Genome-wide identification of the Sec-dependent secretory protease genes in Erwinia amylovora and analysis of their expr
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Zhang et al. / J Zhejiang Univ-Sci B (Biomed & Biotechnol) 2020 21(9):716-726
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology) ISSN 1673-1581 (Print); ISSN 1862-1783 (Online) www.jzus.zju.edu.cn; www.springerlink.com E-mail: [email protected]
Genome-wide identification of the Sec-dependent secretory protease genes in Erwinia amylovora and analysis of their expression during infection of immature pear fruit*# Wang-bin ZHANG§1,2, Hai-lin YAN§1,3, Zong-cai ZHU1,3, Chao ZHANG3, Pei-xiu DU3, Wen-jun ZHAO†‡4, Wei-min LI†‡3 1 2
College of Plant Science, Tarim University, Alar 843300, China
Southern Xinjiang Key Laboratory of Integrated Pest Management, Tarim University, 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 †
E-mail: [email protected]; [email protected]
Received May 28, 2020; Revision accepted July 21, 2020; Crosschecked Aug. 17, 2020
Abstract: The general secretory (Sec) pathway represents a common mechanism by which bacteria secrete proteins, including virulence factors, into the extracytoplasmic milieu. However, there is little information about this system, as well as its associated secretory proteins, in relation to the fire blight pathogen Erwinia amylovora. In this study, data mining revealed that E. amylovora harbors all of the essential components of the Sec system. Based on this information, we identified putative Sec-dependent secretory proteases in E. amylovora on a genome-wide scale. Using the programs SignalP, LipoP, and Phobius, a total of 15 putative proteases were predicted to contain the N-terminal signal peptides (SPs) that might link them to the Sec-dependent pathway. The activities of the predicted SPs were further validated using an Escherichia coli-based alkaline phosphatase (PhoA) gene fusion system that confirmed their extracytoplasmic property. Transcriptional analyses showed that the expression of 11 of the 15 extracytoplasmic protease genes increased significantly when E. amylovora was used to inoculate immature pears, suggesting their potential roles in plant infection. The results of this study support the suggestion that E. amylovora might employ the Sec system to secrete a suite of proteases to enable successful infection of plants, and shed new light on the interaction of E. amylovora with host plants. Key words: Erwinia amylovora; Sec-dependent pathway; Protease; Gene expression; Plant infection https://doi.org/10.1631/jzus.B2000281 CLC number: S432.1
1 Introduction ‡
Corresponding authors The two authors contributed equally to this work * Project supported by the Key Technologies R&D Program of Xinjiang Production and Construction Corps (No. 2018AB038) and the National Natural Science Foundation of China (No. 31970126) # Electronic supplementary materials: The online version of this article (https://doi.org/10.1631/jzus.B2000281) contains supplementary materials, which are available to authorized users ORCI
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