Genetic transformation and regeneration of <Emphasis Type="Italic">Sesbania drummondii</Emphasis> using cotyledonary nodes |
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Authors: | Priya Padmanabhan Shivendra V Sahi |
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Institution: | (1) Department of Biology, Western Kentucky University, 1906 College Heights Blvd. # 11080, Bowling Green, KY 42101-1080, USA |
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Abstract: | Sesbania drummondii (Rydb.) Cory is a source for phytopharmaceuticals. It also hyperaccumulates several toxic heavy metals. Development of an
efficient gene transfer method is an absolute requirement for the genetic improvement of this plant with more desirable traits
due to limitations in conventional breeding methods. A simple protocol was developed for Agrobacterium-mediated stable genetic transformation of Sesbania. Agrobacterium
tumefaciens strain EHA 101 containing the vector pCAMBIA 1305.1 having hptII and GUS plus genes was used for the gene transfer experiments. Evaluation of various parameters was carried out to assess
the transformation frequency by GUS expression analysis. High transformation frequency was achieved by using 7-day-old precultured
cotyledonary node (CN) explants. Further, the presence of acetosyringone (150 μM), infection of explants for 30–45 min and
3 days of cocultivation proved to be critical factors for greatly improving the transformation efficiency. Stable transformation
of S. drummondii was achieved, and putative transgenic shoots were obtained on medium supplemented with hygromycin (25 mg l−1). GUS histochemical analysis of the putative transgenic tissues further confirmed the transformation event. Genomic Southern
blot analysis was performed to verify the presence of transgenes and their stable integration. A transformation frequency
of 4% was achieved for CN explants using this protocol. |
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Keywords: | Sesbania drummondii Genetic transformation Transgenic Cotyledonary nodes |
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