新型大豆皱叶基因CL12的图位克隆及其对产量性状影响

广西、广东等地大豆上出现的一种叶片皱缩症状可导致减产20%~30%，该症状为某种外因所致，可稳定遗传，与已报道皱叶类型不同。前期研究表明该性状由一个显性基因控制，用BSA-seq技术把该基因（CL12）定位于大豆12号染色体末端1.96Mb区间内。为进一步明确该基因对大豆产量性状影响，找出CL12候选基因以期进一步解析皱叶发生机制及找出皱叶诱因，本研究拟（1）用F2群体隐形单株及INDEL、SNP等分子标记在目标基因区间构建高密度连锁图，筛选重组单株，缩小CL12区间；用近等基因系纯系材料（正常叶和皱叶）进行转录组差异分析、候选基因cDNA差异分析等，用RT-qPCR技术对候选基因进行表达验证，结合基因注释和功能分类，确定CL12候选基因并进行因功能验证，最终图位克隆CL12基因；（2）用CL12近等基因系材料研究皱叶对大豆品质、产量性状的影响。本研究可为大豆育种叶片形态发育研究提供依据。

Soybean crinkled leaf disease appeared in Guangxi, Guangdong et al., can reduce soybean yield by 20%~30%. This symptom is caused by some unknown external cause, which can be inherited stably, and is different from that of the known crinkled leaf type. Previous studies showed that the trait was controlled by a dominant gene CL12. And CL12 was located in the range of 1.96Mb at the end of soybean chromosome 12 by using BSA-seq technology. In order to further clarifying the effect of this gene on soybean yield traits, and finding CL12 candidate genes to further analyze the mechanism of corrugation and find out the inducement of crinkle leaf, two main things are expected to carry out. (1) Local high-density linkage map will be constructed in the target gene region by using F2 population recessive single plant, INDEL, SNP and other molecular markers. Development of markers to further screen recombinant strains and narrow the gene interval of CL12. The analysis of genes different expression will be done to crinkled and normal leafs from two homozygous NILs lines by using RNA-seq. together with gene annotation and functional classification and gene expression verification by RT-qPCR, candidate genes will be determined. And functional verification of candidate genes will be done by genetic transformation then. Finally, Map-based cloning of CL12 gene will be done. (2) NILs of CL12 will be constructed to research the effect of crinkled leaf on the quality and yield traits of soybean. This project can provide a basis for the research of leaf morphological development and soybean breeding.