梨火疫病菌两个Sec依赖的外泌纤维素酶鉴定及在侵染梨幼果过程中的基因表达分析

doi:10.19586/j.2095-2341.2020.0084

生物技术进展 ›› 2020, Vol. 10 ›› Issue (5): 517-523.DOI: 10.19586/j.2095-2341.2020.0084

梨火疫病菌两个Sec依赖的外泌纤维素酶鉴定及在侵染梨幼果过程中的基因表达分析

严海璘1,2,3,朱宗财1,2,3,张王斌1,2,杜培秀3,张超3,赵文军4,李为民3*

1.塔里木大学植物科学学院，新疆阿拉尔 843300;

2.塔里木大学，南疆农业有害生物综合治理重点实验室，新疆阿拉尔 843300;

3.中国农业科学院生物技术研究所，北京 100081;

4.中国检验检疫科学研究院，北京 100176

收稿日期:2020-07-24 出版日期:2020-09-25 发布日期:2020-08-05
通讯作者: 李为民 E-mail：liweimin01@caas.cn
作者简介:严海璘 E-mail：yanhailin@ms.xjb.ac.cn
基金资助:
新疆生产建设兵团重点领域科技攻关计划项目(2018AB038)。

Identification of Two Sec-dependent Cellulases in Erwinia amylovora and Gene Expression Analysis During Infection of Immature Pear Fruit

YAN Hailin^,, ZHU Zongcai^,, ZHANG Wangbin^,, DU Peixiu, ZHANG Chao, ZHAO Wenjun⁴, LI Weimin

1.College of Plant Science, Tarim University, Xinjiang Alar 843300, China;

2.Southern Xinjiang Key Laboratory of IPM, Tarim University, Xinjiang Alar 843300, China;

3.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China;

4.Chinese Academy of Inspection and Quarantine, Beijing 100176, China

Received:2020-07-24 Online:2020-09-25 Published:2020-08-05

摘要/Abstract

摘要： 利用生物信息学技术结合基于大肠杆菌的碱性磷酸酯酶（PhoA）检测体系，在梨火疫病菌（Erwinia amylovora）全基因组序列中筛选鉴定了2个依赖Sec途径分泌的纤维素酶：内切葡聚糖酶EAMY_3602和β-葡萄糖苷酶EAMY_1236。将梨火疫病菌DSM17948接种酥梨幼果，分别于接种后0、8和24 h定量检测基因表达，结果显示EAMY_3602编码基因(Ea3602)在接种后8和24 h的mRNA表达量逐渐降低，但与0 h相比无显著差异；EAMY_1236编码基因(Ea1236)在接种后8和24 h的mRNA表达量逐步提高，较0 h分别提高了2.43和3.26倍，差异具有统计学意义（P<001）。由研究结果推测Sec依赖分泌的内切葡聚糖酶EAMY_3602可能与梨火疫病菌侵染无关，而β-葡萄糖苷酶EAMY_1236可能是梨火疫病菌的毒力因子，在侵染寄主植物过程中发挥作用。

关键词: 梨火疫病菌, Sec依赖分泌蛋白, 纤维素酶, 致病机理

Abstract: In this study, by using bioinformatics analysis and Escherichia coli-based alkaline phosphatase (PhoA) gene fusion system, we identified two Sec-dependent secreted cellulases, an endoglucanase EAMY_3602 and a β-glucosidase EAMY_1236, from the whole genome of Erwinia amylovora. The cell suspension of E. amylovora strain DSM17948 was inoculated into the young pear fruits, quantitative detection of gene expression at 0, 8 and 24 hpi (hours post-inoculation), respectively, which showed that the mRNA expression levels of Ea3602, the coding gene of EAMY_3602, were continuously decreased at 8 and 24 hpi, with no significant difference when compared with that at 0 hpi. The transcript levels of Ea1236, the coding gene of EAMY_1236, at 8 and 24 hpi were 2.43 and 3.26 fold higher than that at 0 hpi. Research results showed that the Sec-dependent secreted endoglucanase EAMY_3602 had no implication in plant infection, but β-glucosidase EAMY_1236 probably acted as a virulence factor that plays role in the process of E. amylovora infecting the immature pears.

Key words: Erwinia amylovora, Sec-dependent secretion, cellulase, pathogenic mechanism

严海璘,,朱宗财,,张王斌,,杜培秀,张超,赵文军4,李为民. 梨火疫病菌两个Sec依赖的外泌纤维素酶鉴定及在侵染梨幼果过程中的基因表达分析[J]. 生物技术进展, 2020, 10(5): 517-523.

YAN Hailin1,2,3, ZHU Zongcai1,2,3, ZHANG Wangbin1,2, DU Peixiu3, ZHANG Chao3, ZHAO Wenjun4, LI Weimin3*. Identification of Two Sec-dependent Cellulases in Erwinia amylovora and Gene Expression Analysis During Infection of Immature Pear Fruit[J]. Curr. Biotech., 2020, 10(5): 517-523.

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梨火疫病菌两个Sec依赖的外泌纤维素酶鉴定及在侵染梨幼果过程中的基因表达分析

Identification of Two Sec-dependent Cellulases in Erwinia amylovora and Gene Expression Analysis During Infection of Immature Pear Fruit

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