Diversity of endophytic actinomycetes from Entire meconopsis sampled in Aba, Sichuan

Abstract:

[Background] At present, the study on the endophytic actinobactreia from Entire meconopsis is relatively rare, the resource of endophytic actinobactreia from Entire meconopsis can be enriched with available culture and non-cultured methods. [Objective] To explore the diversity and community structure of endophytic actinobactreia from Entire meconopsis, a medicinal plant growing at high elevation, samples were collected from a special ecological environment in Aba, Sichuan Province. [Methods] Six different geographical positions were determined to sample and available culture method as well as PCR-DGGE technology was applied to investigate the diversity of endophytic actinobactreia from Entire meconopsis after a thorough surface disinfection. [Results] Based on culture method, the largest amount of isolates from different plant tissues was found in the root, followed in the leaf, stem and flower, respectively. The strains were grouped into 7 genotypes through 16S rDNA-RFLP Restriction fragment length polymorphism analysis, while representative strains were divided into 3 clusters in phylogenic analysis. Streptomyces was the dominant genus accounting for 50%, followed by Kitasatospora and Oerskovia and the greatest abundance of isolates was found in stem. The results of PCR-DGGE showed that the highest diversity index of endophytic actinobactreia was from root, stem and leaf that sampled in Galitai, Songpan and that from flower sampled in Anquxiang, Hongyuan. However, the largest diversity index in the same geographical position was from stem and flower. Additionally, the sequences in phylogenic analysis were classified into 8 genotypes including Rhodococcus, Corynebacterium, Nocardia, Micrococcus, Mycetocola, Microbacterium, Pseudonocardia and Streptomyces, among which Rhodococcus was predominant representing 70%. In general, it was a more effective way of combining available culture with uncultured methods to investigate the diversity of endophytic actinomycetes from Entire meconopsis. [Conclusion] The study revealed a rich diversity from different tissues in Entire meconopsis and laid foundation for further researches in active substances and medicine efficacy.