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Multiplex CRISPR/Cas9-mediated knockout of soybean LNK2 advances flowering time

文献类型： 外文期刊

作者： Li, Zhaobo 1 ; Cheng, Qun 3 ; Gan, Zhuoran 3 ; Hou, Zhihong 4 ; Zhang, Yuhang 3 ; Li, Yongli 3 ; Li, Haiyang 5 ; Nan, Hai 1 ;

作者机构： 1.Chinese Acad Sci, Innovat Acad Seed Design, Northeast Inst Geog & Agroecol, Key Lab Soybean Mol Design Breeding, Harbin 130102, Heilongjiang, Peoples R China

2.Univ Chinese Acad Sci, Beijing 100081, Peoples R China

3.Guangzhou Univ, Innovat Ctr Mol Genet & Evolut, Sch Life Sci, Guangzhou 510006, Guangdong, Peoples R China

4.Heilongjiang Bayi Agr Univ, Agron Coll, Daqing 163319, Heilongjiang, Peoples R China

5.Nanjing Agr Univ, Natl Ctr Soybean Improvement, Jiangsu Collaborat Innovat Ctr Modern Crop Prod, Natl Key Lab Crop Genet & Germplasm Enhancement, Nanjing 210095, Jiangsu, Peoples R China

6.Heilongjiang Acad Agr Sci, Mudanjiang Branch, Mudanjiang 157000, Heilongjiang, Peoples R China

关键词： Soybean; LNK2; CRISPR/Cas9; Genome editing; Flowering time

期刊名称：CROP JOURNAL （ 2020影响因子：4.407； 五年影响因子：5.687 ）

ISSN： 2095-5421

年卷期： 2021 年 9 卷 4 期

页码：

收录情况： SCI

摘要： Flowering time is an important agronomic trait for soybean yield and adaptation. However, the genetic basis of soybean adaptation to diverse latitudes is still not clear. Four NIGHT LIGHT-INDUCIBLE AND CLOCK-REGULATED 2 (LNK2) homeologs of Arabidopsis thaliana LNK2 were identified in soybean. Three single-guide RNAs were designed for editing the four LNK2 genes. A transgene-free homozygous quadruple mutant of the LNK2 genes was developed using the CRISPR (clustered regularly interspaced short palindromic repeats)/Cas9 (CRISPR-associated protein 9). Under long-day (LD) conditions, the quadruple mutant flowered significantly earlier than the wild-type (WT). Quantitative real-time PCR (qRT-PCR) revealed that transcript levels of LNK2 were significantly lower in the quadruple mutant than in the WT under LD conditions. LNK2 promoted the expression of the legume-specific El gene and repressed the expression of FT2a. Genetic markers were developed to identify LNK2 mutants for soybean breeding. These results indicate that CRISPR/Cas9-mediated targeted mutagenesis of four LNK2 genes shortens flowering time in soybean. Our findings identify novel components in flowering-time control in soybean and may be beneficial for further soybean breeding in high-latitude environments. (C) 2021 Crop Science Society of China and Institute of Crop Science, CAAS. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.

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