Effect of Agrobacterium Infection Time, Co-Cultivation and Cell Density on in vitro Response in Hypocotyl of Eggplant (Solanum melongena L.)
DOI:
https://doi.org/10.24154/jhs.v11i1.97Keywords:
Eggplant, Manjarigota, Regeneration, Agrobacterium, PCRAbstract
The present study purports to assess the effect of Agrobacterium infection time, co-cultivation and cell density on in vitro response in hypocotyl explants of eggplant (brinjal) cv. Manjarigota. Agrobacterium(OD600 0.3-0.5) infection for 10-15 minutes (24.44±2.34%) was found to be optimum, while, higher or lower infection-time resulted in reduced callus initiation, shoot regeneration and explant survival. Explants with no (only Agrobacterium infection) or short (1 day) co-cultivation, showed reduced callus-initiation response and turned yellow, with no regeneration. Callus-initiation response increased from Day 1 (96.66±03.33%), and reached a maximum on Day 2 and Day 3 (100±00.00%). It decreased on further increase in co-cultivation time. Explants co-cultivated for three days showed highest regeneration response (30.00±02.96%) which thereafter reduced with further increase in co-cultivation time. Explants infected with Agrobacterium culture at 0.05 OD600 showed hardly any regeneration, and turned yellow and necrotic on the selection medium. Highest regeneration response (28.33±02.33%) was obtained in explants infected with 0.1 OD600 culture, and this gradually reduced as celldensity increased (upto 1.0 OD600), becoming zero in explants treated with cultures at 1.5 OD600 or above. Agrobacterium overgrowth was noticed on explants infected with cultures of 0.5 OD600 and above. Exposure of hypocotyl explants to higher cell-density, longer infection-time and prolonged co-cultivation regime resulted in severe necrosis of explants; time taken for development of Agrobacterium overgrowth was less with increase in the level of these factors. Regenerated shoots were healthy, green, elongated and showed root induction on culture medium containing Kanamycin.
References
Billings, S., Jelenkovic, G., Chin, C.K. and Eberhadt, J. 1997. The effect of growth regulation and antibiotics on eggplant transformation. J. Amer. Soc. Hortl. Sci., 122:158-162
Cardoza, V. and Stewart, C.N. 2003. Increased Agrobacterium-mediated transformation and rooting efficiencies in canola from hypocotyl segment explants. Pl. Cell Rep., 21:599-604
Chakrabarty, R., Viswakarma, N., Bhat, S.R., Kirti, P.B., Singh, B.D. and Chopra, V.L. 2002. Agrobacterium- mediated transformation of cauliflower: optimization of protocol and development of Bt-transgenic cauliflower. J. Biosci., 27:495-502
Fillatti, J.J., Sellmer, J., McCown, B., Haissing, B. and Comai, L. 1987. Agrobacterium mediated transformation and regeneration of hybrid aspen and poplar clones. Pl. Physiol., 93:1110-1116
Hanur, V.S., Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Riaz Mahmood, and Lalitha Anand. 2006. Synergistic use of hypocotyl explants and high BAP pre-conditioning for enhanced transformation frequency in brinjal (Solanum melongena L.). J. Hortl. Sci., 1:116-119
Kumar, V.S. and Rajam, M.V. 2005. Enhanced induction of vir genes results in the improvement of Agrobacterium-mediated transformation of eggplant. J. Biochem. Biotech., 14:89-94
Magioli, C. and Mansur, E. 2005. Eggplant (Solanum melongena L.): tissue culture, genetic transformation and use as an alternative model plant. Acta Bot. Bras., 19:139-148
Magioli, C., Rocha, A.P.M., Pinheiro, M.M., Martins, S.G. and Mansur, E. 2000. Establishment of an efficient Agrobacterium-mediated transformation system for eggplant and study of a potential biotechnologically useful promoter. J. Pl. Biotech., 2:43-49
Murashige, T. and Skoog, F. 1962. A revised medium for rapid growth and bioassays with tissue cultures. Physiol. Plantarum, 15:473-497
Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007a. Effect of explant type and growth regulators in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hortl. Sci., 2:94-98
Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Lalitha Anand and Vageeshbabu S. Hanur. 2007b. Effect of antibiotics and gelling agents in the transformation of brinjal (Solanum melongena L.) cv. Manjarigota. J. Hortl. Sci. 2:19-25
Prakash, D.P., Deepali, B.S., Asokan, R., Ramachandra, Y.L., Shetti, D.L., Lalitha Anand and Vageeshbabu S. Hanur. 2008. Effect of growth regulators on in vitro complete plant regeneration in brinjal (Solanum melongena L.). Ind. J. Hort., 6:112-116
Veluthambi, K., Gupta, K.A. and Sharma, A. 2003. The current status of plant transformation technologies. Curr. Sci., 84:368-379
Zhang, Z., Coyne, D.P. and Mitra, A. 1997. Factors affecting Agrobacterium-mediated transformation of common bean. J. Amer. Soc. Hortl. Sci., 122:300-305
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Copyright (c) 2016 D P Prakash, Y L Ramachandra, Vageeshbabu S Hanur (Author)
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