CRISPR/Cas9-mediated knockout of Ms1 enables the rapid generation of male-sterile hexaploid wheat lines for use in hybrid seed production

被引:131
作者
Okada, Anzu [1 ]
Arndell, Taj [1 ]
Borisjuk, Nikolai [1 ,2 ]
Sharma, Niharika [1 ,3 ]
Watson-Haigh, Nathan S. [1 ]
Tucker, Elise J. [1 ,4 ]
Baumann, Ute [1 ]
Langridge, Peter [1 ]
Whitford, Ryan [1 ]
机构
[1] Univ Adelaide, Sch Agr Food & Wine, Urrbrae, SA, Australia
[2] Huaiyin Normal Univ, Sch Life Sci, Huaian, Peoples R China
[3] New South Wales Dept Primary Ind, Res Excellence, Orange, NSW, Australia
[4] CSIRO, Agr & Food, Urrbrae, SA, Australia
基金
澳大利亚研究理事会;
关键词
CRISPR; Cas9; genome editing; hybrid wheat; Ms1; Triticum aestivum; male sterility; GENOME; ENDONUCLEASE; BINDING; RICE; CAS9; TECHNOLOGIES; SPECIFICITY; DESIGN; SYSTEM; BARLEY;
D O I
10.1111/pbi.13106
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
The development and adoption of hybrid seed technology have led to dramatic increases in agricultural productivity. However, it has been a challenge to develop a commercially viable platform for the production of hybrid wheat (Triticum aestivum) seed due to wheat's strong inbreeding habit. Recently, a novel platform for commercial hybrid seed production was described. This hybridization platform utilizes nuclear male sterility to force outcrossing and has been applied to maize and rice. With the recent molecular identification of the wheat male fertility gene Ms1, it is now possible to extend the use of this novel hybridization platform to wheat. In this report, we used the CRISPR/Cas9 system to generate heritable, targeted mutations in Ms1. The introduction of biallelic frameshift mutations into Ms1 resulted in complete male sterility in wheat cultivars Fielder and Gladius, and several of the selected male-sterile lines were potentially non-transgenic. Our study demonstrates the utility of the CRISPR/Cas9 system for the rapid generation of male sterility in commercial wheat cultivars. This represents an important step towards capturing heterosis to improve wheat yields, through the production and use of hybrid seed on an industrial scale.
引用
收藏
页码:1905 / 1913
页数:9
相关论文
共 35 条
[1]   Easy quantitative assessment of genome editing by sequence trace decomposition [J].
Brinkman, Eva K. ;
Chen, Tao ;
Amendola, Mario ;
van Steensel, Bas .
NUCLEIC ACIDS RESEARCH, 2014, 42 (22)
[2]   Construction of a male sterility system for hybrid rice breeding and seed production using a nuclear male sterility gene [J].
Chang, Zhenyi ;
Chen, Zhufeng ;
Wang, Na ;
Xie, Gang ;
Lu, Jiawei ;
Yan, Wei ;
Zhou, Junli ;
Tang, Xiaoyan ;
Deng, Xing Wang .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2016, 113 (49) :14145-14150
[3]   Targeted mutagenesis of a conserved anther-expressed P450 gene confers male sterility in monocots [J].
Cigan, A. Mark ;
Singh, Manjit ;
Benn, Geoffrey ;
Feigenbutz, Lanie ;
Kumar, Manish ;
Cho, Myeong-Je ;
Svitashev, Sergei ;
Young, Joshua .
PLANT BIOTECHNOLOGY JOURNAL, 2017, 15 (03) :379-389
[4]   A gateway cloning vector set for high-throughput functional analysis of genes in planta [J].
Curtis, MD ;
Grossniklaus, U .
PLANT PHYSIOLOGY, 2003, 133 (02) :462-469
[5]   Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9 [J].
Doench, John G. ;
Fusi, Nicolo ;
Sullender, Meagan ;
Hegde, Mudra ;
Vaimberg, Emma W. ;
Donovan, Katherine F. ;
Smith, Ian ;
Tothova, Zuzana ;
Wilen, Craig ;
Orchard, Robert ;
Virgin, Herbert W. ;
Listgarten, Jennifer ;
Root, David E. .
NATURE BIOTECHNOLOGY, 2016, 34 (02) :184-+
[6]   An Effective Strategy for Reliably Isolating Heritable and Cas9-Free Arabidopsis Mutants Generated by CRISPR/Cas9-Mediated Genome Editing [J].
Gao, Xiuhua ;
Chen, Jilin ;
Dai, Xinhua ;
Zhang, Da ;
Zhao, Yunde .
PLANT PHYSIOLOGY, 2016, 171 (03) :1794-1800
[7]   DNA targeting specificity of RNA-guided Cas9 nucleases [J].
Hsu, Patrick D. ;
Scott, David A. ;
Weinstein, Joshua A. ;
Ran, F. Ann ;
Konermann, Silvana ;
Agarwala, Vineeta ;
Li, Yinqing ;
Fine, Eli J. ;
Wu, Xuebing ;
Shalem, Ophir ;
Cradick, Thomas J. ;
Marraffini, Luciano A. ;
Bao, Gang ;
Zhang, Feng .
NATURE BIOTECHNOLOGY, 2013, 31 (09) :827-+
[8]  
Ishida Y, 2015, METHODS MOL BIOL, V1223, P189, DOI 10.1007/978-1-4939-1695-5_15
[9]   A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity [J].
Jinek, Martin ;
Chylinski, Krzysztof ;
Fonfara, Ines ;
Hauer, Michael ;
Doudna, Jennifer A. ;
Charpentier, Emmanuelle .
SCIENCE, 2012, 337 (6096) :816-821
[10]  
Kovalchuk N, 2014, METHODS MOL BIOL, V1145, P239, DOI 10.1007/978-1-4939-0446-4_18