Poster: Integrated Plant Biology

Abs # 80: Genetic transformation and in vitro regeneration of a lead hyperaccumulator plant

Presenter:	Sahi, Shivendra V., shiv.sahi@wku.edu
Authors	Sahi, Shivendra V. (A)   Cheepala, Satish B. (A)   Sharma, Nilesh C. (A)
Affiliations:	(A): Biotechnology Center, Department of Biology, Western Kentucky University

Sesbania drummondii is a leguminous shrub mostly distributed in the southeastern parts of the USA. Current studies have shown that Sesbania drummondii can hyperaccumulate toxic heavy metal like lead (Pb). This plant is considered a potential source for the phytoremediation of Pb contaminated sites. To develop a transformation system in this species Agrobacterium -mediated transfer of GUS reporter gene was achieved. Agrobacterium (K289) containing pCAMBIA 1305.1 plasmid was used for transformation of cotyledonary explants. Infected explants produced calluses on MS medium containing 0.5mg/L thiadiazuron. Callus when subjected to GUS histochemical assay produced blue spots demonstrating the expression of the GUS gene. PCR analysis of the genomic DNA from transformed tissue using GUS gene specific primers gave 1200 bp band confirming presence of the reporter gene. In vitro regeneration of Sesbania plantlets was also induced from cotyledonary nodes. Attempts are being made to regenerate GUS transformed plants from cotyledonary node segments.