基于RNA-seq和iTRAQ技术对蓖麻耐盐分子机制研究

Castor is a stress-resistant green energy crop and its products are widely used in various industries. Tongliao city, which has a large saline alkali soil, is one of main producing areas for castor in China. In recent years, the castor planting industry is shrinking. Breeding elite varieties is an important method to develop castor planting industry. Molecular breeding, as one of the means, has just been adopted for the research of the creation of salt tolerant varieties. It is of practical significance to explore and utilize the salt tolerant genes and breed salt tolerant varieties by genetic engineering, and this will help promote the utilization of saline-alkali soil and enhance the development of castor planting industries.. This study was performed to identify differentially expressed genes and differential abundance proteins from Zhebi4 and Tongbi5, which showed significant difference in salt tolerance and crop yield. RNA-seq and iTRAQ technologies were used in this study. By conjoint analysis, we will clone the genes which show the same trend between transcription and protein level, construct overexpression vector and RNAi vector, transform them into castor, select homozygous transgenic lines, analyze the salt tolerance phenotype, and finally obtain new salt tolerant castor germplasm. This will lay the foundation for the salt tolerant molecular breeding in castor and enrich salt tolerance germplasm resources.

蓖麻是绿色抗逆能源作物，其产品被广泛应用于众多领域。盐碱地面积较大的通辽地区是我国蓖麻主产区之一，近些年种植业出现萎缩。选育优良品种是发展蓖麻种植业的重要手段，分子育种作为手段之一，专门针对耐盐性状的品种选育研究刚刚起步。挖掘和利用耐盐基因，通过基因工程手段选育耐盐品种，对开发利用盐碱地和促进蓖麻种植业的发展有现实意义。. 本研究拟采用基于RNA-seq与iTRAQ测序技术比较盐胁迫后不同耐盐自交系哲蓖3（耐盐基因型）与通蓖5（感盐基因型）种子萌发阶段的差异表达基因与差异丰度蛋白，通过数据关联分析筛选转录与蛋白表达趋势一致的基因，进而分离目的基因、构建过表达和干扰载体、利用农杆菌介导法对蓖麻进行遗传转化、筛选纯合转基因株系、分析其表型与抗盐性、最终获得蓖麻耐盐新种质，为蓖麻耐盐分子育种的研究奠定基础。

利用 RNA-seq 和 iTRAQ 技术分析耐盐（通蓖5）和感盐（哲蓖3）蓖麻自交系种子萌发的差异表达基因和差异丰度蛋白并进行关联分析，探索种子萌发期蓖麻的耐盐机制，同时筛选选耐盐相关基因。RNA-seq 和iTRAQ 测序完成，数据良好。挖掘盐响应基因5000余条，盐响应蛋白169个。但转录组数据和蛋白组数据关联性不强。筛选耐盐信号通路的一些显著差异的基因，克隆并进行转基因，分析其功能。同时得到耐盐转基因蓖麻种质材料，为进一步培育蓖麻耐盐新品种奠定基础。