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Establishment and application of quantitative detection of Bacillus velezensis HMB26553, a biocontrol agent against cotton damping-off caused by Rhizoctonia

文献类型： 外文期刊

作者： Su, Zhenhe 1 ; Liu, Gaoge 1 ; Li, Cong 1 ; Liu, Xiaomeng 1 ; Guo, Qinggang 1 ; Wang, Peipei 1 ; Dong, Lihong 1 ; Lu, Xiuyun 1 ; Zhao, Weisong 1 ; Zhang, Xiaoyun 1 ; Qu, Yuanghang 1 ; Zhang, Jiaqi 1 ; Mo, Shaojing 1 ; Li, Shezeng 1 ; Ma, Ping 1 ;

作者机构： 1.Hebei Acad Agr & Forestry Sci, Integrated Pest Management Ctr Hebei Prov, Key Lab IPM Crops Northern Reg North China, Minist Agr & Rural Affairs,Plant Protect Inst, Baoding, Peoples R China

2.Hebei Acad Agr & Forestry Sci, Inst Plant Protect, 437 Dongguan St, Baoding 071000, Hebei, Peoples R China

关键词： Bacillus velezensis; detection; quantification; strain-specific primers; TaqMan qPCR; unique gene pB-gene0026

期刊名称：BIOTECHNOLOGY JOURNAL （ 影响因子：4.7； 五年影响因子：4.5 ）

ISSN： 1860-6768

年卷期： 2024 年 19 卷 2 期

页码：

收录情况： SCI

摘要： A highly sensitive quantitative PCR (qPCR) method was developed for detection and quantification of Bacillus velezensis HMB26553 in cotton rhizosphere. The study aimed to develop a quantitative detection method for the strain HMB26553, and explore the relationship between its colonization of the cotton rhizosphere and its control effect. The whole genome sequence of strain HMB26553 was obtained by genome sequencing and a unique specific sequence pB-gene0026 on plasmid plaBV2 was identified by using high-throughput alignment against NCBI. Plasmid plaBV2 could be stably genetically inherited. Based on this sequence, specific primers for amplifying 106 bp and a minor groove binder (MGB) TaqMan probe for enhancing sensitivity were designed. The copy number of plaBV2 in strain HMB26553, which was 2, was confirmed by internal reference primers and the MGB TaqMan probe based on housekeeping gene gyrB. The established detection technique based on these primers and probes had high specificity and sensitivity compared to traditional plate counting method, with a detection limit of 1.5 copy genome. Using this method, the study discovered a likely correlation between the quantity of colonization in cotton rhizosphere and efficacy against cotton damping-off caused by Rhizoctonia after seed soaking and irrigation with strain HMB26553. Thus, this method provides scientific support for the rational application of strain HMB26553 in the future.

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