NDLI: Development of flow cytometric procedures for the efficient isolation of improved lipid accumulation mutants in a Chlorella sp. microalga

Content Provider	Springer Nature Link
Author	Manandhar Shrestha, Kalpana Hildebrand, Mark
Copyright Year	2013
Abstract	The successful development of microalgae-based biofuel production will rely on improvements in the amount and rate of fuel molecule precursor accumulation. Mutagenesis and selection is an attractive approach to improve fuel molecule productivity. Previous screening methods have been laborious, the numbers of mutants isolated have been small, and overall performance of mutants may have been compromised by the presence of deleterious secondary mutations generated by random mutagenesis that affect other cellular processes and growth. We report an improved method of isolating high triacylglycerol (TAG) accumulating mutants of a Chlorella sp., KAS603, using flow cytometric-based selection. In addition to selection for high TAG accumulating strains, the method requires that growth of mutants be competitive with other cells in the population. Not only is growth competitive, but there is improved performance in TAG accumulation with repeated selection, suggesting purification from deleterious secondary mutations. The procedure resulted in the isolation of mutants with far higher efficiency (thousands of fold) that outperformed wild type substantially better (1.8–2.5-fold) than with previous methods. This opens the door to new approaches to the characterization of genes involved in TAG accumulation and other cellular processes.
Starting Page	1643
Ending Page	1651
Page Count	9
File Format	PDF
ISSN	09218971
Journal	Journal of Applied Phycology
Volume Number	25
Issue Number	6
e-ISSN	15735176
Language	English
Publisher	Springer Netherlands
Publisher Date	2013-04-03
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Chlorella Triacylglycerols Mutant Flow cytometry Selection Biofuels Plant Sciences Freshwater & Marine Ecology Plant Physiology Ecology
Content Type	Text
Resource Type	Article
Subject	Plant Science Aquatic Science

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