芽胞杆菌种类脂肪酸鉴定与分子鉴定方法的比较
Comparison of Bacillus Species Identification Based on ITS and Fatty Acid Analysis
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摘要: 基于ITS序列和脂肪酸鉴定方法对从土壤中分离得到的16株芽胞杆菌进行分类研究,与分子鉴定结果相比,脂肪酸种的鉴定准确率可达到99%以上。通过生物软件SPSS和Mega分别对16株菌进行聚类分析,结果表明,脂肪酸比ITS更适合于芽胞杆菌的分类研究,根据脂肪酸成分可以将16株芽胞杆菌准确聚在一起,而ITS则不能。芽胞杆菌特征性脂肪酸为15:0ANTEISO和15:0ISO,但不同种芽胞杆菌的此类脂肪酸含量不同。对芽胞杆菌3大类菌群主要特征性脂肪酸的含量比例(15:0ISO/15:0ANTEISO)进行了分析,其中比较特殊的为蜡状芽胞杆菌类群,蜡状芽胞杆菌主要脂肪酸为15:0ISO,其次为13:0ISO,而15:0ANTEISO的含量很低,而且不是主要类型,15:0ISO/15:0ANTEISO的比值约为5.6~7.9。简单芽胞杆菌群主要脂肪酸为15:0ANTEISO和15:0ISO,15:0ISO/15:0ANTEISO比值约为1/6。巨大芽胞杆菌类群主要脂肪酸15:0ISO和15:0ANTEISO含量相近,其余脂肪酸远远低于这2种主要脂肪酸。巨大芽胞杆菌的15:0ISO/15:0ANTEISO的比值约为4/5~5/4,菌株FJAT-4500为短小芽胞杆菌,其15:0ISO/15:0 ANTEISO比值约为2。短小芽胞杆菌和巨大芽胞杆菌归为一类是因为其脂肪酸类型相近,但含量并不同。枯草芽胞杆菌群主要脂肪酸15:0ISO的含量小于15:0ANTEISO,15:0ISO/15:0ANTEISO的比值为1/3~2/3。Abstract: 16 strains of Bacillus species isolated from soil were studied based on ITS sequences and fatty acids analysis. The accuracy rate of FA identification was 99%. Through biological software of SPSS and Mega cluster analysis, fatty acids were found to be better for Bacillus identification than ITS sequences. In this paper, we could make all the strains cluster together by FA, but ITS not. The major fatty acid of genus Bacillus was 15:0 ANTEISO and 15:0 ISO. The major FA ratio of three group were analyzed. Bacillus cereus group was special, its major FA was 15:0ISO,and 13:0ISO,but 15:0 ANTEISO had very lower profile. The ratio of 15:0ISO/15:0 ANTEISO in Bacillus cereus group was 5.6-7.9. The ratio of 15:0ISO/15:0 ANTEISO in Bacillus simplex group was 1/6. The ratio of 15:0ISO/15:0 ANTEISO in Bacillus megaterium was 4/5-5/4,strain FJAT-4500 was Bacillus pumilus,the ratio of15:0ISO/15:0 ANTEISO was 2. Bacillus pumilus was owed to Bacillus megaterium for similar ratio of 15:0ISO/15:0 ANTEISO, but the couple had different fatty acid profiles. The profile of 15:0ISO was lower than 15:0 ANTEISO in Bacillus subtilis group, and ratio was 1/3-2/3.
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