来自西双版纳3种有毒植物的免培养与纯培养放线菌多样性及生物活性  

Diversity and bioactivity of un-cultured and cultivable actinomycetes from three toxic plants in Xishuangbanna,China

作　　者：陈东波 柳成宾 姜怡[1] 吕元林[3] 姜成林[1] CHEN Dong-Bo1, LIU Cheng-Bin1,2, JIANG Yi1, LYU Yuan-Lin3, JIANG Cheng-Lin1(1. Yunnan Institute of Microbiology, Yunnan University, Kunming, Yunnan 650091, China; 2. College of Life and Health Sciences, Northeastern University, Shenyang, Liaoning 110819, China; 3. Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, Yunnan 650204, Chin)

机构地区：[1]云南大学云南省微生物研究所,云南昆明650091 [2]东北大学生命科学与健康学院,辽宁沈阳110819 [3]中国科学院昆明植物研究所,云南昆明650204

出　　处：《微生物学通报》2018年第5期1100-1111,共12页Microbiology China

基　　金：国家自然科学基金(31460005)~~

摘　　要：【背景】由于土壤放线菌中获得新化合物日益困难,抗生素滥用使致病菌耐药性不断增加,人们转向研究植物内生放线菌以期发现新化合物。【目的】探究西双版纳热带雨林有毒植物内生放线菌的多样性,为开发新药提供具有潜在生物活性的菌株。【方法】通过Illumina Hi Seq高通量测序和纯培养方法分析箭毒木、八角枫、马缨丹3种有毒植物的内生放线菌群落结构组成,利用纸片扩散法筛选抑菌活性,通过PCR扩增检测7类化合物合成基因。【结果】高通量测序的多样性分析和群落结构分析得出:3种有毒植物在门分类水平检测出古菌域的2个门、细菌域的18个门和暂定的Rsa HF231、WD272门;在属分类水平检测出30个属的放线菌,八角枫和马缨丹的微生物群落结构比箭毒木更丰富。纯培养分离获得11个属34株菌,分离自箭毒木的菌株比八角枫和马缨丹的菌株更多,而且大多数高通量检测出的菌种不能通过纯培养获得。抑菌活性检测结果显示:抗菌活性作用明显的菌株以链霉菌属为主。链霉菌属的NRPS和PKS基因的检出率明显高于其他化合物合成基因。【结论】有毒植物内生放线菌多样性非常丰富。有毒植物内生放线菌具有合成次生代谢产物的潜力,可以为生物农药及抗生素开发提供丰富的菌种资源。[Background] People tend to study endophytic actinomycetes to discover new compounds due to the increasing difficulty to obtain new compounds from soil actinomycetes and antibiotic abuse causing increased resistance of pathogens. [Objective] We explored the diversity of endophytic actinomycetes isolated from toxic plants in Xishuangbanna tropical rainforest, and isolated strains with antibacterial potential to develop new drugs. [Methods] We analyzed communities of endophytic actinomycetes from three toxic plants, Antiaris toxicaria Lesch., Alangium chinense（Lour.） Harms and Lantana camara L. by Illumina Hi Seq high-throughput sequencing and culture-dependent approach. Antimicrobial activity was screened by disk diffusion test and seven synthetic genes were detected by PCR amplifications. [Results] Strains from 3 toxic plants were identified at the phylum level with 2 phyla of the Archaea, 18 phyla and the tentative Rsa HF231, WD272 of the bacteria. And 30 genera of actinomycetes were detected at the genus level. Both the microbial community structure of Alangium chinense（Lour.） Harms and Lantana camara L. were more abundant than Antiaris toxicaria Lesch. In total 34 strains of 11 genera were isolated from culture-dependent approach. Most of the genus detected by high-throughput could not be obtained through pure culture. The antibacterial activity results showed that Streptomyces had broad spectrum antimicrobial activity. The detection rates of NRPS and PKS in Streptomyces were significantly higher than other compounds synthetic gene. [Conclusion] The diversity of endophytic actinomycetes in toxic plants is very rich. Actinomycetes in the toxic plants secondary metabolites synthesis have enormous potential to provide an abundant microbial resource for the development of biological pesticides and antibiotics.

关 键 词：有毒植物 放线菌 多样性 生物活性 合成基因 

分 类 号：Q93[生物学—微生物学]