Diversity Of Endophytic Actinobacteria And Rhizosphere Soil Actinobacteria From Meconopsis Punicea Sampled In Aba, Sichuan

Meconopsis punicea as a medicinal plant come from high altitude and there is a special ecological environment in Aba, Sichuan province. This study was to explore diversity and community structure of endophytic actinobactreia and rhizosphrer soil actionbacteria from Meconopsis punicea. After previously surface disinfection use available culture method with PCR-DGGE technology to investigate the diversity of endophytic actinobacteria and rhizosphere soil actinobacteria from Meconopsis punicea in Aba,Sichuan 6 different geographical positions. The results showed that:(1)There were many factors to affect actinomycetes’ distribution. Plant samples have the least number of actinomycetes when soil electric conductivity less than one; Soil organic matter is positively related to the number of actinomycetes within certain limits; Sampling altitude was negatively related to the number of endophytic actinomycetes while positively related to that of rhizospheric actinomycetes. This study showed obvious differences of isolated actinomycetes from 10 difference media:In endophytic actinobactreia, gauze No.1(GA) medium and improvement of proline medium did the maximum number, Chitin medium did the minimum number; In rhizosphrer soil actionbacteria gauze No.1(GA) medium did the maximum number, Chitin medium、SCN medium and humic acid medium did the minimum number. In culture method, the largest amount of isolates from different plant tissues were found in the root (18), then the leaf (17), stem (9) and flower (8).(2)This article made a preliminary study of the diversity of actinomycetes with weighted calculation, based on 7 phenotypic characteristics of them and finally selecting 56 strains in-depth study. ARDRA-RLFP and PCR-BOX analysis of endophytic and rhizospheric actinomycetes indicated that the diversity of 32 endophytic actinomycetes and 24 rhizospheric actinomycetes had some differences. The diversity of actinomycetes was some abundant, but the clustering distribution was uneven. Phylogenetic tree of representative sequenced strains indicated that sequenced strains including Sireptomyces. Kitasatospora, Oerskovia and one just classification to Actinobacteria:SCAUEjDSI. among which Streptomyces was predominant with 78 percent in the insecticides..The best diversity of endophytic actinomycetes isolated from stem in Meconopsis punicea.(3)After PCR-DGGE.in different geographical position.the diversity index of endophytic actinomycetes from root is A>F>E>D>C>B (A:Galitai.Songpan Altitude 3553m; B:Anquxiang,Hongyuan Altitude 3523m; C:Barkam Altitude3894m:D:Xiaojin County Altitude 3789m; E:Xiaojin balang mountain Altitude 4120m:F:Xiaojin County Altitude 4114m, similarly hereinafter):the diversity index of endophytic actinomycetes from stem is A>D>E>B>C>F; the diversity index of endophytic actinomycetes from leaf is A>B>D>C>E>F:the diversity index of endophytic actinomycetes from flower is B>D>E>C>F>A.In the same geographical position, the diversity index of endophytic actinomycetes is stem>leaf>root>flower in Galitai,Songpan, flower>leaf>stem>root in Anquxiang,Hongyuan, leaf> flower> stem> root in Barkam. flower> stem> leaf> root in Xiaojin County (Altitude 3789m), stem> flower> leaf> root in Xiaojin balang mountain, stem> root> leaf> flower in Xiaojin County (Altitude 4114m). In 6 different geographical position, the diversity index of rhizosphrer soil actionbacteria is E>D>A>F>C>B.The DGGE phylogenic analysis classified the representative strains as 8 genotypes, including Rhodococcus, Corynebacterium, Nocardia, Micrococcus, Mycetocola. Microbacterium, Pseudonocardia, Streptomyces, among which Rhodococcus was predominant with 65 percent in the total. The species distribution of endophytic actinomycetes in different plant tissue:stem (5)> root and leaf (3)> flower (2). In 6 different geographical position, the species distribution of endophytic actinomycetes is Anquxiang,Hongyuan (Altitude 3523m), Xiaojin balang mountain (Altitude 4120m) and Xiaojin County (Altitude 4114m) have 3; Galitai.Songpan (Altitude 3553m) and Barkam (Altitudes894m) have 2; Xiaojin County (Altitude 3789m) just have 1.This study showed Meconopsis punicea plant tissues have rich diversity, the distribution of actinomycetes types within the organization and its secondary metabolites has great use value and potential economic value.Research endophytic actinomycetes of Meconopsis punicea in Aba will help us understand its medicinal value, the development of new resources, it also can provide basis for active substance and medicine efficacy research in the future.