National Academy of Agricultural Sciences (NAAS)
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PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
Development of an efficient protocol for genetic transformation in plants requires effective shoot regeneration and antibiotic selection systems. Genetically engineered disarmed Agrobacterium tumefaciens strain containing binary vector pCambia with chiII (fungal resistance gene) and hpt (hygromycin resistance) genes was used for genetic transformation studies. Hygromycin and cefotaxime sensitivity studies were conducted using leaf and petiole explants of lettuce (Lactuca sativa cv.Solan Kriti) to explore the aptness of hygromycin resistance as a selectable marker and cefotaxime in controlling excessive bacterial growth during genetic transformation studies. Explants (leaf and petiole) showed decrease in fresh weight as concentration of the hygromycin increased resulting in full or partial inhibition of shoot regeneration. A negative correlation was observed between the concentration of hygromycin and fresh weight of the explants at different intervals of time. Effect of different concentrations of cefotaxime was studied on the regeneration potential in leaf and petiole explants of lettuce. PCR analysis of genomic DNA using specific designed primers was done to detect the presence of chiII and hpt genes in hygromycin resistant plantlets of lettuce. Out of five randomly selected putative transgenic shoots, four shoots were found positive for the presence/integration of chiII and hpt genes during T-DNA transfer and integration into the plant genome. The results indicate that hygromycin and cefotaxime act as an effective selective agent during genetic transformation studies.
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