Factors influencing <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>-mediated genetic transformation of <Emphasis Type="Italic">Eleusine coracana</Emphasis> (L.) Gaertn |
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| Authors: | Manju Sharma Aditi Kothari-Chajer Swati Jagga-Chugh S L Kothari |
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| Institution: | (1) Experimental Morphogenesis and Plant Tissue Culture Laboratory, Department of Botany, University of Rajasthan, Jaipur, 302 004, India;(2) Centre for Converging Technologies, University of Rajasthan, Jaipur, 302 004, India;(3) College of Agriculture and Veterinary Medicine, Department of Horticulture and Plant Sciences, Jimma University, Jimma, Ethiopia |
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| Abstract: | Agrobacterium-mediated transformation protocol has been developed for Eleusine coracana (var. PR-202) by varying several factors which influence T-DNA delivery. Green nodular regenerative calli with meristematic
nodules of seed origin were used as the target tissue for Agrobacterium
tumefaciens-mediated gene transfer. The highest frequency of transformation (44.4%) was observed when callus was infected, co-cultivated
and incubated at 22°C. Incorporation of higher level of CuSO4 in the regeneration medium had significantly positive effect on the recovery of transformed plants. PCR analysis of T
0 and T
1 generation plants with nptII-specific primers revealed the amplification of nptII gene. Southern blot analysis of six regenerated plants confirmed selectable marker gene integration in three plants. This
is a first report on Agrobacterium-mediated genetic transformation of finger millet and will pave the way for further studies in this and other millet crops. |
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