Ganapathy Selvam, G et al. IRJP 2013, 4 (1)

Page 252

INTERNATIONAL RESEARCH JOURNAL OF PHARMACY www.irjponline.com ISSN 2230 – 8407

Research Article

DEVELOPMENTAL CHANGES IN THE GERMINATION, GROWTH AND CHLOROPHYLLASE ACTIVITY OF

VIGNA MUNGO L. USING SEAWEED EXTRACT OF ULVA RETICULATA FORSSKAL. Ganapathy Selvam, G.*, Balamurugan. M., Thinakaran, T. and Sivakumar, K

Division of Algal Biotechnology, Department of Botany, Faculty of Science, Annamalai University, Annamalainagar-608 002, Tamil Nadu, India

Article Received on: 12/11/12 Revised on: 17/12/12 Approved for publication: 03/01/13

*Email: kshivam69@gmail.com ABSTRACT The effect of seaweed extract prepared from Ulva reticulata on seed germination, seedling growth and chlorophyllase activity of Vigna mungo L. was studied. 100% germination was recorded in the seeds treated with lower concentration of seaweed extract. The V. mungo seeds soaked with lower concentrations of the seaweed extracts showed higher rates of germination, while the higher concentrations of the extracts inhibited the germination. Key words: Ulva reticulata, Vigna mungo, chlorophyllase activity, seed germination INTRODUCTION Naturally occurring organic fertilizers differ from chemical fertilizers, in that; they feed plants while building the soil’s structure. Seaweed fertilizer is a natural bioactive material, water-soluble derived from marine macroalgae. Seaweed fertilizer could be absorbed by plant within several hours after application and safe to humans, animals and the environment. Modern agriculture with use of chemical fertilizers and resulted in an increase of food production. Seaweed liquid fertilizer is widely used as foliar spray to increase yield in many commercial crops1-3. Seaweed and seagrass liquid fertilizer is a bio-organic manure that contains many growth promoting substances like auxins, gibberellins, trace elements, vitamins, amino acids and micronutrients4,5. Seagrasses are the most important and abundant marine resources available and they are not fully exploited6. Seaweeds have been used as manure; cattle feed, food for human consumption as a source of minerals like soda, potash, iodine and also as a source of phycocolloids like agar, alginate and carrageenan7. Seaweed fertilizer was found to be superior to chemical fertilizer because of the high level of organic matter aids in retaining moisture and minerals in the upper soil level available to the roots8. On pepper9,10 and on mung bean indicated that application of seaweed extract significantly increased seed yield and pod weight as well as improved nutrional values of seeds, i-e., protein and carbohydrates11. In recent years the use of natural seaweed products as substitutes to the conventional inorganic fertilizers assumed importance12,13. On the basis of these qualities, the seaweed has been tried in the form of concentrated extracts, which are being marketed under different trade names in Western countries14. In the present study, the developmental changes growth and chlorophyllase activity of Vigna mungo seedling was assessed by using seaweed extract of Ulva reticulata. MATERIALS AND METHODS The seaweed Ulva reticulata was collected from Cinnamuttom coast of Kanyakumari District in Tamil Nadu. It lies Lat.9º11’N,’Long.79º24’E in March 2012. They were handpicked and washed thoroughly with sea water and finally

with fresh water to remove all the unwanted impurities, epiphytes and adhering sand particles. They were shade dried for 4 days followed by oven drying for 24th at 60°C. The oven dried material was taken for the preparation of seaweed Liquid Fertilizer (SLF) as per the method15. Healthy selected black gram seeds (Vigna mungo L.) were soaked in beakers containing equal volume of different concentrations (1%, 2.5%, 5%, 7.5% and 10%) of seaweed extracts for 24 hours. The control and experimental seeds were treated with distilled water regularly for every 24 hours and experimental seeds were treated with seaweed liquid fertilizer of various concentrations (1%, 2.5% 5%, 7.5% and 10%) once in 5 days. The rate of germination, growth, viz. root and shoot length were recorded from the emergence of 3rd leaf upto its senescence. The chlorophyllase activity was assayed by the method16. RESULTS V. mungo showed 78% germination in the control and 100% in the 2.5% seaweed liquid fertilizer treated seeds. A minimum of 76% germination was observed at 10% seaweed liquid fertilizer when compared to all other concentrations of treatment (Table 1). It was found that the treated seeds showed higher percentage of germination than control. The results on root length and shoot length in V. mungo are presented in Table 2 and 3. The root and shoot length gradually increased from 7th day to 42nd day. Maximum root and shoot length was found in V. mungo treated with 2.5% concentration of seaweed liquid fertilizer and minimum in seedling at 10% concentration. In V. mungo, the enzyme chlorophyllase activity showed a decreasing trend upto 21st day and then increased upto 42nd day in seaweed liquid fertilizer treated seedlings. Maximum chlorophyllase activity was found in V. mungo treated with 2.5% concentration of seaweed liquid fertilizer and minimum in seedlings treated with 10% concentration (Table-4). It was found that the 2.5% concentration of seaweed liquid fertilizer showed a significant result on chlorophyllase activity when compared to other concentrations of seaweed liquid fertilizer treatment and control.

Ganapathy Selvam, G et al. IRJP 2013, 4 (1)

Page 253

Table: 1 Effect of Ulva reticulata liquid fertilizer of on germination of Vigna mungo L.

Concentrations (%) Germination (%) Control 78±4.05

1% 95±2.10 2.5% 100±1.20 5% 93±5.15

7.5% 85±4.75 10% 76±3.65

Table: 2 Effect of Ulva reticulata liquid fertilizers on Shoot length of Vigna mungo L.

Days Control Shoot length (cm/Plant at different concentrations) 1% 2.5% 5% 7.5% 10%

7 16.0±.053 19.8±0.31 20.5±0.52 18.5±0.39 17.3±0.04 16.0±0.09 14 16.2±0.30 19.9±0.27 21.7±0.53 18.9±0.40 17.2±0.32 15.7±0.10 21 16.8±0.35 20.0±0.33 24.3±0.51 19.0±0.45 17.8±0.51 16.1±0.35 28 17.2±0.47 20.5±0.43 27.6±0.71 19.3±0.71 17.3±0.48 16.6±0.25 35 18.5±0.60 20.9±0.54 30.5±0.43 19.7±0.67 18.2±0.07 17.8±0.20 42 19.0±0.73 21.5±0.63 32.3±0.97 21.2±0.21 19.7±0.71 18.1±0.15

*day – The day of emergence of 3rd leaf (i.e. the 7th day after germination)

Table: 3 Effect of Ulva reticulata liquid fertilizers on Root length of Vigna mungo L. Days Control Root length (cm/Plant at different concentrations)

1% 2.5% 5% 7.5% 10% 0-7 1.9±0.71 4.0±0.57 4.6±0.71 3.5±0.17 2.3±0.15 1.5±0.41 8-14 2.1±0.65 5.5±0.49 7.2±0.31 4.8±0.21 2.9±0.21 1.9±0.31

15-21 4.5±0.23 7.7±0.50 7.8±0.27 5.5±0.33 3.5±0.13 3.0±0.13 22-28 5.5±0.41 8.3±0.53 8.4±0.23 6.8±0.43 4.5±0.12 4.3±0.20 29-35 6.3±0.32 9.8±0.59 9.8±0.28 7.9±0.57 4.9±0.45 4.3±0.33 36-42 8.0±0.29 10.1±0.72 11.9±0.33 8.8±0.21 7.1±0.53 6.8±0.39

*day – The day of emergence of 3rd leaf (i.e. the 7th day after germination)

Table: 4 Effect of Ulva reticulata liquid fertilizers on Chlorophyllase activity of Vigna mungo L. Days Control Chlorophyllase activity (A663nm) at different concentrations

1% 2.5% 5% 7.5% 10% 0-7 0.531±0.002 0.671±0.001 0.697±0.002 0.678±0.003 0.545±0.003 0.530±0.001

8-14 0.443±0.003 0.652±0.010 0.690±0.003 0.621±0.003 0.508±0.004 0.418±0.002 15-21 0.168±0.004 0.648±0.015 0.410±0.001 0.278±0.002 0.171±0.002 0.138±0.007 22-28 0.313±0.005 0.390±0.018 0.570±0.020 0.413±0.004 0.310±0.004 0.298±0.002 29-35 0.610±0.023 0.570±0.59 0.610±0.030 0.579±0.006 0.477±0.005 0.471±0.004 36-42 0.687±0.028 0.690±0.021 0.750±0.013 0.676±0.007 0.591±0.016 0.517±0.005

DISCUSSION The Vigna mungo seeds soaked with lower concentrations of the seaweed extracts showed higher rates of germination, while the higher concentrations of the extracts inhibited the germination. The increased germination percentage at low concentrations may be due to the presence of some growth promoting substances such as IAA and IBA, Gibberellins (A&B), cytokinins, micronutrients (Fe, Cu, Zn, Co, Mo, Mn, Ni), vitamins and amino acids17. The increased seed germination percentage at low concentrations may be due to the presence of growth promoting substances18. The present finding is in consonance with the earlier results7,19,20. Similar results were recorded in Cajanus cajan red gram19 and Oryza sativa21. Reported that the presence of plant growth regulators, trace elements, vitamins and macronutrients in the seaweed liquid fertilizer enhance the growth of root length and shoot length of Vigna mungo17. The present investigation is in agreement with the earlier studies22,23,24. The higher concentrations showed a decreasing trend. Similar results were recorded in Padina which induced maximum seedling growth at lower concentrations in C. cajan and Vigna radiata and C. cajan19. Reported similar findings with Hypnea musciformis, Spatoglossum asperum, Stoechospermum marginatum and Sargassum on the growth of crops such as green chillies, turnips and pineapple25. The increase in chlorophyllase activity results in lowering of chlorophyll content26. Reported that an enhancement of chlorophyllase activity was more in chlorotic leaf substrate as compared to normal green leaf substrate when iron, copper and zinc were

supplemented to the chlorotic leaf enzyme 27. Reported the decrease in chlorophyll content under salinity is associated with higher activity of enzyme chlorophyllase28. Also reported decrease in the levels of photosynthetic pigments under saline conditions with an increase in chlorophyllase activity29. CONCLUSION The extract of Ulva reticulata found to have promising result by possessing fertilizer activity to enhance the germination and growth of Vigna mungo. Hence, this simple practice of application of ecofriendly seaweed liquid fertilizers to pulses is recommended to the growers for attaining better germination, growth and yield. ACKNOWLEDGEMENT The authors are grateful to Head, Department of Botany, Annamalai University, Annamalainagar for providing necessary facilities to carry out this work and University Grants Commission, New Delhi for financial assistance. REFERENCES 1. Balakrishnan, C. P. Venkataraman Kumarand V.R. Mohan 2007.

Studies on the effect of crude seaweed extract on seedling growth and biochemical parameters in Peninsetum typhoides (Burn. F.) Stapf C.E. Hubbard. Seaweed Res. Utiln., 29(1&2): 89-96.

2. Ramamoorthy, K., K. Sujatha and K. Sivasubramanian 2007. Utilization of seaweedextracts for enhancing yieldin blackgram, Vigna mungo L. Hepper. Seaweed Res. Utiln., 29(1&2): 97-100.

3. Renukabai, N., N.R. Laila Banu, S. Jaquilin Goldi and J.W. Prakash 2008. Effect of seaweed extracts (SLF) on the growth and yield of Phaseolus aureus L. Indian Hydrobiol., 11(1): n 113-119.

Ganapathy Selvam, G et al. IRJP 2013, 4 (1)

Page 254

4. Metting, B., W.J. Zimmerman, I.J. Crouch and J. Van Staden, 1990. Agronomic uses of seaweed and microalgae. In: Introduction to applied Phycology, I.A. Akatuska (ed.): SPB Academic Publishing, The Hague, Netherlands. pp. 269-307

5. Sylvia, S., M. Baluswami, M.D. Vijaya Parthasarathy and V. Krishnamurthy 2005. Effect of liquid seaweed fertilizers extracted from Gracilaria edulis (Gmel.) Silva, Sargassum wightii Greville and Ulva lactuca Linn. on the growth and yield of Abelmoschus esculentus (L) Moench. Indian Hydrobiol., 7: 69-88.

6. Kaliaperumal, N., S. Kalimuthu and J.R. Ramalingam, 2004. Present scenario of seaweed exploitation and industry in India. Seaweed Res. Utiln., 26(1&2): 47-53.

7. Anantharaj, M. and V. Venkatesalu 2001. Effect of seaweed liquid fertilizer on Vigna catajung. Seaweed Res. Utiln., 23 (1& 2): 33-39.

8. Wallen Kemp, J.O., 1955. Treasure from the sea. Organic Gard. F. 2(3), 52–53.

9. Zodape S.T., V. J Kawarkhe, J S Patolia and A.D Warade 2008. Effect of liquid seaweed fertlizer on yield and quality of okra (Abelmoschus esulentus L.) Journal of Scintific and Industrial Research Vol. 67, December, pp. 1115 – 1117.

10. Arthur G D , Stirk W A and Van Staden J , 2003 . Effect of seaweed concentrates On the growth and yield of three varieties of Capsium annuum , South. Afr. J Bot, (69) 207 -211.

11. Zodape S.T.,S. Mukhopadhyay, K.Eswaran,M.P. Reddy and J. Chikara 2010 . Enhanced yield and nutritional in green gram (Phaseolus radiata L ) treated with seaweed (Kappaphycus alvarezii)extract J. of Sci. and Ind. Res. Vol. 69 , June 2010 , pp. 468 -471.

12. Crouch, I.J. and J. Van Staden, 1993. Evidence for the presence of growth regulator in commercial seaweed product. Plant Growth Regulators, 13: 21-29.

13. Gangatharan, M., 1998. Studies on the effect of photo organic compounds on growth and physiological activities of some crop plants, M. Phil., Dissertation, Madurai Kamaraj University, India

14. Stephenson, W. A., 1965. The effect of hydrolyzed seaweed on certain pests and diseases, Fifth International Seaweed Symposium, Halifax.

15. Rama Rao, K., 1990. Preparation of liquid seaweed fertilizer from Sargassum, Seaweed Research and utilization Association Workshop on algal products and Seminar on Phaeophyceae in India, Madras, India, 4-7 June.

16. Holden, M. 1961. The break down of chlorophylls by chlorophyllase. Biochem. J., 78: 359-364.

17. Challen SB, Hemingway JG (1965) Growth of higher plants in response to feeding with seaweed extract. Proc Fifth Int. Seaweed Symp. 5: 359-367

18. Jennings, D.L. and B.M.M. Tulloch 1965. Growth promoting hormones in seaweeds. J. Exp. Bot., 16: 329.

19. Venkataraman Kumar V, Mohan VR, Murugeswari R, Muthusamy M (1993) Effect of crude and commercial seaweed extracts on seed germination and seedling growth in green gram and black gram. Seaweed Res. Utiln. 16 23-27

20. Anantharaj,M. and V.Venkatesalu,2002. Studies on the effect of seaweed extracts on Dolichos erglorus. Seaweed Res. Utiln., 24(1) 129-138.

21. Asir Selin Kumar, 2009. Influence of seagrass Syringodium isoetifolium extract on germination and growth of Oryza sativa. J. Ecotoxicol. Environ. Onit., 19 (2): 157-162.

22. Sobithabai, R., J. Irene Wilsey and R. Asir Selin Kumar 2005. Influence of seagrass liquid fertilizer on cotyledons in cyamosis tetragonoloba Taub. Seaweed Res. Utiln., 27(1& 2): 155-157.

23. Sivasankari, S., M. Chandrasekaran, M.Kannathasan and V. Venkatesalu 2006. Studies on the biochemical constituents of Vigna radiata Linn. treated with seaweed liquid fertilizer. Seaweed Res. Utiln., 28(1): 151-158.

24. Sahaya Anthony Xavier, G.and L. Louis Jesudas 2007. Effect of seaweed extract on cluster bean. Seaweed Res. Utiln., 29(1&2): 85-87

25. Dhargalkar, V.K., Untawale, A.G., 1983. Some observations of the effect of SLF, on higher plants. Indian J. Mar. Sci. 12, 210–214.

26. Lange, O.L. 1975. Progress in Botany. Springer Verlag, Berlin. 27. Srivastava, M.K. and A.K. Shrivastava 2004. Induction of

chlorophyllase activity in normal green and chlorotic leaves of sugarcane. Indian J. Plant Physiol., 9(1): 62-64.

28. Reddy, M.P. and A.B. Vora 1986. Salinity induced changes in pigment composition and chlorophyllase activity of wheat. Indian J. Plant physiol., 29(4): 331-334.

29. Rao, G.G. and G.R. Rao 1981. Pigment composition and chlorophyllase activity in pigeon pea Cajanus indicus spring and gingelly (Sesamum L.) under NaCl salinity. Indian J. Exp. Biol., pp. 768-770.

Source of support: Nil, Conflict of interest: None Declared IRJP is an official publication of Moksha Publishing House. Website: www.mokshaph.com. All rights reserved.