Agrobacterium-mediated transformation of promising oil-bearing marine algae Parachlorella kessleri

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.

Recent Developments on Genetic Engineering of Microalgae for Biofuels and Bio‐Based Chemicals

“…Agrobacterium assisted transfection is widely used in many plants but it is still technically challenging for usage in microalgae, and only have been reported for C. reinhardtii, [56] Haematococcus pluvialis, [57] Chlorella vulgaris, [62] and oil-bearing marine algae Parachlorella kessleri. [63] Except for eukaryotic microalgae, prokaryotic microalgae such as cyanobacteria are amenable to genetic manipulation by conjugation or electroporation. [64] It produced important natural products, such as protein and vitamin or chemical compounds of 3-hydroxypropionic acid [65] and succinate acid [42] through electroporation.…”

Algal Cell Factories: Approaches, Applications, and Potentials

“…In recent years, this method has been successfully applied in algae [52]. Stable nuclear genetic transformation of C. reinhardtii mediated by agrobacterium has been reported [51].…”

“…Stable nuclear genetic transformation of C. reinhardtii mediated by agrobacterium has been reported [51]. Agrobacterium-mediated transformation has shown success in oil-bearing marine algae Parachlorella Kesslerri [52] and Schizochytrium [53], of which the latter is used for commercial production of oil rich in DHA. …”

A Review on Genetic Engineering of Microalgae with Respect to Genomes, Selectable Marker Genes, Promoters and Reporter Genes

“…Several optimization parameters have to be defined such as short life cycle of algae, inexpensive media and high growth rate at different conditions has to be standardized. With respect to these specific characteristics, transformation method needs to be developed or be available for selection of transformed alga (Rathod et al, 2013).…”