番茄SlWDR141基因的克隆及功能研究

发布时间：2018-05-23 16:30

本文选题：植物生理与分子生物学 + WD蛋白　；参考：《中国科技论文》2017年06期

【摘要】：为研究番茄WD40蛋白基因(SlWDR141)的功能,对SlWDR141蛋白进行生物信息学分析,发现SlWDR141蛋白在进化过程中高度保守,与土豆的同源性最高。在克隆SlWDR141基因全长序列的基础上,构建了亚细胞定位和酵母双杂交表达载体,证实SlWDR141蛋白主要定位在细胞核中,与DDB1具有相互作用。实时荧光定量PCR(real-time PCR)分析SlWDR141基因在组织中的表达谱,发现它在各个组织中均有表达,但在根中的表达量最高。为研究该基因的生物学功能,进一步构建了SlWDR141基因的干涉载体,用农杆菌介导法转化番茄得到转基因阳性番茄,荧光定量PCR鉴定出3个表达量下调的独立转基因株系。NaCl胁迫和Pst DC3000侵染野生型和转基因植株发现,转基因植株相对于野生型植株抗盐性和抗病性均下降,表明SlWDR141基因对盐胁迫和细菌侵染均起正调控作用。
[Abstract]:In order to study the function of tomato WD40 protein gene SlWDR141, the bioinformatics analysis of SlWDR141 protein showed that SlWDR141 protein was highly conserved in evolution and had the highest homology with potato. Based on the cloning of full-length SlWDR141 gene, subcellular localization and yeast two-hybrid expression vectors were constructed. It was confirmed that the SlWDR141 protein was mainly located in the nucleus and interacted with DDB1. Real-time fluorescence quantitative PCR(real-time PCR was used to analyze the expression profile of SlWDR141 gene in tissues. It was found that SlWDR141 gene was expressed in all tissues, but the highest expression level was found in roots. In order to study the biological function of the gene, the interference vector of SlWDR141 gene was constructed and transformed into tomato by Agrobacterium tumefaciens to obtain transgenic tomato. Three down-regulated independent transgenic lines. NaCl stress and Pst DC3000 infected wild and transgenic plants were identified by fluorescence quantitative PCR. The results showed that the salt resistance and disease resistance of transgenic plants were decreased compared with wild type plants. The results suggest that SlWDR141 gene plays a positive role in both salt stress and bacterial infection.
【作者单位】：四川大学生命科学学院;生物资源与生态环境教育部重点实验室;水力学与山区河流开发保护国家重点实验室;合肥工业大学食品科学与工程学院;
【基金】：高等学校博士学科点专项科研基金资助项目(20110181130009)
【分类号】：Q943.2;S641.2

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