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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。
OBJECTIVE: To determine the taxonomic status of endophytic fungi P109-4, P116-1b and P312-1 isolated from Acanthopanax senticosus, and to analyze the mode of action to increase E content of Acanthopanax. Methods: The morphological and 18S rDNA sequence analysis methods were used for identification. The bacterial liquid was transferred back to Acanthopanax senticosus leachate for fermentation and the inactivated bacterial liquid was injected into Acanthopanax senticosus. The content of Acanthopanax senticosus was analyzed by HPLC Variety. Results: The morphological characteristics of P109-4, P116-1b and P312-1 were consistent with those of Fusarium oxysporum, 99.28%, 99.76% and 97.23%. The three endophytic fungi in vivo could prolong the existence time of Acanthopanax senticosus E in the extract of Acanthopanax senticosus L. P312-1 could significantly increase the content of Acanthopanax E after inactivation, but P109-4 and P116-1B had no significant effect . Conclusion: P109-4 is Fusarium oxysporum, P116-1b is Botryosphaeria dothidea, and P312-1 is Ceratobasidium spp. The mode of action of P312-1 is to increase the content of acanthopanax E by some kind of substance that is not destroyed by high temperature, and P116-1b and P109-4 can only work on the existing acanthopanax E.