1.烟台大学生命科学学院,山东烟台 264005;2.中国农业科学院作物科学研究所/ 农业农村部粮食作物基因资源评价利用重点实验室,北京 100081
研究方向为生物化学与分子生物学,E-mail: fighting211@163.com
武 晶,研究方向为作物种质资源鉴定、评价与利用,E-mail: wujing@caas.cn
曲江勇,研究方向为基因组适应与进化研究、物种分类学、系统学、进化与生物地理学等,E-mail: qjy@ytu.edu.cn
国家食用豆产业技术体系(CARS-08)
1.College of Life Science, Yantai University, Yantai 264005,Shandong;2.Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/Key Laboratory of Grain Crop Genetic Resources Evaluation and Utilization, Ministry of Agriculture and Rural Affairs, Beijing 100081
Foundation project: China Agriculture Research System of MOF and MARA-Food Legumes (CARS-08)
目前普通菜豆中克隆到的枯萎病抗性基因较少,因此,从种质资源中鉴定挖掘枯萎病抗性基因对于普通菜豆的分子育种具有重要的意义。课题组前期通过全基因组关联分析定位到PvFWR1 (Fusarium Wilt Resistance 1),并且通过单倍型分析推断其可能与普通菜豆枯萎病抗性相关,在此基础上对该基因进行深入研究。在抗病材料土褐腰子豆、感病材料龙芸豆3号中克隆PvFWR1基因序列,经序列比对分析发现,PvFWR1是一个无内含子基因,CDS全长1104 bp,编码367个氨基酸,属于LRR-RLKs家族蛋白。RT-qPCR发现PvFWR1在抗性材料根组织中表达量为感病材料的2.1倍,且在接种枯萎病病原菌后,根组织中表达量明显上升。利用VIGS技术在抗病材料中对PvFWR1沉默,接种病原菌后,植株抗性减弱;同时,基于普通菜豆毛状根转化法在感病材料中对PvFWR1过表达,接种病原菌后,过表达植株抗性显著增强。综合以上分析,认为PvFWR1基因是枯萎病抗性基因。研究结果为普通菜豆枯萎病抗性机制解析及该基因的育种利用奠定了基础。
Identifying and mining resistance genes from germplasm resources is of great significance for molecular breeding of common bean, as few such resistance genes to wilt disease have been deployed in cultivars. By taking advantage of the former study that reported the identification of candidate resistance gene PvFWR1 (Fusarium Wilt Resistance 1) to common bean wilt, using a genome-wide association study and haplotype analysis, this study conducted further molecular characterization of PvFWR1. The PvFWR1 gene sequences from resistant genotype Tuheyaozidou and the susceptible genotype Longyundou No.3 were isolated. This gene harbors one single exon with a complete coding sequence (CDS) of 1104 bp encoding for 367 amino acids, and is annotated as a LRR-RLKs protein family member. Transcriptional analysis using RT-qPCR showed that the PvFWR1 transcripts in the root tissue of resistant genotype were 2.1-fold higher than that of the susceptible material. The expression in the root tissue of infected plants was significantly up-regulated upon pathogen inoculation. Knock-down of PvFWR1 in the resistant material using VIGS technology resulted in decreased resistance to pathogen inoculation, while overexpressing this gene in the susceptible material by deployment of the hairy root transformation method enhanced pathogen disease. Collectively, our results demonstrate that PvFWR1 is a resistance gene against Fusarium wilt in common bean, offering a foundation for elucidating the resistance mechanism and breeding use of this resistance gene.
赵莉莉,常玉洁,王绪敏,等.普通菜豆
