Pleiotropic effects of c-di-GMP content in Pseudomonas syringae

Although the ubiquitous bacterial secondary messenger cyclic diguanylate (c-di-GMP) has important cellular functions in a wide range of bacteria, its function in the model plant pathogen Pseudomonas syringae remains largely elusive. To this end, we overexpressed Escherichia coli diguanylate cyclase...

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Main Authors: Schuster, Stephan Christoph, Wang, Tingting, Cai, Zhao, Shao, Xiaolong, Zhang, Weitong, Xie, Yingpeng, Zhang, Yingchao, Hua, Canfeng, Yang, Liang, Deng, Xin
Other Authors: McBain, Andrew J.
Format: Article
Language:English
Published: 2019
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Online Access:https://hdl.handle.net/10356/105388
http://hdl.handle.net/10220/48683
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-1053882020-09-21T11:35:31Z Pleiotropic effects of c-di-GMP content in Pseudomonas syringae Schuster, Stephan Christoph Wang, Tingting Cai, Zhao Shao, Xiaolong Zhang, Weitong Xie, Yingpeng Zhang, Yingchao Hua, Canfeng Yang, Liang Deng, Xin McBain, Andrew J. School of Biological Sciences Singapore Centre for Environmental Life Sciences and Engineering DRNTU::Science::Biological sciences Pseudomonas Syringae T3SS Although the ubiquitous bacterial secondary messenger cyclic diguanylate (c-di-GMP) has important cellular functions in a wide range of bacteria, its function in the model plant pathogen Pseudomonas syringae remains largely elusive. To this end, we overexpressed Escherichia coli diguanylate cyclase (YedQ) and phosphodiesterase (YhjH) in P. syringae, resulting in high and low in vivo levels of c-di-GMP, respectively. Via genome-wide RNA sequencing of these two strains, we found that c-di-GMP regulates (i) fliN, fliE, and flhA, which are associated with flagellar assembly; (ii) alg8 and alg44, which are related to the exopolysaccharide biosynthesis pathway; (iii) pvdE, pvdP, and pvsA, which are associated with the siderophore biosynthesis pathway; and (iv) sodA, which encodes a superoxide dismutase. In particular, we identified three promoters that are sensitive to elevated levels of c-di-GMP and inserted them into luciferase-based reporters that respond effectively to the c-di-GMP levels in P. syringae; these promoters could be useful in the measurement of in vivo levels of c-di-GMP in real time. Further phenotypic assays validated the RNA sequencing (RNA-seq) results and confirmed the effect on c-di-GMP-associated pathways, such as repressing the type III secretion system (T3SS) and motility while inducing biofilm production, siderophore production, and oxidative stress resistance. Taken together, these results demonstrate that c-di-GMP regulates the virulence and stress response in P. syringae, which suggests that tuning its level could be a new strategy to protect plants from attacks by this pathogen. NMRC (Natl Medical Research Council, S’pore) Published version 2019-06-12T07:38:26Z 2019-12-06T21:50:28Z 2019-06-12T07:38:26Z 2019-12-06T21:50:28Z 2019 Journal Article Wang, T., Cai, Z., Shao, X., Zhang, W., Xie, Y., Zhang, Y., . . . Deng, X. (2019). Pleiotropic effects of c-di-GMP content in Pseudomonas syringae. Applied and Environmental Microbiology, 85(10), e00152-19-. doi:10.1128/AEM.00152-19 0099-2240 https://hdl.handle.net/10356/105388 http://hdl.handle.net/10220/48683 10.1128/AEM.00152-19 en Applied and Environmental Microbiology © 2019 Wang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. 15 p. application/pdf
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences
Pseudomonas Syringae
T3SS
spellingShingle DRNTU::Science::Biological sciences
Pseudomonas Syringae
T3SS
Schuster, Stephan Christoph
Wang, Tingting
Cai, Zhao
Shao, Xiaolong
Zhang, Weitong
Xie, Yingpeng
Zhang, Yingchao
Hua, Canfeng
Yang, Liang
Deng, Xin
Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
description Although the ubiquitous bacterial secondary messenger cyclic diguanylate (c-di-GMP) has important cellular functions in a wide range of bacteria, its function in the model plant pathogen Pseudomonas syringae remains largely elusive. To this end, we overexpressed Escherichia coli diguanylate cyclase (YedQ) and phosphodiesterase (YhjH) in P. syringae, resulting in high and low in vivo levels of c-di-GMP, respectively. Via genome-wide RNA sequencing of these two strains, we found that c-di-GMP regulates (i) fliN, fliE, and flhA, which are associated with flagellar assembly; (ii) alg8 and alg44, which are related to the exopolysaccharide biosynthesis pathway; (iii) pvdE, pvdP, and pvsA, which are associated with the siderophore biosynthesis pathway; and (iv) sodA, which encodes a superoxide dismutase. In particular, we identified three promoters that are sensitive to elevated levels of c-di-GMP and inserted them into luciferase-based reporters that respond effectively to the c-di-GMP levels in P. syringae; these promoters could be useful in the measurement of in vivo levels of c-di-GMP in real time. Further phenotypic assays validated the RNA sequencing (RNA-seq) results and confirmed the effect on c-di-GMP-associated pathways, such as repressing the type III secretion system (T3SS) and motility while inducing biofilm production, siderophore production, and oxidative stress resistance. Taken together, these results demonstrate that c-di-GMP regulates the virulence and stress response in P. syringae, which suggests that tuning its level could be a new strategy to protect plants from attacks by this pathogen.
author2 McBain, Andrew J.
author_facet McBain, Andrew J.
Schuster, Stephan Christoph
Wang, Tingting
Cai, Zhao
Shao, Xiaolong
Zhang, Weitong
Xie, Yingpeng
Zhang, Yingchao
Hua, Canfeng
Yang, Liang
Deng, Xin
format Article
author Schuster, Stephan Christoph
Wang, Tingting
Cai, Zhao
Shao, Xiaolong
Zhang, Weitong
Xie, Yingpeng
Zhang, Yingchao
Hua, Canfeng
Yang, Liang
Deng, Xin
author_sort Schuster, Stephan Christoph
title Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
title_short Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
title_full Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
title_fullStr Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
title_full_unstemmed Pleiotropic effects of c-di-GMP content in Pseudomonas syringae
title_sort pleiotropic effects of c-di-gmp content in pseudomonas syringae
publishDate 2019
url https://hdl.handle.net/10356/105388
http://hdl.handle.net/10220/48683
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