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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>KNO3>Ca(N03)2>NH4N03>(NH4)2HPO4>(NH4)2SO4>尿素;最适培养温度为25～30℃;最适pH为5～7。
Abstract：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.
[1]Wani MC,Taylor HL,Wall ME,et al.Plant antitumor agents.VI.The isolation and structure of taxol,a novel antilekemic and antitumor agent fromTaxus brevifolia[J].Journal of the American Chemical Society,1971,93(9):2325-2327.
[2]陈未名.红豆杉属植物的化学成分和生理活性[J].药物学报,1990,259(3):236.
[3]Caplow M,Shanks J,Ruhlen R.Howtaxol modulates microtubule disassembly[J].Journal of Biological Chemistry,1994,269(38):23399-23402.
[4]Jordan MA,Wilson L.Microtubules and actinfilmnents:dynamic targets for cancer chemotherapy[J].Current Opinion in Cell Biolo-gy,1998,10(1):123-130.
[5]王玉震,仝川,柯春婷.红豆杉植株紫杉醇含量研究进展(综述)[J].亚热带植物科学,2008,37(4):59-63.
[6]Stilerle A,Strobel G,Stierle D.Taxol and taxane production by Taxomyces andreanae,an endophytic fungus of pacific yew[J].Science,1993,260(5105):214-216.
[7]何颖,谈锋,谢建平.红豆杉内生真菌产紫杉醇研究进展[J].天然产物研究与开发,2006,18(3):519-523,513.
[8]Strobel G A,Hess WM,Li J Y,et al.Pestalotiopsis guepinii,a taxol producing endophyte of the Wollemi pine,Wollemia nobilis[J].Australian Journal of Botany,1997,45(6):1073-1082.
[9]周东坡,平文祥,孙剑秋,等.紫杉醇产生菌的分离研究[J].微生物学杂志,2001,21(1):18-20.
[10]陈毅坚,张硕,王艳,等.云南红豆杉内生真菌中产紫杉醇真菌的筛选[J].生物技术,2003,13(2):10-11.
[11]Zhou X W,Wang Z N,Jiang KJ,et al.Screening of taxol-pro-ducing endophytic fungi fromTaxus chinensisvar.mairei[J].Applied Biochemistry and Microbiology,2007,43(4):439-443.
[12]Chakravarthi BVSK,Das P,Surendranath K,et al.Production of paclitaxel byFusariumsolaniisolated fromTaxus celebica[J].Journal of Biosciences,2008,33(2):259-267.
[13]Zhang P,Zhou P P,Yu LJ.An endophytic taxol-producingfun-gus fromtaxus media,Cladosporium cladosporioidesMD2[J].Current Microbiology,2009,59(3):227-232.
[14]Hou Y H,Wang TH,Long H,et al.Novel cold-adaptivePeni-cilliumstrain FS010secretingthermo-labile xylanase isolated from YellowSea[J].Acta Biochimica et Biophysica Sinica,2006,38(2):142-149.
[15]宋益民,印建峰.银杏链格孢菌生物学特性研究[J].长江大学学报:自然科学版,2008,5(3):1-4.
[16]刘卫,鲁海菊,张举成,等.产生红色素的竹红菌生物学特性研究[J].安徽农业科学,2008,36(2):412,474.
[17]张春霞,李加智,明霞.两种橡胶炭疽病菌生物学特性的比较[J].西南农业学报,2008,21(3):667-670.
基本信息:
DOI：10.13341/j.jfr.2010.01.009
中图分类号:Q93
引用信息:
[1]郑文龙,朱慧芳,刘璐等.产紫杉醇内生真菌EFY-21的鉴定与生物学特性研究[J].菌物研究,2010,8(01):35-40.DOI:10.13341/j.jfr.2010.01.009.
基金信息:
上海市科委中药现代化专项基金项目(07DZ19724);; 国家高新技术发展计划(863)项目(2007AA021501)