Pratheesh P. T., Shonima G. M., Jiji Thomas, Abraham C. I. and Muraleedhara Kurup G.
The objective of this study was to determine the efficacy of three commonly used Agrobacterium strains in genetic transformation of the microalga Chlamydomonas reinhardtii. Three different strains of Agrobacterium tumefaciens (EHA 101, EHA 105, LBA 4404) and pCAMBIA 1304 binary vector hosting the genes coding for GUS (β- glucuronidase) and hpt (hygromycin phosphotransferase) were used for the present study. Transformation processes using Agrobacterium were evaluated without the addition of acetosyringone or by wounding the cells. Colonies resistant to hygromycin at 10 mg/l were selected. GUS gene expression in transformed cells was detected with GUS histochemical assay. PCR and RT-PCR analysis confirmed the integration and expression of hpt (610 bp) and GUS (640 bp) genes in transgenic Chlamydomonas reinhardtii. Agrobacterium strain EHA 105 was found to be the most efficient with a higher rate of transformation frequency.
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