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分离的铜绿假单胞菌菌株 VIH2 及其对青枯雷尔氏菌的拮抗特性。

Isolated Pseudomonas aeruginosa strain VIH2 and antagonistic properties against Ralstonia solanacearum.

机构信息

Soil Ecology Lab, College of Resources and Environmental Sciences, Nanjing Agricultural University, No. 6 TongWei Road, Nanjing 210095, Jiangsu Province, People's Republic of China.

Soil Ecology Lab, College of Resources and Environmental Sciences, Nanjing Agricultural University, No. 6 TongWei Road, Nanjing 210095, Jiangsu Province, People's Republic of China.

出版信息

Microb Pathog. 2017 Oct;111:519-526. doi: 10.1016/j.micpath.2017.08.020. Epub 2017 Aug 25.

Abstract

The aim of this study was to isolates with antagonist activity against R. solanacearum. Thirty-two bacterial isolates were obtained from samples, and they were screened for potential antagonistic activity against R. Solanacearum. Using the agar spot method, ten out of the 21 tested bacteria showed antilisterial activity. VIH2 had the highest inhibitory effect on the growth of R. Solanacearum. Based on 16S rDNA and Biolog test analysis, the strain VIH2 was identified as Pseudomonas aeruginosa. Single-factor and Response Surface Methodology experiments were used to optimize the culture medium and conditions. This study was to explore whether the hemolysin-co-regulated protein secretion island I (HSI-I)-encoded type VI secretion system (T6SS) in Pseudomonas can be used as a biological control approach against Ralstonia solanacearum under field conditions. Bacterial competition assay showed that the HSI-I type T6SS of strain VIH2 exhibited dramatic antibacterial killing activity against R. solanacearum. The HSI-I T6SS of P. aeruginosa was regulated by the ppKA gene. We disrupted the gene ppKA in VIH2 by a single crossover to yield the VIH2 (ΔppKA) mutant. The antagonism of VIH2 was significantly decreased by ppKA gene disruption. In conclusion, our data supported the idea that HSI-I T6SS plays a crucial role in the antagonistic action of strain VIH2 against R. solanacearum. This alternative approach for antagonism against R. solanacearum might help develop attenuated strains of engineered bacteria for biological control.

摘要

本研究的目的是分离对茄青枯雷尔氏菌具有拮抗活性的菌株。从样品中获得了 32 个细菌分离株,并对其潜在的拮抗活性进行了筛选。采用琼脂点法,从 21 株供试细菌中筛选出 10 株具有抑菌活性的细菌。VIH2 对茄青枯雷尔氏菌的生长抑制作用最强。根据 16S rDNA 和 Biolog 试验分析,菌株 VIH2 被鉴定为铜绿假单胞菌。采用单因素和响应面法优化了培养基和培养条件。本研究旨在探索铜绿假单胞菌溶血素协同调控蛋白分泌岛 I (HSI-I)编码的 VI 型分泌系统 (T6SS)是否可作为田间防治茄青枯雷尔氏菌的生物防治方法。细菌竞争试验表明,VIH2 的 HSI-I 型 T6SS 对茄青枯雷尔氏菌具有显著的杀菌活性。铜绿假单胞菌的 HSI-I T6SS 受 ppKA 基因调控。我们通过单交换敲除 VIH2 中的基因 ppKA,得到 VIH2 (ΔppKA) 突变体。ppKA 基因敲除后,VIH2 的拮抗作用显著降低。综上所述,我们的数据支持 HSI-I T6SS 在 VIH2 拮抗茄青枯雷尔氏菌中的关键作用的观点。这种拮抗茄青枯雷尔氏菌的替代方法可能有助于开发用于生物防治的工程菌减毒株。

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