三株提高刺五加苷E 含量内生真菌的鉴定及其作用方式分析

目的:确定自刺五加分离的内生真菌P109-4、P116-1b和P312-1的分类地位,并初步分析其提高刺五加苷E含量的作用方式。方法:应用形态学及18S rDNA序列分析方法进行鉴定,将菌液回接刺五加浸出液进行发酵培养和将灭活后的菌液注射刺五加后,HPLC法分析刺五加苷含量E的变化。结果:P109-4、P116-1b和P312-1分别与尖孢镰孢菌、葡萄座腔菌和角担菌的形态学特征相符,与各对应种属真菌的18S rDNA序列同源性分别高达99.28%、99.76%和97.23%。活体的3株内生真菌能延长刺五加浸出液中刺五加苷E的存在时间,灭活后P312-1可显著提高刺五加苷E的含量,P109-4和P116-1B无显著作用。结论:P109-4为Fusarium oxysporum,P116-1b为Botryosphaeria dothidea,P312-1为Ceratobasidium spp.。P312-1的作用方式为通过菌体某种不被高温破坏的物质提高刺五加苷E的含量,P116-1b和P109-4仅能对已存在的刺五加苷E发挥作用。

Identification of Three Endophytic Fungus that Increase Eleutheroside E and Analysis on its Function Mechanism

Objective: To confirm the evolutionary station of endogenous fungus P109-4,P116-1b and P312-1 isolated from Eleutherococcus senticosus,and to preliminarily investigate its mechanism of improving content of eleutheroside.Methods: Morphological characterization and ITS sequencing were used for identification of the fungus.The fungus were cultured in extraction of E.senticosus and then the inactivated fungus were injected into E.senticosus.The content of eleutheroside E was detected by HPLC.Results: P109-4,P116-1b and P312-1 correspond to morphological characteristics of Fusarium oxysporum,Botryosphaeria dothidea and Ceratobasidium spp.respectively.And the 18S rDNA similarity with each genus was as high as 99.28 %,99.76 % and 97.23 % respectively.The three fungus prolonged the existence of eleutheroside E,and inactivated P312-1 increased content of eleutheroside E significantly.However the other inactivated fungus didn’t show such significant action.Conclusion: P109-4,P116-1b and P312-1 was Fusarium oxysporum,Botryosphaeria dothidea and Ceratobasidium spp.respectively.The mechanism of P312-1 in improving the content of eleutheroside E was due to some materials which cannot be destroyed by high temperature,whereas P116-1b and P109-4 worked on the existent eleutheroside E.