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调节2P24中2,4-二乙酰基间苯三酚的产生和群体感应系统。

regulates 2,4-DAPG production and quorum sensing system in 2P24.

作者信息

Wei Yarui, Dong Baozhu, Wu Xiaogang, Zhao Mingmin, Wang Dong, Li Na, Zhang Qian, Zhang Liqun, Zhou Hongyou

机构信息

College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot, Inner Mongolia, China.

College of Agriculture, Guangxi University, Nanning, China.

出版信息

Front Microbiol. 2023 May 12;14:1160913. doi: 10.3389/fmicb.2023.1160913. eCollection 2023.

Abstract

INTRODUCTION

2P24 was isolated from soil of natural decay associated with wheat take-all and it can effectively control soil-borne diseases caused by a variety of plant pathogens. 2,4-diacetylphloroglucinol (2,4-DAPG), is produced by 2P24 and plays an important role in the prevention and control of plant diseases. To understand the resistant mechanism, in this study, we conducted experiments to explore the regulation role of in the synthesis of the antibiotic 2,4-DAPG and regulation of QS system.

METHODS

A random mini-Tn5 mutagenesis procedure was used to screen regulators for transcription in stain PM901, which containing a phlA∷lacZ transcriptional fusion reporter plasmid. We identified 12 insertion mutants could significantly change gene expression. By analyzing the amino acid sequences of the interrupted gene, we obtained a mutant strain Aa4-29 destroyed the gene, which encodes the omiga subunit. We constructed the plasmid of mutant (pBBR-△rpoZ) transformed into competent cells of 2P24 by electro-transformation assay. The strains of 2P24/pBBR, 2P24-△rpoZ/pBBR, 2P24-△rpoZ/pBBR-rpoZ were used to evaluate the regulation role of in 2,4-DAPG production and quorum sensing system.

RESULTS

According to β-galactosidase activity, we found that positively regulated the expression of (a synthesis gene of 2,4-DAPG) and (a synthesis gene of PcoI/PcoR QS signal system) at the transcriptional level. The production of 2,4-DAPG antibiotic and signal molecule AHL was influenced by . Further, was involved in regulating expression. also has a certain regulatory effect on transcription, but no effect on the transcription of , and . According to the biocontrol assay, 2P24 strains with showed obvious antagonism ability against the in cotton, while the mutant strain of lost the biocontrol effect. had a significant effect on the swimming and biofilm formation in 2P24.

CONCLUSION

Our data showed that was an important regulator of QS system, 2,4-DAPG in 2P24. This may imply that has evolved different regulatory features to adapt to different environmental threats.

摘要

引言

2P24是从与小麦全蚀病相关的自然腐烂土壤中分离出来的,它能有效控制由多种植物病原体引起的土传病害。2,4-二乙酰基间苯三酚(2,4-DAPG)由2P24产生,在植物病害防治中起重要作用。为了解其抗性机制,在本研究中,我们进行了实验以探究其在抗生素2,4-DAPG合成及群体感应系统调控中的作用。

方法

采用随机mini-Tn5诱变程序,在含有phlA∷lacZ转录融合报告质粒的PM901菌株中筛选调控转录的因子。我们鉴定出12个插入突变体可显著改变基因表达。通过分析中断基因的氨基酸序列,我们获得了一个破坏编码ω亚基基因的突变菌株Aa4-29。我们构建了突变体质粒(pBBR-△rpoZ),通过电转化法将其导入2P24感受态细胞。利用2P24/pBBR、2P24-△rpoZ/pBBR、2P24-△rpoZ/pBBR-rpoZ菌株评估其在2,4-DAPG产生和群体感应系统中的调控作用。

结果

根据β-半乳糖苷酶活性,我们发现其在转录水平上正向调控2,4-DAPG合成基因phlA和PcoI/PcoR群体感应信号系统合成基因pcoI的表达。2,4-DAPG抗生素和信号分子AHL的产生受其影响。此外,其参与调控phlD表达。它对phlB转录也有一定调控作用,但对phlC、phlE和phlF转录无影响。根据生防测定,野生型2P24菌株对棉花立枯丝核菌表现出明显的拮抗能力,而突变菌株失去了生防效果。其对2P24的游动性和生物膜形成有显著影响。

结论

我们的数据表明其是2P24中群体感应系统、2,4-DAPG的重要调控因子。这可能意味着其已进化出不同的调控特征以适应不同的环境威胁。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e9c0/10213339/bf9f11add313/fmicb-14-1160913-g001.jpg

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