Abstract
The heterotrophic microalga Crypthecodinium cohnii is well known for its lipid accumulation, with a high proportion of docosahexaenoic acid (DHA). In this study, we report a novel screening approach to obtain mutants of C. cohnii with high growth and lipid content using the acetyl-CoA carboxylase (ACCase) inhibitor sethoxydim. C. cohnii mutants were generated using atmospheric and room-temperature plasma (ARTP) and then screened for two rounds in media supplemented with sethoxydim. These efforts led to the identification of mutant M-1-2, which had 24.32% higher growth and 7.05% higher lipid content than the wild type, demonstrating the effectiveness of the sethoxydim-based screening. Consistently, the M-1-2 mutant displayed a 16.15% increase in ACCase enzymatic activity and 1.53-fold upregulation of its ACCase-encoding gene based on comparative ACCase activity analysis and transcriptomic analysis, respectively. In addition, transcriptomic analysis showed that transcripts involved in fatty acid biosynthesis, energy, central carbohydrate, and amino acid metabolism were upregulated in the mutant compared to the wild type.
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Acknowledgments
The authors would like to thank Prof. Hao Song for the help with the ARTP experiments. This study was funded by grants from the National High-tech R&D Program (“863” program, project No. 2012AA02A707), the National Basic Research Program of China (“973” program, project No. 2014CB745101), the Doctoral Program of Higher Education of China (Nos. 20120032110020 and 20130032120022), the National Natural Science Foundation of China (No. 21621004), the Tianjin Municipal Science and Technology Commission (No. 15JCZDJC32500), and Zaoneng Biotechnology Inc.
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Liu, J., Pei, G., Diao, J. et al. Screening and transcriptomic analysis of Crypthecodinium cohnii mutants with high growth and lipid content using the acetyl-CoA carboxylase inhibitor sethoxydim. Appl Microbiol Biotechnol 101, 6179–6191 (2017). https://doi.org/10.1007/s00253-017-8397-z
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DOI: https://doi.org/10.1007/s00253-017-8397-z