暗褐网柄牛肝菌(Phlebopus portentosus)是第一个栽培成功的牛肝菌目(Boletales)真菌,一般采用覆土栽培模式,覆土微生物对子实体的发育具有重要作用,目前的研究主要集中在覆土微生物上,但缺少对子实体内生菌的研究.为揭开暗褐网柄牛肝菌子实体内生菌的组成,本研究基于Illumina MiSeq平台,利用16S rRNA V3～V4区高通量测序进行细菌群落结构分析.结果表明子实体(fruiting body,FB)内部存在很高丰度的细菌,主要以变形菌门(Proteobacteria)为主,在属水平上以贪铜菌属(Cupriavidus)(64.98％)、不动杆菌属(Acine-tobacter)(7.62％)、苍白杆菌属(Ochrobactrum)(6.02％)为主.与覆土(casing soil,CS)基质固有的细菌种群进行比较,VENN分析显示子实体内有468个OTUs为独有,占总序列的26.65％,分类上隶属于贪铜菌属、不动杆菌属和Delftia.FB和CS差异性分析以及Lefse标志性微生物分析均显示贪铜菌属、苍白杆菌属、不动杆菌属、嗜糖假单胞菌属(Pelomonas)等为FB特异性、标志性类群.利用Picrust(Phylogenetic investigation of com-munities by reconstruction of unobserved states)进行 Kyoto Encyclopedia of Genes and Genomes(KEGG)通路预测发现,子实体内生菌群的主要代谢途径为膜传输、氨基酸代谢、碳水化合物代谢、外源生物降解与代谢,与覆土菌群有20个代谢通路的差异,并且外源生物降解与代谢、信号转导、其他氨基酸代谢、脂质代谢、细胞过程和信号5个通路的比例要高于覆土,微生物类群的差异可能是导致产生这些功能差异的原因.
Bacterial Community Structure in the Fruiting-Bodies of Cultivated Phlebopus portentosus
As the first successfully cultivated fungus belonging to Boletales,casing soil(CS)is usually used to im-prove the cultivation.Microorganisms play important roles in the development of fruiting bodies(FB).Present resear-ches mainly focus on microorganisms in casing soil,but lack of exploring endophytic bacteria in fruiting bodies.In or-der to pierce the bacterial components in the FB of Phlebopus portentosus(Berk.& Broome)Boedijn,16S rRNA based on Illumina MiSeq platrform high throughput sequencing targeting V3-V4 regions was used to analyze the bacte-rial population structure in P.portentosus.The results indicated that there were high abundance of bacterial species inside the fruitification of P.portentosus,mainly predominated by Proteobacteria.Cupriavidus,Acinetobacter,Ochrobactrum,Pelomonas,Delftia and Aquabacterium were the most abundant genus.A total of 468 unique OTUs were detected in FB,which occupied 26.65％of the total sequences.Their classification belonged to the genus of Cu-priavidus,Acinetobacter and Delftia.In addition,both results of FB and CS difference analysis and Lefse iconic micro-bial analysis all showed that the genera of Cupriavidus,Acinetobacter,Ochrobactrum,Pelomonas,Delftia and Aquabacterium were the specificity and iconic groups.The results of using Picrust(Phylogenetic investigation of com-munities by reconstruction of unobserved states)to carried out KEGG(Kyoto Encyclopedia of Genes and Genomes)access prediction found that the main metabolic pathway in FB endophytic bacteria was membrane transmission,amino acid metabolism,carbohydrate metabolism,xenobiotics biodegradation and metabolism,and there were 20 differences of metabolic accesses,moreover,xenobiotics biodegradation and metabolism,signal conduction,other amino acid me-tabolism,lipid metabolism,cellular process and signal the proportion of these 5 accesses were higher than in CS.The difference of microbial groups may be the reason of these performances differences.
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