Chinese Journal OF Rice Science >
Directed Knockout of SD1 Gene Improves Lodging Resistance and Blast Resistance of Rice
Received date: 2022-11-24
Revised date: 2023-02-08
Online published: 2023-07-17
【Objective】 Huai 119 is a high-yielding japonica rice variety. To improve its unfavorable traits, especially high plant height and poor resistance to rice blast, we used the CRISPR/Cas9 gene editing technology to knock out its ‘Green Revolution’ gene SD1.【Method】 We selected the SD1 gene as the target to construct the CRISPR/Cas9 gene knockout vector, which was then transformed into Huai 119 by Agrobacterium-mediated transformation method, and the homozygous sd1 knockout line without transgenic insertion was obtained. Subsequently, we compared and analyzed the plant height, rice blast resistance, and nitrogen uptake and utilization efficiency of the sd1 and wild-type lines.【Result】 We successfully isolated a homozygous sd1 knockout line without transgenic insertion in the background of Huai 119. In field paddy, it was found that over 60% of the planting area of wild-type Huai 119 was lodging, while the sd1 homozygous mutant population effectively avoided lodging in the later stages of growth due to its shorter plant height. In addition, after treatment with different concentrations of GA (0.01-1.00 μmol/L), the plant height increase of wild-type plants was significantly greater than that of the sd1 line, indicating that the sensitivity of sd1 line to exogenous GA treatment was reduced. The identification of rice blast resistance shows that knocking out SD1 also contributed to the improvement of rice blast resistance of Huai 119. However, the absorption and utilization efficiency of nitrogen decreased due to the knockout of SD1. 【Conclusion】 The knockout of SD1 gene in the rice variety Huai 119 not only improves lodging resistance by reducing plant height, but also enhances the resistance to rice blast.
Key words： CRISPR/Cas9; gene editing; SD1; rice blast; lodging resistance; rice
LI Gang, GAO Qingsong, LI Wei, ZHANG Wenxia, WANG Jian, CHEN Baoshan, WANG Di, GAO Hao, XU Weijun, CHEN Hongqi, JI Jianhui . Directed Knockout of SD1 Gene Improves Lodging Resistance and Blast Resistance of Rice[J]. Chinese Journal OF Rice Science, 2023 , 37(4) : 359 -367 . DOI: 10.16819/j.1001-7216.2023.221113
| [1] | Doebley J F, Gaut B S, Smith B D. The molecular genetics of crop domestication[J]. Cell, 2006, 127(7): 1309-1321. |
| [2] | Cho S W, Kim S, Kim J M, Kim J S. Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease[J]. Nature Biotechnology, 2013, 31(3): 230-232. |
| [3] | Cong L, Ran F A, Cox D, Lin S L, Barretto R, Habib N, Hsu P D, Wu X B, Jiang W Y, Marraffini L A, Zhang F. Multiplex genome engineering using CRISPR/Cas systems[J]. Science, 2013, 339(6121): 819-823. |
| [4] | Jinek M, East A, Cheng A R, Lin S, Ma E B, Doudna J. RNA-programmed genome editing in human cells[J]. Elife, 2013, 2: e00471. |
| [5] | Mali P, Aach J, Stranges P B, Esvelt K M, Moosburner M, Kosuri S, Yang L H, Church G M. CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering[J]. Nature Biotechnology, 2013, 31(9): 833-838. |
| [6] | Li Y, Li W J, Li J. The CRISPR/Cas9 revolution continues: From base editing to prime editing in plant science[J]. Journal of Genetics and Genomics, 2021, 48(8): 661-670. |
| [7] | Chen K L, Wang Y P, Zhang R, Zhang H W, Gao C X. CRISPR/Cas genome editing and precision plant breeding in agriculture[J]. Annual Review of Plant Biology, 2019, 70: 667-697. |
| [8] | 沈兰, 李健, 付亚萍, 王俊杰, 华宇峰, 焦晓真, 严长杰, 王克剑. 利用CRISPR/Cas9系统定向改良水稻粒长和穗粒数性状[J]. 中国水稻科学, 2017, 31(3): 223-231. |
| [8] | Shen L, Li J, Fu Y P, Wang J J, Hua Y F, Jiao X Z, Yan C J, Wang K J. Orientation improvement of grain length and grain number in rice by using CRISPR/Cas9 system[J]. Chinese Journal of Rice Science, 2017, 31(3): 223-231. (in Chinese with English abstract) |
| [9] | Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush G S, Kitano H, Matsuoka M. Green revolution: A mutant gibberellin-synthesis gene in rice[J]. Nature, 2002, 416(6882): 701-702. |
| [10] | Peng Y L, Hu Y G, Qian Q, Ren D Y. Progress and prospect of breeding utilization of green revolution gene SD1 in rice[J]. Agriculture, 2021, 11(7): 611. |
| [11] | Spielmeyer W, Ellis M H, Chandler P M. Semidwarf(sd-1), "green revolution" rice, contains a defective gibberellin 20-oxidase gene[J]. Proceedings of the National Academy of Sciences of the United States of America, 2002, 99(13): 9043-9048. |
| [12] | Monna L, Kitazawa N, Yoshino R, Suzuki J, Masuda H, Maehara Y, Tanji M, Sato M, Nasu S, Minobe Y. Positional cloning of rice semidwarfing gene, sd-1: Rice "green revolution gene" encodes a mutant enzyme involved in gibberellin synthesis[J]. DNA Research, 2002, 9(1): 11-17. |
| [13] | 李铮友, 师常俊, 李晓艾, 林贤国. 香软米滇屯502的选育[J]. 云南农业大学学报, 1999, 14(1): 27-31. |
| [13] | Li Z Y, Shi C J, Li X A, Ling X G. Breeding of Diantun 502: Fragrant and soft rice[J]. Journal of Yunnan Agricultural University, 1999, 14(1): 27-31. (in Chinese with English abstract) |
| [14] | Biswas S, Tian J Q, Li R, Chen X F, Luo Z J, Chen M J, Zhao X X, Zhang D B, Persson S, Yuan Z, Shi J X. Investigation of CRISPR/Cas9-induced SD1 rice mutants highlights the importance of molecular characterization in plant molecular breeding[J]. Journal of Genetics and Genomics, 2020, 47(5): 273-280. |
| [15] | Hu X, Cui Y, Dong G, Feng A, Wang D, Zhao C, Zhang Y, Hu J, Zeng D, Guo L, Qian Q. Using CRISPR-Cas9 to generate semi-dwarf rice lines in elite landraces[J]. Scientific Reports, 2019, 9(1): 19096. |
| [16] | 胡雪娇, 杨佳, 程灿, 周继华, 牛付安, 王新其, 张美良, 曹黎明, 储黄伟. 利用CRISPR/Cas9系统定向编辑水稻SD1基因[J]. 中国水稻科学, 2018, 32(3): 219-225. |
| [16] | Hu X J, Yang J, Cheng C, Zhou J H, Niu F A, Wang X Q, Zhang M L, Cao L M, Chu H W. Targeted editing of rice SD1 gene Using CRISPR/Cas9 system[J]. Chinese Journal of Rice Science, 2018, 32(3): 219-225. (in Chinese with English abstract) |
| [17] | 黎起秦, 陈育新, 韦绍兴. 赤霉素对稻秆抗瘟性影响的分析[J]. 广西农学院学报, 1990, 9(1): 19-26. |
| [17] | Li Q Q, Chen Y X, Wei S X. An analysis on effect of bibberellin on resistance of rice stalk to rice stalk blast[J]. Journal of Guangxi Agricultual College, 1990, 9(1): 19-26. (in Chinese with English abstract) |
| [18] | Qin X, Liu J H, Zhao W S, Chen X J, Guo Z J, Peng Y L. Gibberellin 20-oxidase gene OsGA20ox3 regulates plant stature and disease development in rice[J]. Molecular Plant-Microbe Interactions, 2013, 26(2): 227-239. |
| [19] | Tanaka N, Matsuoka M, Kitano H, Asano T, Kaku H, Komatsu S. gid1, a gibberellin-insensitive dwarf mutant, shows altered regulation of probenazole-inducible protein (PBZ1) in response to cold stress and pathogen attack[J]. Plant Cell & Environment, 2006, 29(4): 619-631. |
| [20] | Yang D L, Li Q, Deng Y W, Lou Y G, Wang M Y, Zhou G X, Zhang Y Y, He Z H. Altered disease development in the eui mutants and Eui overexpressors indicates that gibberellins negatively regulate rice basal disease resistance[J]. Molecular Plant, 2008, 1(3): 528-537. |
| [21] | Liu H, Ding Y D, Zhou Y Q, Jin W Q, Xie K B, Chen L L. CRISPR-P 2.0: An improved CRISPR-Cas9 tool for genome editing in plants[J]. Molecular Plant, 2017, 10(3): 530-532. |
| [22] | Liu W Z, Xie X R, Ma X L, Li J, Chen J H, Liu Y G. DSDecode: A web-based tool for decoding of sequencing chromatograms for genotyping of targeted mutations[J]. Molecular Plant, 2015, 8(9): 1431-1433. |
| [23] | 杨光, 雷海霞, 乔利. 稻瘟病室内产孢及人工接种鉴定[J]. 种业导刊, 2018(12): 23-24. |
| [23] | Yang G, Lei H X, Qiao L. Identification of indoor spore production and inoculation of rice blast[J]. Seed Industry Tribune, 2018(12): 23-24. (in Chinese) |
| [24] | 中华人民共和国农业部. 水稻品种试验稻瘟病抗性鉴定与评价技术规程: NY/T 2646-2014[S]. 北京: 中国标准出版社, 2014: |
| [24] | Ministry of Agriculture of the People’s Republic of China. Technical Specification for Identification and Evaluation of Blast Resistance in Rice Variety Region Test: NY/T 2646-2014[S]. Beijing: Standards Press of China, 2014. (in Chinese) |
| [25] | Arnon D I. Copper enzymes in isolated chloroplasts: Polyphenoloxidase in Beta vulgaris[J]. Plant Physiology, 1949, 24(1): 1-15. |
| [26] | Gao Z Y, Wang Y F, Chen G, Zhang A P, Yang S L, Shang L G, Wang D Y, Ruan B P, Liu C L, Jiang H Z, Dong G J, Zhu L, Hu J, Zhang G H, Zeng D L, Guo L B, Xu G H, Teng S, Harberd N P, Qian Q. The indica nitrate reductase gene OsNR2 allele enhances rice yield potential and nitrogen use efficiency[J]. Nature Communication, 2019, 10(1): 5207. |
| [27] | 王新, 韩悦, 冯璇, Nawaz G, 罗亮, 刘芳, 覃宝祥, 刘耀光, 李容柏. 应用 CRISPR-Cas9 基因编辑技术改良传统优质糯稻品种[J]. 分子植物育种, 2019, 17(19): 6332-6342. |
| [27] | Wang X, Han Y, Feng X, Nawaz G, Luo L, Liu F, Qin B X, Liu Y G, Li R B. Improvement of a traditional high-quality glutinous rice variety by CRISPR-Cas9 gene editing system[J]. Molecular Plant Breeding, 2019, 17(19): 6332-6342. (in Chinese with English abstract) |
| [28] | Eshed Y, Lippman Z B. Revolutions in agriculture chart a course for targeted breeding of old and new crops[J]. Science, 2019, 366(6466): eaax0025. |
| [29] | Wang B L, Wei H F, Zhang H, Zhang W H. Enhanced accumulation of gibberellins rendered rice seedlings sensitive to ammonium toxicity[J]. Journal of Experimental Botany, 2020, 71(4): 1514-1526. |
| [30] | Li S, Tian Y H, Wu K, Ye Y F, Yu J P, Zhang J Q, Liu Q, Hu M Y, Li H, Tong Y P, Harberd N P, Fu X D. Modulating plant growth-metabolism coordination for sustainable agriculture[J]. Nature, 2018, 560(7720): 595-600. |
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