利用CRISPR/Cas9技术编辑AFP1基因提高水稻耐逆性
收稿日期: 2020-05-03
修回日期: 2020-05-27
网络出版日期: 2021-01-10
基金资助
国家重点研发计划资助项目(2016YFD0101100);湖南省自然科学基金资助项目(2019JJ40208);湖南省科技人才托举工程资助项目(2019TJ-Q08)
CRISPR/Cas9-mediatedEditing of AFP1Improves Rice Stress Tolerance
Received date: 2020-05-03
Revised date: 2020-05-27
Online published: 2021-01-10
关键词： 水稻; CRISPR/Cas9基因编辑技术; AFP1; 耐逆性
周天顺, 余东, 刘玲, 欧阳宁, 袁贵龙, 段美娟, 袁定阳 . 利用CRISPR/Cas9技术编辑AFP1基因提高水稻耐逆性[J]. 中国水稻科学, 2021 , 35(1) : 11 -18 . DOI: 10.16819/j.1001-7216.2021.0503
【Objective】It is important to reveal the functions of rice AFP1 in response to abiotic stress, and createnewstress-resistant rice germplasm in innovative ways.【Method】Excellent indica restoring line Huazhan was used as transgenic receptor material to generate afp1 mutants usingCRISPR/Cas9 system, and abiotic stress tolerances of afp1 knockout lines were preliminaryanalyzed. 【Results】The editing efficiency of AFP1 in two targets were 66.67% and 75.00%, respectively.In all mutant plants, only insertions and deletions occurred.About 90% of mutant bases were small fragments,with a lengthless than 5bp.Six homozygous mutants without genetically modified ingredients were generated.Under normal conditions, plant height and seed setting rate of afp1 mutants were reduced, but effective tiller number per plant and panicle length were significantly increased. Interestingly, the yield per plant of mutants changed between-4.06%and +11.75%. Compared with the wild type, the ABA sensitivity and water loss rate of afp1 mutants were obviously decreased. Drought, heat and osmotic stress resistances were significantly improved.【Conclusion】MutationofAFP1genecouldimprovemultiple abiotic stress resistances in rice.
Key words： rice; CRISPR/Cas9 gene editing technology; AFP1; abiotic stresstolerance
| [1] | Akram R, Fahad S, Masood N, Fahad S, Masood N, Rasool A, Ijaz M, Ihsan M Z, Maqbool M M, Ahmad S, Hussain S, Ahmed M, Kaleem S, Sultana S R, Mubeen M, Saud S, Kamran M, Nasim W.Advances in Rice Research for Abiotic Stress Tolerance[M]. Cambridge: Woodhead Publishing, 2019: 69-85. |
| [2] | Boyer J S.Plant productivity and environment[J]. Science, 1982, 218: 443-448. |
| [3] | Pauwels L, Barbero G F, Geerinck J, Tilleman S, Grunewald W, Pérez A C, Chico J M, Bossche R V, Sewell J, Gil E, García-Casado G, Witters E, Inzé D, Long J A, De Jaeger G, Solano R, Goossens A.NINJA connects the co-repressor TOPLESS to jasmonatesignaling[J]. Nature, 2010, 464(7289): 788-791. |
| [4] | Huang M D, Wu W L.Overexpression of TMAC2, a novel negative regulator of abscisic acid and salinity responses, has pleiotropic effects in Arabidopsis thaliana[J]. Plant Molecular Biology, 2007, 63(4): 557-569. |
| [5] | Chang G X, Wang C T, Kong X X, Chen Q, Yang Y P, Hu X Y.AFP2 as the novel regulator breaks high-temperature-induced seeds secondary dormancy through ABI5 and SOM in Arabidopsis thaliana[J]. Biochemical and Biophysical Research Communications, 2018, 501(1): 232-238. |
| [6] | Chang G X, Yang W J, Zhang Q L, Huang J L, Yang Y P, Hu X Y. ABI5-BINDING PROTEIN2coordinates CONSTANS to delay flowering by recruiting the transcriptional corepressor TPR2[J]. Plant Physiology, 2019, 179(2): 477-490. |
| [7] | 安敏敏, 杨立明, 罗玉明. AFP2基因调控拟南芥茉莉酸合成与开花时间的分析[J]. 分子植物育种, 2019, 17(10): 3259-3266. |
| [7] | An M M, Y L M, Luo Y M. Analysis of the role of AFP2gene in regulating jasmonic acid biosynthesis and flowering time of Arabidopsis[J]. Molecular Plant Breeding, 2019, 17(10): 3259-3266. (in Chinese with English abstract) |
| [8] | Tang N, Ma S Q, Zong W, Yang N, Lv Y, Yan C, Guo Z, Li J, Li X, Xiang Y, Song H Z, Xiao J H, Li X H, Xiong L Z.MODD mediates deactivation and degradation of OsbZIP46 to negatively regulate ABA signaling and drought resistance in rice[J]. The Plant Cell, 2016, 28(9): 2161-2177. |
| [9] | Ma S, Tang N, Li X, Xie Y J, Xiang D H, Fu J, Shen J Q, Yang J, Tu H F, Li X H, Hu H H, Xiong L Z.Reversible histone H2BMonoubiquitinationfine-tunes abscisic acid signaling and drought response in rice[J]. Molecular Plant, 2019, 12(2): 263-277. |
| [10] | Liu X, Baird W V.Identification of a novel gene, HaABRC5, from Helianthus annuus (Asteraceae) that is upregulated in response to drought, salinity, and abscisic acid[J]. American Journal of Botany, 2004, 91(2): 184-191. |
| [11] | Wu J, Seng S S, Carianopol C, Sui J J, Yang Q Y, Zhang F Q, Jiang H R, He J N, Yi M F.Cloning and characterization of a novel Gladiolus hybridusAFP family gene (GhAFP-like) related to corm dormancy[J]. Biochemical and Biophysical Research Communications, 2016, 471(1): 198-204. |
| [12] | Miao C B, Xiao L H, Hua K, Zou C S, Zhao Y, Bressan R A, Zhu J K.Mutations in a subfamily of abscisic acid receptor genes promote rice growth and productivity[J]. Proceedings of the National Academy of Sciences, 2018, 115(23): 6058-6063. |
| [13] | Lou D J, Wang H P, Liang G, Yu D Q.OsSAPK2 confers abscisic acid sensitivity and tolerance to drought stress in rice[J]. Frontiers in Plant Science, 2017, 8: 993. |
| [14] | Cui Y T, Hu X M, Liang G H, Feng A H, Wang F M, Ruan S, Dong G J, Shen L, Zhang B, Chen D D,Zhu L,Hu J,Lin Y J,Guo L B,Matsuoka M,Qian Q. Production of novel beneficial alleles of a rice yield-related QTL by CRISPR/Cas9[J]. Plant Biotechnology Journal, 2020, 18(10): 1987-1989. |
| [15] | Zhou T S, Yu D, Dong H, Sun Z Z, Tan Y N, Sun X W, Sheng X B, Duan M J, Yuan D Y.Genome-Wide Identification and Expression Profile of NINJA and AFP Genes in Rice[J]. International Journal of Advances in Biology, 2020, 23(1): 171-182. |
| [16] | Ma X L, Zhang Q Y, Zhu Q L, Liu W, Chen Y, Qiu R, Wang B, Yang Z F, Li H Y, Lin Y R, Xie Y Y, Shen R X, Chen S F, Wang Z, Chen Y L, Guo J X, Chen L T, Zhao X C, Dong Z C, Liu Y G.A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants[J]. Molecular Plant, 2015, 8(8): 1274-1284. |
| [17] | Liang G, Zhang H M, Lou D J, Yu D Q.Selection of highly efficient sgRNAs for CRISPR/Cas9-based plant genome editing[J]. Scientific Reports, 2016, 6: 21451. |
| [18] | Wang Y P, Cheng X, Shan Q W, Zhang Y, Liu J X, Gao C X.Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew[J]. Nature Biotechnology, 2014, 32(9): 947-951. |
| [19] | Li J Y, Sun Y W, Du J L, Zhao Y D, Xia L Q.Generation of targeted point mutations in rice by a modified CRISPR/Cas9 system[J]. Molecular Plant, 2017, 10(3): 526-529. |
| [20] | 王影, 李相敢, 邱丽娟. CRISPR/Cas9基因组定点编辑中脱靶现象的研究进展[J]. 植物学报, 2018, 53(4): 521-541. |
| [20] | Wang Y, Li X G, Qiu L J.Research progress in off-target in CRISPR/Cas9 genome editing[J]. Chinese Bulletin of Botany, 2018, 53(4): 521-541. (in Chinese with English abstract) |
| [21] | Garcia M E, Lynch T, Peeters J, Snowden C, Finkelstein R.A small plant-specific protein family of ABI five binding proteins (AFPs) regulates stress response in germinating Arabidopsis seeds and seedlings[J]. Plant Molecular Biology, 2008, 67(6): 643-658. |
| [22] | Lopez-Molina L, Mongrand S, Kinoshita N, Chua N H.AFP is a novel negative regulator of ABA signaling that promotes ABI5 protein degradation[J]. Genes & Development, 2003, 17(3): 410-418. |
/
| 〈 |
|
〉 |