普通菜豆FWR1基因克隆及功能验证
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1.中国农业科学院作物科学研究所;2.烟台大学生命科学学院

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国家现代农业产业技术体系


Cloning and functional validation of FWR1 gene in Common bean
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China Agriculture Research System-Food Legumes

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    摘要:

    镰孢菌枯萎病是普通菜豆生产中危害最为严重的土传病害之一。本课题组前期通过全基因组关联分析定位到PvFWR1,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,在此基础上本研究对其进行深入研究。首先,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;然后,qRT-PCR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;最后,利用VIGS技术在抗病材料对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。

    Abstract:

    Fusarium wilt is one of the most serious soil-borne diseases that affects common bean production. In the previous study, our research group identified PvFWR1 through a genome-wide association study and inferred that it might be related to resistance to common bean wilt through haplotype analysis. In this study, we conducted further research on PvFWR1. First, we cloned the PvFWR1 gene sequence from resistant material Tuheyaozidou and the susceptible material Longyundou No.3, and found that PvFWR1 is an intronless gene with a CDS length of 1104 bp and 367 amino acids, belonging to the LRR-RLKs protein family. Then, qRT-PCR showed that the expression level of PvFWR1 in the root tissue of resistant material was 2.1 times that of the susceptible material, and the expression level in the root tissue of the infected plants was significantly higher after inoculation with the pathogen. Finally, we used VIGS technology to silence PvFWR1 in the resistant material, and the resistance of the plants was weakened after inoculation with the pathogen. At the same time, we used the common bean hairy root transformation method to overexpress PvFWR1 in the susceptible material, and the resistance of the overexpression plants was significantly increased after inoculation with the pathogen. Based on these analyses, we concluded that PvFWR1 is a resistance gene against Fusarium wilt. The research results lay a foundation for elucidating the resistance mechanism of common bean wilt and the genetic breeding and utilization of PvFWR1.

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历史
  • 收稿日期:2024-10-11
  • 最后修改日期:2024-11-09
  • 录用日期:2024-12-05
  • 在线发布日期: 2025-01-13
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