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Research Article

volume 52 issue 1 (february 2018) : 66-70,   Doi: 10.18805/IJARe.A-4874

Three step protocol for regeneration of plantlets in Indian garlic varieties using root meristem

A
Ashwini P. Benke
P
Punam Shelke
V
V. Mahajan
1ICAR-Directorate of Onion and Garlic Research, Rajgurunagar-410 505, Pune, Maharashtra, India
Cite article:- Benke P. Ashwini, Shelke Punam, Mahajan V. (2018). Three step protocol for regeneration of plantlets in Indian garlic varieties using root meristem. Indian Journal of Agricultural Research. 52(1): 66-70. doi: 10.18805/IJARe.A-4874.

ABSTRACT

Protocol for plantlets regeneration through callus induction using root meristem was developed in present investigation. Different mediums were standardized for each step vis-a-via callus induction, callus proliferation and shoot regeneration. In first stage, B5 medium added with 0.7mg/l 2, 4-D and 1mg/l BAP showed 100% callusing while reduction in both growth regulators up to level of 0.5 mg/l 2,4-D and 0.1mg/l BAP results in significant callus proliferation. In last step, shoot regeneration was found to good and healthier in B5 medium supplemented with 0.1 mg/l 2, 4-D only. After regeneration normal growth in plantlets observed under plain B-5 medium, however no abnormality has been noted in developed population. Present protocol will be efficiently useful for chemical mutagen induction studies through callus, observing somaclonal variations and transgenic studies.

REFERENCES

  1. Abo El-Nil, M. M. (1977). Organogenesis and embryogenesis in callus cultures of garlic (Allium sativum L.). Plant Science Letters. 9:259–264.
  2. Ali, A. M. and Bahman, F. N. (2012). In vitro culture of Allium scoroprasum spp. Rodundam callus induction, somatic embryogenesis and direct bulblet formation. International Journal. Of Agriculture Crop Science, 4 (1): 1-7.
  3. Chen, Wu. L. J. and Long, Y. J. (2009). Long analysis of ultrastructure and reactive oxygen species of hyperhydric garlic (Allium sativum L.) shoots. In Vitro Cellular Developmental Biology- Plant 45 (4): 483-490.
  4. De Klerk, G.J., Arnholdt-Schmitt, B., Lieberei, R., Neumann, K.H. (1997). Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biologia Plantarum 39:53–66.
  5. Haque, H.M., Wada, T., Hattori, K. (2003). Shoot regeneration and bulblets formation from root and shoot meristem of Garlic Cv Bangladesh Local Asia. Journal of Plant Science.2 (1):23-27.
  6. Havra, A. P. and Novak, F. J. (1973). The bud formation in callus cultures of Allium sativum L. Z. Pflanzenphysiol. 68:308- 318.
  7. Kehr, A. E., and Schaeffer, G. W. (1976). Tissue culture and differentiation of garlic. Horticutural Science 11:422-423.
  8. Khan, N., Alam, M. S., Nath, U. K. (2004) In vitro Regeneration of Garlic through Callus Culture Journal of Biological Sciences 4 (2): 189-191.
  9. Kondo, T., Hasegawa, H., Suzuki, M. (2000). Transformation and regeneration of garlic (Allium sativum L.) by Agrobacterium-mediated gene transfer. Plant cell reports, 19(10), 989-993.
  10. Lee, S. Y., Kim, H. H., Kim, Y. K., Park, N., Park, S. U. (2009) Plant regeneration of garlic (Allium sativum L.) via somatic embryogenesis. Scientific Research and Essay. 4 (13): 1569-1574.
  11. Luciani, G, F Æ Ana K. Mary Æ Cecilia Pellegrini Æ N. R. Curvetto (2006). Effects of explants and growth regulators in garlic callus formation and plant regeneration. Plant Cell Tissue Organ Culture. 87:139–143.
  12. Mehta, J., Sharma, A., Sharma, N., Megwal, S., Sharma, G., Gehlot P., Naruka R. (2013) An improved method for callus culture and In vitro propagation of garlic (Allium sativum L.) International Journal of. Pure and Applied Bioscience. 1 (1): 1-6
  13. Nasim, S. A., Mujib, A., Kapoor, R., Fatima, S., Aslam, J., Mahmooduzzafar (2010) Somatic embryogenesis in Allium sativum L. (cv. Yamuna Safed 3): Improving embryo maturation and germination with PGRs and carbohydrates. Anales de Biología. 32: 1-9.
  14. Pandey, A., Misra, P., Chandrashekar, K., Trivedi, P. (2012). Development of AtMYB12-expressing transgenic tobacco callus culture for production of rutin with biopesticidal potential. Plant Cell Report, 31(10): 1867–1876 DOI: 10.1007/s00299-012-1300-6
  15. Rashid, U. and Shaukat, A. (2009). Establishment of an efficient callus induction and plant regeneration system in Pakistani wheat (Triticum aestivum) cultivars. Electronic Journal of Biotechnology. 12(3). DOI: 10.2225/vol12-issue3-fulltext-1
  16. Scotton, D. C., Benedito, V. A., Molfetta1 J. B., Rodrigues, B. I., Tulmann-Neto, A., Figueira, A. (2013) Response of root explants to in vitro cultivation of marketable garlic cultivars. Horticultura Brasileira 31: 80-85.
  17. Toaima, N., Novak, E., Schumann, G. (2003). Callus induction from different explants of commercial cultivars of leek, Allium ampeloprasum var. porrum L. Acta Horticuture. (ISHS) 597:303-309.
  18. Valk, P. V. D., Scholten, O. E., Verstappen, F., Jansen, R. C., Dons, J. J. M.(1992). High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species. Plant Cell Tissue Organ Culture.30 (3):181-    191. DOI: 10.1007/BF00040020
  19. Wang, Y. M., Wang, J. B., Luo d., Jia J. F. (2001) Regeneration of plants from callus cultures of roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi. Cell Research. 11:279–284. DOI: 10.1038/sj.cr.7290097
  20. Zheng, S. B., Henken, E., Sofiari, E., Jacobsen, F. A., Krens Kik, C. (1998).Factors influencing induction, propagation and regeneration of mature zygotic embryo-derived callus from Allium cepa. Plant Cell, Tissue Organ Culture. 53: 99–105.
  21. Zheng, S. J., Henken, B., Ahn, Y. K., Krens, F. A., Kik, C (2004) The development of a reproducible Agrobacterium tumefaciens transformation system for garlic (Allium sativum L.) and the production of transgenic garlic resistant to beet armyworm (Spodoptera exigua Hübner) Molecular Breeding, 14(3):293–307.
  22. Zheng, S.J., Henken B., KrensFa., Kik, C. (2003). The development of an efficient cultivar-independent plant regeneration system from callus derived from both apical and non-apical root segments of garlic (Allium sativum L.). In Vitro Cellular and Developmental Biology- Plant 39: 288-292. 
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Indian Journal of Agricultural Research
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    Research Article

    volume 52 issue 1 (february 2018) : 66-70,   Doi: 10.18805/IJARe.A-4874

    Three step protocol for regeneration of plantlets in Indian garlic varieties using root meristem

    A
    Ashwini P. Benke
    P
    Punam Shelke
    V
    V. Mahajan
    1ICAR-Directorate of Onion and Garlic Research, Rajgurunagar-410 505, Pune, Maharashtra, India
    Cite article:- Benke P. Ashwini, Shelke Punam, Mahajan V. (2018). Three step protocol for regeneration of plantlets in Indian garlic varieties using root meristem. Indian Journal of Agricultural Research. 52(1): 66-70. doi: 10.18805/IJARe.A-4874.

    ABSTRACT

    Protocol for plantlets regeneration through callus induction using root meristem was developed in present investigation. Different mediums were standardized for each step vis-a-via callus induction, callus proliferation and shoot regeneration. In first stage, B5 medium added with 0.7mg/l 2, 4-D and 1mg/l BAP showed 100% callusing while reduction in both growth regulators up to level of 0.5 mg/l 2,4-D and 0.1mg/l BAP results in significant callus proliferation. In last step, shoot regeneration was found to good and healthier in B5 medium supplemented with 0.1 mg/l 2, 4-D only. After regeneration normal growth in plantlets observed under plain B-5 medium, however no abnormality has been noted in developed population. Present protocol will be efficiently useful for chemical mutagen induction studies through callus, observing somaclonal variations and transgenic studies.

    REFERENCES

    1. Abo El-Nil, M. M. (1977). Organogenesis and embryogenesis in callus cultures of garlic (Allium sativum L.). Plant Science Letters. 9:259–264.
    2. Ali, A. M. and Bahman, F. N. (2012). In vitro culture of Allium scoroprasum spp. Rodundam callus induction, somatic embryogenesis and direct bulblet formation. International Journal. Of Agriculture Crop Science, 4 (1): 1-7.
    3. Chen, Wu. L. J. and Long, Y. J. (2009). Long analysis of ultrastructure and reactive oxygen species of hyperhydric garlic (Allium sativum L.) shoots. In Vitro Cellular Developmental Biology- Plant 45 (4): 483-490.
    4. De Klerk, G.J., Arnholdt-Schmitt, B., Lieberei, R., Neumann, K.H. (1997). Regeneration of roots, shoots and embryos: physiological, biochemical and molecular aspects. Biologia Plantarum 39:53–66.
    5. Haque, H.M., Wada, T., Hattori, K. (2003). Shoot regeneration and bulblets formation from root and shoot meristem of Garlic Cv Bangladesh Local Asia. Journal of Plant Science.2 (1):23-27.
    6. Havra, A. P. and Novak, F. J. (1973). The bud formation in callus cultures of Allium sativum L. Z. Pflanzenphysiol. 68:308- 318.
    7. Kehr, A. E., and Schaeffer, G. W. (1976). Tissue culture and differentiation of garlic. Horticutural Science 11:422-423.
    8. Khan, N., Alam, M. S., Nath, U. K. (2004) In vitro Regeneration of Garlic through Callus Culture Journal of Biological Sciences 4 (2): 189-191.
    9. Kondo, T., Hasegawa, H., Suzuki, M. (2000). Transformation and regeneration of garlic (Allium sativum L.) by Agrobacterium-mediated gene transfer. Plant cell reports, 19(10), 989-993.
    10. Lee, S. Y., Kim, H. H., Kim, Y. K., Park, N., Park, S. U. (2009) Plant regeneration of garlic (Allium sativum L.) via somatic embryogenesis. Scientific Research and Essay. 4 (13): 1569-1574.
    11. Luciani, G, F Æ Ana K. Mary Æ Cecilia Pellegrini Æ N. R. Curvetto (2006). Effects of explants and growth regulators in garlic callus formation and plant regeneration. Plant Cell Tissue Organ Culture. 87:139–143.
    12. Mehta, J., Sharma, A., Sharma, N., Megwal, S., Sharma, G., Gehlot P., Naruka R. (2013) An improved method for callus culture and In vitro propagation of garlic (Allium sativum L.) International Journal of. Pure and Applied Bioscience. 1 (1): 1-6
    13. Nasim, S. A., Mujib, A., Kapoor, R., Fatima, S., Aslam, J., Mahmooduzzafar (2010) Somatic embryogenesis in Allium sativum L. (cv. Yamuna Safed 3): Improving embryo maturation and germination with PGRs and carbohydrates. Anales de Biología. 32: 1-9.
    14. Pandey, A., Misra, P., Chandrashekar, K., Trivedi, P. (2012). Development of AtMYB12-expressing transgenic tobacco callus culture for production of rutin with biopesticidal potential. Plant Cell Report, 31(10): 1867–1876 DOI: 10.1007/s00299-012-1300-6
    15. Rashid, U. and Shaukat, A. (2009). Establishment of an efficient callus induction and plant regeneration system in Pakistani wheat (Triticum aestivum) cultivars. Electronic Journal of Biotechnology. 12(3). DOI: 10.2225/vol12-issue3-fulltext-1
    16. Scotton, D. C., Benedito, V. A., Molfetta1 J. B., Rodrigues, B. I., Tulmann-Neto, A., Figueira, A. (2013) Response of root explants to in vitro cultivation of marketable garlic cultivars. Horticultura Brasileira 31: 80-85.
    17. Toaima, N., Novak, E., Schumann, G. (2003). Callus induction from different explants of commercial cultivars of leek, Allium ampeloprasum var. porrum L. Acta Horticuture. (ISHS) 597:303-309.
    18. Valk, P. V. D., Scholten, O. E., Verstappen, F., Jansen, R. C., Dons, J. J. M.(1992). High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species. Plant Cell Tissue Organ Culture.30 (3):181-    191. DOI: 10.1007/BF00040020
    19. Wang, Y. M., Wang, J. B., Luo d., Jia J. F. (2001) Regeneration of plants from callus cultures of roots induced by Agrobacterium rhizogenes on Alhagi pseudoalhagi. Cell Research. 11:279–284. DOI: 10.1038/sj.cr.7290097
    20. Zheng, S. B., Henken, E., Sofiari, E., Jacobsen, F. A., Krens Kik, C. (1998).Factors influencing induction, propagation and regeneration of mature zygotic embryo-derived callus from Allium cepa. Plant Cell, Tissue Organ Culture. 53: 99–105.
    21. Zheng, S. J., Henken, B., Ahn, Y. K., Krens, F. A., Kik, C (2004) The development of a reproducible Agrobacterium tumefaciens transformation system for garlic (Allium sativum L.) and the production of transgenic garlic resistant to beet armyworm (Spodoptera exigua Hübner) Molecular Breeding, 14(3):293–307.
    22. Zheng, S.J., Henken B., KrensFa., Kik, C. (2003). The development of an efficient cultivar-independent plant regeneration system from callus derived from both apical and non-apical root segments of garlic (Allium sativum L.). In Vitro Cellular and Developmental Biology- Plant 39: 288-292. 
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