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采用NA培养基分离纯化细菌,通过形态特征、生理生化指标结合16S r DNA序列同源性分析,对引起化血胆组培苗污染的内生细菌进行鉴定。结果表明,引起化血胆组培苗污染的内生细菌为HXD5菌株,其形态特征及22项生理生化指标与枯草芽孢杆菌相符;16S rDNA序列分析发现,HXD5与B.amyloliquefaciens(HQ727971)、B.vallismortis(FJ386541)、B.licheniformis(EF644414)、B.subtilis(AY583216)在同一系统发育分支,同源性为99.9%~100%。因此确定引起化血胆组培苗污染的内生细菌HXD5为枯草芽孢杆菌。抑菌试验表明,氯霉素对HXD5的抑菌效果要优于链霉素。
The bacteria were isolated and purified by NA medium. The endophytic bacteria which caused the pollution of chemotactic blood vessel were identified by morphological, physiological and biochemical indexes and 16S r DNA sequence homology analysis. The results showed that the endophytic bacteria that caused the pollution of chemotactic blood group were HXD5 strain, and its morphological characteristics and 22 physiological and biochemical indexes were consistent with those of Bacillus subtilis. 16S rDNA sequence analysis showed that HXD5 and B. amyloliquefaciens (HQ727971), B .vallismortis (FJ386541), B.licheniformis (EF644414), B.subtilis (AY583216) in the same phylogenetic tree, with homology of 99.9% -100%. Therefore, the endophytic bacterium HXD5 which is the cause of the contamination of chemotactic blood group was identified as Bacillus subtilis. Antibacterial tests showed that chloramphenicol HXD5 antibacterial effect better than streptomycin.