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CRISPR-Based Assessment of Gene Specialization in the Gibberellin Metabolic Pathway in Rice.
- Source :
-
Plant physiology [Plant Physiol] 2019 Aug; Vol. 180 (4), pp. 2091-2105. Date of Electronic Publication: 2019 Jun 03. - Publication Year :
- 2019
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Abstract
- Gibberellin (GA) functions as an essential natural regulator of growth and development in plants. For each step of the GA metabolic pathway, different copy numbers can be found in different species, as is the case with the 13 genes across four enzymatic steps in rice ( Oryza sativa ). A common view is that such gene duplication creates homologs that buffer organisms against loss-of-function (LOF) mutations. Therefore, knockouts of any single homolog might be expected to have little effect. To test this question, we generated clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) knockouts for these homologs and measured effects on growth and reproduction. Surprisingly, we report here that there is consistently one or more essential gene at each enzymatic step, for which LOF mutation induces death or sterility-suggesting that the GA pathway does not have a redundancy route and that each gene family is essential for GA metabolism. In most of these genes from the same gene family, we observed defects in plant height and infertility, suggesting that the duplicated members retain functions related to GA synthesis or degradation. We identified both subfunctionalization of the three recently diversified homologs OsKO1 , OsKO2 , and OsKO5 and neofunctionalization in OsKO3 and OsKO4 Thus, although the function of each step is conserved, the evolution of duplicates in that step is diversified. Interestingly, the CRISPR/Cas9 lines at the SD1 locus were typically sterile, whereas the natural sd1 mutants, related to the "Green Revolution" in rice, show normal setting rates. Collectively, our results identify candidates for control of GA production and provide insight into the evolution of four critical gene families in plants.<br /> (© 2019 American Society of Plant Biologists. All Rights Reserved.)
- Subjects :
- Gene Editing methods
Mutation genetics
Plant Proteins genetics
Plant Proteins metabolism
Plants, Genetically Modified genetics
Plants, Genetically Modified metabolism
CRISPR-Cas Systems genetics
Clustered Regularly Interspaced Short Palindromic Repeats genetics
Gibberellins metabolism
Oryza genetics
Oryza metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1532-2548
- Volume :
- 180
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Plant physiology
- Publication Type :
- Academic Journal
- Accession number :
- 31160507
- Full Text :
- https://doi.org/10.1104/pp.19.00328