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用于控制参与葡萄树干病害的真菌病原体葡萄生褐座壳孢的拮抗细菌的筛选及其作用模式

Screening and modes of action of antagonistic bacteria to control the fungal pathogen Phaeomoniella chlamydospora involved in grapevine trunk diseases.

作者信息

Haidar Rana, Roudet Jean, Bonnard Olivier, Dufour Marie Cécile, Corio-Costet Marie France, Fert Mathieu, Gautier Thomas, Deschamps Alain, Fermaud Marc

机构信息

SAVE, INRA, Institut National de Recherche Agronomique, Bordeaux Sciences Agro, ISVV, 33882 Villenave d'Ornon, France; Tichreen University, Faculty of Science, Biology Department, P.O. Box 2231, Latakia, Syrian Arab Republic.

SAVE, INRA, Institut National de Recherche Agronomique, Bordeaux Sciences Agro, ISVV, 33882 Villenave d'Ornon, France.

出版信息

Microbiol Res. 2016 Nov;192:172-184. doi: 10.1016/j.micres.2016.07.003. Epub 2016 Jul 18.

DOI:10.1016/j.micres.2016.07.003
PMID:27664735
Abstract

The antagonistic activity of 46 bacterial strains isolated from Bordeaux vineyards were evaluated against Phaeomoniella chlamydospora, a major grapevine pathogen involved in Esca. The reduction of the necrosis length of stem cuttings ranged between 31.4% and 38.7% for the 8 most efficient strains. Two in planta trials allowed the selection of the two best strains, Bacillus pumilus (S32) and Paenibacillus sp. (S19). Their efficacy was not dependent on application method; co-inoculation, prevention in the wood and soil inoculation were tested. The involvement of antibiosis by the secretion of diffusible and/or volatile compounds in the antagonistic capacity of these two strains was assessed in vitro. Volatile compounds secreted by B. pumilus (S32) and Paenibacillus sp. (S19) were identified by gas chromatography/mass spectroscopy (GC/MS). The volatile compounds 1-octen-3-ol and 2,5-dimethyl pyrazine were obtained commercially and tested, and they showed strong antifungal activity against P. chlamydospora, which suggested that these compounds may play an important role in the bacterial antagonistic activity in planta. Furthermore, the expression of 10 major grapevine defense genes was quantified by real-time polymerase chain reaction, which demonstrated that the two strains significantly affected the grapevine transcripts four days after their application on the plants. High expression levels of different genes associated with P. chlamydospora infection in B. pumilus pre-treated plants suggests that this strain induces systemic resistance in grapevine. For the first time, we demonstrated the ability of two bacterial strains, B. pumilus and Paenibacillus sp., isolated from grapevine wood, to control P. chlamydospora via direct and/or indirect mechanisms.

摘要

对从波尔多葡萄园分离出的46株细菌菌株针对葡萄座腔菌(Phaeomoniella chlamydospora)进行了拮抗活性评估,葡萄座腔菌是导致葡萄枝干病害(Esca)的一种主要葡萄病原体。对于8株最有效的菌株,茎段插条坏死长度的减少幅度在31.4%至38.7%之间。两项田间试验选出了两株最佳菌株,短小芽孢杆菌(Bacillus pumilus,S32)和类芽孢杆菌属(Paenibacillus sp.,S19)。它们的功效不依赖于施用方法;测试了共接种、木质部预防接种和土壤接种。在体外评估了这两种菌株通过分泌可扩散和/或挥发性化合物产生的抗菌作用在拮抗能力中的作用。通过气相色谱/质谱联用仪(GC/MS)鉴定了短小芽孢杆菌(S32)和类芽孢杆菌属(S19)分泌的挥发性化合物。商业获取并测试了挥发性化合物1-辛烯-3-醇和2,5-二甲基吡嗪,它们对葡萄座腔菌表现出很强的抗真菌活性,这表明这些化合物可能在植物体内的细菌拮抗活性中发挥重要作用。此外,通过实时聚合酶链反应对10个主要葡萄防御基因的表达进行了定量,结果表明这两种菌株在施用于植物四天后显著影响了葡萄转录本。短小芽孢杆菌预处理的植物中与葡萄座腔菌感染相关的不同基因的高表达水平表明该菌株诱导了葡萄的系统抗性。我们首次证明了从葡萄木质部分离出的两种细菌菌株,短小芽孢杆菌和类芽孢杆菌属,能够通过直接和/或间接机制控制葡萄座腔菌。

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