T T M M M

0.1 0.2 0.1 0.1 0.1 0.1 0.8 1.6 0.8 0.5 1.4 0.8 2.9

0.4 0.6 0.9 0.2 0.2 - 1.9 0.9 - 0.8 4.2 -

-

T T

0.1 0.2 0.1 - 0.3

-

-

T T T T T T T

2.6 1.9 0.1 -

0.2 0.4 0.9 0.2 0.2

-

-

-

M

0.1 0.4

-

-

-

M M M

- 4.4 4.0 0.6 0.4 0.5 0.4 -

T T T T T T T T

- 1.1 1.5 9.6 - 7.2 0.6 1.5 4.7 - 3.3 16.4 - 0.6 0.1 2.1 2.5 1.6 -

M

5.6

-

-

-

-

M

-

0.2

-

-

-

M M M T

5.5 4.0 6.3 8.2 3.4 3.6 3.0

-

-

-

M M M

- 2.5 21.3 0.9 - 2.5 4.6 6.8 0.5 2.1 -

-

Amphiprion ocellaris Amphiprion sandaracinos Chrysiptera unimaculata Chromis analis Chromis atripectoralis Chromis delta Chromis dimidiate Chromis nitida Chromis verater Chromis viridis Chrysiptera cyanea Chrysiptera hemicyanea Chrysiptera rollandi Chrysiptera springeri Dascyllus aruanus Dascyllus carneus Dascyllus melanurus Dascyllus trimaculatus Dischistodus melanotus Dischistodus perspicillatus Dischistodus prosopotaenia Hemiglyphidodon plagiometopon Neopomacentrus azysron Neopomacentrus cyanomos Neopomacentrus xanthurus Plectroglyphidodon lacrymatus Pomacentrus alexandrae Pomacentrus amboinensis Pomacentrus brachialis Pomacentrus lepidogenys Pomacentrus milleri (juv.)

Pomacentrus nigromarginatus Pomacentrus tripunctatus Stegastes aureus Scaridae Chlorurus microrhinos Scarus globiceps Scarus melanurus Scarus niger Scarus rivulatus Scarus rubroviolaceus Scarus sp.

Serranidae Cephalopholis cyanostigma Epinephelus fasciatus Epinephelus spilotoceps Plectropomus leopardus Pseudanthias bimaculatus Pseudanthias pleurotaenia Pseudanthias randalli Pseudanthias squamipinnis Pseudanthias tuka Siganidae Siganus puellus Siganus virgatus Tetraodontidae Canthigaster compressa Theraponidae Terapon jarbua Zanclidae Zanclus cornutus Number of individual Species richness Target (T) Major (M) Indicator (I)

149 M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M M

0.4 0.1 0.4 0.2 0.6 0.2 0.2 0.4 0.9 0.1 0.1 0.1 2.2 0.4 0.1 0.2 0.5 0.4 0.7 1.3 1.4

0.2 1.3 0.2 0.2 0.2 1.7 4.7 1.7 1.3 0.4 2.1 0.6 0.4 5.5

T T T T T T T

0.1 1.2 2.7 - 1.7 0.5 3.4 0.1 0.2 -

T T T T T T T T T

10.2 0.6 1.4 1.9 1.7 2.8 8.5 0.6 -

0.3

-

- 4.6 1.1 - 0.3 0.4 1.9 2.6 0.2 1.6 1.1 0.4 -

-

17.8 -

T T

- 0.3 0.1 1.9 -

-

-

T

0.2 0.2

-

-

-

T

0.8 0.4

-

-

-

M

2.5 0.2 2.5 2.1 690 86 40 39 7

±163 ±16 ±8 ±7 ±1

-

- 9.6 - 1.4 1.2 1.5 4.1 1.2 2.1 0.3 7.0 -

255 22 7 13 3

±92 ±7 ±2 ±4 ±2

-

B I O D I V E R S IT A S 13 (3): 145-150, July 2012

150 CONCLUSION

The study observed that the diversity of fish species in the reef slope was higher than in the lagoon. This shows that the characteristics of the habitat were instrumental in shaping the fish community. Habitats in a tropical lagoon are an important area for various fish species of coral reef ecosystems cite. Despite the complexity of the lagoon habitat characteristics are not as high in coral reef ecosystems, but also provides a location for a few juvenile fish to grow and evolve, which is also vital for the conservation of species. Therefore, management strategies and conservation of coral reef ecosystems should include a lagoon habitat as an important and integrated part.

ACKNOWLEDGEMENTS The research is part of the project ‘Action plan for the outer islands of East Kalimantan’ by BAPPEDA East Kalimantan Province year 2006 and ‘Inventory of Lagoon ecosystem’ by the Ministry of Marine Affairs and Fisheries year 2005.

REFERENCES Allen GR. 2000. Marine fishes of South-East Asia. Periplus. Singapore.

Allen GR. 2008. Conservation hotspots of biodiversity and endemism for Indo-Pacific coral reef fishes. Aquatic Conservation: Mar Freshw Ecosyst 18: 541-556.

Allen GR, Adrim M. 2003. Coral reef fishes of Indonesia. Zool Stud 42: 1-72.

Allen GR, Werner TB. 2002. Coral reef fish assessment in the ‘Coral Triangle’ of Southeastern Asia. Environ Biol Fish 65: 209-214 Alwany M, Thaler E, Stachowitsch M. 2003. Food selection in two coralivorous butterfly fishes, Chaetodon austriacus and C. trifascialis, in the Northern Red Sea. Mar Ecol 24: 165-167.

Amar EC, Cheong RMT, Cheong MVT. 1996. Small-scale fisheries of coral reefs and the need for community-based resource management in Malalison Island, Philippines. Fish Res 25: 265-277 Birkeland C, Neudecker S. 1981. Foraging behavior of two Caribbean chaetodontids: Chaetodon capistratus and C. aculeatus. Copeia 1: 169-178 Carrasson M, Cartes JE. 2002. Trophic relationships in a Mediterranean deep-sea fish community: partition of food resources, dietary overlap and connections within the benthic boundary layer. Mar Ecol Prog Ser 241: 41-55 Choat JH, Bellwood DR. 1991. Reef fishes: Their history and evolution.

In: Sale PF (ed). The ecology of the fishes on coral reefs. Academic Press. California.

Clarke KR. 1993. Non-parametric multivariate analyses of changes in community structure. Australian J Ecol 18: 117-143.

Clarke KR, Gorley NR. 2001. PRIMER v5: Unser Manual/Tutorial.

PRIMER-E Ltd., Plymouth, UK Clarke KR, Green RH. 1988. Statistical design and analysis for a 'biological effects' study. Mar Ecol Prog Ser 46: 213-226 English S, Wilkinson C, Baker V. 1997. Survey Manual for Tropical Marine Resources. 2nd ed. Australian Institute of Marine Science.

Australia (AIMS), Townsville.

Estradivari, Syahrir M, Susilo N, Yusri S, Timotius S. 2007. Jakarta coral reefs: Long-term monitoring of the Kepulauan Seribu coral reefs (2004-2005). Yayasan Terangi, Jakarta. [Indonesia] Ferse SCA. 2008. Artificial reef structures and coral transplantation: fish community responses and effects on coral recruitment in North Sulawesi/Indonesia. [Dissertation]. University of Bremen, Bremen, Germany.

Field JG, Clarke KR, Warwick RM. 1982. A practical strategy for analysing multispecies distribution patterns. Mar Ecol Prog Ser 8: 3752 Halford AR, Thompson AA. 1994. Visual Census Surveys of Reef Fish.

Long-term monitoring of the Great Barrier Reef. Australian Institute of Marine Science (AIMS), Townsville, Australia Harmelin-Vivien ML, Bouchon-Navaro Y. 1983. Feeding diets and significance of coral feeding among chaetodontid fishes in Moorea (French Polynesia). Coral Reefs 2: 119-127.

Hixon MA. 1991. Predation as a process structuring coral reef fish communities. In: Sale PF (ed). The ecology of fishes on coral reefs.

Academic Press. San Diego.

Hutomo M. 1986. Coral reef fish resources and their relation to reef condition: Some case studies in Indonesian waters. Biotrop spec. publ 19: 67-78.

Hutomo M, Moosa MK. 2005. Indonesian marine and coastal biodiversity: Present status. Indian J Mar Sci 34: 88-97.

Hourigan TF, Tricas TC, Reese ES. 1988. Coral reef fishes as indicators of environmental stress in coral reefs. In: Soule DF, Kleppel GS (Eds.), Marine Organisms as Indicators. Springer-Verlag, New York City, NY, USA, pp. 107-135 Kruskal JB. 1964. Multidimensional scaling by optimizing goodness of fit to a nonmetric hypothesis. Psychometrica 29: 1-27 Lardicci C, Rossi F, Castelli A. 1997. Analysis of macrozoobenthic community structure after severe dystrophic crises in a Mediterranean coastal lagoon. Mar Poll Bull 34: 536–547 Lieske E, Myers R. 2001. Reef fishes of the world: Indo Pacific and Carribean. (Revised edition). Periplus. Singapore.

Ludwig JA, Reynolds JF. 1988. Statistical ecology: A primer on methods and computing. John Wiley and Sons. New York.

Madduppa HH. 2006. Bioecological study on eight band butterfly fish (Chaetodon octofasciatus, Bloch 1787) to detect condition of coral reef ecosystem in East Petondan Island, Seribu Islands, Jakarta. [MSc Thesis]. Bogor Agricultural University, Bogor Madduppa HH, Ferse SCA, Aktani U, Palm HW. 2012. Seasonal trends and fish-habitat associations around Pari Island, Indonesia: setting a baseline for environmental monitoring. Environ Biol Fish. DOI 10.1007/s10641-012-0012-7 Magurran AE. 1988. Ecological diversity and its measurements. Princeton University Press. Princeton.

Markert BA, Breure AM, Zechmeister HG. 2003. Bioindicators and biomonitors: Principles, concepts, and applications. Elsevier, Amsterdam Nybakken JW. 1992. Marine biology: An ecology approach. Gramedia.

Jakarta.

Odum EP. 1971. Fundamental of ecology. 3rd ed. Toppan Co, Ltd. London.

Pratchett MS. 2005. Dietary overlap among coral-feeding butterfly fishes (Chaetodontidae) at Lizard Island, northern Great Barrier Reef. Mar Biol 148: 373-382 Reese ES. 1981. Predation on corals by fishes of the family Chaetodontidae: Implication for conservation and management of coral reef ecosystems. Bull Mar Sci 31: 594-604.

Robertson DR, Sweatman HPA, Fletcher EA, Cleland MG. 1976.

Schooling as a mechanism for circumventing the territoriality of competitors. Ecology 57: 1208-1220.

Sedberry GR, Carter J. 1993. The fish community of a shallow tropical lagoon in Belize, Central America. Estuaries 16: 198-215 Shepard R. 1962. The analysis of proximities: Multidimensional scaling with an unknown distance function. II. Psychometrika 27: 219-246 Smith EP. 2002. Ecological statistics. Vol 2. In: Abdel HE, Walter WP (eds). John Wiley and Sons, Chichester.

Tanner JE, Hughes TP, Connell JH. 1994. Species coexistence, keystone species, and succession - A sensitivity analysis. Ecology 75: 22042219 Tomascik T, Mah AJ, Nontji A, Moosa MK. 1997. The ecology of the Indonesian seas. Part Two. Periplus. Singapore.

Wootton RJ. 1992. Fish ecology (Tertiary Level Biology). Blackie and Son, New York.

Wyrtki K. 1961. Physical oceanography of Southeast Asian waters. In: NAGA report. The University of California, Scripps Institution of Oceanography. LaJolla, California.

B I O D I V E R S IT A S Volume 13, Number 3, July 2012 Pages: 151-160

ISSN: 1412-033X E-ISSN: 2085-4722 DOI: 10.13057/biodiv/d130309

Jernang rattan (Daemonorops draco) management by Anak Dalam Tribe in Jebak Village, Batanghari, Jambi Province IIK SRI SULASMI1,♥, NISYAWATI2, YOHANES PURWANTO3, SITI FATIMAH 1

Conservation Biology, Post Graduate School, Faculty of Mathematic and Natural Sciences, University of Indonesia, Depok, West Java, Indonesia, Tel. +62-21-7270013, email: iik_08@yahoo.com 2 Department of Biology, Faculty of Mathematic and Natural Sciences, University of Indonesia. UI Campus, Depok 16424, West Java, Indonesia 3 Division of Botany, Research Center for Biology, Indonesian Institute of Sciences, Cibinong-Bogor, West Java, Indonesia Manuscript received: 23 June 2012. Revision accepted: 14 July 2012.

ABSTRACT Sulasmi IS, Fatimah S, Nisyawati. 2012. Jernang rattan (Daemonorops draco) management by Anak Dalam Tribe in Jebak Batanghari, Jambi Province. Biodiversitas 13: 151-160. Management of Jernang Rattan (Daemonorops draco Willd.) in Jebak Forest, Batanghari, Jambi is not well documented. It is noted that fruit of D. draco is the best income source for Anak Dalam Jambi people since 1624. They harvest fruit of D. draco as much as they need. The Jebak forest is an open access, so all the people of Anak Dalam Jambi Tribe have the same right and responsibility on the forest. However, almost 60% of Jebak Forest area has been degraded because of illegal conversion into oil palm plantation. This is the reason why people of Suku Anak Dalam, try to cultivate D. draco by growing 40 clumps of this species in their rubber plantation. The aim of their activity is to conserve D. draco at their forest. Based on the recent situation, research study of jernang rattan management in Jebak Forest was conducted. The research method was semi structural interview. All data were analyzed descriptively. The results showed that the management and cultivation of D. draco in Jebak Forest was very difficult because the availability of seeds was not sufficient for root stocks.

Key words: cultivation, income source, Jebak Forest, management, jernang rattan.

INTRODUCTION The people belonging to Anak Dalam Tribe in Jebak Village, Batanghari District, Jambi Province are the refugees who came from South Sumatra. They came to the forest in Jambi province in 1624 because of the war between the Sultanate of Palembang and Jambi Kingdom.

The majority of the populations are moslems. Their daily language is Malay. The people of Anak Dalam Tribe in Jebak forest have yellow skin, ranging in height between 140-160 cm. Their houses are usually made of wood and thatch-roofed (Ministry of Social Affairs 1998).

The people of Anak Dalam Tribe who settle in Jebak forest make use of all forest products as a source of their livelihood. All members of the population have equal opportunity to use forest products. One of uses of jernang rattan is as a producer of red resin called dragon’s blood.

The people of Anak Dalam Tribe who work as seekers of dragon blood are between 40-75 years old. Those under the age of 40 years collect other forest products such as honey, rattan sticks, raman fruit, petai, and kabau and hunt wild bear, snakes, turtles, and birds. According to Muchlas (1975), extraction activities of dragon’s blood are done individually. Extraction of jernang rattan has been done intensively by the people since the 1600s (BKSDA Jambi 2010).

The extraction of jernang rattan in the past was the main income for most people living in forest areas in

Sumatra, especially for people in Jambi, such as the Malay people and Anak Dalam Jambi Tribe. Traditionally, the people of Anak Dalam Jambi Tribe extract dragon’s blood in the surrounding forest, and so do the people of Anak Dalam Tribe who live in Jebak village.

Demand for dragon’s blood continues to increase, causing over-exploitation of jernang rattan. This causes the increase of harvest of jernang rattan without considering its sustainability that is harvesting by cutting the stems. Beside over-exploitation, which threatens the sustainability of the jernang rattan in Batanghari, there is also encroachment and illegal logging. Therefore, the people of Anak Dalam Tribe in the village of Batanghari try hard to find strategies to manage jernang rattan for sustainability and benefit to people's lives and seek to develop and cultivate rattan in the forest of Anak Dalam Tribe in Jebak village.

One of Anak Dalam Tribe’s efforts to conserve jernang rattan is harvesting the fruits by climbing up a tree where the jernang rattan creeps its stems. The people of Anak Dalam Tribe take only the fruits and never cut the stems during harvesting the fruits. Therefore, the harvesting technique done by the Anak Dalam Tribe does not reduce the population of jernang rattan. Although harvesting rules are not institutionalized, but the local people respect and obey the unwritten regulation that has been in effect from the past until now (Winarni et al. 2004; Purwanto et al.

2009b; Soemarna 2009). Based on direct observations and specimen observation in the Herbarium Bogoriense

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B I O D I V E R S IT A S 13 (3): 151-160, July 2012

observations, there are two rattans that produce dragon’s blood in the village of Batanghari. They are jernang rattan (Daemonorops draco) and kelemunting rattan (Daemonorops didymophylla). The jernang rattan can be processed to make modern medicine and high-quality dye (Soemarna and Anwar 1994; Purwanto et al. 2009b).

Development of jernang rattan is very important as one of alternatives to increase the income of Malay people in Tanjung Jabung Barat (Purwanto et al. 2009b; BKSDA Jambi 2010), Anak Dalam Tribe in Sarolangun district, and Anak Dalam Jambi Tribe in Batanghari forest area Jambi (BKSDA Jambi 2010).

The development of jernang rattan can be done by cultivating Daemonorops draco in its natural habitat, especially in the area near the river (Purwanto et al. 2009a).

This species should be chosen as one of the leading crops in agroforestry systems in forest areas of Anak Dalam Jambi Tribe in Jebak village and trade system of dragon’s blood should be regulated by shortening the jernang rattan marketing chain, so there is no monopoly by a producer (Purwanto et al. 2009b; BKSDA Jambi 2010). Jernang rattan cultivation in its natural habitat in the forests area of Anak Dalam Jambi Tribe has some benefits including conservation of biodiversity and reducing the excessive exploitation of nature. The presence of jernang rattan species in the forest will prevent conversion of forest into agricultural land (Soemarna and Anwar 1994; Purwanto et al. 2009b).

The development and the cultivation of jernang rattan can also be done in the area of rubber plantations as intercropping plants which may provide more benefits than converting rubber plantations to oil palm plantations (Purwanto et al. 2009b). This intercropping practice has been done by the people of Anak Dalam Jambi Tribe, in Lumban Sigatal, Sipintun and Sarolangun districts since 2005 (Soemarna 2009), but those in Jebak village of Batanghari have not done this.

The objectives of this study were to examine how the Anak Dalam Tribe manages Daemonorops draco to remain sustainable and beneficial, and to analyze the possibility of the development and cultivation of Daemonorops draco in the forest areas of Anak Dalam Jambi Tribe in order to provide both economic and ecological benefits.

mm/year. The population is 250, consisting of 40 households (BPS Jambi 2010; BKSDA Jambi 2010) (Figure 1).

Procedures The methods used in this study were interviews, direct observation, and literature review. Interview method was done to obtain information about the history of jernang rattan in the forest area of Anak Dalam Jambi Tribe, the use, the habitat, the non-destructive harvesting techniques, the characteristics of fruits that can be harvested, the management, the conservation and the cultivation of jernang rattan by Anak Dalam Jambi Tribe, and the socioeconomic value of dragon’s blood Interview technique used was semi-structural interview that had guidelines in the form of questions, but could be developed, according to the needs in the field. The interviewees were men aged 2075 years. The approach was based on information from key informants from traditional elders, and of ordinary people of Anak Dalam Jambi Tribe as dragon’s blood seekers in the village of Jebak (Rugayah et al. 2004).

Data analysis Data were analyzed descriptively and in conjunction with the data retrieval process. The data analyses consisted of organizing data, sorting data, and drawing conclusions.

RESULTS AND DISCUSSION Anak Dalam Jambi Tribe and jernang rattan Jebak Village has been inhabited by Anak Dalam Jambi Tribe since 1624. But their existence has been definitive since 1970. Prior to 1970, they lived in nomadic thatched house on stilts. Since 1970, they have been localized and given shingle-roofed houses, each measuring 36 m2, and 2 hectares of rubber land near their homes (Figure 2). Jebak village is inhabited by 40 families comprising 250 people: 104 men and 146 women (BPS Jambi 2010).

MATERIALS AND METHODS Time and place The research was carried out in forest areas of Anak Dalam Jambi Tribe in Jebak village, Muara Tembesi, Batanghari, Jambi Province. This selection of site was based upon information that the forest is a habitat of jernang rattan populations (Daemonorops draco).

Jebak Village of Jambi is an old village of Anak Dalam Jambi Tribe. The Jebak village is 60 km from Jambi city, which can be reached within 1 hour road trip. It is located between West Tanjung Jabung, South Sumatra, Muaro Jambi, Sarolangun and Tebo. Geographically, Jebak village is located at 103o 05'-103o 15 E and 01o 40'-01o 50' S, with an altitude of 20 m above sea level, rainfall of 2296

Figure 2. Anak Dalam Jambi Tribe's home in Jebak Village Batanghari, Jambi.

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Indonesia

Jambi Province

Batanghari District

Figure 1. Map of study site (circle) in the village of Jebak, Batanghari (bordered area), Jambi (BKSDA Jambi 2010).

The people of Anak Dalam Jambi Tribe are the descendants of South Sumatran people. Therefore, they are familiar with the Islamic religion, clothing, and food from their ancestors. Most of the people of Anak Dalam Jambi Tribe just graduated from elementary school. Their livelihood is to extract the Non Timber Forest Products (NTFPs), such as balam, jelutung, damar, damar mato kucing, rattan, honey, and dragon’s blood. Because NTFPs are sources of their livelihood, Anak Dalam Tribe in Jambi forest treat them in such a way that prevent damage.

In the 1990s, transmigrants from Java, West Sumatra, South Sumatra and North Sumatra, were located in forest areas of Anak Dalam Jambi Tribe. The first villages for transmigration were Sridadi and Jangga Baru. In 1993 transmigration area was opened in the village of New Bulian, and the last was Mekar Jaya village, which was opened in 1995. Since 1990, the encroachment of forest has grown out of control because of lack of supervision from the government and because of unclearness of the boundary between the forest area and the villages around the area.

(BKSDA Jambi 2010).

Jernang rattan is a plant which is used as a source of family income, because it produces dragon’s blood which is relatively expensive. Jernang rattan has been used by Anak Dalam Jambi Tribe in Jebak village since 1624.

There are two species of rattan producing dragon’s blood in Jebak village, namely jernang rattan g (Daemonorops draco) and kelemunting rattan (Daemonorops didymophylla). According to Rustiami et al. (2004), there are 12 species of rattans producing dragon’s blood, namely Daemonorops acehensis, D. brachystachys, D. didymophylla, Daemonorops draco, D. dracuncula, D. dransfieldii, D. maculata, D. micracantha, D rubra,

D. sekundurensis, D. siberutensis, and D. uschdraweitiana.

The use of dragon blood Dragon’s blood is resin that covers the fruit skin of the jernang rattan. It is red, amorphous, solid, shiny, clear or dull, and having a specific odor (Coppen 1995)(Figure 3).

Dragon’s blood is used for dyes, pharmaceuticals ingredients, perfume ingredient, substitute for incense in religious ritual (Purwanto et al. 2009b), as raw material for varnish, and as medicine to heal a wound (Soemarna 2009).

Since 1624 dragon blood has been used by Anak Dalam Jambi Tribe as a source of income, medicines for injury, diarrhea, headache,, accelerator of parturition and as explosive. Puerperal blood will dry within 3-7 days after using dragon blood pilis.

The main component of dragon blood is alcoholic resin of dragon’s blood, resinolanol draco (56%); when heated it will produce a smell like incense. Because it is red, then it is known as the red incense. Anak Dalam Jambi Tribe communities use the one as a substitute for incense in ritual ceremonies (Table 3). Dragon blood is also used as a power booster in a magical ritual. The burning of incense increases the level of magical spells recited. It is also used to strengthen passion, so it is added to the "love sachets", oil and soap (Purwanto et al. 2009c; Soemarna 2009).

Dragon’s blood contains Draco Resin (11%), Draco Alban (2.5%), amino acid and benzoate (Winarni et al.

2004; Waluyo 2008). Benzopyran serves to stop the bleeding when injury occurs. Benzopyran can be processed as a biopesticide; trierene can cure impotence in men; flavonoids acts as antioxidants; saponins plays a role in neutralizing and clearing toxins (Soemarna and Waluyo 2009), and tannins stops diarrhea (Waluyo 2008).

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Figure 3. A. Jernang rattan fruits that contain lots of dragon blood: Rattan fruit before extraction (left), Rattan fruit after extraction (right). (Waluyo 2008); B. Jernang rattan fruits before extraction. C. Dragon blood

Figure 4. Jernang fruit and the separation process of dragon’s blood (Purwanto 2009)

Extraction activity and production systems Extraction of dragon blood is done every year, but in August and December jernang rattan bears the greatest number of fruits, so Anak Dalam Jambi Tribe call it great harvest. Jernang fruit is harvested when it is not too ripe and not too young. If harvested too ripe or too young, the production of dragon’s blood is not optimal. According to Anak Dalam Jambi Tribe, the thickest dragon blood is obtained from the fruit at the age of 9 months from flowering. In each extraction season (from August to December) every person can obtain 30-50 kg of dragon blood. It also depends on the luck factor. In general, each tree can produce 10-60 kg of fruit, depending on the conditions of growth and soil fertility.

Jernang rattan plants in the forest are an "open access", meaning that every member of the society in the region has the same rights and opportunities to extract jernang fruits.

There is no specific ownership for the jernang fruit in the forest in this region. The harvest principle is: the person who knows first that the jernang rattan is bearing fruit is the one who can harvest it. When he does not harvest it at once, then the next day other community members may harvest the fruit, and the first man cannot claim that the

fruit is his. The harvest system is: who is faster then he gets the fruit (Figure 4).

This situation causes problems, because in the next harvest period, the fruits will be harvested at younger age for fear that other community members will get them.

Some respondents said that if someone finds jernang rattan bearing fruit and then gives a sign, the fruit is generally not harvested by other people. But if someone harvests it, the person who has given the sign can not state his objection.

In the 1990s before the transmigration and oil palm plantation companies entered the region or in the period before the logging of primary forest, more than 15 clumps of jernang rattan could be found in one hectare of forest, it could produce up to 60 kg of fruit at a price of Rp 250,000/kg. In general, these jernang plants grow in the forest area around the river or a forest area that often gets overflow of river water.

In 2009 and 2010, the production of dragon’s blood from logged over forest ranged from 0.1 to 1.5 kg per hectare. The area of forest was 25,000 ha, and the number of community members who extracted dragon’s blood ranged from 3-4 persons from about 40 families.

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The separation of dragon blood is as follows: the fruit of jernang rattan is aerated for 3 days in order that dragon blood attached to the shell of the jernang rattan fruit can be separated easily from the shell. Having been aired for three days, the rattan fruit is then pounded in the filter basket. It produces red powder (dragon’s blood). The dragon’s blood is then sieved to separate the dragon’s blood from its shell.

Then, dragon blood is put in a plastic bag, let stand for 1-2 hours. The dragon’s blood in the form of powder hardens to form lumps. Dragon blood lumps are ready for sale.

Twenty kg of jernang fruit is needed to produce 1 kg of dragon’s blood.

Socio-economic aspects Each jernang rattan plant produced 0.1 kg-1.5 kg of dragon blood each month in 2011. For one month Jebak village was only able to produce 0.4 kg-6 kg of dragon blood. Batanghari district was only able to produce 1.8 kg20 kg dragon blood per month: Bukit 12 produced 0.5 kg-6 kg, Jebak Village 0.4 kg-6 kg, and Batin XXIV 0.9 kg-8 kg. Every month dragon blood collectors were only able to collect 12.3 kg-88 kg of dragon blood from extractors: in Batanghari 1.8 kg-20 kg, in Tebo 3 kg-30 kg, in Sarolangun 5 kg-20 kg, and in Tanjung Jabung 2.5 kg-18 kg (Table 1).

Table 1. The production of dragon blood in each district District Batanghari Tebo Sarolangun Tanjung Jabung

The production of dragon blood production / month 1.8 kg-20 kg 3 kg-30 kg 5 kg-20 kg 2.5 kg-18 kg

According to Soemarna (2009) and the observations, the quality of dragon blood based on the composition of the

Rice cultivation Trade 5% 3%

Jernang 52%

Rubber 15% Resin 1%

Post-harvest handling and trading After the separation process of dragon’s blood is completed, and then the extraction product in the form of powder is put in a plastic bag weighing 0.5 to 1 kg. Thirty minutes-one hour later, the dragon’s blood powder will harden to form lumps. The lump of dragon’s blood is sold in the market or to traders. There is no special treatment while waiting for dragon’s blood to be sold to traders. In general, dragon blood is stored in a safe place, to prevent theft, because it has a high economic value.

Rattan 4%

Balam 3%

Honey 2%

Resin 2%

Mato kucing resin 4% Jelutung 5%

Honey 1%

Balam 2% Rattan Mato kucing 3% resin 3%

mixture can be seen in Table 2. The best dragon blood is the one from Batanghari District, because the result of extraction is pure. Some people mixed dragon’s blood with dammar mato kucing, seeds and rind of jernang rattan fruit, or even brick powder to increase the weight. They cheated because it is hard to find jernang rattan in the forest and the mixture physically looks the same as the pure dragon’s blood.

Dragon’s blood is the main source of Anak Dalam Jambi Tribe’s income, (Figure 5). It gives the largest contribution to the total income of Anak Dalam Jambi Tribe. The condition has decreased since the influx of transmigrants in 1990, because since then the forest of Anak Dalam Jambi Tribe in Jebak village Batanghari has been damaged significantly.

Dragon Blood plays a very important role in the economy of Anak Dalam Jambi Tribe. Eighty percent of the economy of Anak Dalam Jambi Tribe derives from dragon blood extraction (Figure 6). From the extraction of these dragon blood, Anak Dalam Jambi Tribe can meet all their basic needs, such as sending their children to school, supplementing their household needs in order not to be left behind by the outsider community. However, these conditions are hard to come by this time, because the population of jernang rattan in nature has been decreasing.

Service 2%

Hunting 10%

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Jelutung 3%

Figure 5. Income source of Anak Dalam Jambi Tribe in Jebak village

Jernang 80%

Figure 6. Income sources of Anak Dalam Jambi from NTFPs

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Table 2. Quality of dragon blood based on its composition Quality Price/ kg

Market name

Composition

A B

Rp 1.000.000-2.000.000 Jernang (Batanghari) Rp 600.000-Rp 1.200.000 Lulun meson (Sarolangun)

Local Name

Dragon’s blood Dragon’s blood

C

Rp 500.000-Rp 900.000

Lum jernang (Tebo)

Dragon’s blood

D

Rp 350.000-Rp 700.000

Dragon blood (Tanjung Jabung)

Dragon’s blood

Pure dragon blood Mixed with damar mato kucing resin or brick powder (50%-60%) Mixed with jernang rattan fruit and damar mato kucing resin (70%-80%) Mixed with seed, jernang rattan fruit shell and damar mato kucing resin (80%-90%)

Table 3. Results of interviews with eight persons of Anak Dalam Jambi Tribe in Jambi on their knowledge of dragon blood, the use, and the feature of rattan-producing dragon blood.

No 1

Questions Answers Have you known the term of dragon’s blood? Yes, I have

 8

2 3

What is meant by dragon’s blood?

What is the characteristic of dragon’s blood?

8 4 2 6 6 6 4 8 6 6 3 1 8

4 5

6 7 8

9

10 11 12 13

14 15

16

Resin that covers the skin of jernang rattan fruit Reddish black Black Shiny If it is burnt, its smell is like the smell of incense How to extract dragon’s blood?

There are 2 ways, wet and dry What are the uses of dragon’s blood Income source Cure wounds For diarrhea Accelerate the completion of parturition Explosive Headache medicine When has Anak Dalam Jambi Tribe started to Since 1624 make use the dragon’s blood?

Can you distinguish rattan producing dragon Yes, I can blood from the non-producing rattan?

How do you distinguish jernang rattan It can be seen from: stem jernang from other rattans?

Leaves Fruits Thorn What are the differences? ( no 8) Small stem, diameter: 1 cm-3 cm 1 cm-2 cm 1,5 cm-3 cm the young leaves colored reddish green. lots of fruits colored black lots of thorn colored black What species of rattan that produce dragon jernang rattan (Daemonorops draco) and mengkarung/ blood? kelemunting rattan (Daemonorops didymophylla) Which rattan produce the best dragon blood ? jernang rattan (Daemonorops draco) Why? It produces lots of dragon blood What are the characteristics of jernang rattan Lots of fruits that produces high quality Long strands of fruits of dragon blood?

Color of fruits: shiny black Lots of thorn covering stems Stem high: 8 m-15 m Rod segment: 15 cm-20 cm 15 cm-25 cm 20 cm-30 cm Number of individuals in a clump : 5-20 stems When does jernang rattan begin to 3-4 years bear fruits ?

4-5 years What are the characteristics of rattan Light green leaves jernang producing fruits?

Rod segment: 15 cm-20 cm 15 cm-25 cm 20 cm-30 cm Long flower strand Why? There are male and female trees.

Male jernang rattan: flowering no fruit Short flower strand Rod segment: 35 cm-40 cm Number of individuals in a clump: 3-5 stems

6 6 6 6 6 3 1 2 6 6 6 6 6 6 6 6 6 6 6 2 1 3 6 1 5 6 2 1 3 6 6 6 6 1 1

Description Two people not seekers of dragon’s blood

SULASMI et al. – Management of Daemonorops draco by Anak Dalam of Jambi 17 18 19 20

21 22 23

How many jernang rattan fruits can be produced in a clump?

How many times a year is the harvest done?

How to identify the time harvest jernang rattan?

When does the fruits produce lots of resin?

157

3 kg-20 kg 5 kg-20 kg 2 times, August and December Fruit color bright black

2 4 6 6

When it is half ripe

6

Can all fruits be used as seed?

No, they can’t Why ?

Because only the half ripe fruits is taken as seeds.

What is the characteristic of jernang rattan that Reddish black fruits is good for seeds?

One of them added that when fruits are half ripe, it is about 9 months from flowering.

6 6 6

2 persons added that ripe fruits taste sweet and bitter.

Note: Symbol  = Number of individuals who answered the question

Table 4. Utilization, management and conservation of jernang rattan by Anak Dalam Jambi Tribe in Jebak village Batanghari, Jambi.

No 1

Questions What are the uses of jernang rattan for Anak Dalam Jambi Tribe?

Answers For income source.

To take dragon blood

 5 8

2

When did Anak Dalam Jambi start taking use of jernang rattan?

What is the role of jernang rattan in Anak Dalam Jambi Tribe’s economy?

Why?

Since 1624

8

Very large role

8

3 4 5

6 7 8 9 10 11 12 13 14 15

16

17 18

80% Because it produces expensive dragon’s blood , so it can meet the living needs of Anak Dalam Jambi Tribe.

What is the price of dragon Rp 1.5 million blood/kg?

Rp 1 million-Rp 2 million Rp 1.5 million-Rp 2 million In 2009-2010 Rp 3 million What is the percentage of the income of Anak 80% Dalam Jambi Tribe in earning from extracting dragon blood?

Do Anak Dalam Jambi Tribe use jernang It is done purposely rattan arbitrarily or purposely?

How do you harvest jernang rattan?

Climbing trees used to creep rattan plant, then jernang fruits are hooked with poles. why?

In order that jernang rattan doesn’t die and can produce fruits again.

Where is the habitat of jernang rattan?

In river bank, low land, and dry brackish.

How does jernang rattan live?

Creeping on the propagation tree What species of trees can be used as the host All of trees can be used as jernang rattan’s vine. tree?

How many clumps of jernang rattan are there 10 clumps in 2011? < 15 clumps 15 clumps Does the population decline every year?

Yes, it does.

How many clumps of jernang rattan were 25 clumps there before 2011?

15-25 clumps 20-25 clumps 25-30 clumps 30-35 clumps Why? 60 % forest has been encroached to convert as palm oil field.

The forest has damaged, the woods have been logged by transmigants What is the effect of the decline of jernang It’s difficult to look for dragon blood, decrease income. rattan population for Anak Dalam Jambi Tribe?

How is the management of jernang rattan in Maintain together. There are no special rules that bind the forest area of Anak Dalam Jambi Tribe?

Anak Dalam Jambi Tribe. Whatever found in the forests belongs to together and must be maintained together.

Since 1990, when transmigrants began to come in, life Anak Dalam Jambi Tribe communities in Jambi have been more difficult because the forest has been occupied by migrants.

1 8 1 4 3 6 8 8 6 6 6 6 6 1 4 1 6 1 1 1 2 1 5 1 6 6

Description 2 persons are not dragon blood seekers

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19

Is there any special ownership to jernang There is no specific ownership of jernang rattan g in the rattan in the forest area of Anak Dalam Jambi forest of Anak Dalam Jambi Tribe. Anak Dalam Jambi Tribe?

Tribe’s forest area in Jambi is a common property and should be kept together.

20 What efforts have been done by Anak Dalam Harvesting jernang rattan fruit without cutting the trunk.

Jambi Tribe to keep jernang rattan from Thus the jernang rattan tree can bear fruit again. extinction in nature?

21 Are there any persons who are Yes. There are 2 persons, Mr. Suin and Mr. Sudirman. willing to cultivate jernang rattan ?

They grew 40 clumps but there are only 25 clumps left.

22 How to cultivate jernang rattan ?

It is planted by intercropping with rubber trees.

23 How to get the seed of jernang rattan ?

By collecting mature jernang rattan fruit.

Note: Symbol  = Number of individuals who answered the question

The chain of dragon’s blood trade in Jambi is still closed, ie, there are no rules that govern trade and association. Prices are set by the collectors; there is no standardization, depending on them. The flow of dragon blood trade in Jambi is as follows:

Figure 7. Flowchart of dragon’s blood trade in Jambi Province.

The dragon’s blood trade in Jambi province is ruled by a major collector who lives in the town of Jambi (Figure 7).

The major collector has an agent in each district. The district agent usually controls 2-3 collectors in village level. The collectors at the village level are directly related to dragon’s blood seekers. However, it is possible that seekers make direct contact to the major collectors in Jambi, because dragon’s blood is purchased at higher prices than the prices at the village level. If the price is Rp 1 million-2 million/kg at the village level, then it can be Rp 1.2 million-2.2 million/kg at the major collector. The price is lower than the one in 2009-2010 which reached USD 3 million/kg. This is in accordance with the opinion of Soemarna (2009), which stated that the price of dragon’s blood ranged between Rp 2.3-Rp 3 million/kg.

Development and conservation of jernang rattan The decline of population and number of species The forest has been diminishing and even replaced by oil palm plantations, rubber plantations. It is now more and more difficult to find rattan in the forest of Jebak village.

6

6 6 6 6

Damage to the natural habitat of jernang rattan causes a decrease in population size and number of species of rattan-producing dragon blood in the region (Table 4). In fact there are only 8 clumps of jernang rattan in Jebak village. The damage was caused by logging and encroachment. According to Anak Dalam Jambi Tribe, the destruction of forests has reached 60% of forest area (15,830 acres).

The method of harvesting jernang rattan fruit is very good and not against the concept of conservation (Figure 8). This is in accordance with the opinion of Soemarna (2009) which stated that Daemonorops draco harvesting is not done by cutting down trees, but by plucking. The method of harvesting does not damage the crown cover, so it does not disrupt the forest ecosystem (Dali and Soemarna 1985; Sudarmalik et al. 2006). The part used is the resin of fruit shell. According Winarni et al. (2004), Daemonorops draco must be harvested little by little, so it does not directly lead to overexploitation.

Development and conservation efforts The demand of dragon’s blood from China continues to increase every year. The economic value and use value of dragon’s blood are high enough. The development of modern medicine and high-quality staining is the future prospect of dragon’s blood. Thus, the development of jernang rattan is very important, as one alternative to improve the income of Anak Dalam Jambi Tribe, who live in forested areas in Jebak village. What can be done are to cultivate jernang rattan in its natural habitat, develop it as one of the leading crops in the region of agroforestry system and set the trade system of dragon’s blood by shortening the marketing chain of dragon blood, so there is no monopoly which harm producers.

The development and the cultivation of jernang rattan can also be done in the area of rubber plantations as intercropping plants which may provide more benefits than converting rubber plantations into oil palm plantations.

According to Anak Dalam Jambi Tribe, since 2008 they've been doing the cultivation of jernang rattan. That year they tried to plant 40 clumps of jernang rattan under Anak Dalam Jambi Tribe’s rubber trees in Jambi named Sudirman (15 clumps) and Suin (25 clumps). Of those 40 clumps, only 25 clumps still lived in the garden in 2011.

All jernang rattan which grew in Sudirman’s rubber trees died because wild boars ate them. Nowadays people are very difficult to obtain jernang rattan seed in nature

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Figure 8. How to harvest jernang rattan fruit.

because of jernang rattan population decreases in nature and the fruits are not harvested before maturity so the extracted seed can not be germinated.

Planting jernang rattan in a natural habitat in the logged over forests has several benefits, one of them is the conservation of biodiversity. The presence of jernang rattan in the forest will prevent conversion of forest to agricultural land.

Promotion and future role of dragon’s blood Dragon’s blood is produced from the extraction in the forests and not the result cultivation. Excessive extraction can cause damage and population decline of jernang rattan.

In addition, the system of ownership of the "open access" becomes one of triggers for competition in extracting dragon’s blood. Consequently, the method to harvest becomes uncontrolled that causes disruption to the natural population of jernang rattan. Therefore, the strengthening of social institutions in the management of biological resources that involve community participation is very important.

The role of dragon’s blood as a natural product in the future is still needed, although artificial products are being developed. This refers to other natural products such as resins, incense, aloes, which remain important and irreplaceable by their artificial products. The problem is how to conserve the natural habitat of jernang rattan to ensure the sustainability of dragon’s blood production and its benefits to the community.

Consequences of commercialization of dragon’s blood The consequences of commercialization of dragon’s blood can be profitable but also damaging. The advantage is that commercialization will trigger the effort to keep the jernang rattan population and possibly develop it into one of the crops that provides an important role in increasing income of farmers around the forest areas in Jebak village.

The disadvantage is that commercialization will cause over-exploitation, and competition in jernang rattan extraction. Such conditions may accelerate the destruction of jernang rattan population in the region. To prevent this, it is necessary to discover jernang rattan cultivation technique and its application to the public, and to create the dragon’s blood community from forest cultivation.

CONCLUSIONS AND RECOMMENDATIONS Jernang rattan is a source of income for Anak Dalam Tribe because it produces dragon’s blood. The dragon’s blood prices in 2011 ranged from Rp 1 million-Rp 2 million/kg. It is relatively expensive when compared with the price of honey which is between Rp 20,000-Rp 25,000/ kg. Besides, dragon’s blood is utilized by Anak Dalam Jambi Tribe as drug for injuries and migraine headaches and accelerator of parturition. It is also used as substitute for incense in rituals and as explosives. The community has used dragon’s blood since 1624.

Anak Dalam Jambi Tribe Jambi is expected to maintain populations of jernang rattan left in the wild, working in such a way so that supplies of seeds for cultivation is maintained, and They should also maintain the jernang rattan cultivated by Suin. Further research is needed to cultivate jernang rattan through vegetative propagation.

ACKNOWLEDGMENTS The authors thank to Dr. Luthfiralda Sjafirdi, Dr. Kuswata Kartawinata, Dr. Himmah Rustiami, Yana Soemarna, Mega Atria, Wisnu Wardana, and Andrio Ariwibowo, thank you for willing to give time for discussion.

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160 REFERENCES

BKSDA Jambi. 2010. Non-timber forest product of Jambi province.

Department of Forestry, Jambi Area Office, Jambi. [Indonesia] BPS Jambi [Badan Pusat Statistik Provinsi Jambi]. 2010. Jambi in figures.

BPS Provinsi Jambi, Kota Jambi. [Indonesia] Coppen JJW. 1995. Gum, resin, and latex of plant origin. Non wood forest product (6). FAO, Roma.

Dali Y, Soemarna Y. 1985. Potential rattan cultivation. Proceeding of the National Symposium on Rattan. IDRC Canada-Forest Research and Development Agency. Department of Forestry, Jakarta. [Indonesia] Ministry of Social Affairs. 1998. Isolated communities of Anak Dalam Trible from Solea and Melinani hamlets. Department of Social Affairs, Republic of Indonesia, Jakarta. [Indonesia] Purwanto Y, Polosakan R, Susiarti S, Waluyo EB. 2009b. Extractivism of dragon blood (Daemonorops spp.) and its development. In: Purwanto Y, Walujo EB, Wahyudi A. (ed). 2009. Valuation of non-timber forest product after forest logging (in conservation area of PT Wirakarya Sakti Jambi). Indonesian Institute of Sciences (LIPI), Bogor. [Indonesia] Purwanto Y, Walujo EB, Afriastini JJ. 2009a. Analysis of the cultural value of non-timber forest products for the valuation potential and its development. In: Purwanto Y, Walujo EB, Wahyudi A. (ed). 2009.

Valuation of non-timber forest product after forest logging (in conservation area of PT Wirakarya Sakti Jambi). Indonesian Institute of Sciences (LIPI), Bogor. [Indonesia] Rugayah, Widjaya EA, Praptiwi. 2004. Manual of plant diversity collection data. Research Center for Biology, Indonesian Institute of Sciences (LIPI), Bogor. [Indonesia] Rustiami H, Setyowati FM, Kartawinata K. 2004. Taxonomy and uses of Daemonorops draco (Willd.). J Trop Ethnobiol 1 (2): 65-75.

Soemarna Y, Anwar C. 1994. Distribution and ecology of natural forest area of Pasir Tugu, Jasinga Bogor. Bul Kehut 562: 49-61. [Indonesia] Soemarna Y. 2009. Jernang rattan cultivation (Daemonorops draco Willd). J Litbang Kehut 2 (3): 5-10. [Indonesia] Sudarmalik, Rochmayanto Y, Purnomo. 2006. The role of several nontimber forest products (NTFP) in Riau and West Sumatra. Proceeding of Research Seminar of the Center for Research and Development on Forest Engineering and Forest Products Processing, Bogor: 199-219.

[Indonesia] Waluyo T. 2008. Traditional extraction technique and analyzes of jernang characteristic of Jambi. J Penel Hasil Hut 26 (1): 30-40. [Indonesia] Winarni I, Waluyo TK, Hastoeti P. 2004. A review on jernang as a valuable commodity. Proceeding of the forest products, Bogor: 173176. [Indonesia]

GUIDANCE FOR AUTHORS Aims and Scope ”Biodiversitas, Journal of Biological Diversity” or Biodiversitas encourages submission of manuscripts dealing with all biodiversity aspects of plants, animals and microbes at the level of gene, species, and ecosystem.

Article types The journal seeks original full-length research papers, reviews, and scientific feedback (short communication) about material previously published.

Acceptance The only articles written in English (U.S. English) are accepted for publication. Manuscripts will be reviewed by managing editor, communicating editor and invited peer review according to their disciplines.

Authors will generally be notified of acceptance, rejection, or need for revision within 2 to 3 months of receipt. Manuscript is rejected if the content is not in line with the journal scope, dishonest, does not meet the required quality, written in inappropriate format, has incorrect grammar, or ignores correspondence in three months. The primary criteria for publication are scientific quality and biodiversity significance. The accepted papers will be published in a chronological order. This journal periodically publishes in January, April, July, and October.

Uncorrection proofs will be sent to the corresponding author by e-mail as .doc or .docx files for checking and correcting of typographical errors. To avoid delay in publication, proofs should be returned in 7 days.

A charge The cost of each manuscript is IDR 250,000,- plus postal cost or IDR 150,000,- for SIB members. There is free of charge for non Indonesian author(s), but need to pay postal cost for hardcopy.

Reprints Two copies of journal will be supplied to authors; reprint is only available with special request. Additional copies may be purchased by order when sending back the uncorrection proofs by e-mail.

Submission The journal only accepts online submission, through e-mail to the managing editor at unsjournals@gmail.com; better to forward to one of the communicating editor for accelerating evaluation.. The manuscript must be accompanied with a cover letter containing the article title, the first name and last name of all the authors, a paragraph describing the claimed novelty of the findings versus current knowledge, and a list of five suggested international reviewers (title, name, postal address, email address). Reviewers must not be subject to a conflict of interest involving the author(s) or manuscript(s). The editor is not obligated to use any reviewer suggested by the author(s).

Copyright Submission of a manuscript implies that the submitted work has not been published before (except as part of a thesis or report, or abstract); that it is not under consideration for publication elsewhere; that its publication has been approved by all co-authors. If and when the manuscript is accepted for publication, the author(s) agree to transfer copyright of the accepted manuscript to Biodiversitas. Authors shall no longer be allowed to publish manuscript without permission. For the new invention, authors suggested to manage its patent before publishing in this journal.

Open access The journal is committed to free open access that does not charge readers or their institutions for access. Users are entitled to read, download, copy, distribute, print, search, or link to the full texts of the articles, as long as not for commercial purposes. For the new invention, authors are suggested to manage its patent before published.

Preparing the Manuscript 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 re-express 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. Reference to unpublished data and personal communication should not appear in the list but should be cited in the text only (e.g., Rifai MA 2007, personal communication; Setyawan AD 2007, unpublished data).

In the reference list, the references should be listed in an alphabetical order.

Names of journals should be abbreviated according to the ISSN List of Title Word Abbreviations (www.issn.org/2-22661-LTWA-online.php).

APA style in double space is used in the journal reference as follow: Journal: 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

ISSN: 1412-033X E-ISSN: 2085-4722

GENETIC DIVERSTY A comparative phylogenetic analysis of medicinal plant Tribulus terrestris in Northwest India revealed by RAPD and ISSR markers ASHWANI KUMAR, NEELAM VERMA In Silico chloroplast SSRs mining of Olea species ERTUGRUL FILIZ, IBRAHIM KOC SPECIES DIVERSTY Taxonomy of Indonesian giant clams (Cardiidae, Tridacninae) UDHI EKO HERNAWAN The exploration and diversity of red fruit (Pandanus conoideus L.) from Papua based on its physical characteristics and chemical composition MURTININGRUM, ZITA L. SARUNGALLO, NOUKE L. MAWIKERE ECOSYSTEM DIVERSTY Study of biodiversity and limiting factors of Ag-gol Wetland in Hamadan Province, Iran MAHDI REYAHI-KHORAM, VAHID NORISHARIKABAD, HOSHANG VAFAEI Assessment of biodiversities and spatial structure of Zarivar Wetland in Kurdistan Province, Iran MAHDI REYAHI-KHORAM, KAMAL HOSHMAND Diversity of soil macrofauna on different pattern of sloping land agroforestry in Wonogiri, Central Java MARKANTIA ZARRA PERITIKA, SUGIYARTO, SUNARTO Fish biodiversity in coral reefs and lagoon at the Maratua Island, East Kalimantan HAWIS H. MADDUPPA, SYAMSUL B. AGUS, AULIA R. FARHAN, DEDE SUHENDRA, BEGINER SUBHAN ETHNOBIOLOGY Jernang rattan (Daemonorops draco) management by Anak Dalam Tribe in Jebak Village, Batanghari, Jambi Province IIK SRI SULASMI, NISYAWATI, YOHANES PURWANTO, SITI FATIMAH

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Front cover: Tridacna crocea

(PHOTO: UDHI EKO HERNAWAN)

Published four times in one year

PRINTED IN INDONESIA ISSN: 1412-033X

E-ISSN: 2085-4722