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A CRISPR/Cas9‐mediated in situ complementation method for Phytophthora sojae mutants
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A CRISPR/Cas9‐mediated in situ complementation method for Phytophthora sojae mutants
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A CRISPR/Cas9‐mediated in situ complementation method for Phytophthora sojae mutants
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Journal Article
A CRISPR/Cas9‐mediated in situ complementation method for Phytophthora sojae mutants
2021
Overview
Phytophthora sojae is an important model species for oomycete functional genomics research. Recently, a CRISPR/Cas9‐mediated genome‐editing technology has been successfully established in P. sojae, which has been rapidly and widely applied in oomycete research. However, there is an emerging consensus in the biological community that a complete functional gene research system is needed such as developed in the investigations in functional complementation carried out in this study. We report the development of an in situ complementation method for accurate restoration of the mutated gene. We targeted a regulatory B‐subunit of protein phosphatase 2A (PsPP2Ab1) to verify this knockout and subsequent complementation system. We found that the deletion of PsPP2Ab1 in P. sojae leads to severe defects in vegetative hyphal growth, soybean infection, and loss of the ability to produce sporangia. Subsequently, the reintroduction of PsPP2Ab1 into the knockout mutant remedied all of the deficiencies. This study demonstrates the successful implementation of an in situ complementation system by CRISPR/Cas9, which will greatly accelerate functional genomics research of oomycetes in the post‐genomic era.
A regulatory B‐subunit of protein phosphatase 2A was used to demonstrate an in situ complementation method of a mutated gene in Phytophthora sojae, verifying its roles in hyphal growth, soybean infection, and sporangia production.
