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INDEX – GJRMI - Volume 4, Issue 1, January 2015 MEDICINAL PLANTS RESEARCH Bio-Technology INFLUENCE OF GROWTH REGULATORS ON GERMINATION AND GROWTH OF ENDANGERED MEDICINAL PLANT NOTHAPODYTES NIMMONIANA J.GRAHAM UNDER SHADE NET CONDITIONS Anita S Patil, Surendra R Patil, Ankit S Kale
01–09
Agriculture DRUMSTICK FERMENTED LEAF JUICE (DFLJ) - A PROMISING ORGANIC SIGNATURE FOR TOMATO CULTIVATION PACKAGE Rajamani R, Rudresh Kumar Singh, Vinod Kochupillai, Mohit Aggarwal, Sivaraj A K
10–19
COVER PAGE PHOTOGRAPHY: DR. HARI VENKATESH K R, PLANT ID – INFLORESCENCE OF ANJANI – MEMECYLON EDULE ROXB. OF THE FAMILY MELASTOMATACEAE PLACE – KOPPA, CHIKKAMAGALUR DISTRICT, KARNATAKA, INDIA
Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 01–09 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article INFLUENCE OF GROWTH REGULATORS ON GERMINATION AND GROWTH OF ENDANGERED MEDICINAL PLANT NOTHAPODYTES NIMMONIANA J.GRAHAM UNDER SHADE NET CONDITIONS Anita S Patil1*, Surendra R Patil2, Ankit S Kale3 1, 3
Lab no. 106, Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati (M.S) India College of Horticulture, Dr Panjabrao Deshmukh Agriculture University, Akola (M.S) India *Corresponding Author: anitapatil@sgbau.ac.in; Mobile: +91-9881735354 2
Received: 20/11/2014; Revised: 31/12/2014; Accepted: 03/01/2015
ABSTRACT Nothapodytes nimmoniana J. Graham is a tree species found in Western ghats of Maharashtra (India). The plant is one of the most interesting sources of camptothecin (CPT), 9- methoxy camptothecin and other derivatives used as anti-cancer metabolites. Due to overexploitation and low seed germination, the tree currently becomes rare, endangered at present included in Red data books. In the present study, the efforts to increase the seed germination were screened by presoaking them for 24 hours in variable concentration growth regulators (Gibberelic acid 50, 100 and 150 ppm; Potassium nitrate 1.0, 1.5 and 2.0 %) and then compared with control (water soaked). The seeds treated with Gibberelic acid in 150 ppm exhibited superior results in germination percentage, number of days taken for initiation of seed germination and overall seedling growth of N. nimmoniana under shade net conditions. In general pre-soaking treatments to the seeds with different growth regulator and chemical solutions were found to be beneficial to improve the germination as compared to control. KEYWORDS: Nothapodytes nimmoniana J. Graham, Growth regulators, Seed germination, Camptothecin ABBREVATIONS: GA – Gibberellic acid; ppm – Part per million; ERI – Emergence rate index BRI – Bartletts rate index; SVI – Seedling vigour index; KNO3- Pottasium nitrate
Cite this article: Anita S Patil, Surendra R Patil, Ankit S Kale (2015), INFLUENCE OF GROWTH REGULATORS ON GERMINATION AND GROWTH OF ENDANGERED MEDICINAL PLANT NOTHAPODYTES NIMMONIANA J.GRAHAM UNDER SHADE NET CONDITIONS, Global J Res. Med. Plants & Indigen. Med., Volume 4(1): 01–09
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 01–09
INTRODUCTION: Nothapodytes nimmonina Graham (family Icacinaceae) also known as N. foetida (Wight, Sleumer) or Mapia foetida (Miers) is a small sub canopy tree distributed in Western Ghats region of Maharashtra, Goa, Kerala, Karnataka, Assam and Tamilnadu in India. Camptothecin isoquinoline alkaloid is one of the most promising anticancer drug of the twenty-first century (Wall and Wani, 1966; Yan et al., 2003; Lorence and Craig, 2004, Patil et al., 2014) specifically used for treatment of colorectal and ovarian cancer (Lilenbaum et al., 2005) with the highest projected demand in pharmaceutical industry. Camptothecin was initially isolated from the Chinese tree Camptotheca acuminata (Niyssaceae) (Wall and Wani, 1968), later from Merrilliodendron megacarpum (Gunasekera et al., 1979); Ophiorrhiza mungos (Tafuret et al., 1976) and O. pumila, Eravatamia heyneana and Mostuea brunonia (Fulzele, 2003). N. nimmoniana possesses highest concentration of camptothecin 0.3% (w/w) was found out, which was first isolated and chemically defined by Prof. Govinadachari (Govindachari et al., 1972; 1994). Recently, similar explorations in India have led to identification of N. nimmoniana Graham, earlier known as N. foetida (Wight, sleumer) as an alternative source of camptothecin. This plant is distributed in the warmer regions like Indian subcontinent in southern India, Srilanka, Parts of eastern Indian Assam and the Himalayan foothills in north India, Myanmar and Thailand (Padmanabhan et al., 2006). At present, worldwide market demand of camptothecin is very high due to the absence of synthetic derivatives. With an increasing global demand for these alkaloids, there has been a heavy dependence extraction of camptothecin from existing population of N. nimmonina. Due to such essentialities at last decade alone over 20% population of this species has been lost from the western ghats (Gowda et al., 2002), due to which the species has been
declared as endangered (Ciddi, 2000; Suhas, 2007). Conservation efforts are much more required for this plant, as their population is becoming narrow due to habitat destruction and overexploitation (Cragget et al., 1993). The tree growth is slow and propagation is usually achieved by seeds. The seeds are recalcitrant due to high sensitivity of desiccation, freezing and have a short shelf life. Until today, very few tissue culture methods have been developed for the propagation of this plant (Thengane et al., 2001; Rai, 2002). Seeds of N. nimmoniana are of large and intermediate type. The orange-colored receptacle of seeds turns black after drying. The species can be easily recognized by its strong foetid odour during blooming. The fruits (drupes) obliquely ovoid more than 1 cm in girth and 1.5 cm 2.5 cm length with attached nut/ seeds. It has been reported that the cryopreserved zygotic embryo showed 87.67% germination under the controlled conditions. Embryonic axes with cotyledons, having moisture content of 55.7% and presumed to be intermediate in nature, lose their viability within a short period after maturity. But still it is recognized as an effective tool for long term preservation of such plant species those produce recalcitrant /large seeds (Engelmann et al., 1997). Therefore, the major objectives of present work to develop an easy and cost effective method for seed propagation and germination of this rare and endangered medicinal plant N. nimmoniana J. Graham. from Western Ghats of Maharashtra, India. MATERIALS AND METHOD: a. Plant Collection and Identification: The plant saplings and dried plant parts (seeds, bark, leaves) were collected from Chiplun part of Western Ghats, Maharashtra. These saplings were then maintained in green house. The plant was authenticated by Prof. S
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 01–09
R Manik, Dept of Botany, SGBAU, crosschecked with standard flora and stored in the herbarium records at Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati, India as Nothapodytes nimmoniana (J. Graham) with an accession number-BTSGBAU-07. The fruits and seeds were manually collected by hand picking from Chiplun of Western Ghats of Maharashtra between December, 2012 and January, 2013. The seeds are shed dried for a week and later used for the experimentation. b. Preparation of soil bed: The soil bed was prepared in plastic tray using autoclaved fine sand, autoclaved soil and farm yard manure in the proportion of (2:2:1). The soil bed was watered regularly to keep bed moist for the germination of seeds. Seed treatment:
The experiment was carried out adopting Complete Randomized Design with seven treatments replicated thrice. 15 seeds were used per treatment. The treated seeds of N. nimmoniana were sown in the tray filled with sterilized soil (Sand: Soil: Farm yard manner2:2:1) potting mixture. All the seeds were sown at 10 cm row- to- row distance, 5 cm plant-toplant distance at 3 cm deep. Cultural operations like regular watering, weeding and plant protection measures were undertaken. For control of damping of disease, drenching of copper fungicide (Copper oxychloride 1.0%) was done twice at 15-day intervals during the early period of investigation. The emergence rate index was calculated by the formula of Evetts and Burnside (1972). G1 G2 G3 Gn ERI = ---- + ---- + ----- + ----T1 T2 T3 Tn Where, G1 - Per cent of seed germinated at first count T1,
This experiment was conducted under shade net conditions. From the single tree, G2 - Additional percent of seeds germinated at uniformed sized, mature and healthy fruits of second count T2, N. nimmoniana were harvested. The seeds were extracted and washed in water several times G3 - Additional percent of seeds germinated at and dried in shade for a day before sowing. The third count T3, seeds were dipped in distilled water and Gn- Additional percent of seeds germinated at allowed to settle at the bottom of the beaker for final count Tn , a few minutes. The seeds floating on the surface of the water were discarded and those, T1 - Weeks from sowing to first count, T2which settled at the bottom, used for Weeks from sowing to second count, experiment. Selected seeds were divided into 7 T3 - Weeks from sowing to third count, T n lots, each containing 45 seeds. The seed lots Weeks from sowing to last count were treated with different concentrations of The Barlett’s rate index refers to earliness GA3 (50, 100 and 150 ppm), Potassium nitrate of germination and was worked out by the (1.0, 1.5 and 2.0 %) and control (water soaked). following formula (Barlett, 1937) The seeds were soaked for 24 hours in 100 ml of solutions. After imposing the treatments, the seeds were made into 3 groups of 15 seeds each. P1+ (P1+P2) + (P1+P2+P3) +........ + (P1+P2+P3+.......Pn) BRI = ------------------------------------------------------------------------N (P1+P2+P3+.........+ Pn) Where, P1, P2 ...Pn = Germination percent at 1, 2 ...n weeks, respectively. N
= Total number of weeks in the test.
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 01–09
The vigour index was calculated by multiplying percent germination by seedling length in cm. Vigour index = Percent germination × (root length (cm) + shoot length (cm). RESULTS: 1. Germination of seeds under shade net conditions a. Germination percentage (%) In general pre-soaking treatments to the seeds with different growth regulator and chemical solutions were found beneficial to improve the germination percentage compared to control shade net conditions (Table 1). Treatment GA3 150 ppm recorded higher germination (67.33 %) at the end of sixth week after sowing. This treatment was found significantly superior to rest of the treatments,
whereas least germination was found in control (33.00%). b. Days to initiate germination: The seeds treated with GA3 required 11.66 to 13.00 days and KNO3 16.00 to 18.16 days for initiation of germination as compared to control (35.50 days). The GA3 150 ppm treated seeds of N. nimmoniana germinate 23.84 days earlier than control under shade net conditions. c. Emergence rate index (ERI) and Bartlett's rate index (BRI) Treatment GA3 150 ppm recorded the highest ERI (67.711) and BRI (0.619) over rest of the treatments. It has been seen that all concentrations of GA3 and KNO3 increases the ERI and BRI values under shade net conditions over control compared to lower concentrations.
Table 1: Effect of growth regulators and chemicals on germination percentage and earliness of germination in Nothapodytes nimmoniana seeds under shade net conditions
Treatments GA3 50 ppm GA3 100 ppm GA3 150 ppm KNO3 1.0 % KNO3 1.5 % KNO3 2.0 % Control S.E.(m) + CD at 5 %
Germination after Days taken for 6 weeks (%) germination 60.16 (50.97) 62.99 (55.61) 67.33 (59.44) 53.49 (47.00) 53.49 (47.00) 57.33 (49.24) 33.00 (30.78) 1.19 3.55
2. Growth of N. nimmoniana seedlings under shade net conditions: a. Shoot and root length of N. nimmoniana seedling (cm) The results interpreted in Table 2, revealed that, various seed treatments under shade net conditions increased the shoot and root length of N. nimmoniana seedlings. Soaking of seeds in GA3 150 ppm increased shoot (5.33 cm) and root (6.11 cm) length significantly over control.
13.00 12.00 11.66 18.16 16.16 16.00 35.50 0.35 1.12
ERI
51.490 58.468 67.711 42.474 43.703 45.684 35.194 2.04 5.65
Bartlett’s Rate Index (BRI) 0.593 0.601 0.619 0.586 0.588 0.588 0.488 0.0072 0.0260
b. Seedling Vigour Index (SVI) GA3 150 ppm recorded maximum SVI (705.19) as compared to other treatments. The fluctuation in SVI is (517.42 to 705.19) in GA3 and (266.88 to 317.61) in KNO3. Higher values of SVI indicate the good seedling growth. c. Fresh and Dry weight of seedling Seeds pre-soaked in GA3 150 ppm increased the fresh (96.25 mg) and dry (14.68
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mg) weight of N. nimmoniana seedlings considerably over rest of the treatments and control. Further, it is also found that the higher concentrations of growth regulators and chemicals increased the fresh and dry weight of N. nimmoniana seedlings than lower concentrations.
OBSERVATION: It has been observed during the experimentation that the seeds started swelling during pre-germination protocol. A little plumule was seen after approximately 25th day, which further take another 5-6 days for cotyledons to establish properly as shown in Figure 1 and Figure 2.
Table 2: Effect of growth regulators and chemicals on growth of N. nimmoniana seedling under shade net conditions Shoot length Treatments of seedling (cm) GA3 50 ppm GA3 100 ppm GA3 150 ppm KNO3 1.0 % KNO3 1.5 % KNO3 2.0 % Control S.E.(m) + CD at 5 %
3.82 3.96 5.33 2.85 2.95 3.14 2.15 0.68 1.31
Root length of seedling (cm) 4.52 4.71 6.11 3.34 3.48 3.75 2.57 0.95 2.51
No. of leaves
7.76 8.91 9.83 5.89 6.07 6.29 4.88 0.62 1.40
Fresh weight of seedling (mg) 50.85 78.25 96.25 43.05 46.25 50.95 20.50 4.75 13.16
Dry weight of seedling (mg)
9.80 12.80 14.68 6.50 6.70 7.25 2.95 1.49 4.14
Seedling our Index (SVI) 517.42 633.77 705.19 266.88 283.71 317.61 150.90 25.51 73.25
Figure 1: N. nimmoniana dried seeds and preparation of germination bed in plastic tray
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 01–09
Figure 2: Small Sapling, sapling germinated in pots and well grown sapling of N. nimmoniana in nursery
DISCUSSION: N. nimmoniana can be propagated from seeds, but the seeds remain dormant for a long time after sowing, germinate poorly and early seedling is also slower as compared to plants derived from tissue culture (Vasil et al.,1986). Therefore, efficient vegetative propagation is essential for conservation of this plant, which can have many advantages over seedling production. The plant regeneration with indirect somatic embryogenesis, adventitious shoots from immature zygotic embryos and seedling explant sources of N. nimmoniana have been reported previously (Thengane et al., 2001; Fulzele et al., 2003). According to Chen and Park (1973), GA3 acts directly on embryo relieving them from dormancy through promoting protein synthesis and elongation of coleoptile and leaves and also helps in the production of ethylene. This ethylene invokes the synthesis of hydrolases, especially -
amylose, which favors the seed germination (Stewart and Freebairn, 1969). GA3 and KNO3 treatment to the seeds of N. nimmoniana reduces the days required for initiation of seed germination. Similar findings were reported by Jadhav (2003) in Rangpur lime, Kherdekar (2003) in Kagzi lime (Yogananda et al., 2007) in bell pepper. Concentrations of GA3 and KNO3 increases the ERI and BRI values of N. nimmoniana. These results are in agreement with the findings of Tendolkar (1978), who obtained early germination of cracked sapota seeds by presoaking in 200 ppm GA3. Somappa (1979) obtained higher ERI in case of rose wood seeds with GA3 + ethrel each at 100 ppm. Seeds soaked in GA3 increases shoot and root length, which has close proximity with the results obtained by Chaudhari and Chakrawar (1981)
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with GA3 40 ppm in N. nimmoniana. Under shade net conditions, Yogananda et al. (2007) observed higher root length with GA3 200 ppm in bell pepper. GA3 might have promoted more root formation through root cell elongation and more nutrient uptake as suggested by Shanmugavelu (1970). GA3 varies the SVI which is correlated with seedling growth. The results obtained are also in congruence with the findings of Tendolkar (1978) who recorded more SVI when seeds of sapota treated with GA3 400 ppm. Similarly, Yoganandaet al. (2007) got higher SVI (1174) in bell pepper when seeds treated with GA3 200 ppm. Higher seedling vigour index in GA3 treated seeds might be due to the cumulative effect of higher shoot length, root length and germination percentage, which were greatly influenced by gibberellic acid in Rangpur lime under shade net conditions. Fresh and dry weight of plant germinated depends on concentration of GA3 on seed treatment. Similar findings were observed by Gurav (2004) in Rangpur lime seedlings with GA3 80 ppm. Tendolkar (1978) in sapota seedling with GA3 400 ppm, Yogananda et
al.(2007) also recorded more seedling dry weight with GA3 200 ppm seed treatment. CONCLUSION: The study concludes that treatment GA3 in 150 ppm given to N. nimmoniana seeds shown the highest ERI (67.711) and BRI (0.619) over rest of the treatments. Furthermore, this concentration of GA3 increases in extensive growth of the seedling, due to overall assimilation and redistribution of food material results in fast growth and establishment of seedlings. ACKNOWLEDGEMENT: We express our sincere thanks to UGC, New Delhi for providing financial assistance under Major Research Project to Dr Anita Patil and Dr. Surendra Patil (F.No.42-212/2013 (SR). We are also appreciative of Shri Vasant Pusalkar, Aruna Planta Medica for authentication of plant and seed and Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati (M.S) for providing the research facilities.
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Somappa, K. (1979). Studies on improvement of propagation of some forest and plantation crops species (Master’s thesis).Univ. Agri. Sci. Bangalore, Karnataka, India. Stewart, E.R., & Freebairn, H.T. (1969). Ethylene, seed germination and epinasty. Plant Physiolgy, 44, 955–958. Suhas, R., Ramesha, B.T., Ravikanth, G., Gunaga, R.P., Vasudeva, R., Ganeshaiah, K.N., & UmaShaanker, R. (2007). Chemical profiling of Nothapodytes nimmonina populations in the Western Ghats, India for anticancer compound, Camptothecin. Current Science, 92, 1142–1147. Tafur, S., Nelson, J.D., DeLong, D.C., & Svoboda, G.H., (1976). Antiviral components of Ophirrhiza mungos isolation of Camptothecin and 10methoxy camptothecin. Lloydia, 39, 261–262. Tendolkar, S.S.P. (1978). Studies on growth of rootstocks and propagation of sapota (Manilkaraachras(Mill.)Fosberg) (Mastes’s Thesis, University of Agricultural Science, Bangalore, India) Thengane, S.R., Kulkarni, D.K., Shrikhande, V.A., & Krishnamurthy, K.V. (2001). Effect of thidiazuron on adventitious shoot regeneration from seedling explants of Nothapodytes foetida. In
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Conflict of Interest: None Declared
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 10–19 ISSN 2277-4289 | www.gjrmi.com | International, Peer reviewed, Open access, Monthly Online Journal
Research article DRUMSTICK FERMENTED LEAF JUICE (DFLJ) - A PROMISING ORGANIC SIGNATURE FOR TOMATO CULTIVATION PACKAGE Rajamani R1*, Rudresh Kumar Singh2, Vinod Kochupillai3, Mohit Aggarwal4, Sivaraj A K5 1
Research- In charge, Co-Principal Investigator, Sri Sri Institute of Advanced Research (SSIAR), (A Research Division of Ved Vignan Maha Vidya Peeth-VVMVP), Art of Living international centre, 21st Km, Kanakapura, Udayapura, Bangalore-560082, Karnataka, India 2 Scientist-1, Sri Sri Institute of Advanced Research (SSIAR), Art of Living international centre, 21st Km, Kanakapura, Udayapura, Bangalore-560082, Karnataka, India 3 Chair person –Research, Sri Sri Institute of Advanced Research (SSIAR), Art of Living international centre, 21st Km, Kanakapura, Udayapura, Bangalore-560082, Karnataka, India 4 Research Scholar- Sri Sri Institute of Advanced Research (SSIAR), Art of Living international centre, 21st Km, Kanakapura, Udayapura, Bangalore-560082, Karnataka, India 5 QA-Manager- Sri Sri Ayurveda Trust (SSAT), Art of Living international centre, 21st Km, Kanakapura, Udayapura, Bangalore-560082, Karnataka, India *Corresponding author email: raj19112002@gmail.com; Mob: +91- 9343820040.
Received: 10/12/2014; Revised: 15/01/2015; Accepted: 26/01/2015
ABSTRACT The present investigation was carried out to study the performance of tomato under organic pot culture containing coco-peat, red soil and cow dung. Three dosage of Drumstick Fermented Leaf Juice treatment with Jeevamrit, Humic acid and consortium and one control without Drumstick Fermented Leaf Juice supply were compared. A single transplanted plant per pot was raised with manual and natural irrigation in open area. The result showed that the plant performance with respect to biomass components such as number of flowers, fresh shoot and root weight, dry shoot and root weight, shoot length and root volume were significantly influenced by Drumstick Fermented Leaf Juice testing. The total plant biomass was higher with Drumstick Fermented Leaf Juice , applied at 45 ml/plant along with Jeevamrit 50 ml/plant, consortium 3 gm/plant and Humic acid 5 ml/ plant as compared to other organic input such as Jeevamrit, consortium and Humic acid combination without Drumstick Fermented Leaf Juice. Similar trend was noticed with respect to number of flower and root volume; It was significantly higher (20.13and 79.38cm3) with plant treatment receiving dose of 45ml/plant followed by 60 ml/plant (16.67 and 55 cm3) and 75 ml/plant (11.50 and 51.25 cm3). It can be concluded that production of organic tomato with Drumstick Fermented Leaf Juice as an organic supplement is a natural, cheap and adaptable for farmers to grow all types of crops. KEY WORDS: Drumstick Fermented Leaf Juice (DFLJ), Jeevamrit, Humic acid, consortium, Organic cultivation, Tomato, Moringa leaf extract
Cite this article: Rajamani R, Rudresh Kumar Singh, Vinod Kochupillai, Mohit Aggarwal, Sivaraj A K (2015), DRUMSTICK FERMENTED LEAF JUICE (DFLJ) - A PROMISING ORGANIC SIGNATURE FOR TOMATO CULTIVATION PACKAGE, Global J Res. Med. Plants & Indigen. Med., Volume 4(1): 10–19
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INTRODUCTION: For most of our history, agriculturists have been blinded by overzealous imperative of more, more in agriculture-growing more crops, clearing more land to grow crops, using more synthetic fertilizers to increase yield and to help meet demands and natural resources. Farming of both livestock and crops is the largest human endeavor on earth using more than 19.4 million square miles (36%) from the total earth surface area of 196.9 million square miles to feed the current world population. In India 89% crop produce are still consumed directly by humans (Jonathan Foley, 2014). It would be far easier to feed nine billion people by 2050 if more of the crops we grew ended up in human stomachs by tackling of wastage effectively (Jonathan Foley, 2014). The more scientifically challenging questions here are; Is it possible to get a good yield and manage the insects and pests without using chemical fertilizer and synthetic pesticide? Will a shift to organic farming affect our food security? Will Organic cultivation still be profitable for farmers? These are the questions often asked by farmers when problems of modern agriculture are being discussed. Though practically proved and organic cultivation technologies are available to farmers; the main issue is lack of proper communication and dissemination of information to them. Many farmers still practice traditional organic agriculture and are either not aware or not prepared to make use of proper organic inputs to boost their yields. Many traditional farmers have been striving hard for several years to indigenously develop many articulate organic inputs to improve the reliability of organic cultivation and to take traditional cultivation to greater heights (Masanobu Fukuoka, 1978, Binbumathi Mohan,2008).The successful reentry of different commercial bio organic inputs like bio fertilizer, humic acid, sea weed extract and non commercial products such as Panchagavya, Amirt pani, and Jeevamrit (prepared from cow by product) were the greatest land mark achievement in agriculture footprint(Binbumathi Mohan,2008).
Among myriad of natural plants, Moringa oleifera is called miracle vegetable because it is a medicinal as well as functional food and it possesses highly therapeutic and pharmacological values (Paliwal R et al., 2011). The leaves are rich in starch, minerals, iron, vitamins A, B6, C, Riboflavin, Calcium, Magnesium and Protein (Fisher E, 2011). With a weight for weight comparison, it has four times the vitamin A in carrot, seven times the vitamin C in oranges and four times the calcium in milk (www.villagevolunteers.org, Jed W.Fahey, 2005 & Satya Prakash Mishra et al, 2012) M. oleifera is one such alternative, being investigated to ascertain its effect on growth and yield of crops and thus can be promoted among farmers as a possible supplement or substitute to inorganic fertilizer (Phiri, 2010). Although M. oleifera plant extract is known to possess diverse medicinal and biological activity on human and animals, little is known scientifically about its effect as bioorganic fertilizer on the hormonal, metabolic and antioxidant potential on plants (Mona M. Abdalla, 2013). The study presented here focuses on the establishment of the vitality of Drum stick fermented leaf juice (DFLJ) of M. oleifera leaves on tomato plant. This liquid fermented leaf juice has consistently improved transplant growth, survival and yield on a variety of crops in combination with Panchagavya, Humic acid, Jeevamrit and consortium. Objectives of this research were to: (1) monitor the DFLJ applied as aqueous organic input to potting culture at seedling and at various times during the period of cultivation of tomato. (2) Determine treatment effect and assess the optimum dosage of DFLJ along with Jeevamrit, Humic acid and consortium for Tomato cultivation. MATERIALS AND METHODS:The seeds of Tomato were obtained from Perma culture, division of Sri Sri Institute of Agricultural Science and Technology Trust
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(SSIAST), Art of living International center, Bangalore, Karnataka, India. Preparation of Soil Mixture:-
Preparation of Nursery Bed:The seeds of tomato were sown in plastic pots filled with equal amount of red soil, cocopeat and cow dung. Twenty days old seedlings were transplanted into experimental pots.
To evaluate DFLJ application, a specially designed soil mixture was prepared. The composition of soil mixture contains 3 Kg of Preparation of Drumstick fermented leaf Coco-peat, 4 Kg of red soil and 1Kg of cow Juice (DFLJ) and Jeevamrit. dung (3:4:1), mixed thoroughly and filled in DFLJ and Jeevamrit solution was prepared the experimental pots (1×1×1 ft). 32 as per methodology cited by Vijayan Pillai experimental pots were arranged for four (2012), & Rajamani et al. (2014) (Table-1). treatments and each treatment contains eight replicates. Table:-1.component of Drumstick fermented leaf Juice (DFLJ) and Jeevamrit.
Sr.No 1 2 3 4 -
DFLJ Inputs Drum stick leaves Sea salt (raw) Tamarind Water -
Quantity 3Kg 3g 3g 3Lit. -
Experimental Design:Twenty days old seedlings of tomato were transplanted to the pot containing soil mixture along with different quantity of bioorganic inputs. Each set of treatment had eight plants. The first set of eight plants of tomato were treated with 15 ml of DFLJ/plant, second set 20 ml/plant and third set 25 ml/plant at the time of transplantation. The same treatment was repeated again on 15th day and 30th day on tomato plants along with 10 ml of Jeevamrit solution and 1 gm of bacterial and fungal consortium / plant (Product obtained from Indian institute of Horticulture Research (IIHR), Bangalore). Irrigation was done by common agricultural practice throughout the experiment. 5 ml of Humic acid / plant was applied to the soil mixture during the transition period, 10 ml of Jeevamrit/ plant was added at every ten days interval. Analysis of Biomass: For biomass parameter analysis eighty days old tomato plants were uprooted from the pot
Sr.No 1 2 3 4 5 6
Jeevamrit Inputs Cow dung Cow urine Besan flour Jaggery Native soil Water
Quantity 10 Kg 10 Lit 2 Kg 2 Kg 200 g 200 Lit.
carefully and washed with tap water to remove the soil. For fresh and dry biomass analysis, the root and shoot were separated and placed in a paper envelop after weighing the fresh individual plants and measuring the root volume. The root and shoot were kept in oven at 60ο C for three days to remove complete inbound moisture (Gamalero, 2004). The dry weight of root and shoot of individual plants was recorded. The other parameters like number of leaves, flowers, root length and shoot length were also recorded. SPSS-19 version has been used to analyze the data. RESULTS The influence of DFLJ application on tomato attributes had significantly differed between the dosages. Invariably all the parameters had responded well for the treatment in combination of DFLJ, Jeevamrit, consortium and humic acid in each individual plant. The result showed that the plant treated with 45 ml DFLJ along with 3gm consortium, 50 ml Jeevamrit and 5 ml Humic acid significantly increased number of leaves
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Global J Res. Med. Plants & Indigen. Med. | Volume 4, Issue 1 | January 2015 | 10–19
(164.63), flowers (20.13), fresh shoot weight (162.125 gm), dry shoot weight (25.49 gm), dry root weight (14.71 gm) and root volume (79.38 cm3) than control (Fig-1). The analysis of data showed that the plant tested with different dosage of DFLJ did not
affect the mean fresh root weight and root length. A highly significant increase in number of leaves, fresh shoot weight, dry shoot weight and dry root weight were recorded in case of plant treated with 45 ml followed by 60 ml and 75 ml DFLJ along with Jeevamrit, consortium and Humic acid than control (Table -2 and 3).
Fig: - 1. Graph of Vegetative and reproductive growth in Tomato.
Table:-2. Effect of various dosage of DFLJ on Biomass of Tomato. Treatment groups
Control Dose45ml Dose60ml Dose75ml
Mean no leaves
Number of flowers
Fresh shoot. Wt (gm)
103.57 164.63
7 20.13
96.286 162.125
142.33
16.67
140.75
11.50
Fresh root Wt (gm)
Dry shoot Wt (gm)
Dry root Wt (gm)
64.71 81.5
15.6386 25.4975
7.0971 14.7138
105.29 124.5
51.71 62.71
58.57 79.38
138.667
59.08
23.7583
7.7383
97
50
55
110.563
46.43
17.4763
6.2688
141.25
48.88
51.25
Similarly the per cent increase in total biomass both fresh and dry in combined application at lower concentration over the control was 51.3% (45 ml), 22.8 % (60 ml) and −2.48 % (75 ml) (plates-1-8). The total dry biomass was 23 % (45 ml), 3.81 % (60 ml) and 4.44 % (75 ml). The plant treated with 75 ml DFLJ showed fresh biomass (156.99gm) and dry biomass (23.75gm) were on par with
Root length (cm)
Shoot length (cm)
Root volume
(cm3)
control (160.99 and 22.74 gm).The inbound water (cytosol) of plant system in each treatment were analyzed and compared with control. It showed that the highest inbound water holding capacity in 45 ml/plant was 47.13% followed by 20.26% in 60 ml/plant, −3.62% in 75 ml/plant and −3.75% in control (Table-4).
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Table:-3. Analytical Data of various dosage of DFLJ on Biomass of Tomato. Treatments
Control
DFLJ DOSE 45ml/plant
DFLJ DOSE 60ml/plant
DFLJ DOSE 75 ml/ plant
Biomass Parameter
Mean
Mean
p-value
Mean
p-value
Mean
p-value
No. of leaves
103.57
164.63
0.000**
142.33
0.03*
140.75
0.008*
No. of flowers
7.00
20.13
0.006*
16.67
.000*
11.50
0.190
Fresh shoot wt (gm).
96.286
162.125
0.003**
138.667
0.074
110.563
0.363
Fresh root wt (gm).
64.71
81.50
0.181
59.08
0.401
46.43
0.315
Dry shoot wt (gm).
15.6386
25.4975
0.004**
23.7583
0.053
17.4763
0.325
Dry root wt (gm)
7.0971
14.7138
0.002**
7.7383
0.431
6.2688
0.365
Root length (cm)
105.29
124.50
0.122
97.00
0.129
141.25
0.620
Shoot length (cm)
51.71
62.71
0.023*
50.00
0.734
48.88
0.453
Root Volume (cm3)
58.57
79.38
0.055*
55.00
0.646
51.25
0.328
Test applied: Independent t test (Testing of means),*Statistically Significant at 5% level of Significance (p<0.05) **Highly Significant at 5% level of Significance (p<0.005).
Plates: Effect of DFLJ on Areial growth of Tomato
Plate 1: - 30 Days- Control Plant.
Plate 2:- 30 Days old- 45 ml DFLJ/ Plant.
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Plate 3:- 30 Days old- 60 ml DFLJ/ Plant.
Plate 4:- 30 Days old- 75 ml DFLJ/ Plant.
Effect of DFLJ on Root Architecture of Tomato:
Plate 5:- Control DFLJ Tomato Plant
Plate: 7- 60 ml DFLJ/ Tomato Plant
Plate 6:- 45ml DFLJ/ Tomato Plant
Plate: 8 -75 ml DFLJ/ Tomato Plant
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Table: - 4.Differential Effect of DFLJ on Biomass of Tomato Plant. Treatment groups
Total fresh shoot & Root Wt (gm)
Differences in wet biomass Wt (gm)
Differences in wet biomass %
Total Dry shoot& Root Wt (gm)
Differences in dry biomass Wt (gm)
Differences in dry biomass %
Inbound Water
Control Dose-
Differences inbound water %
160.99 243.63
0 82.64
0 51.3
22.74 40.21
0 17.47
0 23
138.25 203.42
0 47.13
197.75
36.76
22.8
31.49
8.75
3.814
166.26
20.26
156.99
−4
−2.48
23.75
1.01
4.44 13
133.24
−3.62
45 ml
Dose60 ml
Dose75 ml
DISCUSSION Owing to the reported potential performance of fresh drum stick leaf juice on several crops and, DFLJ in B. olerarcea plant (Rajamani et al., 2014), the present study was carried out in an attempt to elucidate its effect on vegetative and reproductive growth of pot cultured tomato plant. The composition of Drumstick fermented leaf juice was first developed and applied on tomato and papaya at AL Khaly farm in UAE to withstand temperature and better yield was observed (Vijayan Pillai, 2012). DFLJ showed significant and consistent improvement in vegetative growth and also significantly increased the number of flowers and number of leaves at different intervals throughout the period of experiments as compared to the control indicating its potent growth stimulating activity. It is well recognized that the foliar application of fresh Drumstick leaf and twigs juice along with Panchagavya, Humic acid, and de oiled seed cake can positively influence the plant biomass parameters (Prabhu, 2010; Balakumbahan and Rajamani, 2010; Emmanuel 2011a & b). It has been reported that 30 times diluted Moringa leaf extract significantly increased seed and seedling vigour in wheat (Afzal et al., 2008), Maize (Basra et al., 2011) and many grass species including Cenchrus ciliaris, Panicum antidotala and Echinochola crusgalli (Nouman
et al., 2012a). Moringa leaf extract spray increased the yield in crops like peanut (5319 kg/hec), onion (4194 kg /hec) and black bean (1194 kg / hec) compared to their respective control (Foidl et al., 2001). Several experiments were sustained that the increased growth and yield parameters by foliar application of Moringa leaf extracts at different percentage and combined with Panchagavya and Humic acid. The mixture of all different organic substances are economically viable for producing higher dry herbage yield in sacred basil - Ocimum sanctum L., (Prabhu et al., 2010). The same impacts of higher value of growth and yield were recorded in Senna crop Cassia angustifolia var.KKM.1 (Balakumbahan and Rajamani, 2010). The field investigation established the fact that application of M. oleifera de-oils seed cake without pre-decomposition as an organic fertilizer on a maize farm achieved significant improvement on soil nutrient as well as the plant yield, as compared to the control (Emmanuel et al., 2011a & b). Moringa leaf extract (MLE) is a plant growth enhancer that improves seedling emergence of rangeland grasses, seedling vigor and growth as compared to other seed priming techniques (Nouman et al., 2012 a & b). Wheat (Triticum aestivum) seeds priming with Moringa oleifera leaf extract had induced the antioxidative system together with
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increased chlorophyll contents, ascorbic acid and soluble phenolics contents (Yesmeen et al., 2013).
might be contributed to free microbial metabolites of Jeevamrit solution and beneficial inoculum of consortium.
The foliar spray of aqueous extracts of 2% of leaf and 3% of twig of Moringa oleifera on Rocket plants (Eruca vesicaria) potentially increased all measured growth criteria of Biomass and physiological activities like photosynthetic rates, stomatal conductance, the Bio-molecules of chlorophyll a and b, carotenoids, total sugar, total protein, phenols, ascorbic acid, N, P, K, Ca, Mg, Fe as well as growth promoting hormones-Auxins, gibberellins and cytokinins. The extracts at all concentrations applied, negatively reduced the level of lipid peroxidation and the activities of antioxidant enzymes (Catalase, proxidase, superoxide, dismutase). Thus it is concluded that Moringa oleifera leaf and twig extracts can be used as bio-organic fertilizer for various crops due to its high productivity, high nutritive value, antioxidant effect, and easy preparation, low cost and environmental friendly nature. (Mona M. Abdalla, 2013).
The result is in accordance with the previous results conducted with DFLJ and aqueous extract of drum stick leaves further suggesting that the available form of active principles (Not analyzed) in DFLJ and higher inbound water in plant system (45 ml / plant) might be contributing in the regulation of plant hormones. Taken together, it can be summarized as DFLJ might possess both vegetative and reproductive stimulation mechanisms in it actions and such apparent dual action of DFLJ would be more advantageous to the other existing commercial organic inputs.
In our previous study we showed the empirical evidence that the treatment of DFLJ caused significant as well as moderate improvement on biomass level of Brassica olerarcea .L, (Rajamani et al., 2014) further sustaining its potential in tomato. Since DFLJ effectively improves the root surface area, it causes early flowering in tomato. The significant growth perhaps might include the establishment of favorable microclimate niche around the rhizosphere for consortium and Jeevamrit microbial community to enhance feeding root growth. These evidences tempt us to speculate that apart from the aforementioned probable DFLJ action, the other possible mechanism i.e. development of disease resistant in tomato
CONCLUSION The present study demonstrates that DFLJ along with consortium, Jeevamrit and Humic acid at the dose level tested exhibits potent increased wet and dry plant biomass, root volume and number of flowers in tomato. The organic agent with multiple advantageous properties and less preparation cost would be greater organic cultivation package for farmers. ACKNOWLEDGEMENT The authors are grateful to Mr. Prasanth S Nair, Director, VVMVP Trust, Art of Living International Center, Bengaluru, India for generously supporting this work. We wise to thank Mr.Vinay.R .G, (VVMVP Trust), Binay Kumar Singh, Perma culture-supply of seeds (SSIAST) and Karthik Krishna (ABC-Editing ) for providing facilities and showing keen interest for completion of the work. This entire work was supported by grants from VVMVP Trust, Art of Living International Center, Bengaluru, India.
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Basra SMA, Iftikhar MN, and Afzal I (2011) Potential of Moringa (Moringa oleifera) leaf extract as priming agent for hybrid maize seeds. International Journal of Agriculture and Biology. 13; 1006– 1010. Emmanuel S.A, Zaku S.G and Thomas S.A (2011a). Biodiversity and agricultural productivity enhancement in Nigeria: Application of processed Moringa oleifera seeds for improved organic farming. Agric. Biol. J. N. Am. 2(5): 867–871. Emmanuel S.A, Zaku S.G, Adedirin s.o,Tafida M and Thomas S.A (2011b). Moringa oleifera seed cake, alternative biodegradable and biocompatibility organic fertilizer for modern farming. Agric. Biol. J. N. Am. 2(9): 1289–1292. Fisher E (2011). Nutritional value: Amazing facts about Moringa oleifera (Adaptogen) Zeal for life wellness formula. Available on: http:myzealdrink.ericfisher.com. Foidl, N., H.P.S. Makkar and K. Becker, 2001. The potential of Moringa oleifera for agricultural and industrial uses. International workshop on “Development potential for Moringa products”. October 29th to November 2nd, 2001. Fuglie, L. New Uses of Moringa Studied in Nicaragua. ECHO Development Notes#68,June,2000.<http://www.echot ech.org/network/modules.php?name=N ews&file=article&sid=194> Gamalero, E., Martinnoti, A. Trotta, P. Lemanceau and G. Berta, (2004). Morphogenetic modification induced by Pseudomonas Fluorescence A6RI and Glomus mossseae BEG12 in the root system according to plant growth conditions. New Phytol.155:293–300.
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Prabhu M, Ramesh Kumar A, Rajamani K (2010). Influence of different organic substances on growth and herb yield of sacred basil (Ocimum sanctum L.,). Ind. J. Agric. Res. 44(1): 48–52.
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Source of Support: VVMVP Trust, Art of Living International Center, Bengaluru, India.
www.villagevolunteers.org Yesmeen A, Basra SMA, Wahid A,Nouman W, Rehman H (2013) Exploring the potential of Moringa oleifera leaf extract (MLE) as a seed priming agent in improving performance.Turk.J.Bot, 37:512–520.
Conflict of Interest: None Declared
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