CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress
Pectin is one of the constituents of the cell wall, distributed in the primary cell wall and middle lamella, affecting the rheological properties and the cell wall stickiness. Pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) are the most important factors for modifying methyl e...
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/24/16082 |
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| author | Zhaoyang Li Muhammad Junaid Rao Jiaying Li Yanting Wang Peng Chen Hua Yu Chongjian Ma Lingqiang Wang |
| author_facet | Zhaoyang Li Muhammad Junaid Rao Jiaying Li Yanting Wang Peng Chen Hua Yu Chongjian Ma Lingqiang Wang |
| author_sort | Zhaoyang Li |
| collection | DOAJ |
| description | Pectin is one of the constituents of the cell wall, distributed in the primary cell wall and middle lamella, affecting the rheological properties and the cell wall stickiness. Pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) are the most important factors for modifying methyl esterification. In this study, 45 <i>PMEI</i> genes from rice (<i>Oryza sativa</i> L.) were screened by bioinformatics tools, and their structure, motifs, cis-acting elements in the promoter region, chromosomal distribution, gene duplication, and phylogenetic relationship were analyzed. Furthermore, CRISPR/Cas9 was used to edit the <i>OsPMEI12</i> (<i>LOC_Os</i>03G01020) and two mutant <i>pmei12</i> lines were obtained to explore the functions of <i>OsPMEI</i> in plant growth and development, and under cadmium (Cd) stress. Compared to wild type (WT) Nipponbare, the second inverted internodes of the mutant plants shortened significantly, resulting in the reduction in plant height at mature stage. The seed setting rate, and fresh and dry weights of the mutants were also decreased in mutant plants. In addition, the pectin methylation of <i>pmei12</i> lines is decreased as expected, and the pectin content of the cell wall increased at both seedling and maturity stages; however, the cellulose and hemicellulose increased only at seedling stage. Interestingly, the growth of the <i>pmei12</i> lines was better than the WT in both normal conditions and under two phytohormone (GA<sub>3</sub> and NAA) treatments at seedling stage. Under Cd stress, the fresh and dry weights were increased in <i>pmei12</i> lines. These results indicated that <i>OsPMEI12</i> was involved in the regulation of methyl esterification during growth, affected cell wall composition and agronomic traits, and might play an important role in responses to phytohormones and stress. |
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| spelling | doaj.art-aacf96e7bd6f428a9aeaff1fd8d87fa92023-11-24T15:32:38ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123241608210.3390/ijms232416082CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal StressZhaoyang Li0Muhammad Junaid Rao1Jiaying Li2Yanting Wang3Peng Chen4Hua Yu5Chongjian Ma6Lingqiang Wang7College of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaState Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Agriculture, Guangxi University, Nanning 530004, ChinaCollege of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaCollege of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaHenry Fok School of Biology and Agriculture, Shaoguan University, Shaoguan 512005, ChinaCollege of Plant Science and Technology, Biomass and Bioenergy Research Center, Huazhong Agricultural University, Wuhan 430070, ChinaPectin is one of the constituents of the cell wall, distributed in the primary cell wall and middle lamella, affecting the rheological properties and the cell wall stickiness. Pectin methylesterase (PME) and pectin methylesterase inhibitor (PMEI) are the most important factors for modifying methyl esterification. In this study, 45 <i>PMEI</i> genes from rice (<i>Oryza sativa</i> L.) were screened by bioinformatics tools, and their structure, motifs, cis-acting elements in the promoter region, chromosomal distribution, gene duplication, and phylogenetic relationship were analyzed. Furthermore, CRISPR/Cas9 was used to edit the <i>OsPMEI12</i> (<i>LOC_Os</i>03G01020) and two mutant <i>pmei12</i> lines were obtained to explore the functions of <i>OsPMEI</i> in plant growth and development, and under cadmium (Cd) stress. Compared to wild type (WT) Nipponbare, the second inverted internodes of the mutant plants shortened significantly, resulting in the reduction in plant height at mature stage. The seed setting rate, and fresh and dry weights of the mutants were also decreased in mutant plants. In addition, the pectin methylation of <i>pmei12</i> lines is decreased as expected, and the pectin content of the cell wall increased at both seedling and maturity stages; however, the cellulose and hemicellulose increased only at seedling stage. Interestingly, the growth of the <i>pmei12</i> lines was better than the WT in both normal conditions and under two phytohormone (GA<sub>3</sub> and NAA) treatments at seedling stage. Under Cd stress, the fresh and dry weights were increased in <i>pmei12</i> lines. These results indicated that <i>OsPMEI12</i> was involved in the regulation of methyl esterification during growth, affected cell wall composition and agronomic traits, and might play an important role in responses to phytohormones and stress.https://www.mdpi.com/1422-0067/23/24/16082<i>Oryza sativa</i> L.pectin methylesterase inhibitorcell wallplant growth regulatorsCd stress |
| spellingShingle | Zhaoyang Li Muhammad Junaid Rao Jiaying Li Yanting Wang Peng Chen Hua Yu Chongjian Ma Lingqiang Wang CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress International Journal of Molecular Sciences <i>Oryza sativa</i> L. pectin methylesterase inhibitor cell wall plant growth regulators Cd stress |
| title | CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress |
| title_full | CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress |
| title_fullStr | CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress |
| title_full_unstemmed | CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress |
| title_short | CRISPR/Cas9 Mutant Rice <i>Ospmei12</i> Involved in Growth, Cell Wall Development, and Response to Phytohormone and Heavy Metal Stress |
| title_sort | crispr cas9 mutant rice i ospmei12 i involved in growth cell wall development and response to phytohormone and heavy metal stress |
| topic | <i>Oryza sativa</i> L. pectin methylesterase inhibitor cell wall plant growth regulators Cd stress |
| url | https://www.mdpi.com/1422-0067/23/24/16082 |
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