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A method for the production and expedient screening of CRISPR/Cas9-mediated non-transgenic mutant plants

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成果类型:
期刊论文
作者:
Chen, Longzheng;Li, Wei;Katin-Grazzini, Lorenzo;Ding, Jing;Gu, Xianbin;...
通讯作者:
Li, Yi
作者机构:
[Chen, Longzheng; Wang, Ren; Gu, Xianbin; Li, Wei; Li, Yanjun; Li, Yi; McAvoy, Richard J.; Katin-Grazzini, Lorenzo; Lin, Xinchun] Univ Connecticut, Dept Plant Sci & Landscape Architecture, Storrs, CT 06269 USA.
[Chen, Longzheng] Jiangsu Acad Agr Sci, Inst Vegetable Crops, Nanjing, Jiangsu, Peoples R China.
[Gu, Tingting; Li, Yi; Ding, Jing] Nanjing Agr Univ, Coll Hort, Nanjing, Jiangsu, Peoples R China.
[Gu, Tingting; Li, Yi; Ding, Jing] Nanjing Agr Univ, State Key Lab Crop Genet & Germplasm Enhancement, Nanjing, Jiangsu, Peoples R China.
[Lin, Xinchun] Zhejiang Agr & Forestry Univ, State Key Lab Subtrop Silviculture, Hangzhou, Zhejiang, Peoples R China.
通讯机构:
[Li, Yi] U
[Li, Yi] N
Univ Connecticut, Dept Plant Sci & Landscape Architecture, Storrs, CT 06269 USA.
Nanjing Agr Univ, Coll Hort, Nanjing, Jiangsu, Peoples R China.
Nanjing Agr Univ, State Key Lab Crop Genet & Germplasm Enhancement, Nanjing, Jiangsu, Peoples R China.
语种:
英文
关键词:
Genetic engineering;High-throughput screening;Molecular engineering in plants;Mutation;Next-generation sequencing;Gene editing
期刊:
园艺研究(英文)
ISSN:
2662-6810
年:
2018
卷:
5
期:
1
页码:
1-12
基金类别:
We thank the financial support from the USDA National Institute of Food and Agriculture SCRI (grant no. 2015-70016-23027), the Florida Citrus Development Foundation (2016-001) and the Genetically Modified Organisms Breeding Major Projects of China (2014ZX0801008B-001). The Connecticut-Storrs Agriculture Experimental Station, the Priority Academic Program Development of Jiangsu Higher Education Institutions and the Innovative Research Project of JAAS (ZX-17-2006) also contributed financially to some experiments presented in this manuscript. Y.L. holds a 2-month/-year-visiting professor position at Nanjing Agricultural University, China.
机构署名:
本校为其他机构
院系归属:
园艺园林学院
摘要:
Developing CRISPR/Cas9-mediated non-transgenic mutants in asexually propagated perennial crop plants is challenging but highly desirable. Here, we report a highly useful method using an Agrobacterium-mediated transient CRISPR/Cas9 gene expression system to create non-transgenic mutant plants without the need for sexual segregation. We have also developed a rapid, cost-effective, and high-throughput mutant screening protocol based on Illumina sequencing followed by high-resolution melting (HRM) analysis. Using tetraploid tobacco as a model species and the phytoene desaturase (PDS) gene as a tar...

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