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

Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

Journal Article · Tue Jun 21 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]

University of Maryland Center for Environmental Science, Baltimore, MD (United States)
University of Maryland Baltimore County (UMBC), Baltimore, MD (United States)
University of Maryland Center for Environmental Science, Baltimore, MD (United States); University of Maryland Baltimore County (UMBC), Baltimore, MD (United States)

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 Δvtc1 mutant, in which PDAT was upregulated under stress, resulting in a 90% increase in TAG content. Knockout of PDAT in the Δvtc1 background induced a 12.8-fold upregulation of DGTT1 and a 272.3% increase in TAG content in Δvtc1/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:: University of Maryland Baltimore County (UMBC), Baltimore, MD (United States); University of Maryland Center for Environmental Science, Baltimore, MD (United States); University of Maryland, College Park, MD (United States)

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

Grant/Contract Number:: FE0031914

OSTI ID:: 1881522

Alternate ID(s):: OSTI ID: 1881523
OSTI ID: 1904840
OSTI ID: 3013940

Report Number(s):: DOE-UMCES--31914

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

Publisher:: Society for Experimental BiologyCopyright Statement

Country of Publication:: United States

Language:: English

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

Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii

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