摘要
目的筛选产生物碱和石杉碱甲的内生真菌。方法以分离自健康蛇足石杉植株组织中的14株内生真菌为材料,采用酸性染料比色法筛选产生物碱的菌株;在此基础上,对内生真菌发酵液进行HPLC测定,分析是否产生石杉碱甲;并利用形态学和rDNA ITS序列分析进行菌种的初步鉴定。结果筛选出8株产生物碱的菌株,并从中筛选出一株产生石杉碱甲的内生真菌SY-02,初步鉴定内生真菌SY-02为1株炭角菌。结论在分离自蛇足石杉的内生真菌中,能产生生物碱的真菌比例较高,内生真菌菌株SY-02能在发酵条件下产生石杉碱甲。以产生物碱的菌株为出发菌株,结合活性及结构检测,可以使产石杉碱甲内生真菌的筛选更具有方向性。
OBJECTIVE To screen endophytic fungal strains isolated from Huperzia serrata, which can produce alkaloid and huperzine A. METHODS Fourteen endophytic fungal strains were obtained from H. serrata. Alkaloid production was assayed with acid dye colorimetry. Then, the crude extracts of alkaloid-producing fungal strains were analyzed by HPLC for the presence of huperzine A （HupA）. Identification of endophytic fungi producing HupA was based on morphology and ITS1-5.8S-ITS2 sequence analysis. RE- SULTS Eight alkaloid-producing endophytic fungal strains were determined based on acid dye colorimetry, among which, strain SY- 02 generated＇ HupA in vitro. Morphology and ITS1-5. 8S-ITS2 sequence analysis showed that strain SY-02 was a Xylarialern species. CONCLUSION A high percentage of fungal strains isolated from H. serrata have alkaloid-producing potentials. It is the first report that Xylarialern species produced huperzine A. , The screening of endophytic fungi producing huperzine A based on alkaloid-producing potential, activity and structure identification is more directive.
出处
《中国药学杂志》
CAS
CSCD
北大核心
2011年第19期1477-1481,共5页
Chinese Pharmaceutical Journal
基金
福建省自然科学基金资助项目(B0620005)
福建师范大学本科生课外科技计划项目资助(BKL2010-066),福建师范大学生物学国家级实验教学示范中心资助项目(2009LS013)
福建省大学生创新性实验计划资助项目(Fjnu2010-012)
福建省科技厅重点项目(2010N0027)
