Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

Lee, Yi‐Ying; Park, Rudolph; Miller, Stephen M.; Li, Yantao

doi:10.1111/tpj.15874

Title: Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

Journal Article · Wed Jul 06 00:00:00 EDT 2022 · The Plant Journal

DOI:https://doi.org/10.1111/tpj.15874· OSTI ID:1881522

Lee, Yi‐Ying ^[1]; Park, Rudolph ^[2]; Miller, Stephen M. ^[2];

^[3]

Institute of Marine and Environmental Technology University of Maryland Center for Environmental Science Baltimore MD 21202 USA
Department of Biological Sciences University of Maryland, Baltimore County Baltimore MD 21250 USA
Institute of Marine and Environmental Technology University of Maryland Center for Environmental Science Baltimore MD 21202 USA, Department of Marine Biotechnology University of Maryland, Baltimore County Baltimore MD 21202 USA

SUMMARY Genetic compensation has been proposed to explain phenotypic differences between gene knockouts and knockdowns in several metazoan and plant model systems. With the rapid development of reverse genetic tools such as CRISPR/Cas9 and RNAi in microalgae, it is increasingly important to assess whether genetic compensation affects the phenotype of engineered algal mutants. While exploring triacylglycerol (TAG) biosynthesis pathways in the model alga Chlamydomonas reinhardtii , it was discovered that knockout of certain genes catalyzing rate‐limiting steps of TAG biosynthesis, type‐2 diacylglycerol acyltransferase genes ( DGTTs ), triggered genetic compensation under abiotic stress conditions. Genetic compensation of a DGTT1 null mutation by a related PDAT gene was observed regardless of the strain background or mutagenesis approach, for example, CRISPR/Cas 9 or insertional mutagenesis. However, no compensation was found in the PDAT knockout mutant. The effect of PDAT knockout was evaluated in a Δ vtc1 mutant, in which PDAT was upregulated under stress, resulting in a 90% increase in TAG content. Knockout of PDAT in the Δ vtc1 background induced a 12.8‐fold upregulation of DGTT1 and a 272.3% increase in TAG content in Δ vtc1 / pdat1 cells, while remaining viable. These data suggest that genetic compensation contributes to the genetic robustness of microalgal TAG biosynthetic pathways, maintaining lipid and redox homeostasis in the knockout mutants under abiotic stress. This work demonstrates examples of genetic compensation in microalgae, implies the physiological relevance of genetic compensation in TAG biosynthesis under stress, and provides guidance for future genetic engineering and mutant characterization efforts.

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Research Organization:: Univ. of Maryland, College Park, MD (United States)

Sponsoring Organization:: USDOE Office of Fossil Energy (FE); National Science Foundation (NSF)

Grant/Contract Number:: FE‐0031914; FE0031914; CBET-1511939; NSF 1332344

OSTI ID:: 1881522

Alternate ID(s):: OSTI ID: 1881523; OSTI ID: 1904840

Journal Information:: The Plant Journal, Journal Name: The Plant Journal Vol. 111 Journal Issue: 4; ISSN 0960-7412

Publisher:: Wiley-BlackwellCopyright Statement

Country of Publication:: United Kingdom

Language:: English

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59 BASIC BIOLOGICAL SCIENCES
genetic compensation
triacylglycerol biosynthesis
Chlamydomonas reinhardtii
CRISPR
gene knockout

Title: Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

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References (51)

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