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棉花黄萎病拮抗细菌H14的筛选鉴定及其拮抗机理分析

引用本文：李全胜,谢宗铭,刘政,张国丽,武冬梅,田英.棉花黄萎病拮抗细菌H14的筛选鉴定及其拮抗机理分析.植物保护学报,2018,45(6):1204-1211

DOI：10.13802/j.cnki.zwbhxb.2018.2017091

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作者	单位	E-mail
李全胜	新疆农垦科学院生物技术研究所, 作物种质创新与基因资源利用兵团重点实验室, 石河子 832003
谢宗铭	新疆农垦科学院生物技术研究所, 作物种质创新与基因资源利用兵团重点实验室, 石河子 832003	xiezmchy@163.com
刘政	新疆农垦科学院植物保护研究所, 石河子 832003
张国丽	新疆农垦科学院生物技术研究所, 作物种质创新与基因资源利用兵团重点实验室, 石河子 832003
武冬梅	新疆农垦科学院生物技术研究所, 作物种质创新与基因资源利用兵团重点实验室, 石河子 832003
田英	兵团农业技术推广总站, 乌鲁木齐 830011

中文摘要:为筛选对大丽轮枝菌Verticillium dahliae Kleb.具有拮抗活性的根际细菌，以大丽轮枝菌为靶菌，分离获得棉花根际细菌，利用平板对峙法和琼脂扩散法筛选具有较高拮抗活性的菌株，采用室内盆栽法测定筛选所得菌株对棉花黄萎病的防效，并通过形态特征、生理生化特性和16S rDNA基因序列分析确立其分类地位，采用底物降解法和抗菌肽基因克隆法检测其产生水解酶和抗菌肽的能力。结果显示，试验共分离获得182株棉花根际细菌，筛选得到3株对大丽轮枝菌抑菌率大于50.00%且抑菌圈直径大于15 mm的菌株，其中菌株H14的抑菌率和抑菌圈直径最大，分别为54.25%和18.10 mm。该菌能在0~9% NaCl和pH 4.5~9.0的NB培养基上生长；经形态特征观察、生理生化试验及16S rDNA基因序列分析，最终将其鉴定为莫哈韦芽胞杆菌Bacillus mojavensis；菌株H14对大丽轮枝菌孢子萌发的抑制率和对棉花黄萎病的盆栽防效分别为89.55%和74.57%；菌株H14能合成蛋白酶，含有srfAA、fenD、bacA脂肽类抗生素合成基因。表明莫哈韦芽胞杆菌菌株H14能够合成蛋白酶和脂肽类拮抗物质，具有良好的抑菌和防病能力。

中文关键词:棉花黄萎病生物防治拮抗细菌拮抗机理

Screening and identification of antagonistic bacterium H14 against Verticillium dahliae Kleb. and its antagonistic mechanisms

Author Name	Affiliation	E-mail
Li Quansheng	Xinjiang Production & Construction Group Key Laboratory of Crop Germplasm Enhancement and Gene Resources Utilization, Biotechnology Research Institute, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832003, Xinjiang Uygur Autonomous Region, China
Xie Zongming	Xinjiang Production & Construction Group Key Laboratory of Crop Germplasm Enhancement and Gene Resources Utilization, Biotechnology Research Institute, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832003, Xinjiang Uygur Autonomous Region, China	xiezmchy@163.com
Liu Zheng	Plant Protection Institute, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832003, Xinjiang Uygur Autonomous Region, China
Zhang Guoli	Xinjiang Production & Construction Group Key Laboratory of Crop Germplasm Enhancement and Gene Resources Utilization, Biotechnology Research Institute, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832003, Xinjiang Uygur Autonomous Region, China
Wu Dongmei	Xinjiang Production & Construction Group Key Laboratory of Crop Germplasm Enhancement and Gene Resources Utilization, Biotechnology Research Institute, Xinjiang Academy of Agricultural and Reclamation Sciences, Shihezi 832003, Xinjiang Uygur Autonomous Region, China
Tian Ying	Production & Construction Corps General Station for Popularization of Agricultural Technology, Urumqi 830011, Xinjiang Uygur Autonomous Region, China

Abstract:To screen the antagonistic rhizosphere bacteria with strong antifungal activity against Verticillium dahliae Kleb., the strain with higher fungicidal activity against cotton Verticillium wilt was obtained with the dual culture and agar diffusion methods. The control efficiency of the strain against cotton Verticillium wilt was evaluated with pot experiments. Its taxonomic status was defined using morphological identification, physiological characterization, and 16S rDNA sequencing. Additionally, the hydrolase activity and antimicrobial peptide of the strain were tested with substrate degradation and gene cloning methods. The results showed that 182 rhizosphere bacteria were isolated. There were three strains with inhibition rates higher than 50.00% and inhibition zones greater than 15 mm. Stain H14 had the highest inhibition rate and the largest inhibition zone diameter (54.25% and 18.10 mm, respectively). Strain H14 could grow on nutrient broth medium containing 0-9% NaCl with pH ranging from 4.5 to 9.0. Based on morphological observation, physiological assay and 16S rDNA sequence analysis, strain H14 was identified as Bacillus mojavensis. The inhibition rate of its fermentation liquid on conidial germination was 89.55%, and the control efficiency of strain H14 against cotton Verticillium wilt was 74.57%. Strain H14 could synthesize protease and had srfAA, fend, bacA genes. The results indicated that B. mojavensis H14 could synthesize protease, produce lipopeptide antagonists, and control diseases.

keywords:cotton Verticillum wilt biological control antagonistic bacterium antagonistic mechanism

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