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目的从杜仲Eucommia ulmoides叶和果实中分离具有抑菌作用的内生真菌。方法采用平板培养法从杜仲叶片和果实中分离内生真菌,通过对峙法和生长速率法考察这些内生真菌的抑菌活性,并通过HPLC法考察其具有代谢产生杜仲有效活性成分的能力。对具有良好抑菌效果和能够代谢杜仲活性成分的内生真菌进行ITS序列测定,鉴定其归属。结果从健康叶片和果实中分离到52株内生真菌,其中2个菌株的提取液中含有绿原酸,11个菌株至少对4种病原菌具有抑制作用,其中29号菌株(链格孢属Alternaria sp.)对芽孢杆菌Fusarium graminearum的抑制效果最好,其抑制率高达82.6%。通过与Gen Bank数据比对,这些内生真菌分别属于链格孢属Alternaria、黑孢属Niqrospora和座囊菌纲(Dothideomycetes)。结论从杜仲叶和果实中分离到11株具有抑菌作用的内生真菌,这些具有抑菌作用的内生真菌可用于生物制药和农业产业。
OBJECTIVE To isolate endophytic fungi with inhibitory effect on Eucommia ulmoides leaves and fruits. Methods Endophytic fungi were isolated from the leaves and fruits of Eucommia ulmoides by plate culture method. The antibacterial activities of these endophytic fungi were investigated by confrontation method and growth rate method, and their bioactivities were assayed by HPLC. Endophytic fungi with good antibacterial activity and ability to metabolize Eucommia ulmoides were sequenced and identified. Results 52 endophytic fungi were isolated from healthy leaves and fruits. Chlorogenic acid was found in the extracts of 2 strains and 11 strains at least for 4 pathogens. Among them, strain 29 (Alternaria Alternaria sp.) had the best inhibitory effect on Fusarium graminearum with the inhibition rate as high as 82.6%. By comparison with Gen Bank data, these endophytic fungi belong to the genus Alternaria, the genus Niqrospora, and the genus Dothideomycetes, respectively. Conclusion Eleven endophytic fungi with antibacterial activity were isolated from Eucommia ulmoides leaves and fruits. These endophytic fungi with antibacterial activity could be used in biopharmaceutical and agricultural industries.