Cao Fengming, Shen Delong, Li Jun, Guan Dawei, Jiang Xin, Li Li, Feng Ruihua, Yang Xiaohong, Chen Huijun, Ge Yifan
Institute of Agricultural Resources and Regional Planning, Chinese Academy of Agricultural Sciences, Beijing 10081, China.
Wei Sheng Wu Xue Bao. 2008 May;48(5):651-6.
To discriminate the strains of Bacillus subtilis group including B. subtilis, B. amyloliquefaciens, B. licheniformis, and B. pumilus, a rapid and accurate distinguishing method is essential for the identification of the target strains to ensure the quality and safety of microbial fertilizers.
By analyzing unique nucleotide sequences of the rpoA, gyrA and 16S rDNA genes, 4 pairs of species-specific primers were optimized and the multiplex PCR was developed to discriminate and identify B. subtilis, B. amyloliquefaciens, B. licheniformis and B. pumilus.
Thirty-three reference strains belonging to three genera of Bacillus, Paenibacillus and Brevibacillus were tested and the anticipated results appeared except for four species with cross amplification results with the primers of B. pumilus. However, the four species can be easily discriminated by morphology characters. In addition, the multiplex-PCR results of 23 strains of B. subtilis group isolated from MF products were identical with the biochemical assay.
The newly constructed multiplex-PCR assay is species-specific and effective. This method can be used to detect and identify the strains of B. subtilis group from microbial fertilizers products.
为鉴别枯草芽孢杆菌属的菌株，包括枯草芽孢杆菌、解淀粉芽孢杆菌、地衣芽孢杆菌和短小芽孢杆菌，一种快速准确的区分方法对于目标菌株的鉴定至关重要，以确保微生物肥料的质量和安全性。
通过分析rpoA、gyrA和16S rDNA基因的独特核苷酸序列，优化了4对种特异性引物，并开发了多重PCR来鉴别和鉴定枯草芽孢杆菌、解淀粉芽孢杆菌、地衣芽孢杆菌和短小芽孢杆菌。
对属于芽孢杆菌属、类芽孢杆菌属和短芽孢杆菌属三个属的33株参考菌株进行了检测，除4个物种与短小芽孢杆菌引物有交叉扩增结果外，均出现预期结果。然而，这4个物种可以通过形态特征很容易地鉴别出来。此外，从微生物肥料产品中分离出的23株枯草芽孢杆菌属菌株的多重PCR结果与生化分析结果一致。
新构建的多重PCR检测方法具有种特异性且有效。该方法可用于检测和鉴定微生物肥料产品中的枯草芽孢杆菌属菌株。
