产紫杉醇内生真菌EFY-21的鉴定与生物学特性研究

利用PCR技术克隆了产紫杉醇内生真菌EFY-21的18S rDNA序列,通过同源性分析,初步确定该菌与拟盘多毛孢属(Pestalotiopsis)有较高的同源性,相似性为99%。为了进一步了解EFY-21的有关生物学特性,分别选用PDA、PSA、查氏、玉米粉琼胶、牛肉膏蛋白胨5种培养基,按照常规方法培养,用十字法测量菌落直径;同时选用查氏培养基为基本培养基,分别观察不同碳源葡萄糖、甘露醇、麦芽糖、果糖、可溶性淀粉,不同氮源KNO3、Ca(N03)2、(NH4)2SO4、NH4N03、(NH4)2HPO4、蛋白胨、尿素,不同培养温度10,15,20,25,28,30,37℃,不同pH值4,5,6,7,8,9对内生真菌菌丝的影响。试验结果表明:EFY-21在PDA培养基上生长最快,生长状况最好;供试的碳氮源中,对EFY-21菌丝生长影响的大小顺序为葡萄糖甘露醇果糖麦芽糖可溶性淀粉;蛋白胨KNO3Ca(N03)2NH4N03(NH4)2HPO4(NH4)2SO4尿素;最适培养温度为25～30℃;最适pH为5～7。

Identification and Biological Characteristic Research of the Taxol-Producing Endophytic Fungus EFY-21

By analyzing 18S rDNA sequences,the taxol-producing endophytic fungus 21（EFY-21）was determined to have higher homology with Pestalotiopsis.Experiments were carried out to explore EFY-21 culture characters,including suitable medium,carbon（C）source,nitrogen（N）source,temperature and primary pH.Five different kinds of culture media,including PDA,PSA,Czapek＇s medium,beef paste and peptone medium and cornmeal medium,were selected to study their effects on the growth of mycelia.In the same way,five different kinds of C sources,seven kinds of N sources,seven levels of temperature and six levels of pH were selected to screen the optimum conditions.The results showed that EFY-21 grew faster and best on PDA medium.Among tested C sources,EFY-21 grew best on the medium using mannitol as carbon source and secondary glucose.Among tested N sources,EFY-21 grew best on the medium using polypeptone.The optimum temperature was 25—30℃.The optimum pH value was 5—7.This study will establish foundation for exploration and application of EFY-21.