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Creation of Early Flowering Germplasm of Soybean by CRISPR/Cas9 Technology

文献类型： 外文期刊

作者： Han, Jianan 1 ; Guo, Bingfu 3 ; Guo, Yong 1 ; Zhang, Bo 4 ; Wang, Xiaobo 5 ; Qiu, Li-Juan 1 ;

作者机构： 1.Chinese Acad Agr Sci, Inst Crop Sci, Natl Key Facil Crop Gene Resources & Genet Improv, Beijing, Peoples R China

2.Chinese Acad Agr Sci, Inst Crop Sci, MOA Key Labs Crop Germplasm & Soybean Biol Beijin, Beijing, Peoples R China

3.Jiangxi Acad Agr Sci, Crop Res Inst, Nanchang, Jiangxi, Peoples R China

4.Virginia Polytech Inst & State Univ, Sch Plant & Environm Sci, Blacksburg, VA 24061 USA

5.Anhui Agr Univ, Coll Agron, Hefei, Anhui, Peoples R China

关键词： soybean; CRISPR; Cas9; gene editing; E1; Agrobacterium-mediated transformation

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2019 年 10 卷

页码：

收录情况： SCI

摘要： Soybean is an important economic crop and a typical short-day crop, sensitive to photoperiod, and has narrow geographical adaptative region, which limit the creation of transgenic materials and reduce the breeding efficiency of new varieties. In addition, the genetic transformation efficiency of soybean is lower than that of many other crops, and the available receptor genotypes are limited. In this study, Agrobacterium-mediated transformation were used to introduce the CRISPR/Cas9 expression vector into soybean cultivar Jack and generated targeted mutants of E1 gene controlling soybean flowering. We obtained two novel types of mutations, 11 bp and 40 bp deletion at E1 coding region, respectively, and frameshift mutations produced premature translation termination codons and truncated E1 proteins, causing obvious early flowering under long day condition. In addition, no off-target effects were observed by predicting and analyzing the potential off-target sites of E1 targets. Significant decreased E1 gene expression of two novel mutants showed that the truncated E1 protein disinhibited GmFT2a/5a and increasing GmFT2a/5a gene expressions resulted obvious early flowering. Homozygous trans-clean mutants without T-DNA elements were also obtained and showed early flowering under long day condition. The photo-insensitive soybean transformation receptor we created laid a foundation for breeding excellent transgenic receptors suitable for high latitudes.

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