Agrobacterium-Mediated Transformation of Three Freshwater Microalgal Strains
Mary Sanitha, Radha Sudhakar , Aliya F. Anwar, Gayathri D. Selvaraju, Ramya Mohandass Pol J Microbiol 2014; 63 (4): ICID: 1130748 Article type: Original article IC™ Value: 10.00
Abstract provided by Publisher
Microalgal transformation has gained interest in recent years. Agrobacterium-mediated transformation remains as the most efficient method for the development of transgenic plants and microalgae due to its wide host range, inexpensive procedure and transfer of large segments of DNA. In the present study, freshwater samples were collected from different places of Tamil Nadu, India and screening and isolation was carried out using BG-11 medium. Nine different species were isolated and identified based on the morphological characteristics and ITS-2 region amplification. Agrobacterium-mediated transformation efficiency was tested for all the isolates. Among them, three species such as Chlorella sp, Ankistrodesmus sp and Scenedesmus bajacalifornicus showed positive results for transformation. Gene integration and expression was confirmed by PCR amplification of hpt and GUS histochemical assay. A. tumifaciens contamination was checked by amplification of npt II gene (kanamycin resistant) which lies outside of the T-border. Based on GUS assay, transformation efficiencies were found to be 12.25% for Chlorellasp 2.96% for Scenedesmus bajacalifornicus and 3.5% for Ankistrodesmus sp.