中文摘要
籽粒大小不仅是菜用豌豆产量构成的重要因素,而且直接影响其商品性。因此鉴定大粒相关优质基因已成为菜用豌豆优质高产育种的关键问题之一。本项目拟采用复合型基因捕获策略高效鉴定菜用豌豆大粒相关优质基因。以菜用豌豆自然群体为材料,利用RAD-seq技术获取SNP,整合本项目组已开发的分子标记,与籽粒表型数据结合进行全基因组关联分析,筛选籽粒大小相关候选基因∕区域;选择粒型差异显著材料开展高通量转录组测序,筛选差异表达基因;同时通过同源克隆获取已报道的籽粒大小相关基因,作为基因发掘的重要补充;采用高通量小RNA测序与降解组分析,鉴定差异表达miRNA及其靶基因,从转录后水平发掘籽粒大小相关基因。最后,综合所有候选基因分析其结构、时空表达模式及代谢途径,广泛全面地从基因表达与转录后调控水平获得菜用豌豆籽粒大小重要调控基因,进而解析籽粒大小分子调控机制,为菜用豌豆大粒型分子设计育种奠定基础。
英文摘要
Seed size is not only a major component of yield, but aslo a key determinant of market values in the vegetable pea. Identification and characterization of key regulatory related to seed size remains a challenge in the current breeding program. In this project,we will employ the composite gene trapping strategy to discovery the key genes associated with seed size. Frist, we will conduct RAD-seq technology to identify SNPs in the natural populations of vegetable pea, and use GWAS to discover the seed size-related genes by relating SNPs as well as our developed EST-SSR markers to phenotypic data. Meanwhile, the transcriptome sequencing will be constructed and applied for screening differential expression of genes and finding their genetic metabolic pathway in vegetable pea cultivars with different seed size. As an important supplement, we will clone the seed size-related genes which reported in other plants by homologous sequence analysis. Subsequently, in order to explore the seed size-related gene from the post-transcriptional regulation level, a combined approach of high-throughput sequencing and degradome analysis will be applied to reveal the different expression miRNA and their target genes. The spatial-temporal dynamic expressions and metabolic pathway of all the candidate genes∕miRNAs will be further analyzed by qRT-PCR and bioinformatic analyses. Finally, integrating all the results of mining genes methods, we will explore the key genes related to seed size from the gene expression and post-transcriptional regulation levels, and establish molecular regulation mechanism for seed size in the vegetable pea, which will be helpful for the molecular breeding of the vegetable pea.
