Endophytic Bacterial Diversity In Glycyrrhiza Inflata Bat. From Xinjiang By Culture-independent Method

We investigated endophytic bacterial diversity in root of Glycyrrhiza inflata Bat. from Xingjian by culture-independent method,aimed to lay a foundation for furthering to tap.Glycyrrhiza roots were sampled from Ziniquan, Kuerle, in XinJiang province, the original habitat. Samples were strictly surface-sterilized followed by CTAB (Hexadecyl trimethyl ammonium Bromide) procedure with some modifications. A pair of bacterial PCR primers was used for endophytic bacterial 16S rDNA gene amplification and a clone library was constructed for the Glycyrrhiza DNA samples. Thirty two OTUs, including 150 positive clones, were assigned in the clone library after amplifying rDNA restriction analysis (ARDRA). The representatives of each OTU were sequenced and compared with the sequences in GenBank of National Center for Biotechnology Information (NCBI), followed constructed Neighbor-Joining tree using MEGA3.1.The result of blast suggested that 74% of the clones showed high similarity to the 16S rDNA gene sequence of known cultured bacteria,and Sequence analysis revealed diverse phyla of bacteria in the 16S rDNA library, which consisted of alpha, gamma subclasses of the Proteobacteria, Bacteroides, Firmicutes, Actinobacteria, and Uncultured bacteria; 74% of the clones were highly related to the known bacteria in 14 genus including Sphingobium, Phyllobacterium, Hyphomonas, Agrobacterium etc (>96% sequence similarity); 17% of the clones showed lower affiliation with known bacteria in the genus Promicromonospora,Nocardia,Legionella and Novosphingobium (93%-95% sequence similarity); still 9% of the clones showed lower affiliation with unknown genus (<96% sequence similarity) and may represent novel taxa.There was abundant endophytic bacterial diversity in Glycyrrhiza inflata Bat.from Xinjiang as well as many unknown organisms.