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Journal of Integrative Agriculture  2026, Vol. 25 Issue (6): 2475-2484    DOI: 10.1016/j.jia.2024.09.031
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(p)ppGpp interacts with TrmE and regulates the PcoI/PcoR quorum-sensing system of Pseudomonas fluorescens 2P24

Ruijing Shang1, Shihai Liang1, Qing Yan2Bingxin Wang3, Guoliang Qian3Lang Yang4, Xiaogang Wu1*

1 Guangxi Key Laboratory of Agro-Environment and Agro-Product Safety/College of Agriculture, Guangxi University, Nanning 530004, China

2 Department of Plant Sciences and Plant Pathology, Montana State University, Bozeman 59717, USA 

3 College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China

4 Institute of Plant Protection, Guangxi Academy of Agricultural Sciences, Nanning 530007, China

 Highlights 
RelA and SpoT exert opposite effects on the PcoI/PcoR quorum-sensing (QS) system in Pseudomonas fluorescens 2P24.
(p)ppGpp regulates the PcoI/PcoR QS system by influncing the function of the tRNA modification GTPase TrmE.

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摘要  

[目的]植物有益假单胞菌(Pseudomonas spp.)在植物根围的有效定殖是其发挥生防功能的先决条件,而前期研究表明荧光假单胞菌(P. fluorescens2P24中PcoI/PcoR群体感应(quorum sensing;QS)系统正调控其在植物根围的定殖。定殖过程中,菌株2P24会受到植物来源活性氧和其他环境胁迫的多重影响,但不利环境条件影响QS系统的分子机制尚不清楚。[方法]以菌株2P24为出发菌,通过同源重组技术构建了relA突变体、spoT突变体和relA spoT双突变体。利用转录融合等技术检测了(p)ppGpp合成酶RelA和(p)ppGpp合成酶/水解酶SpoT对QS系统的调控作用;微量热泳动技术检测了(p)ppGpp与TrmE蛋白互作关系。[结果]菌株2P24中同时缺失spoTrelA基因可显著提高QS系统中信号合成基因pcoI的表达,而缺失spoT基因则显著抑制pcoI基因的表达。遗传学和微量热泳动试验表明TrmE蛋白(一种tRNA修饰GTP酶)可结合(p)ppGpp且是(p)ppGpp调控QS系统所必需的遗传因子。此外,spoTrelA基因还参与菌株2P24中游动性、生物膜形成、氧化应激、渗透耐受性和根围定殖等多种生防相关性状。[结论]菌株2P24中relA基因和spoT基因介导的(p)ppGpp信号通路对QS系统的功能起重要调控作用,并对研究生防菌株通过(p)ppGpp调控通路影响其定殖能力的分子机制具有重要意义。



Abstract  

The efficient colonization of plant-beneficial Pseudomonas spp. is a prerequisite for their biocontrol capacity.  Prior work revealed that the PcoI/PcoR quorum-sensing (QS) system plays a pivotal role in the root colonization of Pfluorescens 2P24.  During the colonization, strain 2P24 has faced diverse impacts from plant-derived reactive oxygen species and other environmental stress.  However, the molecular mechanism by which the PcoI/PcoR QS system is regulated under unfavored conditions remains unclear.  Thus, in this study, the role of the (p)ppGpp synthetase RelA and the bifunctional (p)ppGpp synthase/hydrolase SpoT in the PcoI/PcoR QS system of Pfluorescens was investigated.  Our data indicated that the deficiency of relA and spoT genes remarkably improved the expression of the pcoI gene, whereas the mutation of the spoT gene significantly repressed the expression of the pcoI gene.  We further demonstrated that the regulation of the PcoI/PcoR QS system by (p)ppGpp was dependent on the function of the trmE gene, which encodes a tRNA modification GTPase.  Furthermore, the mutation of relA, spoT, or both significantly influenced the motility, biofilm formation, oxidative stress, osmotic tolerance, and rhizosphere colonization.  Collectively, our data indicated that the (p)ppGpp signaling pathway mediated by the relA gene and spoT gene was important to the function of the PcoI/PcoR QS system and had important implications for the understanding of the molecular mechanism of (p)ppGpp in epiphytic fitness via TrmE of Pfluorescens.

Keywords:  Pseudomonas fluorescens        quorum sensing        relA        spoT        trmE  
Received: 12 June 2024   Accepted: 11 August 2024 Online: 26 September 2024  
Fund: 
The authors would like to thank Dr. Liqun Zhang from China Agricultural University, for sharing bacteria strains.  This work was supported by the Natural Science Foundation of Guangxi, China (2021GXNSFDA220006) and the Guangxi Key Laboratory of Agro-Environment and Agro-Product Safety Open Projects Fund, China (GKLAEAPS2023-04).
About author:  #Correspondence Xiaogang Wu, Tel/Fax: +86-771-3235643, E-mail: wux@gxu.edu.cn

Cite this article: 

Ruijing Shang, Shihai Liang, Qing Yan, Bingxin Wang, Guoliang Qian, Lang Yang, Xiaogang Wu. 2026. (p)ppGpp interacts with TrmE and regulates the PcoI/PcoR quorum-sensing system of Pseudomonas fluorescens 2P24. Journal of Integrative Agriculture, 25(6): 2475-2484.

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