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Na⁺/苹果酸同向转运蛋白MaeN对胡萝卜软腐果胶杆菌致病力的影响

引用本文：梁葆文,许自龙,刘培刚,林天宝,朱燕,吕志强,周旭东,魏佳.Na⁺/苹果酸同向转运蛋白MaeN对胡萝卜软腐果胶杆菌致病力的影响.植物保护学报,2025,52(2):379-389

DOI：10.13802/j.cnki.zwbhxb.2025.2024122

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作者	单位	E-mail
梁葆文	浙江农林大学林业与生物技术学院, 省部共建亚热带森林培育国家重点实验室, 杭州 311300
许自龙	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021
刘培刚	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021
林天宝	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021
朱燕	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021
吕志强	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021
周旭东	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021	xudong.zhou@zafu.edu.cn
魏佳	浙江省农业科学院, 蚕桑与茶叶研究所, 杭州 310021	weijia@zaas.ac.cn

中文摘要:为鉴定胡萝卜软腐果胶杆菌Pectobacterium carotovorum侵染桑树Morus alba的关键致病基因，定量分析本课题组前期通过致病力筛选获得的致病力减弱突变体m400和野生型菌株18#侵染桑树时的致病力，对突变体m400的突变基因进行定位并分析其编码蛋白质的理化性质与结构特征，利用实时荧光定量PCR技术测定突变基因及其同源基因在胡萝卜软腐果胶杆菌侵染宿主桑树时的表达量，探索突变基因在胡萝卜软腐果胶杆菌侵染宿主时的功能。结果表明：与野生型菌株18#相比，突变体m400的致病力在侵染进程中显著减弱；突变体m400中的突变基因为gene0047，该基因编码1个2-羟基羧酸盐转运蛋白家族的Na⁺/苹果酸同向转运蛋白（Na⁺/malate symporter，MaeN）。maeN基因及其同源基因的表达量在胡萝卜软腐果胶杆菌侵染桑树时显著上调；此外，桑叶中的苹果酸含量在胡萝卜软腐果胶杆菌侵染过程中升高，且maeN基因突变体m400对苹果酸高敏感，桑树组织受侵染后苹果酸浓度升高到1.25 mg/mL时足以抑制突变体m400的生长，但对野生型菌株18#生长的抑制能力有限。表明maeN基因可能通过增强胡萝卜软腐果胶杆菌对苹果酸的耐受性来适应宿主植物细胞内的苹果酸胁迫环境，进而维持其致病力。

中文关键词:苹果酸转运蛋白胡萝卜软腐果胶杆菌桑细菌性病害突变体苹果酸致病力

Effect of Na⁺/malic acid co-transporter MaeN on the pathogenicity of soft rot bacterium Pectobacterium carotovorum

Author Name	Affiliation	E-mail
Liang Baowen	State Key Laboratory of Subtropical Silviculture, Zhejiang A&F University, Hangzhou 311300, Zhejiang Province, China
Xu Zilong	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China
Liu Peigang	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China
Lin Tianbao	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China
Zhu Yan	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China
Lü Zhiqiang	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China
Zhou Xudong	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China	xudong.zhou@zafu.edu.cn
Wei Jia	Sericulture and Tea Research Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, Zhejiang Province, China	weijia@zaas.ac.cn

Abstract:In order to identify key pathogenic genes of Pectobacterium carotovorum that infect mulberry tree Morus alba, research was conducted using the mutant m400 with reduced pathogenicity, which was previously obtained through pathogenicity screening. This study quantitatively compared the pathogenicity of m400 and the wild-type strain 18# during the infection of M. alba. The mutated gene in m400 was localized and the physicochemical properties and structural characteristics of the protein encoded by the mutated gene were analyzed. We also used real-time fluorescent quantitative technology to measure the expression levels of the mutated gene and its homologous genes during the infection of M. alba by P. carotovorum, and explored the function of the mutated gene during the infection process. The results showed that the pathogenicity of the mutant m400 was significantly reduced compared with the wild-type strain during the infection process. The mutated gene was identified as gene0047, which encodes a Na⁺/malate symporter (MaeN) belonging to the 2-hydroxycarboxylate transporter family. The expression levels of the maeN gene and its homologous genes were significantly upregulated during the infection of mulberry trees by P. carotovorum. Additionally, the concentration of malic acid in mulberry leaves increased during the infection process, and the maeN gene mutant m400 was highly sensitive to malic acid. The increased concentration of malic acid (1.25 mg/mL) in the infected mulberry tissues was sufficient to inhibit the growth of m400, but had limited inhibitory effect on the wild-type strain 18#. These results suggest that the maeN gene may enhance the tolerance of P. carotovorum to malic acid, helping the bacterium adapt to the malic acid stress environment within host plant cells during infection, thereby maintaining its pathogenicity.

keywords:malic acid transporter Pectobacterium carotovorum mulberry bacterial diseases mutant malic acid pathogenicity

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