Bacterial Diversity In The Application Of Pcr-dgge Technology Research Marijuana Retting System

In order to find the dominant flora in the hemp retting, and lay the fundament for breeding hemp gumming strains with bacteria retting technology, the bacterium in hemp retting water were used to analysis for biodiversity.24 bacteria strains were isolated from hemp retting water of 10 different times by traditional cultivation. The strains were identified as members of 11 generas and 15 species according to the morphological, physiological, biochemical characteristics and the sequencing of 16S rDNA: Bacillus megaterium, Bacillus cereus, Bacillus licheniformis, Bacillus anthracis, Shigella sonnei,Shigella dysenteriae, Pseudomonas argentinensis, Exiguobacterium sp., Escherichia coli, Paenibacillus sp., Klebsiella pneumoniae subsp., Kurthia gibsonii, Enterobacteriaceae, Lactococcus lactis subsp., Pantoea agglomerans.The V3 domain sequences of 16S rDNA from bacterium in 10 different times in fermentetion liquid of hemp retting were amplified by PCR. The optimum condition of DGGE was finally confirmed that electrophoreses were run for 8h at 150V on a 8.0% acrylamide/bis- acrylamide (37.5:1) gel with 40%-70% denaturant gradient. All DGGE analysis was run at a constant temperature of 60℃. DGGE sequencing analysis show that the dominant bands were: Chryseobacterium sp., Listeria grayi, Pantoea agglomerans, Serratia marcescens, Klebsiella pneumoniae, Enterobacter sp., Stenotrophomonas maltophilia, Pediococcus pentosaceus, Clostridium sp., Lactococcus lactis subsp.and uncultured bacterium. DGGE fingerprint was analyzed by Gel-Pro Analyzer 4.5. Diversity index revealed that the dominant flora were more obvious during 12h,72h,108h and 120h. The dominant flora were Clostridium sp. and uncultured bacteria during the whole fermentetion process.