农业生物技术学报
農業生物技術學報
농업생물기술학보
JOURNAL OF AGRICULTURAL BIOTECHNOLOGY
2014年
11期
1441-1446
,共6页
陈倩倩%刘波%关雄%唐建阳
陳倩倩%劉波%關雄%唐建暘
진천천%류파%관웅%당건양
金线莲%内生菌%宏基因组
金線蓮%內生菌%宏基因組
금선련%내생균%굉기인조
Anoectochilus roxburghii (Wall.) Lindl%Endophytes%Metagenome
金线莲(Anoectochilus roxburghii (Wall.) Lindl)是中国传统中药,具有重要的药用价值,现代医学用来治疗糖尿病和癌症。金线莲作为兰科植物,与内生菌在长期的生长过程中建立起和谐的共生关系,内生菌产生多种次生代谢产物,帮助宿主植物抗菌防虫。现有实验条件下,环境中绝大多数的微生物不能培养,而宏基因组学是认识环境中不可培养微生物的重要手段。本研究对福建金线莲根部内生菌富集进行探索,提取混合内生菌样品宏基因组DNA,并对宏基因组DNA纯化,PCR扩增微生物16S rDNA片段,克隆至大肠杆菌(Escherichia coli)并测序。根据16S rDNA信息初步对富集的内生菌群落分析,内生菌主要是不可培养细菌(uncultured bacterium)、不可培养堆肥细菌(uncultured compost bacterium)、肠杆菌科(enterobacteriaceae)、类芽胞杆菌属(Paenibacillus sp.)、芽胞杆菌属(Bacillus sp.)和短芽胞杆菌属(Brevibacillus sp.)的微生物。本研究为挖掘金线莲内生菌资源提供基础资料。
金線蓮(Anoectochilus roxburghii (Wall.) Lindl)是中國傳統中藥,具有重要的藥用價值,現代醫學用來治療糖尿病和癌癥。金線蓮作為蘭科植物,與內生菌在長期的生長過程中建立起和諧的共生關繫,內生菌產生多種次生代謝產物,幫助宿主植物抗菌防蟲。現有實驗條件下,環境中絕大多數的微生物不能培養,而宏基因組學是認識環境中不可培養微生物的重要手段。本研究對福建金線蓮根部內生菌富集進行探索,提取混閤內生菌樣品宏基因組DNA,併對宏基因組DNA純化,PCR擴增微生物16S rDNA片段,剋隆至大腸桿菌(Escherichia coli)併測序。根據16S rDNA信息初步對富集的內生菌群落分析,內生菌主要是不可培養細菌(uncultured bacterium)、不可培養堆肥細菌(uncultured compost bacterium)、腸桿菌科(enterobacteriaceae)、類芽胞桿菌屬(Paenibacillus sp.)、芽胞桿菌屬(Bacillus sp.)和短芽胞桿菌屬(Brevibacillus sp.)的微生物。本研究為挖掘金線蓮內生菌資源提供基礎資料。
금선련(Anoectochilus roxburghii (Wall.) Lindl)시중국전통중약,구유중요적약용개치,현대의학용래치료당뇨병화암증。금선련작위란과식물,여내생균재장기적생장과정중건립기화해적공생관계,내생균산생다충차생대사산물,방조숙주식물항균방충。현유실험조건하,배경중절대다수적미생물불능배양,이굉기인조학시인식배경중불가배양미생물적중요수단。본연구대복건금선련근부내생균부집진행탐색,제취혼합내생균양품굉기인조DNA,병대굉기인조DNA순화,PCR확증미생물16S rDNA편단,극륭지대장간균(Escherichia coli)병측서。근거16S rDNA신식초보대부집적내생균군락분석,내생균주요시불가배양세균(uncultured bacterium)、불가배양퇴비세균(uncultured compost bacterium)、장간균과(enterobacteriaceae)、류아포간균속(Paenibacillus sp.)、아포간균속(Bacillus sp.)화단아포간균속(Brevibacillus sp.)적미생물。본연구위알굴금선련내생균자원제공기출자료。
Anoectochilus roxburghii (Wall.) Lindl is a traditional Chinese medicine, which has been used for diabetes and tumor treatment. The wild A. roxburghii and its endophytes live together to form a balanced symbiotic system. The endophytes produce various secondary metabolites to protect the host from being attacked by fungi and pests. Endophytes are rich bio-resources. Because of a major limitation of isolation methods and cultivation media, the diversity endophytes still remain undiscovered. Metagenomics simultaneous study of endophytes genomes from environmental samples may avoid the limitations of culture-dependent methods. We enriched the microbiota from the root of A. roxburghii and extracted and purified the metagenomic DNA.16S rDNA fragments were amplified by PCR and cloned to Escherichia coli DH5α to sequence. The endophytes were mainly uncultured bacterium, uncultured compost bacterium, Enterobacteriaceae, Paenibacillus sp., Bacillus sp. and Brevibacillus sp., according to 16S rDNA information. This research provided the basis for mining endophytes resources of A. roxburghii. Analyzed the groups of endophytes by partial sequence of 16S rDNA. This work opens further insight into the great potential of A. roxburghii microbiota.