CRISPR/Cas9-mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean.
CRISPR/Cas9-mediated targeted mutagenesis of GmFT2a delays flowering time in soya bean.
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CRISPR/Cas9 介导的 GmFT2a 定向诱变延迟了大豆的开花时间。
DOI：
10.1111/pbi.12758
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发表时间：
2018-01
影响因子：
13.8
通讯作者：
Hou W
中科院分区：
文献类型：
--
作者：
Cai Y;Chen L;Liu X;Guo C;Sun S;Wu C;Jiang B;Han T;Hou W
开花是大豆从营养生长向生殖生长过渡的标志，对大豆的生活史有相当大的影响。在这项研究中，我们利用CRISPR/Cas9系统特异性地诱导大豆光周期开花途径中的整合子GmFT2a的定向突变。利用根癌农杆菌介导法，将3个针对内源GmFT2a不同位点的sgRNA/Cas9载体转化大豆品种JACK。通过DNA测序分析，发现所有靶点都发生了定点突变。在北京，中国(N39°58‘，E116°20’)，通过1个碱基插入或短缺失突变的GmFT2a移码突变缺失零等位基因纯合的T1代大豆植株在自然条件下(夏季)表现出晚开花。我们还发现，定向突变在下一代T2代中稳定可遗传，纯合子GmFT2a突变体在长日照和短日照条件下都表现出延迟开花。我们鉴定了一些转基因干净的大豆植株，它们是内源GmFT2a零等位基因的纯合子，没有来自T1和T2世代的任何转基因元素。这些转基因干净的GmFT2a突变体可能为更深入地研究GmFT2a的功能和大豆光周期反应的分子机制提供材料。他们还将为大豆育种和区域引种做出贡献。
Flowering is an indication of the transition from vegetative growth to reproductive growth and has considerable effects on the life cycle of soya bean (Glycine max). In this study, we employed the CRISPR/Cas9 system to specifically induce targeted mutagenesis of GmFT2a, an integrator in the photoperiod flowering pathway in soya bean. The soya bean cultivar Jack was transformed with three sgRNA/Cas9 vectors targeting different sites of endogenous GmFT2a via Agrobacterium tumefaciens‐mediated transformation. Site‐directed mutations were observed at all targeted sites by DNA sequencing analysis. T1‐generation soya bean plants homozygous for null alleles of GmFT2a frameshift mutated by a 1‐bp insertion or short deletion exhibited late flowering under natural conditions (summer) in Beijing, China (N39°58′, E116°20′). We also found that the targeted mutagenesis was stably heritable in the following T2 generation, and the homozygous GmFT2a mutants exhibited late flowering under both long‐day and short‐day conditions. We identified some ‘transgene‐clean’ soya bean plants that were homozygous for null alleles of endogenous GmFT2a and without any transgenic element from the T1 and T2 generations. These ‘transgene‐clean’ mutants of GmFT2a may provide materials for more in‐depth research of GmFT2a functions and the molecular mechanism of photoperiod responses in soya bean. They will also contribute to soya bean breeding and regional introduction.
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影响因子：
27.5
作者：
Ma, Xingliang;Zhang, Qunyu;Liu, Yao-Guang
通讯作者：
Liu, Yao-Guang
影响因子：
17.3
作者：
Gaj, Thomas;Gersbach, Charles A.;Barbas, Carlos F., III
通讯作者：
Barbas, Carlos F., III
影响因子：
56.9
作者：
Jinek, Martin;Chylinski, Krzysztof;Charpentier, Emmanuelle
通讯作者：
Charpentier, Emmanuelle
影响因子：
56.9
作者：
Kobayashi, Y;Kaya, H;Araki, T
通讯作者：
Araki, T
影响因子：
9.2
作者：
Jaeger, Katja E.;Wigge, Philip A.
通讯作者：
Wigge, Philip A.