Construction of a high-density mutant population of Chinese cabbage facilitates the genetic dissection of agronomic traits
Construction of a high-density mutant population of Chinese cabbage facilitates the genetic dissection of agronomic traits
复制标题
大白菜高密度突变体群体的构建有利于农艺性状的遗传解析
DOI：
10.1016/j.molp.2022.02.006
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发表时间：
2022-05-02
期刊：
影响因子：
27.5
通讯作者：
Zhao, Jianjun
中科院分区：
文献类型：
--
作者：
Sun, Xiaoxue;Li, Xing;Zhao, Jianjun
白菜(Brassica Rapa SSP.)北京)是一种重要的经济作物，在世界各地，特别是在亚洲。大白菜可以获得高质量的基因组序列，但基因功能研究仍然具有挑战性。为促进大白菜功能基因组学研究，我们以大白菜双单倍体自交系A03为亲本，构建了一个与8000株M-2植株相似的甲基磺酸乙酯突变体群体。对A03的基因组进行了测序，并作为高通量全基因组水平上基因突变功能鉴定的参考。对300个M-2到M-5 EMS突变进行了表型筛选和测序，发现了750 629个SNPs和46 272个Indel突变，覆盖了A03基因组所有预测基因的98.27%。一种正向遗传学方法被成功地用于鉴定两个具有叶绿体相关功能的基因，这两个基因与黄叶突变性状有关。反向遗传学方法也被用来确定硫代葡萄糖苷生物合成途径的五个基因突变与突变植株中硫代葡萄糖苷含量变化之间的联系。此外，我们还建立了大白菜细胞质雄性不育突变数据库(CCEMD，www.bialinformaticslab.cn/EMS/home)，以提高这一突变群体资源的可用性。综上所述，我们在表型和基因水平上对一个结球白菜EMS突变体进行了大规模的筛选，这将为大白菜的基因挖掘提供便利，也可能对其他大白菜作物的研究有所帮助。
Chinese cabbage (Brassica rapa ssp. pekinensis) is an economically important vegetable crop throughout the world, especially in Asia. High-quality genome sequences are available for Chinese cabbage, but gene functional studies remain challenging. To promote functional genomic studies of Chinese cabbage, we generated an ethyl methane sulfonate (EMS) mutant population of similar to 8000 M-2 plants using the double haploid inbred line A03 as the parent. The genome of A03 was sequenced and used as a reference for high-throughput functional characterization of gene mutations at the whole-genome level. A total of 300 M-2 to M-5 EMS mutants were phenotypically screened and then sequenced, revealing 750 629 SNPs and 46 272 InDel mutations that cover 98.27% of all predicted genes in the A03 genome. A forward-genetics approach was successfully used to identify two genes with chloroplast-related functions that are responsible for the yellow leaf mutant trait. A reverse-genetics approach was also used to identify associations between mutations in five genes of the glucosinolate biosynthetic pathway and variations in glucosinolate content of the mutant plants. In addition, we built the Chinese cabbage EMS mutation database (CCEMD, www.bioinformaticslab.cn/EMSmutation/home) to increase the usability of this mutant population resource. In summary, we performed large-scale screening of a heading Chinese cabbage EMS mutant collection at the phenotypic and genotypic levels, which will facilitate gene mining of Chinese cabbage and might also be useful for the study of other Brassica crops.