肿节风内生真菌的分离鉴定及拮抗真菌筛选

Isolation and Identification of Endophytic Fungi from Medicinal Plants of Sarcandra glabra(Thunb.)and Screening of Antagonistic Fungi

【目的】分析药用植物肿节风的内生真菌多样性,筛选具有较强抗菌活性的菌株,为寻找新的抗菌药物资源打下基础。【方法】采用平板对峙法从肿节风的不同组织中分离出具有抗菌活性的内生真菌,结合形态特征和ITS序列对其进行种类鉴定,筛选出抗菌活性强的内生真菌菌株。【结果】从肿节风的不同组织中分离、纯化得到111株内生真菌,选择42株不同形态类型的菌株进行分子鉴定,分属于9目20属28种。筛选出抗菌活性强的RJ-1和JJ-1菌株[分别鉴定为橡胶生拟茎点霉菌(Phomopsis heveicola)和新壳梭孢菌(Neofusicoccum parvum)],对15种病原真菌具有较强的抑制作用,其中对三七灰霉病菌(Botrytis sp.)、三七根腐病菌(Fusarium solani)、三七炭疽病菌(Colltetorrichum gloeosporioides)、香蕉炭疽病菌(Colltetorrichum sp.)、艾纳香褐斑病菌(Phoma sp.)、苦玄参叶斑病菌(Alternaria sp.)和肿节风炭疽病菌(Colletotrichum dematium)等7种病原菌的抑制率均在80%以上,具有强的抑菌广谱性。【结论】肿节风存在丰富的内生真菌,其中的RJ-1和JJ-1菌株对15种病原真菌具有较强的抑制作用,可作为抗菌活性菌株开展深入研究。

【Objective】The diversity of endophytic fungi of medicinal plants of Sarcandra glabra was analyzed,and the strains with strong antifungal activity were screened to lay a foundation for the search for new antimicrobial resources.【Method】Endophytic fungi with strong anti-microbial activities were isolated from different tissues of S.glabra by plate confrontation method,and their species was identified by combining morphological characteristics and ITS sequence to screen out endophytic fungi with strong antimicrobial activities.【Result】One hundred and eleven of various endophytes were obtained from the different organizations of S.glabra,forty-two strains of different morphological types were selected for molecular identification,which belonged to nine orders and twenty genera,twenty-eight species.The RJ-1 and JJ-1 had re-markable antimicrobial activities to fifteen pathogenic fungi.Morphological and molecular identification showed that RJ-1 and JJ-1 were Phomopsis heveicola and Neofusicoccum parvum,respectively.Their antifungal activity to seven pathogens including Botrytis sp.,Fusarium solani,Colltetorrichum gloeosporioides of Panax notoginseng,Colltetorrichum sp.of Musa,Phoma sp.of Blumea balsamifera,Alternaria sp.of Picria felterrae,Colletotrichum dematium of S.glabra was over 80%,with strong antifungal broad-spectrum.【Conclusion】The endophytic fungi in S.glabra were diverse and rich.Some strains such as RJ-1 and JJ-1 deserve further research as an alternative antimicrobial resource.