代谢组分析亚洲兰茂牛肝菌原基发育的潜在调控物质

Metabolomic analyses of potential regulatory substances ofprimordium development of Lanmaoa asiatica

亚洲兰茂牛肝菌Lanmaoaasiatica为云南珍稀的食药用真菌,属于外生菌根类真菌,在纯培养基上其部分菌丝能扭结形成原基。为了揭示调控原基发育的潜在物质,本研究运用核磁共振(1H-NMR)、气相质谱(GC-MS)和液相质谱(LC-MS)3种代谢组学技术,结合SMICA-P软件,定性及定量分析了纯培养35 d后菌丝体与原基的差异物质。结果表明,菌丝体与原基比较,3种方法共检测到谷氨酸、天冬氨酸、色氨酸、4-氨基丁酸、甜菜碱和海藻糖等96个显著或极显著上调的差异物质;调控通路分析表明,这些差异物质在原基发育中可能涉及丙氨酸、天冬氨酸和谷氨酸、精氨酸和脯氨酸、丁酸等22个通路;KEGG富集分析表明,谷氨酸涉及P<0.05的所有调控通路,推测谷氨酸在原基发育中起着重要的调控作用;在培养基中加入0.16 g/L谷氨酸可促进原基的萌发与生长。上述研究为亚洲兰茂牛肝菌的人工繁育技术研发奠定了基础。

Lanmaoa asiatica is a rare edible and medicinal fungus in Yunnan Province,which belongs to ectomycorrhizal fungi,and part of its mycelia could kink together to form primordia on pure media.In order to reveal the potential substances regulating the development of primordia,three metabolomics techniques,1H-NMR,GC-MS and LC-MS,were used to analyze qualitatively and quantitatively the differential substances between the mycelia and the primordia cultured for 35 days,in combination with SMICA-P software.The results showed that 96 significantly or extremely significantly up-regulated substances such as glutamic acid,aspartic acid,tryptophan,4-aminobutyric acid,betaine,trehalose and so on were detected in mycelia;analysis of the regulatory pathways showed these differential substances might involve 22 important pathways in the development of primordia.According to KEGG enrichment analysis,glutamate is involved in all regulatory pathways with P<0.05,so it is speculated that glutamate plays an important role in the regulation of primordium development.By adding 0.16 g/L glutamate in the media,the germination and growth of primordia were promoted.The above results provide reference for development of artificial breeding of Lanmaoa asiatica.