Identification, characterization, and rescue of CRISPR/Cas9 generated wheat SPO11-1 mutants.
Identification, characterization, and rescue of CRISPR/Cas9 generated wheat SPO11-1 mutants.
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DOI：
10.1111/pbi.13961
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发表时间：
2023-02
影响因子：
13.8
通讯作者：
Edwards, Keith J.
中科院分区：
文献类型：
--
作者：
Hyde, Lucy;Osman, Kim;Winfield, Mark;Sanchez-Moran, Eugenio;Higgins, James D.;Henderson, Ian R.;Sparks, Caroline;Franklin, F. Chris H.;Edwards, Keith J.
通过植物育种提高作物产量既费时又费力，等位基因的新组合的产生受到染色体连锁阻断和连锁阻力的限制。减数分裂重组对于通过亲本等位基因的改组来产生新的遗传变异是必不可少的。同源染色体之间的遗传信息交换发生在交叉（CO）位点，但CO频率通常较低且分布不均匀。这种偏差在重组“冷”区域产生了连锁阻力的问题，其中不期望的变异仍然与有用的性状相关联。在植物中，由SPO 11复合物催化的程序性减数分裂特异性DNA双链断裂启动重组途径，尽管只有约5%导致CO的形成。为了研究SPO 11 - 1在小麦减数分裂中的作用，作为操作的前奏，我们使用CRISPR/Cas9在六倍体小麦的所有三个SPO 11 - 1同源物中生成编辑。后代品系的表征显示，所有六个SPO 11 - 1拷贝缺陷的植物不能经历染色体联会，缺乏CO并且是不育的。相比之下，携带三种野生型同源物中任何一种的单拷贝的品系在营养生长和育性方面与未编辑的植物在表型上是不可区分的。然而，对编辑过的植物的细胞遗传学分析表明，同源物在产生CO的能力和突触的动力学方面存在差异。此外，我们还发现，携带6个经编辑的SPO 11 - 1拷贝和TaSPO 11 - 1B基因的小麦突变体的转化，恢复了突触、CO形成和生育力，从而为这种具有重要农业意义的作物的改良重组开辟了一条途径。
Increasing crop yields through plant breeding is time consuming and laborious, with the generation of novel combinations of alleles being limited by chromosomal linkage blocks and linkage‐drag. Meiotic recombination is essential to create novel genetic variation via the reshuffling of parental alleles. The exchange of genetic information between homologous chromosomes occurs at crossover (CO) sites but CO frequency is often low and unevenly distributed. This bias creates the problem of linkage‐drag in recombination ‘cold’ regions, where undesirable variation remains linked to useful traits. In plants, programmed meiosis‐specific DNA double‐strand breaks, catalysed by the SPO11 complex, initiate the recombination pathway, although only ~5% result in the formation of COs. To study the role of SPO11‐1 in wheat meiosis, and as a prelude to manipulation, we used CRISPR/Cas9 to generate edits in all three SPO11‐1 homoeologues of hexaploid wheat. Characterization of progeny lines shows plants deficient in all six SPO11‐1 copies fail to undergo chromosome synapsis, lack COs and are sterile. In contrast, lines carrying a single copy of any one of the three wild‐type homoeologues are phenotypically indistinguishable from unedited plants both in terms of vegetative growth and fertility. However, cytogenetic analysis of the edited plants suggests that homoeologues differ in their ability to generate COs and in the dynamics of synapsis. In addition, we show that the transformation of wheat mutants carrying six edited copies of SPO11‐1 with the TaSPO11‐1B gene, restores synapsis, CO formation, and fertility and hence opens a route to modifying recombination in this agronomically important crop.
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影响因子：
5.6
作者：
Osman K;Algopishi U;Higgins JD;Henderson IR;Edwards KJ;Franklin FCH;Sanchez-Moran E
通讯作者：
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