Diversity Analysis Of Endogenous Actinomycetes Of Xinjiang Characteristic Medicinal Plants And Multi-phase Classification Of Two New Streptomyces Species

Xinjiang's unique geographical ecological environment contains abundant medicinal plant endophytic actinomycetes and its secondary metabolite resources,and is a treasure trove of drug research and development.In this paper,10 different isolation media were used to isolate and identify 834 endophytic actinomycetes from 6 Xinjiang special medicinal plants included Glycyrhiza uralense,Apocynum venetum,Lycium ruthenicum,Cynanchumc hinense,Bupleurum chinense and Nigella sativa.The 834 endophytic actinomycetes were distributed in 5 orders,6 families and 9 genera including Streptomyces,Kitatotospora,Actinomadura,Nocardiopsis,Nocardioides,Mycoplasma,Nocardia,Promicromonspora and Kocuria.Among them,the dominant genus is Streptomyces,accounting for 60.89%of the total number of fragments separated.The largest number of actinomycetes isolated and identified from Apocynum venetum was 391strains,accounting for 46.89%;followed by Glycyrhiza uralense,124 strains were isolated and identified,accounting for 14.87%.Through multi-phase classification,the new endophytic actinomycete TRM 66187 isolated from the roots of Lycium barbarum and the new endophytic actinomycete TRM 66233 of the roots of Apocynum venetum were identified,and their taxonomic classification was determined according to their source of separation.They were named Streptomyces lycii and Streptomyces apocyni.The genome of strain TRM66187 was approximately 8.2 Mb in size with DNA G+C content of 71.8 mol%,The whole-cell sugar pattern of TRM 66187 consist of galactose,glucose and ribose.The predominant menaquinones are MK-9?H₄?and MK-9?H₆?.,the G+C content is 71.8 mol%.Major cellular fatty acids are iso-C_14:0,iso-C_15:0,anteiso-C_15:0 and anteiso-C_16:0.The polar lipids detected diphosphatidylglycerol?DPG?,phosphatidylcholine?PC?,phosphatidylethanolamine?PE?,phosphatidylinositolsphos?PI?,phatidylinositol mannoside?PIM?and an unidentified polar lipid.Optimum growth was observed at 37°C,pH 7.0 and with1%NaCl.The deposit number was TRM 66187=CCTCC AA 2018094=LMG31493.The genome of strain TRM 66233 was approximately 7.03 Mb in size with DNA G+C content of 70.35 mol%,The whole-cell sugar pattern of TRM 66233 consist of glucose and ribose.The predominant menaquinones are MK-9?H₂??MK-9?H₆??MK-9?H₈?and MK-9(H₁₀).Major cellular fatty acids are iso-C_15:0,anteiso-C_15:0,iso-C_16:0,C_16:0 and iso-C_17:0.The polar lipids detected diphosphatidylglycerol?DPG?,phosphatidylethanolamine?PE?,phosphatidylinositolsphos?PI?and four unidentified polar lipids L1-L4.Optimum growth was observed at 32°C,pH 7.0 and with 1%NaCl.The deposit number was TRM 66233=CCTCC AA 2019056=LMG31559.After screening the fermentation medium of the screw TRM 66187 with four kinds of fermentation medium,the corn starch medium was selected for fermentation.The fermentation broth was extracted by macroporous resin to obtain extract A.One compound was isolated from extract A using silica gel column chromatography,gel column chromatography,and efficient preparation of liquid phase.The structure of the compound was analyzed using NMR technology,and the compound was finally determined to be Thienodolin.