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苹果树腐烂病菌VmMFS1~VmMFS3基因的功能分析
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引用本文:张琼,刘昭阳,高承宇,杜旋,冯浩,黄丽丽.苹果树腐烂病菌VmMFS1~VmMFS3基因的功能分析.植物保护学报,2024,51(1):249-260
DOI:10.13802/j.cnki.zwbhxb.2024.2022119
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作者单位E-mail
张琼 西北农林科技大学作物抗逆与高效生产全国重点实验室, 陕西 杨凌 712100  
刘昭阳 西北农林科技大学植物保护学院, 陕西 杨凌 712100  
高承宇 西北农林科技大学植物保护学院, 陕西 杨凌 712100  
杜旋 西北农林科技大学植物保护学院, 陕西 杨凌 712100  
冯浩 西北农林科技大学作物抗逆与高效生产全国重点实验室, 陕西 杨凌 712100
西北农林科技大学植物保护学院, 陕西 杨凌 712100 
xiaosong04005@163.com 
黄丽丽 西北农林科技大学作物抗逆与高效生产全国重点实验室, 陕西 杨凌 712100
西北农林科技大学植物保护学院, 陕西 杨凌 712100 
huanglili@nwsuaf.edu.cn 
中文摘要:为绿色持久防控苹果树腐烂病,该研究分析苹果树腐烂病菌Valsa mali的3个主要协同转运蛋白超家族(major facilitator superfamily,MFS)编码基因的氨基酸序列特征,利用实时荧光定量PCR (quantitative real-time PCR,RT-qPCR)技术分析这3个基因在苹果树腐烂病菌侵染阶段的表达水平,通过构建这3个基因的缺失突变体和回补菌株分析其在病原菌营养生长、致病力和非生物胁迫应答等方面的功能。结果表明,这3个基因的氨基酸序列均具有MFS保守结构域,将其命名为VmMFS1~VmMFS3; VmMFS1和VmMFS2的进化距离较近,均与VmMFS3的进化距离较远;在苹果树腐烂病菌侵染过程中VmMFS1~VmMFS3基因表达均显著上调;与野生型03-8菌株相比,VmMFS1~VmMFS3基因缺失突变体的菌落形态无明显差异,但生长速度下降; VmMFS1~VmMFS3基因缺失突变体的致病力均显著降低; VmMFS1~VmMFS3基因缺失突变体对H2O2胁迫的敏感性无明显变化,但对NaCl胁迫更敏感;基因回补后基因缺失突变体的表型缺陷能恢复到野生型菌株的水平。
中文关键词:非生物胁迫  主要协同转运蛋白超家族  苹果黑腐皮壳菌  致病力  H2O2胁迫  NaCl胁迫
 
Functional analysis of three facilitator superfamily protein genes VmMFS1-VmMFS3 in apple Valsa canker pathogen Valsa mali
Author NameAffiliationE-mail
Zhang Qiong State Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Northwest A&F University, Yangling 712100, Shaanxi Province, China  
Liu Zhaoyang College of Plant Protection, Northwest A&F University, Yangling 712100, Shaanxi Province, China  
Gao Chengyu College of Plant Protection, Northwest A&F University, Yangling 712100, Shaanxi Province, China  
Du Xuan College of Plant Protection, Northwest A&F University, Yangling 712100, Shaanxi Province, China  
Feng Hao State Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Northwest A&F University, Yangling 712100, Shaanxi Province, China
College of Plant Protection, Northwest A&F University, Yangling 712100, Shaanxi Province, China 
xiaosong04005@163.com 
Huang Lili State Key Laboratory for Crop Stress Resistance and High-Efficiency Production, Northwest A&F University, Yangling 712100, Shaanxi Province, China
College of Plant Protection, Northwest A&F University, Yangling 712100, Shaanxi Province, China 
huanglili@nwsuaf.edu.cn 
Abstract:For integrated and durable control of apple Valsa canker, the amino acid sequence characteristics of three MFS-coding genes of pathogen V. mali were analyzed. Quantitative real-time PCR (RTqPCR) technology was used to analyze the expression levels of these three genes during the infection of V. mali. Additionally, the function of these three MFS genes in vegetative growth, pathogenicity, and abiotic stress responses was analyzed using gene knockout and complementation approaches. The results showed that the deduced amino acid sequences of polyproteins encoded by all the three genes possess a conserved MFS domain, named as VmMFS1-VmMFS3. VmMFS1 and VmMFS2 are closely related in evolution, whereas they are distant from VmMFS3 based on the phylogenetic analysis. The expression of VmMFS1-VmMFS3 during V. mali infection was further determined and all were upregulated significantly. Compared with that of wild-type strain 03-8, the deletion mutants of VmMFS1-VmMFS3 show no noticeable difference in colony morphology, while the growth rate of deletion mutants was reduced to some extent. Importantly, the pathogenicity of VmMFS1-VmMFS3 deletion mutants was significantly reduced. In addition, VmMFS1-VmMFS3 deletion mutants exhibited no significant variation in their tolerance to H2O2 stress; however, they were more susceptible to NaCl stress. The gene complementation test showed that the complemented strains could restore the phenotypic defect of the gene deletion mutants to the level of the wild-type strain 03-8.
keywords:abiotic stress  major facilitator superfamily  Valsa mali  pathogenicity  H2O2 stress  NaCl stress
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