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Abstract                 Volume:4  Issue-1  Year-2017          Original Research Articles

IJCRBP is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCRBP Articles.

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A Modified Protocol for Agrobacterium Mediated Transformation of Tomato Plants
Pranav Kumar Mishra1, H. P. Jai Shanker Pillai2, Herman Kopa2, Megha Gupta1 and Selvam Arjunan2*
1Center for Research and PG studies, Department of Biotechnology, Indian Academy Degree College (Autonomous), Kalyan Nagar, Bangalore, Karnataka-560 043, India
2Center for Research and PG studies, Department of Life Sciences, Kristu Jayanti College (Autonomous), K. Narayanapura, Kothanur (P.O), Bangalore, Karnataka-560 077, India
*Corresponding author

An improved protocol for Agrobacterium-mediated transformation was developed for tomato plant. In this study, transformation was carried out using disarmed A. tumefaciens strain LBA4404 harboring a binary vector pGPTV-kan (13400bp) containing the CP-gene, was used in this study. Factors that affect transformation/regeneration protocols were optimized in a series of experiments. Results indicated that exposure of cotyledonary/leaf explants to Agrobacterium inoculums of 0.7 OD at 600nm for 10 mins, co-cultivation in MS-liquid media, selection on kanamycin (kanamycin 50mg/ml), Augmentin, and cefotaxime (250mg/ml)-containing medium and subsequent regeneration on MS medium supplemented with 2.5mg/L BAP and 0.1 mg/L IAA resulted in transformation efficiency of 24.50%. Coat protein (CP) expression was observed in transformed tomato shoots but never in the control plants. PCR amplification of DNA extracted from the transformed tissues demonstrated the generation of the expected amplicon, corresponding to CP gene. This result strongly verifies the successful transformation of tomato plant. Moreover, this protocol may pave the way for problem solving-applications encompassing other Indian crops of economic importance.

Keywords: Agrobacterium tumefaciens, Coat protein, Transformation efficiency
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How to cite this article:

Mishra, P. K., Jai Shanker Pillai, H. P., Kopa, H., Gupta, M., Arjunan, S., 2017. A modified protocol for Agrobacterium mediated transformation of tomato plants.Int.J.Curr.Res.Biosci.Plantbiol. 4(1): 123-130. doi: http://dx.doi.org/10.20546/ijcrbp.2017.401.016
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