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Research Article

volume 40 issue 6 (december 2017) : 1020-1027,   Doi: 10.18805/lr.v0iOF.9097

In vitro differentiation and plant regeneration from root and other explants of juvenile origin in pea (Pisum sativum L.)

S
Shikha Sharma
G
Geetika Gambhir
D
D.K. Srivastava
1Department of Biotechnology, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan-173 230, Himachal Pradesh, India
Cite article:- Sharma Shikha, Gambhir Geetika, Srivastava D.K. (2017). In vitro differentiation and plant regeneration from root and other explants of juvenile origin in pea (Pisum sativum L.). Legume Research. 40(6): 1020-1027. doi: 10.18805/lr.v0iOF.9097.

ABSTRACT

In vitro regeneration of pea explants (Pisum sativum L. var. ‘Lincon’) was done in 49 different combinations and concentrations of BAP, BAP and NAA, BAP and IBA, TDZ, TDZ and Adenine for shoot regeneration from hypocotyl, root, leaf and cotyledonary node. High frequency shoot regeneration was obtained in hypocotyl (81.43%), root(83.53%) and cotyledonary node(72.76%) on MS medium supplemented with 4.50 mg/l BAP and 1.86mg/l NAA, 2.00mg/l TDZ and4.50 mg/l BAP and 1.86mg/l NAA respectively. No shoot regeneration was obtained from leaf explants on any of the combination used. Shoot elongation was observed on the same medium used for shoot regeneration respectively.MS medium supplemented with 0.20 mg/l IBA was found best for root regeneration from in vitro raised shoots. The plantlets were able to regenerate within 6-7 weeks. The regenerated plantlets were acclimatized in pre-sterilized cocopeat. 

REFERENCES

  1. Boehmer, P., Meyer, B. and Jacobsen, H. J. (1995). Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast-derived pea callus. Plant Cell Reports. 66: 26-29.
  2. Cherian, S. K., Prutki, I. S. and Tandon, G. L. (1995). Recent advances in the canning of processed peas. Indian Food Packing. 9(3): 25-30.
  3. Christianson, M., Warnick, D.A., Carlson, P.S.A. (1983). A morphogenetically competent soybean suspension culture. Science 222:632-634.
  4. Christianson, M. and Warnick. (1985). Temporal requirement for phytohormone balance in the control of organogenesis. In vitro Developmental Biology. 112: 494-497.
  5. Doorne, L. E., Marshall, G. and Kirkwood. (1995). Somatic embryogenesis in pea (Pisum sativum L.): effect of explant, genotype and culture conditions. Annals of Applied Biology. 126(1): 169-179.
  6. Emmanouil, N. T., Marjan, B., Krit, R., Vincken, J. P., Lammeren, A. V. and Visser, R. (2004). Regeneration of pea (Pisum sativum L.) by a cyclic organogenic system. Plant Cell Reports. 23(7): 453-460.
  7. Esposito, M. A., Almiron, P., Gatti, I., Cravero, V. P., Anido, F. and Cointry, E. L. (2012). Rapid method to increase the number of F1 plants in pea (Pisum sativum) breeding programs. Genetics and Molecular Research. 11(3): 2729-2732.
  8. FAO. (2012) (http://faostat.fao.org/)
  9. Franklin, G., Pius, P. K. and Ignacimuthu, S. (2000). Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.). Euphytica. 115(1): 65-74.
  10. Gill, R. and Saxena, P K. (1993). Somatic embryogenesis in Nicotiana tabaccum L.: induction by thidiazuron of direct embryo differentiation from cultured leaf discs. Plant Cell Reports.12: 154-159. 
  11. Gomez, K. A. and Gomez, A. A. (1984). Statistical Procedures for Agricultural Research. John Willey and Sons. New York:
  12. Griga, M., Stejskal, J. and Beber, K. (1995). Analysis of tissue culture derived variation in pea (Pisum sativum L.). Euphytica. 85 (1): 335-339. 
  13. Hartmut, E. S., Andrea, H. S., Terese, W. R., Donald, S. and Thomas, J. V. (1993).Transformation and regeneration of two cultivars of pea (Pisum sativum L.). Plant Physiology. 101:751-757
  14. Huetteman, C. A. and Preece, J. E. (1993). Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Culture. 33: 105-119.
  15. Jackson, J. A. and Hobbs, S. L. A. (1990). Rapid multiple shoot production from cotyledonary node explants of pea (Pisum sativum L.). In Vitro Cellular and Developmental Biology Plant. 26(8): 835-838.
  16. Kosturkova, G., Mehandjiev, A., Dobreva, I. and Tzvetkova, V. (1997). Regeneration systems for immature embryos of Bulgarian pea genotypes. Plant Cell Tissue Organ Culture. 139-142.
  17. Malik, K. A. and Saxena, P. K. (1992). In vitro regeneration of grain legumes: novel approach. In Vitro. 28 (3): 92-99.
  18. Mroginski, L. A. and Kartha, K. K. (1981). Regeneration of pea (Pisum sativum L. cv. Century) plants by in vitro culture of immature leaflets. Plant Cell Reports. 1(2): 64-66.
  19. Murashige, T. and Skoog, F. (1962). Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-497.
  20. Natali, L. and Cavallini, A. (1987a). Nuclear cytology of callus and plantlets regenerated from pea (Pisum sativum L.) meristems. Protoplasma. 143:121-25.
  21. Natali, L. and Cavallini, A. (1987b). Regeneration of pea (Pisum sativum L.) plantlets by in vitro culture of immature embryos. Plant Breeding. 99 (2): 172-176.
  22. Pniewski, T., Wachowiak, J., Kapusta, J. and Legocki, A. B. (2003). Organogenesis and long term micropropagation of polish pea cultivars. Acta Societatis Botanicorum Poloniae. 72(4): 295-302.
  23. Rana, U., Sharma, A., Paul, Y. S. and Sharma, K. D. (2009). Survey for pea diseases and identification of fungi associated with wilt/    root rot complex in Himachal Pradesh, a major off season pea growing state. Journal of Mycology and Plant Pathology. 39(3): 416-421.
  24. Rubluo, A., Kartha, K. K., Mroginski, L. A. and Dyck, J. (1984). Plant regeneration from pea leaflets cultured in vitro and genetic stability of regenerants. Plant Physiology. 117(2): 119-130.
  25. Sharma, K. D. 2000. Management of pea powdery mildew in trans- Himalayan region. Indian Journal of Agricultural Sciences. 70(1): 50-52.
  26. Sharma, R. and Kaushal, R. P. (2004). Generation and characterization of pea (Pisum sativum) somaclones for resistance to Ascochyta blight and powdery mildew. Indian Journal of Biotechnology. 3(3): 400-408.
  27. Srivastava, D.K. (2003). Genetic transformation and crop improvement. In: Marwaha, S.S. and Arora J.K,Eds. Biotechnology strategies in agro-processing. New Delhi: Eds Asia Tech Publisher. pp.18-34
  28. Szasz, A., Nervo, G. and Fari, M. (1995). Screening for in vitro shoot forming capacity of seedling explants in bell pepper (Capsicum annum L.) genotypes and efficient plant regeneration using thidiazuron. Plant Cell Reports. 14(10): 666-669. 
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    Research Article

    volume 40 issue 6 (december 2017) : 1020-1027,   Doi: 10.18805/lr.v0iOF.9097

    In vitro differentiation and plant regeneration from root and other explants of juvenile origin in pea (Pisum sativum L.)

    S
    Shikha Sharma
    G
    Geetika Gambhir
    D
    D.K. Srivastava
    1Department of Biotechnology, Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan-173 230, Himachal Pradesh, India
    Cite article:- Sharma Shikha, Gambhir Geetika, Srivastava D.K. (2017). In vitro differentiation and plant regeneration from root and other explants of juvenile origin in pea (Pisum sativum L.). Legume Research. 40(6): 1020-1027. doi: 10.18805/lr.v0iOF.9097.

    ABSTRACT

    In vitro regeneration of pea explants (Pisum sativum L. var. ‘Lincon’) was done in 49 different combinations and concentrations of BAP, BAP and NAA, BAP and IBA, TDZ, TDZ and Adenine for shoot regeneration from hypocotyl, root, leaf and cotyledonary node. High frequency shoot regeneration was obtained in hypocotyl (81.43%), root(83.53%) and cotyledonary node(72.76%) on MS medium supplemented with 4.50 mg/l BAP and 1.86mg/l NAA, 2.00mg/l TDZ and4.50 mg/l BAP and 1.86mg/l NAA respectively. No shoot regeneration was obtained from leaf explants on any of the combination used. Shoot elongation was observed on the same medium used for shoot regeneration respectively.MS medium supplemented with 0.20 mg/l IBA was found best for root regeneration from in vitro raised shoots. The plantlets were able to regenerate within 6-7 weeks. The regenerated plantlets were acclimatized in pre-sterilized cocopeat. 

    REFERENCES

    1. Boehmer, P., Meyer, B. and Jacobsen, H. J. (1995). Thidiazuron-induced high frequency of shoot induction and plant regeneration in protoplast-derived pea callus. Plant Cell Reports. 66: 26-29.
    2. Cherian, S. K., Prutki, I. S. and Tandon, G. L. (1995). Recent advances in the canning of processed peas. Indian Food Packing. 9(3): 25-30.
    3. Christianson, M., Warnick, D.A., Carlson, P.S.A. (1983). A morphogenetically competent soybean suspension culture. Science 222:632-634.
    4. Christianson, M. and Warnick. (1985). Temporal requirement for phytohormone balance in the control of organogenesis. In vitro Developmental Biology. 112: 494-497.
    5. Doorne, L. E., Marshall, G. and Kirkwood. (1995). Somatic embryogenesis in pea (Pisum sativum L.): effect of explant, genotype and culture conditions. Annals of Applied Biology. 126(1): 169-179.
    6. Emmanouil, N. T., Marjan, B., Krit, R., Vincken, J. P., Lammeren, A. V. and Visser, R. (2004). Regeneration of pea (Pisum sativum L.) by a cyclic organogenic system. Plant Cell Reports. 23(7): 453-460.
    7. Esposito, M. A., Almiron, P., Gatti, I., Cravero, V. P., Anido, F. and Cointry, E. L. (2012). Rapid method to increase the number of F1 plants in pea (Pisum sativum) breeding programs. Genetics and Molecular Research. 11(3): 2729-2732.
    8. FAO. (2012) (http://faostat.fao.org/)
    9. Franklin, G., Pius, P. K. and Ignacimuthu, S. (2000). Factors affecting in vitro flowering and fruiting of green pea (Pisum sativum L.). Euphytica. 115(1): 65-74.
    10. Gill, R. and Saxena, P K. (1993). Somatic embryogenesis in Nicotiana tabaccum L.: induction by thidiazuron of direct embryo differentiation from cultured leaf discs. Plant Cell Reports.12: 154-159. 
    11. Gomez, K. A. and Gomez, A. A. (1984). Statistical Procedures for Agricultural Research. John Willey and Sons. New York:
    12. Griga, M., Stejskal, J. and Beber, K. (1995). Analysis of tissue culture derived variation in pea (Pisum sativum L.). Euphytica. 85 (1): 335-339. 
    13. Hartmut, E. S., Andrea, H. S., Terese, W. R., Donald, S. and Thomas, J. V. (1993).Transformation and regeneration of two cultivars of pea (Pisum sativum L.). Plant Physiology. 101:751-757
    14. Huetteman, C. A. and Preece, J. E. (1993). Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Culture. 33: 105-119.
    15. Jackson, J. A. and Hobbs, S. L. A. (1990). Rapid multiple shoot production from cotyledonary node explants of pea (Pisum sativum L.). In Vitro Cellular and Developmental Biology Plant. 26(8): 835-838.
    16. Kosturkova, G., Mehandjiev, A., Dobreva, I. and Tzvetkova, V. (1997). Regeneration systems for immature embryos of Bulgarian pea genotypes. Plant Cell Tissue Organ Culture. 139-142.
    17. Malik, K. A. and Saxena, P. K. (1992). In vitro regeneration of grain legumes: novel approach. In Vitro. 28 (3): 92-99.
    18. Mroginski, L. A. and Kartha, K. K. (1981). Regeneration of pea (Pisum sativum L. cv. Century) plants by in vitro culture of immature leaflets. Plant Cell Reports. 1(2): 64-66.
    19. Murashige, T. and Skoog, F. (1962). Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum. 15: 473-497.
    20. Natali, L. and Cavallini, A. (1987a). Nuclear cytology of callus and plantlets regenerated from pea (Pisum sativum L.) meristems. Protoplasma. 143:121-25.
    21. Natali, L. and Cavallini, A. (1987b). Regeneration of pea (Pisum sativum L.) plantlets by in vitro culture of immature embryos. Plant Breeding. 99 (2): 172-176.
    22. Pniewski, T., Wachowiak, J., Kapusta, J. and Legocki, A. B. (2003). Organogenesis and long term micropropagation of polish pea cultivars. Acta Societatis Botanicorum Poloniae. 72(4): 295-302.
    23. Rana, U., Sharma, A., Paul, Y. S. and Sharma, K. D. (2009). Survey for pea diseases and identification of fungi associated with wilt/    root rot complex in Himachal Pradesh, a major off season pea growing state. Journal of Mycology and Plant Pathology. 39(3): 416-421.
    24. Rubluo, A., Kartha, K. K., Mroginski, L. A. and Dyck, J. (1984). Plant regeneration from pea leaflets cultured in vitro and genetic stability of regenerants. Plant Physiology. 117(2): 119-130.
    25. Sharma, K. D. 2000. Management of pea powdery mildew in trans- Himalayan region. Indian Journal of Agricultural Sciences. 70(1): 50-52.
    26. Sharma, R. and Kaushal, R. P. (2004). Generation and characterization of pea (Pisum sativum) somaclones for resistance to Ascochyta blight and powdery mildew. Indian Journal of Biotechnology. 3(3): 400-408.
    27. Srivastava, D.K. (2003). Genetic transformation and crop improvement. In: Marwaha, S.S. and Arora J.K,Eds. Biotechnology strategies in agro-processing. New Delhi: Eds Asia Tech Publisher. pp.18-34
    28. Szasz, A., Nervo, G. and Fari, M. (1995). Screening for in vitro shoot forming capacity of seedling explants in bell pepper (Capsicum annum L.) genotypes and efficient plant regeneration using thidiazuron. Plant Cell Reports. 14(10): 666-669. 
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