Precise editing of CLAVATA genes in Brassica napus L. regulates multilocular silique development.
Precise editing of CLAVATA genes in Brassica napus L. regulates multilocular silique development.
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甘蓝型油菜 CLAVATA 基因的精确编辑调节多房长角果的发育
DOI：
10.1111/pbi.12872
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发表时间：
2018-07
影响因子：
13.8
通讯作者：
Zhou Y
中科院分区：
文献类型：
--
作者：
Yang Y;Zhu K;Li H;Han S;Meng Q;Khan SU;Fan C;Xie K;Zhou Y
多室单倍体是一种理想的农业性状，在培育芸苔属高产品种方面具有很大的潜力。到目前为止，在甘蓝型油菜中还没有自发或诱导的多室突变的报道，这可能反映了甘蓝型油菜的异源四倍体性质，以及具有功能冗余的多个基因拷贝同时随机突变的可能性极低。在这里，我们提供了证据，证明使用CRISPR/Cas9系统有效地敲除CLAVATA3 （CLV3）的油菜籽同源物，用于CLV信号通路中的分泌肽及其相关受体CLV1和CLV2，并实现了突变在三代之间的稳定传播。每个BnCLV基因在两个亚基因组中有两个拷贝。多室表型只能在每个BnCLV基因的两个拷贝的敲除突变中恢复，说明通过CRISPR/Cas9诱变同时改变多个基因拷贝在油菜籽中产生重要的农艺突变具有很大的潜力。不同的单导rna （sgRNA）在T0中的诱变效率从0%到48.65%不等，这表明sgRNA的选择对油菜籽中有效产生索引很重要。与野生型和单突变植物相比，BnCLV3的双突变产生了更多的叶片和多室角果，每个角果的种子数量和种子重量显著增加，可能有助于增加种子产量。我们还通过授粉评估了Cas9/gRNA磁带的水平转移效率。我们的发现揭示了植物育种策略的潜力，以提高目前栽培的油菜籽品种的产量性状。
Multilocular silique is a desirable agricultural trait with great potential for the development of high‐yield varieties of Brassica. To date, no spontaneous or induced multilocular mutants have been reported in Brassica napus, which likely reflects its allotetraploid nature and the extremely low probability of the simultaneous random mutagenesis of multiple gene copies with functional redundancy. Here, we present evidence for the efficient knockout of rapeseed homologues of CLAVATA3 (CLV3) for a secreted peptide and its related receptors CLV1 and CLV2 in the CLV signalling pathway using the CRISPR/Cas9 system and achieved stable transmission of the mutations across three generations. Each BnCLV gene has two copies located in two subgenomes. The multilocular phenotype can be recovered only in knockout mutations of both copies of each BnCLV gene, illustrating that the simultaneous alteration of multiple gene copies by CRISPR/Cas9 mutagenesis has great potential in generating agronomically important mutations in rapeseed. The mutagenesis efficiency varied widely from 0% to 48.65% in T0 with different single‐guide RNAs (sgRNAs), indicating that the appropriate selection of the sgRNA is important for effectively generating indels in rapeseed. The double mutation of BnCLV3 produced more leaves and multilocular siliques with a significantly higher number of seeds per silique and a higher seed weight than the wild‐type and single mutant plants, potentially contributing to increased seed production. We also assessed the efficiency of the horizontal transfer of Cas9/gRNA cassettes by pollination. Our findings reveal the potential for plant breeding strategies to improve yield traits in currently cultivated rapeseed varieties.
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影响因子：
56.9
作者：
Fletcher, LC;Brand, U;Meyerowitz, EM
通讯作者：
Meyerowitz, EM
影响因子：
13.8
作者：
Jia H;Zhang Y;Orbović V;Xu J;White FF;Jones JB;Wang N
通讯作者：
Wang N
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：
12.3
作者：
Lawrenson T;Shorinola O;Stacey N;Li C;Østergaard L;Patron N;Uauy C;Harwood W
通讯作者：
Harwood W
影响因子：
7.4
作者：
Braatz, Janina;Harloff, Hans-Joachim;Jung, Christian
通讯作者：
Jung, Christian