Cargando…
Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins
Abscisic acid (ABA) is involved in regulating drought tolerance, and pyrabactin resistance-like (PYL) proteins are known as ABA receptors. To elucidate the role of one of the ABA receptors in rice, OsPYL9 was mutagenized through CRISPR/Cas9 in rice. Homozygous and heterozygous mutant plants lacking...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7660227/ https://www.ncbi.nlm.nih.gov/pubmed/33113937 http://dx.doi.org/10.3390/ijms21217854 |
_version_ | 1783608967265517568 |
---|---|
author | Usman, Babar Nawaz, Gul Zhao, Neng Liao, Shanyue Liu, Yaoguang Li, Rongbai |
author_facet | Usman, Babar Nawaz, Gul Zhao, Neng Liao, Shanyue Liu, Yaoguang Li, Rongbai |
author_sort | Usman, Babar |
collection | PubMed |
description | Abscisic acid (ABA) is involved in regulating drought tolerance, and pyrabactin resistance-like (PYL) proteins are known as ABA receptors. To elucidate the role of one of the ABA receptors in rice, OsPYL9 was mutagenized through CRISPR/Cas9 in rice. Homozygous and heterozygous mutant plants lacking any off-targets and T-DNA were screened based on site-specific sequencing and used for morpho-physiological, molecular, and proteomic analysis. Mutant lines appear to accumulate higher ABA, antioxidant activities, chlorophyll content, leaf cuticular wax, and survival rate, whereas a lower malondialdehyde level, stomatal conductance, transpiration rate, and vascular bundles occur under stress conditions. Proteomic analysis found a total of 324 differentially expressed proteins (DEPs), out of which 184 and 140 were up and downregulated, respectively. The OsPYL9 mutants showed an increase in grain yield under both drought and well watered field conditions. Most of the DEPs related to circadian clock rhythm, drought response, and reactive oxygen species were upregulated in the mutant plants. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that DEPs were only involved in circadian rhythm and Gene Ontology (GO) analysis showed that most of the DEPs were involved in response to abiotic stimulus, and abscisic acid-activated signaling pathways. Protein GIGANTEA, Adagio-like, and Pseudo-response regulator proteins showed higher interaction in protein–protein interaction (PPI) network. Thus, the overall results showed that CRISPR/Cas9-generated OsPYL9 mutants have potential to improve both drought tolerance and the yield of rice. Furthermore, global proteome analysis provides new potential biomarkers and understandings of the molecular mechanism of rice drought tolerance. |
format | Online Article Text |
id | pubmed-7660227 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-76602272020-11-13 Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins Usman, Babar Nawaz, Gul Zhao, Neng Liao, Shanyue Liu, Yaoguang Li, Rongbai Int J Mol Sci Article Abscisic acid (ABA) is involved in regulating drought tolerance, and pyrabactin resistance-like (PYL) proteins are known as ABA receptors. To elucidate the role of one of the ABA receptors in rice, OsPYL9 was mutagenized through CRISPR/Cas9 in rice. Homozygous and heterozygous mutant plants lacking any off-targets and T-DNA were screened based on site-specific sequencing and used for morpho-physiological, molecular, and proteomic analysis. Mutant lines appear to accumulate higher ABA, antioxidant activities, chlorophyll content, leaf cuticular wax, and survival rate, whereas a lower malondialdehyde level, stomatal conductance, transpiration rate, and vascular bundles occur under stress conditions. Proteomic analysis found a total of 324 differentially expressed proteins (DEPs), out of which 184 and 140 were up and downregulated, respectively. The OsPYL9 mutants showed an increase in grain yield under both drought and well watered field conditions. Most of the DEPs related to circadian clock rhythm, drought response, and reactive oxygen species were upregulated in the mutant plants. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that DEPs were only involved in circadian rhythm and Gene Ontology (GO) analysis showed that most of the DEPs were involved in response to abiotic stimulus, and abscisic acid-activated signaling pathways. Protein GIGANTEA, Adagio-like, and Pseudo-response regulator proteins showed higher interaction in protein–protein interaction (PPI) network. Thus, the overall results showed that CRISPR/Cas9-generated OsPYL9 mutants have potential to improve both drought tolerance and the yield of rice. Furthermore, global proteome analysis provides new potential biomarkers and understandings of the molecular mechanism of rice drought tolerance. MDPI 2020-10-23 /pmc/articles/PMC7660227/ /pubmed/33113937 http://dx.doi.org/10.3390/ijms21217854 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Usman, Babar Nawaz, Gul Zhao, Neng Liao, Shanyue Liu, Yaoguang Li, Rongbai Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title | Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title_full | Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title_fullStr | Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title_full_unstemmed | Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title_short | Precise Editing of the OsPYL9 Gene by RNA-Guided Cas9 Nuclease Confers Enhanced Drought Tolerance and Grain Yield in Rice (Oryza sativa L.) by Regulating Circadian Rhythm and Abiotic Stress Responsive Proteins |
title_sort | precise editing of the ospyl9 gene by rna-guided cas9 nuclease confers enhanced drought tolerance and grain yield in rice (oryza sativa l.) by regulating circadian rhythm and abiotic stress responsive proteins |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7660227/ https://www.ncbi.nlm.nih.gov/pubmed/33113937 http://dx.doi.org/10.3390/ijms21217854 |
work_keys_str_mv | AT usmanbabar preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins AT nawazgul preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins AT zhaoneng preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins AT liaoshanyue preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins AT liuyaoguang preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins AT lirongbai preciseeditingoftheospyl9genebyrnaguidedcas9nucleaseconfersenhanceddroughttoleranceandgrainyieldinriceoryzasativalbyregulatingcircadianrhythmandabioticstressresponsiveproteins |