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Research Article

volume 53 issue 5 (october 2019) : 589-593,   Doi: 10.18805/IJARe.A-5169

An efficient protocol for rapid plant regeneration from deembryonated cotyledons of black gram [Vigna mungo (L.) Hepper]

R
R. Anandan
T
T. Deenathayalan
R
R. Bhuvaneshwari
M
M. Merlin Monisha
M
M. Prakash
1Department of Genetics and Plant Breeding, Faculty of Agriculture, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India. 
Cite article:- Anandan R., Deenathayalan T., Bhuvaneshwari R., Monisha Merlin M., Prakash M. (2019). An efficient protocol for rapid plant regeneration from deembryonated cotyledons of black gram [Vigna mungo (L.) Hepper]. Indian Journal of Agricultural Research. 53(5): 589-593. doi: 10.18805/IJARe.A-5169.

ABSTRACT

Here an efficient protocol for micropropagation of black gram [Vigna mungo (L.) Hepper] cv. VBN 3 is reported. The deembryonated cotyledonary explants were cultured on MS medium containing different concentrations of plant growth regulators. The maximum frequency (72%) of direct shoot regeneration (devoid of callus phase), multiple shoot induction and shoot elongation was achieved from culturing the explants on MS medium containing 3.0 mg/l of 6-benzylaminopurine (BAP). Up to 65% of the regenerated shoots were rooted on MS medium containing 0.25 mg/l of á-naphthalene acetic acid (NAA) within 3 weeks after subculturing. The in vitro-raised plantlets were successfully hardened first under culture room conditions with 62% survival rate and then in greenhouse. The identified regeneration system could be efficiently used in various in vitro manipulation studies in black gram as well.

REFERENCES

  1. Abhijeeta, K. N. and Rajesh, B. M. (2018). In vitro plant regeneration in pigeon pea [Cajanus cajan (L.) MILL Sp.] using various explants. Legume Research. 41(2): 226-229. 
  2. Adlinge, P. M., Samal, K. C., Swamy, R. K., Rout, G. R. (2014). Rapid in vitro plant regeneration of black Gram [Vigna mungo (L.) Hepper] Var. Sarala, an important legume crop. Proceedings of the National Academy of Sciences, India Section B. 84(3): 823-827.
  3. Anand, R. P., Ganapathi, A., Vengadesan, G., Selvaraj, N., Anbazhagan, V. R., Kulothungan, S. (2001). Plant regeneration from immature cotyledon derived callus of Vigna ungiculata (L.). Current Science. 80: 671-674.
  4. Anandan , R., Prakash, M., Deenadhayalan, T., Nivetha, R., Sumanth Kumar, N. (2018). Efficient in vitro plant regeneration from cotyledon-derived callus cultures of sesame (Sesamum indicum L.) and genetic analysis of True-to-Type regenerants using RAPD and SSR markers. South African Journal of Botany. 119: 244-251.
  5. Anandan, R., M. Prakash, B. Sunilkumar & T. Deenathayalan. (2019). In vitro transverse thin cell layer culture in mung bean [Vigna radiata (L.) Wilczek]. Indian Journal of Experimental Biology. 57:324-329.
  6. Anonymous, (2016). Agricultural Statistics at a Glance, Directorate of Economics and Statistics, Ministry of Agriculture, Govt. of India (Website http://www.dacnet.nic.in/ean).
  7. Anwar, F., Alghamdi, S. S., Ammar, M. H., Siddique, K. H. M. (2011). An efficient in vitro regeneration protocol for faba bean (Vicia faba L.). Journal of Medicinal Plants Research. 5: 6460-6467.
  8. Chakraborti, D., Sarkar, A., Das, S. (2006). Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.). Plant Cell Tissue and Organ Culture. 86: 101-105.
  9. Chandra, A. and Pental, D. (2003). Regeneration and genetic transformation of grain legumes: an overview. Current Science. 84: 381-387.
  10. Chopra, R. and Saini, R. (2014). Transformation of blackgram [Vigna mungo (L.) Hepper] by barley chitinase and ribosome-inactivating protein genes towards improving resistance to Corynespora leaf spot fungal disease. Biotechnology and Applied Biochemistry. 174(8): 2791-2800.
  11. Das, D, K, Shivaprakash, N., Sarin, N,B. (2004). An efficient regeneration system of black gram (Vigna mungo L.) through organogenesis. Plant Science. 134: 199-206.
  12. Delic, D., Stajkovic, O., Kuzmanovic, D., Rasulic, N., Knezevic-Vukcevic, J., Milicic, B. (2009). The effects of rhizobial inoculation on growth and yield of Vigna mungo L. in Serbian soils. Biotechnology in Animal Husbandry. 25: 1197– 1202.
  13. Duncan, D. B. (1955). Multiple range and multiple F-tests. Biometrics. 11: 1-42.
  14. Eapen, S. (2008). Advances in development of transgenic pulse crops. Biotechnology Advances. 26: 162-168.
  15. Gill, R., Eapen, S., Rao, P.S. (1987). Morphogenic studies of cultured cotyledons of urd bean [Vigna mungo (L.) Hepper]. Journal of Plant Physiology. 139: 1-5.
  16. Ignacimuthu, S., Franklin, G., Melchias, G. (1997). Multiple shoot formation and in vitro fruiting from cotyledonary nodes of Vigna mungo (L.) Hepper. Current Science. 3: 733-735.
  17. Karamany, E.L. (2006). Double purpose (forage and seed) of mungbean production. 1-Effect of plant density and forage cutting date on forage and seed yields of mungbean [Vigna radiata (L.) Wilczeck]. Research Journal of Agriculture and Biological Sciences. 2: 162-165.
  18. Kamboj, R. and Nanda, V. (2018). Proximate composition, nutritional profile and health benefits of legumes – A review. Legume Research. 41(3): 325-332. 
  19. Mony, S. A., Haque, M. S., Alam, M. M., Hasanuzzaman, M., Nahar, K. (2010). Regeneration of black gram (Vigno mungo L.) on changes of hormonal condition. Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 38(3): 140-145.
  20. Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-497.
  21. Muruganantham, M., Ganapathi, A., Amutha, S., Vengadesan, G., Selvaraj, N. (2005) Shoot regeneration from immature cotyledonary nodes in black gram [Vigna mungo (L.) Hepper]. Indian Journal of Biotechnology. 4: 551–555.
  22. Muthukumar, B., Mariamma, M., Veluthambi, K., Gnanam, A. (1996). Genetic transformation of cotyledon explants of cowpea [Vigna unguiculata (L.) Walp] using Agrobacterium tumefaciens. Plant Cell Reports. 15: 980-985.
  23. Prakash, M., Elangaimannan, R., Sunilkumar, B., Narayanan, G. S. (2018). Evaluation of black gram genotypes for drought tolerance based on root dynamics and gas exchange parameters. Legume Research. 41(3): 384-391. 
  24. Rani, V. and Raina, S. N. (2000). Genetic fidelity of organized meristem derived micropropagated plants: a critical reappraisal. In vitro Cellular & Developmental Biology. 36: 319-330.
  25. Rao, C,R. and Chand, P. (2006). Impact of different mating approaches in generating variability in blackgram [Vigna mungo (L.) Hepper]. Legume Research. 24: 174-177.
  26. Sahoo, L., Sugla, T., Jaiwal, P.K. (2002). In vitro regeneration and genetic transformation of Vigna species. In Biotechnology for the Improvement of Legumes [PK Jaiwal, RP Singh, eds], Kluwer, Dordrecht, pp 1-48.
  27. Sainger, M., Chaudhary, D., Dahiya, S., Jaiwal, R., Jaiwal, P. K. (2015). Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume black gram [Vigna mungo (L.) Hepper]. Physiology and Molecular Biology of Plants. 21(4): 505-517.
  28. Saini,l R. and Jaiwal, P.K. (2002). Age, position in mother seedling, orientation, and polarity of the epicotyl segments of black gram [Vigna mungo (L.) Hepper] determines its morphogenic response. Plant Science. 163: 101-109.
  29. Sarma, K. K. and Borah, B.K. (2004). Record of parasitoids of legume pod borer Maruca testulalis (Geyer) in Assam. Insect Environment. 10: 77-78.
  30. Suneja, Y., Kaur, S., Gupta, A. K., and Kaur, N. (2011). Levels of nutritional constituents and antinutritional factors in black gram [Vigna mungo (L.) Hepper]. Food Research International. 44: 621-628.
  31. Thallapathy, S. (2017). In vitro micropropogation of black gram through shoot tip explants by Thidiazuron (TDZ). European Journal of Biomedical and Pharmaceutical Sciences. 4(11): 288-293. 
Published In
Indian Journal of Agricultural Research
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    Research Article

    volume 53 issue 5 (october 2019) : 589-593,   Doi: 10.18805/IJARe.A-5169

    An efficient protocol for rapid plant regeneration from deembryonated cotyledons of black gram [Vigna mungo (L.) Hepper]

    R
    R. Anandan
    T
    T. Deenathayalan
    R
    R. Bhuvaneshwari
    M
    M. Merlin Monisha
    M
    M. Prakash
    1Department of Genetics and Plant Breeding, Faculty of Agriculture, Annamalai University, Annamalai Nagar-608 002, Tamil Nadu, India. 
    Cite article:- Anandan R., Deenathayalan T., Bhuvaneshwari R., Monisha Merlin M., Prakash M. (2019). An efficient protocol for rapid plant regeneration from deembryonated cotyledons of black gram [Vigna mungo (L.) Hepper]. Indian Journal of Agricultural Research. 53(5): 589-593. doi: 10.18805/IJARe.A-5169.

    ABSTRACT

    Here an efficient protocol for micropropagation of black gram [Vigna mungo (L.) Hepper] cv. VBN 3 is reported. The deembryonated cotyledonary explants were cultured on MS medium containing different concentrations of plant growth regulators. The maximum frequency (72%) of direct shoot regeneration (devoid of callus phase), multiple shoot induction and shoot elongation was achieved from culturing the explants on MS medium containing 3.0 mg/l of 6-benzylaminopurine (BAP). Up to 65% of the regenerated shoots were rooted on MS medium containing 0.25 mg/l of á-naphthalene acetic acid (NAA) within 3 weeks after subculturing. The in vitro-raised plantlets were successfully hardened first under culture room conditions with 62% survival rate and then in greenhouse. The identified regeneration system could be efficiently used in various in vitro manipulation studies in black gram as well.

    REFERENCES

    1. Abhijeeta, K. N. and Rajesh, B. M. (2018). In vitro plant regeneration in pigeon pea [Cajanus cajan (L.) MILL Sp.] using various explants. Legume Research. 41(2): 226-229. 
    2. Adlinge, P. M., Samal, K. C., Swamy, R. K., Rout, G. R. (2014). Rapid in vitro plant regeneration of black Gram [Vigna mungo (L.) Hepper] Var. Sarala, an important legume crop. Proceedings of the National Academy of Sciences, India Section B. 84(3): 823-827.
    3. Anand, R. P., Ganapathi, A., Vengadesan, G., Selvaraj, N., Anbazhagan, V. R., Kulothungan, S. (2001). Plant regeneration from immature cotyledon derived callus of Vigna ungiculata (L.). Current Science. 80: 671-674.
    4. Anandan , R., Prakash, M., Deenadhayalan, T., Nivetha, R., Sumanth Kumar, N. (2018). Efficient in vitro plant regeneration from cotyledon-derived callus cultures of sesame (Sesamum indicum L.) and genetic analysis of True-to-Type regenerants using RAPD and SSR markers. South African Journal of Botany. 119: 244-251.
    5. Anandan, R., M. Prakash, B. Sunilkumar & T. Deenathayalan. (2019). In vitro transverse thin cell layer culture in mung bean [Vigna radiata (L.) Wilczek]. Indian Journal of Experimental Biology. 57:324-329.
    6. Anonymous, (2016). Agricultural Statistics at a Glance, Directorate of Economics and Statistics, Ministry of Agriculture, Govt. of India (Website http://www.dacnet.nic.in/ean).
    7. Anwar, F., Alghamdi, S. S., Ammar, M. H., Siddique, K. H. M. (2011). An efficient in vitro regeneration protocol for faba bean (Vicia faba L.). Journal of Medicinal Plants Research. 5: 6460-6467.
    8. Chakraborti, D., Sarkar, A., Das, S. (2006). Efficient and rapid in vitro plant regeneration system for Indian cultivars of chickpea (Cicer arietinum L.). Plant Cell Tissue and Organ Culture. 86: 101-105.
    9. Chandra, A. and Pental, D. (2003). Regeneration and genetic transformation of grain legumes: an overview. Current Science. 84: 381-387.
    10. Chopra, R. and Saini, R. (2014). Transformation of blackgram [Vigna mungo (L.) Hepper] by barley chitinase and ribosome-inactivating protein genes towards improving resistance to Corynespora leaf spot fungal disease. Biotechnology and Applied Biochemistry. 174(8): 2791-2800.
    11. Das, D, K, Shivaprakash, N., Sarin, N,B. (2004). An efficient regeneration system of black gram (Vigna mungo L.) through organogenesis. Plant Science. 134: 199-206.
    12. Delic, D., Stajkovic, O., Kuzmanovic, D., Rasulic, N., Knezevic-Vukcevic, J., Milicic, B. (2009). The effects of rhizobial inoculation on growth and yield of Vigna mungo L. in Serbian soils. Biotechnology in Animal Husbandry. 25: 1197– 1202.
    13. Duncan, D. B. (1955). Multiple range and multiple F-tests. Biometrics. 11: 1-42.
    14. Eapen, S. (2008). Advances in development of transgenic pulse crops. Biotechnology Advances. 26: 162-168.
    15. Gill, R., Eapen, S., Rao, P.S. (1987). Morphogenic studies of cultured cotyledons of urd bean [Vigna mungo (L.) Hepper]. Journal of Plant Physiology. 139: 1-5.
    16. Ignacimuthu, S., Franklin, G., Melchias, G. (1997). Multiple shoot formation and in vitro fruiting from cotyledonary nodes of Vigna mungo (L.) Hepper. Current Science. 3: 733-735.
    17. Karamany, E.L. (2006). Double purpose (forage and seed) of mungbean production. 1-Effect of plant density and forage cutting date on forage and seed yields of mungbean [Vigna radiata (L.) Wilczeck]. Research Journal of Agriculture and Biological Sciences. 2: 162-165.
    18. Kamboj, R. and Nanda, V. (2018). Proximate composition, nutritional profile and health benefits of legumes – A review. Legume Research. 41(3): 325-332. 
    19. Mony, S. A., Haque, M. S., Alam, M. M., Hasanuzzaman, M., Nahar, K. (2010). Regeneration of black gram (Vigno mungo L.) on changes of hormonal condition. Notulae Botanicae Horti Agrobotanici Cluj-Napoca. 38(3): 140-145.
    20. Murashige, T. and Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-497.
    21. Muruganantham, M., Ganapathi, A., Amutha, S., Vengadesan, G., Selvaraj, N. (2005) Shoot regeneration from immature cotyledonary nodes in black gram [Vigna mungo (L.) Hepper]. Indian Journal of Biotechnology. 4: 551–555.
    22. Muthukumar, B., Mariamma, M., Veluthambi, K., Gnanam, A. (1996). Genetic transformation of cotyledon explants of cowpea [Vigna unguiculata (L.) Walp] using Agrobacterium tumefaciens. Plant Cell Reports. 15: 980-985.
    23. Prakash, M., Elangaimannan, R., Sunilkumar, B., Narayanan, G. S. (2018). Evaluation of black gram genotypes for drought tolerance based on root dynamics and gas exchange parameters. Legume Research. 41(3): 384-391. 
    24. Rani, V. and Raina, S. N. (2000). Genetic fidelity of organized meristem derived micropropagated plants: a critical reappraisal. In vitro Cellular & Developmental Biology. 36: 319-330.
    25. Rao, C,R. and Chand, P. (2006). Impact of different mating approaches in generating variability in blackgram [Vigna mungo (L.) Hepper]. Legume Research. 24: 174-177.
    26. Sahoo, L., Sugla, T., Jaiwal, P.K. (2002). In vitro regeneration and genetic transformation of Vigna species. In Biotechnology for the Improvement of Legumes [PK Jaiwal, RP Singh, eds], Kluwer, Dordrecht, pp 1-48.
    27. Sainger, M., Chaudhary, D., Dahiya, S., Jaiwal, R., Jaiwal, P. K. (2015). Development of an efficient in vitro plant regeneration system amenable to Agrobacterium- mediated transformation of a recalcitrant grain legume black gram [Vigna mungo (L.) Hepper]. Physiology and Molecular Biology of Plants. 21(4): 505-517.
    28. Saini,l R. and Jaiwal, P.K. (2002). Age, position in mother seedling, orientation, and polarity of the epicotyl segments of black gram [Vigna mungo (L.) Hepper] determines its morphogenic response. Plant Science. 163: 101-109.
    29. Sarma, K. K. and Borah, B.K. (2004). Record of parasitoids of legume pod borer Maruca testulalis (Geyer) in Assam. Insect Environment. 10: 77-78.
    30. Suneja, Y., Kaur, S., Gupta, A. K., and Kaur, N. (2011). Levels of nutritional constituents and antinutritional factors in black gram [Vigna mungo (L.) Hepper]. Food Research International. 44: 621-628.
    31. Thallapathy, S. (2017). In vitro micropropogation of black gram through shoot tip explants by Thidiazuron (TDZ). European Journal of Biomedical and Pharmaceutical Sciences. 4(11): 288-293. 
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