Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators
Type-B response regulator proteins in rice contain a conserved receiver domain, followed by a GARP DNA binding domain and a longer C-terminus. Some type-B response regulators such as RR21, RR22 and RR23 are involved in the development of rice leaf, root, flower and trichome. In this study, to evalua...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/22/14165 |
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| author | Chuanhong Li Chenbo Gong Jiemin Wu Linfeng Yang Lei Zhou Bian Wu Liang Gao Fei Ling Aiqing You Changyan Li Yongjun Lin |
| author_facet | Chuanhong Li Chenbo Gong Jiemin Wu Linfeng Yang Lei Zhou Bian Wu Liang Gao Fei Ling Aiqing You Changyan Li Yongjun Lin |
| author_sort | Chuanhong Li |
| collection | DOAJ |
| description | Type-B response regulator proteins in rice contain a conserved receiver domain, followed by a GARP DNA binding domain and a longer C-terminus. Some type-B response regulators such as RR21, RR22 and RR23 are involved in the development of rice leaf, root, flower and trichome. In this study, to evaluate the application potential of type-B response regulators in rice genetic improvement, thirteen type-B response regulator genes in rice were respectively knocked out by using CRISPR/Cas9 genome editing technology. Two guide RNAs (gRNAs) were simultaneously expressed on a knockout vector to mutate one gene. T<sub>0</sub> transformed plants were used to screen the plants with deletion of large DNA fragments through PCR with specific primers. The mutants of CRISPR/Cas9 gene editing were detected by <i>Cas9</i> specific primer in the T<sub>1</sub> generation, and homozygous mutants without <i>Cas9</i> were screened, whose target regions were confirmed by sequencing. Mutant materials of 12 <i>OsRRs</i> were obtained, except for <i>RR24</i>. Preliminary phenotypic observation revealed variations of various important traits in different mutant materials, including plant height, tiller number, tillering angle, heading date, panicle length and yield. The <i>osrr30</i> mutant in the T<sub>2</sub> generation was then further examined. As a result, the heading date of the <i>osrr30</i> mutant was delayed by about 18 d, while the yield was increased by about 30%, and the chalkiness was significantly reduced compared with those of the wild-type under field high temperature stress. These results indicated that <i>osrr30</i> has great application value in rice breeding. Our findings suggest that it is feasible to perform genetic improvement of rice by editing the type-B response regulators. |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T18:16:47Z |
| publishDate | 2022-11-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-8ea5b472205c4762b8e3788de078aa932023-11-24T08:39:39ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-11-0123221416510.3390/ijms232214165Improvement of Rice Agronomic Traits by Editing Type-B Response RegulatorsChuanhong Li0Chenbo Gong1Jiemin Wu2Linfeng Yang3Lei Zhou4Bian Wu5Liang Gao6Fei Ling7Aiqing You8Changyan Li9Yongjun Lin10National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaHubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, ChinaHubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaHubei Key Laboratory of Food Crop Germplasm and Genetic Improvement, Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, ChinaType-B response regulator proteins in rice contain a conserved receiver domain, followed by a GARP DNA binding domain and a longer C-terminus. Some type-B response regulators such as RR21, RR22 and RR23 are involved in the development of rice leaf, root, flower and trichome. In this study, to evaluate the application potential of type-B response regulators in rice genetic improvement, thirteen type-B response regulator genes in rice were respectively knocked out by using CRISPR/Cas9 genome editing technology. Two guide RNAs (gRNAs) were simultaneously expressed on a knockout vector to mutate one gene. T<sub>0</sub> transformed plants were used to screen the plants with deletion of large DNA fragments through PCR with specific primers. The mutants of CRISPR/Cas9 gene editing were detected by <i>Cas9</i> specific primer in the T<sub>1</sub> generation, and homozygous mutants without <i>Cas9</i> were screened, whose target regions were confirmed by sequencing. Mutant materials of 12 <i>OsRRs</i> were obtained, except for <i>RR24</i>. Preliminary phenotypic observation revealed variations of various important traits in different mutant materials, including plant height, tiller number, tillering angle, heading date, panicle length and yield. The <i>osrr30</i> mutant in the T<sub>2</sub> generation was then further examined. As a result, the heading date of the <i>osrr30</i> mutant was delayed by about 18 d, while the yield was increased by about 30%, and the chalkiness was significantly reduced compared with those of the wild-type under field high temperature stress. These results indicated that <i>osrr30</i> has great application value in rice breeding. Our findings suggest that it is feasible to perform genetic improvement of rice by editing the type-B response regulators.https://www.mdpi.com/1422-0067/23/22/14165rice breedingCRISPR/Cas9type-B response regulatorsheading dateyieldquality |
| spellingShingle | Chuanhong Li Chenbo Gong Jiemin Wu Linfeng Yang Lei Zhou Bian Wu Liang Gao Fei Ling Aiqing You Changyan Li Yongjun Lin Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators International Journal of Molecular Sciences rice breeding CRISPR/Cas9 type-B response regulators heading date yield quality |
| title | Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators |
| title_full | Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators |
| title_fullStr | Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators |
| title_full_unstemmed | Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators |
| title_short | Improvement of Rice Agronomic Traits by Editing Type-B Response Regulators |
| title_sort | improvement of rice agronomic traits by editing type b response regulators |
| topic | rice breeding CRISPR/Cas9 type-B response regulators heading date yield quality |
| url | https://www.mdpi.com/1422-0067/23/22/14165 |
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