Agrobacterium-mediated transformation of promising oil-bearing marine algae Parachlorella kessleri |
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Authors: | Jayant Pralhad Rathod Gunjan Prakash Reena Pandit Arvind M. Lali |
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Affiliation: | 1. DBT-ICT Centre for Energy Biosciences, Institute of Chemical Technology [University Under Section-3 of UGC Act 1956], Maharashtra Govt.’s Elite Status and Centre of Excellence, Grade ‘A’ by MHRD, Matunga, Mumbai, India 2. Department of Chemical Engineering, Institute of Chemical Technology [University Under Section-3 of UGC Act 1956], Maharashtra Govt.’s Elite Status and Centre of Excellence, Grade ‘A’ by MHRD, Matunga, Mumbai, India
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Abstract: | Parachlorella kessleri is a unicellular alga which grows in fresh as well as marine water and is commercially important as biomass/lipid feedstock and in bioremediation. The present study describes the successful transformation of marine P. kessleri with the help of Agrobacterium tumefaciens. Transformed marine P. kessleri was able to tolerate more than 10 mg l?1 hygromycin concentration. Co-cultivation conditions were modulated to allow the simultaneous growth of both marine P. kessleri and A. tumefaciens. For co-cultivation, P. kessleri was shifted from Walne’s to tris acetate phosphate medium to reduce the antibiotic requirement during selection. In the present study, the transfer of T-DNA was successful without using acetosyringone. Biochemical and genetic analyses were performed for expression of transgenes by GUS assay and PCR in transformants. Establishment of this protocol would be useful in further genetic modification of oil-bearing Parachlorella species. |
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