Breeding of Indica glutinous cytoplasmic male sterile line WX209A via CRISPR/Cas9 mediated genomic editing

https://doi.org/10.17221/197/2017-CJGPBCitation:Wang X., Han Y., Feng X., Li Y., Qin B., Luo J., Wei Z., Qiu Y., Liu F., Li R. (2019): Breeding of Indica glutinous cytoplasmic male sterile line WX209A via CRISPR/Cas9 mediated genomic editing. Czech J. Genet. Plant Breed., 55: 93-100.
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Glutinous cytoplasmic male sterile (CMS) line is necessary to select hybrid glutinous rice combination with high yield and quality. To develop glutinous CMS with low amylose content, in this study, we firstly knocked out the granule-bound starch synthase OsWaxy in 209B using CRISPR/Cas9 mediated genome editing technology and successfully obtained a glutinous maintainer line WX209B. Comparing with maintainer line 209B, WX209B showed decreased amylose contents and similar agronomic characters. And then, through one generation of hybridization and two generations of backcrossing with WX209B as the male parent and 209A as the female parent, the glutinous CMS line WX209A was successfully achieved. Our study provides a strategy to efficiently breed for the glutinous cytoplasmic male sterile line by combining CRISPR/Cas9-mediated gene editing technology with conventional backcross breeding method in a short period, which prepares the ground for further breeding of hybrid glutinous rice variety.

 

References:
Cong L., Ran F. A., Cox D., Lin S., Barretto R., Habib N., Hsu P. D., Wu X., Jiang W., Marraffini L. A., Zhang F. (2013): Multiplex Genome Engineering Using CRISPR/Cas Systems. Science, 339, 819-823 https://doi.org/10.1126/science.1231143
 
DiCarlo James E., Norville Julie E., Mali Prashant, Rios Xavier, Aach John, Church George M. (2013): Genome engineering in Saccharomyces cerevisiae using CRISPR-Cas systems. Nucleic Acids Research, 41, 4336-4343 https://doi.org/10.1093/nar/gkt135
 
Feng Zhengyan, Zhang Botao, Ding Wona, Liu Xiaodong, Yang Dong-Lei, Wei Pengliang, Cao Fengqiu, Zhu Shihua, Zhang Feng, Mao Yanfei, Zhu Jian-Kang (2013): Efficient genome editing in plants using a CRISPR/Cas system. Cell Research, 23, 1229-1232 https://doi.org/10.1038/cr.2013.114
 
Hiei Yukoh, Ohta Shozo, Komari Toshihiko, Kumashiro Takashi (1994): Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. The Plant Journal, 6, 271-282 https://doi.org/10.1046/j.1365-313X.1994.6020271.x
 
Hwang Woong Y, Fu Yanfang, Reyon Deepak, Maeder Morgan L, Tsai Shengdar Q, Sander Jeffry D, Peterson Randall T, Yeh J-R Joanna, Joung J Keith (2013): Efficient genome editing in zebrafish using a CRISPR-Cas system. Nature Biotechnology, 31, 227-229 https://doi.org/10.1038/nbt.2501
 
Jiang Wenzhi, Zhou Huanbin, Bi Honghao, Fromm Michael, Yang Bing, Weeks Donald P. (2013): Demonstration of CRISPR/Cas9/sgRNA-mediated targeted gene modification in Arabidopsis, tobacco, sorghum and rice. Nucleic Acids Research, 41, e188-e188 https://doi.org/10.1093/nar/gkt780
 
Li M., Li X.X., Zhou Z.J., Wu P.Z., Fang M.C., Pan X.P., Lin Q.P., Luo W.B., Wu G.L., Li H.Q. (2016): Reassessment of the four yield-related genes Gn1a, DEP1, GS3, and IPA1 in rice using a CRISPR/Cas9 system. Frontiers in Plant Science, 7: 377.
 
Liang Zhen, Zhang Kang, Chen Kunling, Gao Caixia (2014): Targeted Mutagenesis in Zea mays Using TALENs and the CRISPR/Cas System. Journal of Genetics and Genomics, 41, 63-68 https://doi.org/10.1016/j.jgg.2013.12.001
 
Ma Xingliang, Zhang Qunyu, Zhu Qinlong, Liu Wei, Chen Yan, Qiu Rong, Wang Bin, Yang Zhongfang, Li Heying, Lin Yuru, Xie Yongyao, Shen Rongxin, Chen Shuifu, Wang Zhi, Chen Yuanling, Guo Jingxin, Chen Letian, Zhao Xiucai, Dong Zhicheng, Liu Yao-Guang (2015): A Robust CRISPR/Cas9 System for Convenient, High-Efficiency Multiplex Genome Editing in Monocot and Dicot Plants. Molecular Plant, 8, 1274-1284 https://doi.org/10.1016/j.molp.2015.04.007
 
Miao Jin, Guo Dongshu, Zhang Jinzhe, Huang Qingpei, Qin Genji, Zhang Xin, Wan Jianmin, Gu Hongya, Qu Li-Jia (2013): Targeted mutagenesis in rice using CRISPR-Cas system. Cell Research, 23, 1233-1236 https://doi.org/10.1038/cr.2013.123
 
Perez C. M., Juliano B. O. (1978): Modification of the Simplified Amylose Test for Milled Rice. Starch - Stärke, 30, 424-426 https://doi.org/10.1002/star.19780301206
 
Rowland L.J., Nguyen B. (1993): Use of polyethylene glycol for purification of DNA from leaf tissue of woody plants. Biotechniques, 14: 734–736.
 
Shan Qiwei, Wang Yanpeng, Li Jun, Zhang Yi, Chen Kunling, Liang Zhen, Zhang Kang, Liu Jinxing, Xi Jianzhong Jeff, Qiu Jin-Long, Gao Caixia (2013): Targeted genome modification of crop plants using a CRISPR-Cas system. Nature Biotechnology, 31, 686-688 https://doi.org/10.1038/nbt.2650
 
Shan Qiwei, Wang Yanpeng, Li Jun, Gao Caixia (2014): Genome editing in rice and wheat using the CRISPR/Cas system. Nature Protocols, 9, 2395-2410 https://doi.org/10.1038/nprot.2014.157
 
Terada Rie, Nakajima Midori, Isshiki Masayuki, Okagaki Ron J., Wessler Susan R., Shimamoto Ko (2000): Antisense Waxy Genes with Highly Active Promoters Effectively Suppress Waxy Gene Expression in Transgenic Rice. Plant and Cell Physiology, 41, 881-888 https://doi.org/10.1093/pcp/pcd008
 
Wang Haoyi, Yang Hui, Shivalila Chikdu S., Dawlaty Meelad M., Cheng Albert W., Zhang Feng, Jaenisch Rudolf (2013): One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering. Cell, 153, 910-918 https://doi.org/10.1016/j.cell.2013.04.025
 
Wang Tim, Wei Jenny J., Sabatini David M., Lander Eric S. (2014): Genetic Screens in Human Cells Using the CRISPR-Cas9 System. Science, 343, 80-84 https://doi.org/10.1126/science.1246981
 
Wang Yanpeng, Cheng Xi, Shan Qiwei, Zhang Yi, Liu Jinxing, Gao Caixia, Qiu Jin-Long (2014): Simultaneous editing of three homoeoalleles in hexaploid bread wheat confers heritable resistance to powdery mildew. Nature Biotechnology, 32, 947-951 https://doi.org/10.1038/nbt.2969
 
Wang Zong-yang, Wu Zhi-liang, Xing Yan-yan, Zheng Fei-gin, Guo Xiao-li, Zhang Wei-guo, Hong Meng-min (1990): Nucleotide sequence of rice waxy gene. Nucleic Acids Research, 18, 5898-5898 https://doi.org/10.1093/nar/18.19.5898
 
Xu Rong-Fang, Li Hao, Qin Rui-Ying, Li Juan, Qiu Chun-Hong, Yang Ya-Chun, Ma Hui, Li Li, Wei Peng-Cheng, Yang Jian-Bo (2015): Generation of inheritable and “transgene clean” targeted genome-modified rice in later generations using the CRISPR/Cas9 system. Scientific Reports, 5, - https://doi.org/10.1038/srep11491
 
Zhang Hui, Zhang Jinshan, Wei Pengliang, Zhang Botao, Gou Feng, Feng Zhengyan, Mao Yanfei, Yang Lan, Zhang Heng, Xu Nanfei, Zhu Jian-Kang (2014): The CRISPR/Cas9 system produces specific and homozygous targeted gene editing in rice in one generation. Plant Biotechnology Journal, 12, 797-807 https://doi.org/10.1111/pbi.12200
 
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