Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii
Abstract
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 geneticmore »
- Authors:
-
[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
- Publication Date:
- Research Org.:
- Univ. of Maryland, College Park, MD (United States)
- Sponsoring Org.:
- USDOE Office of Fossil Energy (FE); National Science Foundation (NSF)
- OSTI Identifier:
- 1881522
- Alternate Identifier(s):
- OSTI ID: 1881523; OSTI ID: 1904840
- Grant/Contract Number:
- FE‐0031914; FE0031914; CBET-1511939; NSF 1332344
- Resource Type:
- Published Article
- Journal Name:
- The Plant Journal
- Additional Journal Information:
- Journal Name: The Plant Journal Journal Volume: 111 Journal Issue: 4; Journal ID: ISSN 0960-7412
- Publisher:
- Wiley-Blackwell
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; genetic compensation; triacylglycerol biosynthesis; Chlamydomonas reinhardtii; CRISPR; gene knockout
Citation Formats
Lee, Yi‐Ying, Park, Rudolph, Miller, Stephen M., and Li, Yantao. Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii. United Kingdom: N. p., 2022.
Web. doi:10.1111/tpj.15874.
Lee, Yi‐Ying, Park, Rudolph, Miller, Stephen M., & Li, Yantao. Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii. United Kingdom. https://doi.org/10.1111/tpj.15874
Lee, Yi‐Ying, Park, Rudolph, Miller, Stephen M., and Li, Yantao. Wed .
"Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii". United Kingdom. https://doi.org/10.1111/tpj.15874.
@article{osti_1881522,
title = {Genetic compensation of triacylglycerol biosynthesis in the green microalga Chlamydomonas reinhardtii},
author = {Lee, Yi‐Ying and Park, Rudolph and Miller, Stephen M. and Li, Yantao},
abstractNote = {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.},
doi = {10.1111/tpj.15874},
journal = {The Plant Journal},
number = 4,
volume = 111,
place = {United Kingdom},
year = {Wed Jul 06 00:00:00 EDT 2022},
month = {Wed Jul 06 00:00:00 EDT 2022}
}
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