Research On The Soil Actinomycetes Diversity,Bioactivitv Of Nujiang Prefecture In Nujiang Grand Canyon

Recent years,the development and utilization of the microbial medicine have turn into a soft patch.It takes huge manpower and material resouces in order to find new secondary metabolite which also have effective bioactivity,and the traditional isolation sources of the microbial medicine have been researchedrepeatedly.So,it's necesarry to discover the new reseachful actinomycete strians by exploiting the uninvestigated environment.As we know,soil environment is always a great treasury of actinomycetes resource.Although it has been investigated by many researchers,base on the widely distribution and various physicochemical properties,remaining some unknown soil environment sample waiting people to research.In this study,Nujiang Prefecture in Yunnan Province of Nujiang Grand Canyon is chosen by high diversity of plants and unique landform.Meanwhile,this soil sample is rarely reported in terms of actinomycetes resource.In order to comprehensive investigate the actinomycete diversity and bioactivity ofit,the research would be carried out as following procedure:1.High throughput sequencing was emploied to investigate the actinomycete culture-independent diversity of the seven soil samples.A total of 474203 bacteria gene and 15671 OTUs were obtained,the results destributed in 47 phylums,90 classes,170 orders,320 families,561 genus.Specific to the actinomycete phylum,high throughput sequencing detected 9 class,53 families,80 genus.We used several bioinformatical figures to making comparison of the differences in actinomycetes community composition structure among the seven samples.Combined with the soil physical and chemical indicators,we analyzed its influence on the bacterial and actinomycetes community composition structure of different samples on genus level.2.The suited culture-dependent isolation methods were determined by tested different pre-treatment methods,undesired bacteria inhibators and multiple isolation media.The results showed that 351 actinomycete strains were obtained from seven soil samples.It belonged to 8 orders,14 families,26 genus.YIM171 media showed the greatest isolation results.The sample of BH exhibited the most rich actinomycetes diversity obviously.3.In this study,we also screened the bioactivity,which comprised antimicrobial activity,fuctional genes,xylanase activity and urease activity,among the isolated actinomycete strains.Against to the indicator strains,the positive rate of the tested strains is 9%,7%,21%,14%,3%,10%.The positive rate for functional genes was 26%?36%?44%?16%?12%.We also detected 38 xylanase positive strains and 42 urease positive strains.4.Based on the isolation methods derived from the culture-dependent sector,we attempt to probe the antibiotic selective isolation method.In general,experiment divided into two parts,which comprised two samples,six types antibiotic and three type media.The finnal result showed that Micromonospora was isolated solely,using the pretreated sample of Bingzhongluo,which spreading on the YIM171 media contained 80 mg/L apramyin.