天麻内生真菌的分离鉴定及药物化学的初步研究
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摘要
目前,关于药用植物资源保护的研究中,药用植物的内生真菌已经成为药用植物资源研究的一个新热点。由于内生真菌的次级代谢产物中含有与药用植物相同或相似的生物活性物质,所以研究药用植物内生真菌多样性,利用内生真菌发酵生产药物具有相当的优势。
本文首先对内生真菌的国内外研究概况及其研究意义进行了综述。从内生真菌的分布、生物多样性、代谢物质的多样性到内生真菌的生物学作用做了介绍,并且对天麻及应用价值做了简单概述。
本文以我国传统中药——天麻(Gastrodia elata B1)为实验材料,对天麻内生真菌的菌群组成及活性成分进行初步研究。取得以下结果：
1、内生真菌的分离鉴定：采用组织印迹培养法,对产自贵州、四川、陕西、云南等地天麻的不同组织进行内生真菌的分离,并确定如何区分组织内生真菌与表面附生菌的方法。实验结果表明,天麻内生真菌分离的表面消毒以75%酒精和0.1%升汞结合的方法较好。在所试条件下共计分离得到94株内生真菌。对其中分离纯化的64株真菌,经形态学与分子生物学鉴定结果结合鉴定,分属于4个亚门、6纲、8目、11科、15属。鉴定结果在一定程度上反应了天麻内生真菌的生物多样性。采集于不同地区的天麻中均分离到了一定数量、不同类群的内生真菌,表明内生真菌在天麻体内广泛存在。
天麻不同组织部位内生真菌的种类存在着一定的差异。块茎中鉴定的43株真菌分布于12个属(占27.9%),花茎中鉴定的10株菌分布于5个属(占50%),花中鉴定的7株菌分布于4个属(57.1%),果实中鉴定的4株菌分布于3个属(75%)。
本试验还对同一季节不同地点来源的天麻块茎内生真菌进行了研究,其数量和种类存在着差异。数量上贵州大方的优势菌群为刺盘孢属(Colletotrichum sp.),占总数的16%；贵州毕节和四川广元的优势菌群是链格孢属(Alternaria sp.),分别是4株和5株,占总数的16%和20%。种类上,四川广元的内生真菌种类所占比例最高,共鉴定9株分布于5个属,占55.6%。
2、产天麻素内生真菌的筛选：本文采用薄层层析TLC方法与高效液相色谱HPLC结合,从3株内生真菌DF101 (Chaetomium sp.)、BY302 (Fusarium sp.)、SC108 (Armillaria sp.)的发酵产物中检出天麻素,DF101为0.057mg/g, BY302为0.089mg/g, SC108为0.184 mg/g；对照品天麻中天麻素含量为1.038 mg/g,约为最高含量SC108的6倍。这3株内生真菌有2株来自块茎,1株来自花茎。
At present, for the protection of medicinal plant resources, the endophytic fungi of medicinal plant has become a new hot spot for research.Reasearch results in recent years show that endophytic fungi can produce some similar physiological active products to the host plant. So studies on endophytic fungi diversity of the medicinal plants and potential bioactive compounds from endophytic fungi has become important.
In this thesis, studies on endophytic fungi in China and oversea were summarized. Gastrodia elata Bl, a chinese medicinal plant, was selected for the study. The content of the study includes distribution, diversity and function of endophytic fungi in G elata Bl, and ecological characteristics and worthness in application of G elata Bl. The distribution of endophytic fungi in G elata B1 tissues and isolation method were studied.
The results obtained were as following:
1 Isolation and identification of endophytic fungi:A method named tissue print culture was used to isolate endophytic fungi in G. elata B1 from Guizhou, Sichuan, Shanxi, Yunnan. The result indicated that, the superficial disinfection for endophytic fungi isolation was better with the method of 75% ethanol combined with 0.1% HgCl2. A total of 94 endophytic fungi islates were isolated, of which 64 islates were identified with morphology and the molecular biology. They belong to 15 genera,11 families,8orders,6 class and 4 phylum which showed the biodiversity of endophytic fungi in G elata Bl. Endophytic fungi were isolated from G. elata B1 from all different origins which demonstrated the widespread existence of endophytic fungi in G elata Bl.
The population differences of endophytic fungi exist in different tissues or positions of G. elata Bl. In the tubers, scapes, flowers and fruits 43 fungal isolates of 12 genera (27.9%),10 isolates of 5 genera (50%),7 isolates of 4 genera (57.1%) and 4 isolates of 3 genera (75%) were isolated and identified respectively.
In the research the endophytic fungi of G elata B1 from different origins at the same season were also studied. The dominant groups of Guizhou DaFang, Guizhou BiJie and Sichuan were Colletotrichum and Alternaria.
2 Gastrodin production of the endophytic fungi :Thin layer chromatography(TLC) method and highly effective liquid chromatography(HPLC) were employed to detect metabolites of DF101 (Chaetomium sp.), BY302 (Fusarium sp.), SC108 (Armillaria sp.). The results indicated that the mycelia of these 3 isolates contained gastrodin and similar compounds. The gastrodin contents of DF101, BY302 and SC108 were 0.057mg/g,0.089mg/g and 0.184 mg/g respectively.
本文首先对内生真菌的国内外研究概况及其研究意义进行了综述。从内生真菌的分布、生物多样性、代谢物质的多样性到内生真菌的生物学作用做了介绍,并且对天麻及应用价值做了简单概述。
本文以我国传统中药——天麻(Gastrodia elata B1)为实验材料,对天麻内生真菌的菌群组成及活性成分进行初步研究。取得以下结果：
1、内生真菌的分离鉴定：采用组织印迹培养法,对产自贵州、四川、陕西、云南等地天麻的不同组织进行内生真菌的分离,并确定如何区分组织内生真菌与表面附生菌的方法。实验结果表明,天麻内生真菌分离的表面消毒以75%酒精和0.1%升汞结合的方法较好。在所试条件下共计分离得到94株内生真菌。对其中分离纯化的64株真菌,经形态学与分子生物学鉴定结果结合鉴定,分属于4个亚门、6纲、8目、11科、15属。鉴定结果在一定程度上反应了天麻内生真菌的生物多样性。采集于不同地区的天麻中均分离到了一定数量、不同类群的内生真菌,表明内生真菌在天麻体内广泛存在。
天麻不同组织部位内生真菌的种类存在着一定的差异。块茎中鉴定的43株真菌分布于12个属(占27.9%),花茎中鉴定的10株菌分布于5个属(占50%),花中鉴定的7株菌分布于4个属(57.1%),果实中鉴定的4株菌分布于3个属(75%)。
本试验还对同一季节不同地点来源的天麻块茎内生真菌进行了研究,其数量和种类存在着差异。数量上贵州大方的优势菌群为刺盘孢属(Colletotrichum sp.),占总数的16%；贵州毕节和四川广元的优势菌群是链格孢属(Alternaria sp.),分别是4株和5株,占总数的16%和20%。种类上,四川广元的内生真菌种类所占比例最高,共鉴定9株分布于5个属,占55.6%。
2、产天麻素内生真菌的筛选：本文采用薄层层析TLC方法与高效液相色谱HPLC结合,从3株内生真菌DF101 (Chaetomium sp.)、BY302 (Fusarium sp.)、SC108 (Armillaria sp.)的发酵产物中检出天麻素,DF101为0.057mg/g, BY302为0.089mg/g, SC108为0.184 mg/g；对照品天麻中天麻素含量为1.038 mg/g,约为最高含量SC108的6倍。这3株内生真菌有2株来自块茎,1株来自花茎。
At present, for the protection of medicinal plant resources, the endophytic fungi of medicinal plant has become a new hot spot for research.Reasearch results in recent years show that endophytic fungi can produce some similar physiological active products to the host plant. So studies on endophytic fungi diversity of the medicinal plants and potential bioactive compounds from endophytic fungi has become important.
In this thesis, studies on endophytic fungi in China and oversea were summarized. Gastrodia elata Bl, a chinese medicinal plant, was selected for the study. The content of the study includes distribution, diversity and function of endophytic fungi in G elata Bl, and ecological characteristics and worthness in application of G elata Bl. The distribution of endophytic fungi in G elata B1 tissues and isolation method were studied.
The results obtained were as following:
1 Isolation and identification of endophytic fungi:A method named tissue print culture was used to isolate endophytic fungi in G. elata B1 from Guizhou, Sichuan, Shanxi, Yunnan. The result indicated that, the superficial disinfection for endophytic fungi isolation was better with the method of 75% ethanol combined with 0.1% HgCl2. A total of 94 endophytic fungi islates were isolated, of which 64 islates were identified with morphology and the molecular biology. They belong to 15 genera,11 families,8orders,6 class and 4 phylum which showed the biodiversity of endophytic fungi in G elata Bl. Endophytic fungi were isolated from G. elata B1 from all different origins which demonstrated the widespread existence of endophytic fungi in G elata Bl.
The population differences of endophytic fungi exist in different tissues or positions of G. elata Bl. In the tubers, scapes, flowers and fruits 43 fungal isolates of 12 genera (27.9%),10 isolates of 5 genera (50%),7 isolates of 4 genera (57.1%) and 4 isolates of 3 genera (75%) were isolated and identified respectively.
In the research the endophytic fungi of G elata B1 from different origins at the same season were also studied. The dominant groups of Guizhou DaFang, Guizhou BiJie and Sichuan were Colletotrichum and Alternaria.
2 Gastrodin production of the endophytic fungi :Thin layer chromatography(TLC) method and highly effective liquid chromatography(HPLC) were employed to detect metabolites of DF101 (Chaetomium sp.), BY302 (Fusarium sp.), SC108 (Armillaria sp.). The results indicated that the mycelia of these 3 isolates contained gastrodin and similar compounds. The gastrodin contents of DF101, BY302 and SC108 were 0.057mg/g,0.089mg/g and 0.184 mg/g respectively.
引文
[1]曹晓冬,周立刚,徐利剑,谈满良,汪冶.滇重楼内生真菌的生物学性状及其甾体化合物含量.西北农林科技大学学报(自然科学版),2005,33：125-128.
[2]陈世苹,高玉仪,梁宇,等.水分胁迫下内生真菌感染对黑麦草叶内游离脯氨酸和脱落酸含量的影响[J].生态学报,2001,21(12)：1965-1972
[3]陈贤兴,陈析丰,南旭阳,何献武.喜树果内生真菌的分离与鉴定.河南科学,2003,21(4)：431-433.
[4]邓世贤,乙酰天麻素的镇痛及对心脏的作用[J],云南医药,1985,6(4)：252-253
[5]冯孝章、陈玉武,天麻化学成分的研究[J],化学学报,1979,37(3)：177-181
[6]郭良栋.内生真菌研究进展[J].菌物系统,2001,20(1)：148-152.
[7]郭顺星,徐锦堂.促进石斛等兰科药用植物种子萌发的真菌分离与培养[J].中草药.1990,21(6)：30-31.
[8]谷苏,邵华,蒋晓华,等.药用植物内生真菌多样性及其活性成分的潜在应用价值[J].中国药学杂志,2001,36(1)：14-15.
[9]高微微,郭顺星.内生真菌菌丝及其代谢物对铁皮石斛及金线莲生长的影响[J].中国医学科学院学报,2001,23(6)：556-559.
[10]高玉葆,陈世苹,梁宇,等.干早胁迫下内生真菌感染对黑麦草实验种群光合、蒸腾和水分利用的影响[J].植物生态学报,2001,25(5)：537-543
[11]高南南、等,天麻对老龄大鼠学习记忆的改善作用[J],中国药学杂志,1995,9(20)：562-563
[12]郭仕平,蒋斌,苏莹珍等.2001.川八角莲内生真菌产鬼臼毒素类似物的初步研究.生物技术,14(2)：55-57.
[13]胡谷平,余志刚,吴耀文等.南海海洋红树林内生真菌抱外多糖的研究.中山大学学报(自然科学版),2002,41(1)：121-122.
[14]黄俊华、等,天麻注射液及天麻苷药理作用的初步研究[J],中国医学科学院学报,1985,7(5)：399-401
[15]黄丽亚.天麻注射液上调抗氧化酶表达作用的实验研究[J].陕西中医,2006,27(2)：242—243.
[16]郝小燕、谭宁华,黔产天麻的化学成分[J],云南植物研究,2000,22(1)：81-84
[17]刘国卿、等,天麻成分香荚兰醇的神经药理研究[J],中草药通讯,1974,5：33
[18]刘建新、等,天麻对大鼠大脑胶质细胞影响的实验研究[J],中国中医基础医学杂志,1997,3(6)：23-25
[19]刘蕴哲.喜树生态系内生真菌的分子鉴定及药物化学研究.贵州大学硕士学位论文,2006
[20]刘芸,殷红,彭辉等植物内生真菌—潜在的天然药物新来源中国现代中西医杂志,2005年第三卷第五期
[21]刘中华,胡海涛,冯改丰,等.天麻素对Aβ25—35诱导的AIZhimer病细胞模型的保护作用.四川大学学报,2005,36(4)：
[22]刘仕平,曾松荣,郭仕平等.中华山荷叶内生真菌的分离及其发酵产生药用成分的初步研究.中国药物与临床,2003,3(3)：227-228.
[23]黎万奎,胡之璧.2005.内生菌与天然药.中国天然药物,3(4)：193-199.
[24]罗海、孙中吉,天麻素的基础药理研究和临床应用,武警医学院学报,2007,16(4)：465-468
[25]梁宇,高玉葆.内生真菌对植物生长发育及抗逆性的影响[[J].植物学通报,2000 17(1)：52-59
[26]兰琪,姜广华,吴文君,等.植物内生真菌及其应用研究植物农药与药剂毒理学研究进展[M].北京：中国农业科学技术出版社,2002,87-91
[27]兰琪,姬志勤,顾爱国等.苦皮藤内生真菌中杀虫杀菌活性物质的初步研究.西北农林科技大学学报(自然科学版),2004,32(10)：79-84.
[28]李强,刘军,周东坡.植物内生菌的开发与研究进展[J].生物技术通报,2006,(3)：33～37.
[29]李海燕等.一种桃儿七内生真菌的分离初报.云南大学学报(自然科学版),1999,21(3)：243
[30]李桂玲,王建锋,黄耀坚,等.植物内生真菌抗肿瘤活性菌株的筛选[J].菌物系统,2001,20(3)：387-391.
[31]李时珍,本草纲目[M],北京：人民卫生出版社,1977,第1版,730-733
[32]李茂进,胡俊峰,李国珍,等.天麻对铅所致大鼠海马损害的拮抗作用[J].中华劳动卫生职业病杂志,2002,20(5)：331—333.
[33]李雪玲,徐成东两种八角莲属植物内生与根际真菌的多样性初探大理学院学报第3卷第3期2004年5月
[34]李强,刘军,周东坡.植物内生菌的开发与研究进展[J].生物技术通报,2006,(3)：33-37.
[35]莫云强,天麻的药理研究[J],云南医药,1983,4(6)：388
[36]马天有等.从植物分离产紫杉醇的内生真菌的研究.西北大学学报(自然科学版),1999,29(1)：47-49
[37]曲淑梅、等,黄芪、天麻对老年心脑血管患者血中LPO水平的影响[J],锦州医学院学报,1994,15(1)：28-29
[38]任安芝,高玉葆.植物内生真菌一类应用前景广阔的资源微生物[J].微生物学通报,2001,28(6)：90-93.
[39]邱德有等.一种云南红豆杉真菌的分离.真菌学报,1994,13(4)：314-316
[40]塞冬、等,黄芪、天麻、首乌的延缓衰老作用的实验研究[J],锦州医学院学报,1991,12(4)：71
[41]沈书庆,殷红,刘芸,等.产杜仲黄酮内生真菌的初步研究[J].吉林农业大学学报,2008,6(1)：46-48.
[42]王莹,赵华斌,郝家胜.分子系统学的理论方法及展望[J],安徽师范大学学报(自然科学版),2005,28(1)：85-88
[43]王伟等.南方红豆杉内真菌及紫杉烷类产物的初步鉴定.中山大学学报(自然科学版),1999,38(3)：116-118
[44]王永中,肖亚中.植物内生菌及其活性代谢产物.生物学杂志,2004,21(4)：1-5.
[45]王军,林永成,吴雄宇,等.从南海红树林内生真菌No.2533分离出新的异香豆素[J].中山大学学报(自然科学版),2001,40(1)：127-128.
[46]王晓晨,徐庆,陈章良.天麻中一种抗真菌蛋白基因的克隆[J].植物学报,1999,41(10)：1041～1045
[47]吴静萍,郑师章.密花石解菌根菌分离鉴定及其代谢产物的测定[J].复旦学
报,1994,33(5)：547～552.
[48]文才艺,吴元华,田秀玲.植物内生菌研究进展及其存在的问题[J].生态学杂志,2004,23(2)：86-91
[49]肖盛元,郭顺星,杨俊山.粉红粘帚霉菌丝体化学成分研究[J].菌物系统,2001,20(4)：536～538.
[50]肖光,黄芪、天麻、首乌对小白鼠血SOD活力的影响[J],锦州医学院学报,1992,13(4)：15-16
[51]薛美芹.石斛内生真菌的形态和分子鉴定及其细胞毒活性研究.贵州大学硕士学位论文,2006
[52]袁志林,戴传超,史央,等.内生真菌B3促进水稻生长的机理研究[J].江苏农业科学,2004,2：10-13.
[53]袁崇文,中国天麻[M],贵阳：贵州科技出版社,2002,第1版
[54]易润华,朱西儒,周而勋.简化CTAB法快速微量提取丝状真菌DNA[J].湛江海洋大学学,2003,23(6)：72-73
[55]于龙顺、等,天麻抗炎效应的研究[J],中草药,1989,20(5)：211-213
[56]杨箐、等,黄芪、天麻对老年心脑血管疾病患者血中SOD的影响[J],锦州医学院学报,1995,16(1)：15
[57]俞斌,熊健.草坪草抗虫性的研究进展[[J].江西植保,2002.25(4)：127-131
[58]张集慧,王春兰,郭顺星,等.兰科药用植物的5种内生真菌产生的植物激素[J].中国医学科学学院学报,1999,21(6)：460-465.
[59]张玲琪等.发酵产鸢尾酮真菌的分离鉴定及产香特性的初步研究.菌物系统.1999,18(1)：49
[60]张玲琪,王海昆,邵华,等.美登木内生真菌产抗癌物质球毛壳甲素的分离及鉴定[J].中国药学杂志,2002,37(3)：172—175.
[61]张维经.天麻与其真菌营养[J].西北植物学报,1987,7(3)：215-221.
[62]周俊、杨雁宾、浦湘渝,新鲜天麻的酚类成分[J],云南植物研究,1980,2(1)：370
[63]赵国举、等,人培与野生天麻中枢药理作用的比较研究[J],武汉医学院学报,1980,9(1)：91
[64]钟裕国、等,天麻苷元同系物与同型物的合成及其中枢镇静抗惊活性[J],四川医学院学报,1984,15(1)：17
[65]全国中草药汇编[M],北京：人民卫生出版社,1996,169-170
[66]国家药典委员会.中华人民共和国药典[M].
[67]邹容.天麻共生蜜环菌及同属真菌的基因系谱学及药用成分的研究.贵州大学硕士学位论文,2005
[68]曾松荣,徐倩雯,叶保童,卿野,方白玉,黄晓敏.虎杖内生真菌的分离及产抗菌活性物质的筛选.菌物研究,2005,3(2)：24-26
[69]魏景超.真菌鉴定手册[M].上海：上海科学技术出版社,1979.
[70]巴尼特H L,亨特B B.半知菌属图解[M].沈崇尧,译.北京：科学出版社,1977.
[71]Bradly SF, Singh MP,,Janso,JE, et al. Cytoskyrins A and B, new BIA active hisanthraquinones isolated from an endophytic fungus[J].Org Lett,2000 Dec,14, 2(25):4047-4049.
[72]Carrol G.C. The biology of endophytism in plants with particular reference to woody perennials In Fokkema N.J. and van den Heuvel J. Eds Microbiology of the Phyllosphere. Cambridge:Cambridge University Press.1986.205-222.
[73]Guo L D,Hyde K D,Liew E C Y. Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences [J].New Phytology, 2000,147(3):617-630.
[74]Guo B. Dai,JR, Ng S, et al. Cytonic:acids A and B:novel tridepside inhibitors of hCMV protease from the endophytic:fungus Cytonaema species[J].J Nat Prod, 2000, May,63(5):602-604.
[75]HUANG Yaojian,WANG Jianfeng,LIGuiling,et al.Antitumor and antifungal activities in endophytic fungi isolated from pharmaceutical plants Taxus mairei,Cephalataxus fortunei and Torreya grandis[J].FEMSI mmunology and Medical Microbiology,2001,31 (2):163-167.
[76]Hao XY, Tan NH, Zhou J. Anewdi-(p-hydroxybenzyl)hydroxyhmine from Gastrodiaelata B1[J]. Chinese Chemical Letters(中国化学快报),1999,10(6):
467.
[77]Hsieh MT, Wu CR, Chen CF. Gastrodin and p-hydroxybenzyl alchol facilitate moemory consolidation and retrieval, but not acquisition, on the passive avoidance task in rata[J]. J of Ethnopharmacology,1997,56(1):45—54.
[78]Jia-yao Li,et al. Etal. Endophytic taxol-producing fungi from bald cypress,Taxodium distichum. Microbiology.1996,142:2223-2226
[79]Judy F. Shearer.2002. The Potential Role of an Endophytic Fungus in the Decline of Stressed Eurasian Watermilfoil. J. Aquat. Plant Manage,40:76-78.
[80]Kim HJ, Moon KD, Lee DS, et al. Ethyl ether fraction of Gastrodia elata Blume protects amyloid betapeptideinduced cell death[J]. J Ethnopharmacol,2003,84 (1):95-98.
[81]L i J Y, Strobel G A. Jesterone and hydroxyl-jesterone antioomycete cyclohexenone epoxides from the endophytic fungus Pestalotiop sis jesteri[J]. Phytochemistry,2001,57:261～265.
[82]Li J Y,Strobel G A, Harper J,et al. Cryptocin,a potent tetramic acid antimycotic from the endophytic fungus Cryptosporiopsis cf.quercina[J].Org Lett,2000,2:767～770.
[83]Lu H, Zou W X, Meng J C,et al. New bioactive metabolites produced by Colletotrichum sp.,an endophytic fungus in Artemisia annua[J].Plant Sci,2000,151:67～73
[84]Petrini O. Fungal endophytes of tree leaves [M] New York, Springer-Verlag, Andrews J H, Hirano S S. eds,1991,179-197.
[85]Schutz B.2001.Endophytic fungi:a source of novel biologically active secondary metabolites. British Mycological Society, International Symposium Proceedings, Bioactive Fungal Metabolites-Impact and Exploitation, University of Wales, Swansea.
[86]Siegel MR,Johnson MC,Varney DR,et al.1984.A fungal endo-phyte in tall fescue: incidence and dissemination[J].Phytopatho-logy,74:932～937.
[87]Southcott K A,Johnson J A. Isolation of endophytes from two Species of palm,from Bermuda [J].Canadian Journal of microbiology,1997,43(4):789-792.
[88]Strobel G,DaisyB,Castillo U,et al.Naturalproductsfrom endophytic microorganisms.[J].JNat prod.2004,67(2):257～68
[89]Strobel G A.2003. Endophytes as sources of bioactive products. Microbes and Infection,5:535-544.
[90]Strobel G A, Miller R V, Martinez-Miller C,et al.Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina [J]. Mcrobiology,1999,145:1919～1926.
[91]Strobel Qet al. Taxomyces andreanae,A proposed new Taxon for aBubilliferous hyphomyceteassociated with pa-cific yew(Taxus brevifolia).Mycotaxon,1993,X Ⅳ Ⅱ:71～80
[92]Stierle A,et al.Taxol and taxane production by Taxomyces and reanal,an endophytic fangus of pacific yew comments. Science,1993,260:214～216
[93]Stierle A, Strobel G, Stierle D.Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew [J].Science,1993,260:214-216.
[94]Sutton B C. The Coelomycetes[M]. Kew:Commonwealth Mycological Institute,1980.
[95]Taxon for a Bubilliferous hyphomyceteassociated with pa-cific yew(Taxus brevifolia).Mycotaxon,1993, ⅩⅣⅡ:71～80
[96]Zou W X, Meng J C, Lu H,et al. Metabolites of Colletotrichum gloeosporioides, an endophytic fungus in Artemisia mongolica [J]. J Nat Prod,2000,63:1529 ～1530.
[97]Wang Yu,Guo liang-Dong,Morphological and Molecular indentification of an endophytic fungus Epicoccum Nigrum,Mucosystema,2004,23(4):2004.
[98]Wagenaar MM, Clardy J.Dicerandrols, new antibiotic and cytotoxic:dimmers produce by the fungus Phomcpsis Longicolla isolated from an endangered mint[J].J Nat Prod.2001, Aug,64(8):1006-1009.
[2]陈世苹,高玉仪,梁宇,等.水分胁迫下内生真菌感染对黑麦草叶内游离脯氨酸和脱落酸含量的影响[J].生态学报,2001,21(12)：1965-1972
[3]陈贤兴,陈析丰,南旭阳,何献武.喜树果内生真菌的分离与鉴定.河南科学,2003,21(4)：431-433.
[4]邓世贤,乙酰天麻素的镇痛及对心脏的作用[J],云南医药,1985,6(4)：252-253
[5]冯孝章、陈玉武,天麻化学成分的研究[J],化学学报,1979,37(3)：177-181
[6]郭良栋.内生真菌研究进展[J].菌物系统,2001,20(1)：148-152.
[7]郭顺星,徐锦堂.促进石斛等兰科药用植物种子萌发的真菌分离与培养[J].中草药.1990,21(6)：30-31.
[8]谷苏,邵华,蒋晓华,等.药用植物内生真菌多样性及其活性成分的潜在应用价值[J].中国药学杂志,2001,36(1)：14-15.
[9]高微微,郭顺星.内生真菌菌丝及其代谢物对铁皮石斛及金线莲生长的影响[J].中国医学科学院学报,2001,23(6)：556-559.
[10]高玉葆,陈世苹,梁宇,等.干早胁迫下内生真菌感染对黑麦草实验种群光合、蒸腾和水分利用的影响[J].植物生态学报,2001,25(5)：537-543
[11]高南南、等,天麻对老龄大鼠学习记忆的改善作用[J],中国药学杂志,1995,9(20)：562-563
[12]郭仕平,蒋斌,苏莹珍等.2001.川八角莲内生真菌产鬼臼毒素类似物的初步研究.生物技术,14(2)：55-57.
[13]胡谷平,余志刚,吴耀文等.南海海洋红树林内生真菌抱外多糖的研究.中山大学学报(自然科学版),2002,41(1)：121-122.
[14]黄俊华、等,天麻注射液及天麻苷药理作用的初步研究[J],中国医学科学院学报,1985,7(5)：399-401
[15]黄丽亚.天麻注射液上调抗氧化酶表达作用的实验研究[J].陕西中医,2006,27(2)：242—243.
[16]郝小燕、谭宁华,黔产天麻的化学成分[J],云南植物研究,2000,22(1)：81-84
[17]刘国卿、等,天麻成分香荚兰醇的神经药理研究[J],中草药通讯,1974,5：33
[18]刘建新、等,天麻对大鼠大脑胶质细胞影响的实验研究[J],中国中医基础医学杂志,1997,3(6)：23-25
[19]刘蕴哲.喜树生态系内生真菌的分子鉴定及药物化学研究.贵州大学硕士学位论文,2006
[20]刘芸,殷红,彭辉等植物内生真菌—潜在的天然药物新来源中国现代中西医杂志,2005年第三卷第五期
[21]刘中华,胡海涛,冯改丰,等.天麻素对Aβ25—35诱导的AIZhimer病细胞模型的保护作用.四川大学学报,2005,36(4)：
[22]刘仕平,曾松荣,郭仕平等.中华山荷叶内生真菌的分离及其发酵产生药用成分的初步研究.中国药物与临床,2003,3(3)：227-228.
[23]黎万奎,胡之璧.2005.内生菌与天然药.中国天然药物,3(4)：193-199.
[24]罗海、孙中吉,天麻素的基础药理研究和临床应用,武警医学院学报,2007,16(4)：465-468
[25]梁宇,高玉葆.内生真菌对植物生长发育及抗逆性的影响[[J].植物学通报,2000 17(1)：52-59
[26]兰琪,姜广华,吴文君,等.植物内生真菌及其应用研究植物农药与药剂毒理学研究进展[M].北京：中国农业科学技术出版社,2002,87-91
[27]兰琪,姬志勤,顾爱国等.苦皮藤内生真菌中杀虫杀菌活性物质的初步研究.西北农林科技大学学报(自然科学版),2004,32(10)：79-84.
[28]李强,刘军,周东坡.植物内生菌的开发与研究进展[J].生物技术通报,2006,(3)：33～37.
[29]李海燕等.一种桃儿七内生真菌的分离初报.云南大学学报(自然科学版),1999,21(3)：243
[30]李桂玲,王建锋,黄耀坚,等.植物内生真菌抗肿瘤活性菌株的筛选[J].菌物系统,2001,20(3)：387-391.
[31]李时珍,本草纲目[M],北京：人民卫生出版社,1977,第1版,730-733
[32]李茂进,胡俊峰,李国珍,等.天麻对铅所致大鼠海马损害的拮抗作用[J].中华劳动卫生职业病杂志,2002,20(5)：331—333.
[33]李雪玲,徐成东两种八角莲属植物内生与根际真菌的多样性初探大理学院学报第3卷第3期2004年5月
[34]李强,刘军,周东坡.植物内生菌的开发与研究进展[J].生物技术通报,2006,(3)：33-37.
[35]莫云强,天麻的药理研究[J],云南医药,1983,4(6)：388
[36]马天有等.从植物分离产紫杉醇的内生真菌的研究.西北大学学报(自然科学版),1999,29(1)：47-49
[37]曲淑梅、等,黄芪、天麻对老年心脑血管患者血中LPO水平的影响[J],锦州医学院学报,1994,15(1)：28-29
[38]任安芝,高玉葆.植物内生真菌一类应用前景广阔的资源微生物[J].微生物学通报,2001,28(6)：90-93.
[39]邱德有等.一种云南红豆杉真菌的分离.真菌学报,1994,13(4)：314-316
[40]塞冬、等,黄芪、天麻、首乌的延缓衰老作用的实验研究[J],锦州医学院学报,1991,12(4)：71
[41]沈书庆,殷红,刘芸,等.产杜仲黄酮内生真菌的初步研究[J].吉林农业大学学报,2008,6(1)：46-48.
[42]王莹,赵华斌,郝家胜.分子系统学的理论方法及展望[J],安徽师范大学学报(自然科学版),2005,28(1)：85-88
[43]王伟等.南方红豆杉内真菌及紫杉烷类产物的初步鉴定.中山大学学报(自然科学版),1999,38(3)：116-118
[44]王永中,肖亚中.植物内生菌及其活性代谢产物.生物学杂志,2004,21(4)：1-5.
[45]王军,林永成,吴雄宇,等.从南海红树林内生真菌No.2533分离出新的异香豆素[J].中山大学学报(自然科学版),2001,40(1)：127-128.
[46]王晓晨,徐庆,陈章良.天麻中一种抗真菌蛋白基因的克隆[J].植物学报,1999,41(10)：1041～1045
[47]吴静萍,郑师章.密花石解菌根菌分离鉴定及其代谢产物的测定[J].复旦学
报,1994,33(5)：547～552.
[48]文才艺,吴元华,田秀玲.植物内生菌研究进展及其存在的问题[J].生态学杂志,2004,23(2)：86-91
[49]肖盛元,郭顺星,杨俊山.粉红粘帚霉菌丝体化学成分研究[J].菌物系统,2001,20(4)：536～538.
[50]肖光,黄芪、天麻、首乌对小白鼠血SOD活力的影响[J],锦州医学院学报,1992,13(4)：15-16
[51]薛美芹.石斛内生真菌的形态和分子鉴定及其细胞毒活性研究.贵州大学硕士学位论文,2006
[52]袁志林,戴传超,史央,等.内生真菌B3促进水稻生长的机理研究[J].江苏农业科学,2004,2：10-13.
[53]袁崇文,中国天麻[M],贵阳：贵州科技出版社,2002,第1版
[54]易润华,朱西儒,周而勋.简化CTAB法快速微量提取丝状真菌DNA[J].湛江海洋大学学,2003,23(6)：72-73
[55]于龙顺、等,天麻抗炎效应的研究[J],中草药,1989,20(5)：211-213
[56]杨箐、等,黄芪、天麻对老年心脑血管疾病患者血中SOD的影响[J],锦州医学院学报,1995,16(1)：15
[57]俞斌,熊健.草坪草抗虫性的研究进展[[J].江西植保,2002.25(4)：127-131
[58]张集慧,王春兰,郭顺星,等.兰科药用植物的5种内生真菌产生的植物激素[J].中国医学科学学院学报,1999,21(6)：460-465.
[59]张玲琪等.发酵产鸢尾酮真菌的分离鉴定及产香特性的初步研究.菌物系统.1999,18(1)：49
[60]张玲琪,王海昆,邵华,等.美登木内生真菌产抗癌物质球毛壳甲素的分离及鉴定[J].中国药学杂志,2002,37(3)：172—175.
[61]张维经.天麻与其真菌营养[J].西北植物学报,1987,7(3)：215-221.
[62]周俊、杨雁宾、浦湘渝,新鲜天麻的酚类成分[J],云南植物研究,1980,2(1)：370
[63]赵国举、等,人培与野生天麻中枢药理作用的比较研究[J],武汉医学院学报,1980,9(1)：91
[64]钟裕国、等,天麻苷元同系物与同型物的合成及其中枢镇静抗惊活性[J],四川医学院学报,1984,15(1)：17
[65]全国中草药汇编[M],北京：人民卫生出版社,1996,169-170
[66]国家药典委员会.中华人民共和国药典[M].
[67]邹容.天麻共生蜜环菌及同属真菌的基因系谱学及药用成分的研究.贵州大学硕士学位论文,2005
[68]曾松荣,徐倩雯,叶保童,卿野,方白玉,黄晓敏.虎杖内生真菌的分离及产抗菌活性物质的筛选.菌物研究,2005,3(2)：24-26
[69]魏景超.真菌鉴定手册[M].上海：上海科学技术出版社,1979.
[70]巴尼特H L,亨特B B.半知菌属图解[M].沈崇尧,译.北京：科学出版社,1977.
[71]Bradly SF, Singh MP,,Janso,JE, et al. Cytoskyrins A and B, new BIA active hisanthraquinones isolated from an endophytic fungus[J].Org Lett,2000 Dec,14, 2(25):4047-4049.
[72]Carrol G.C. The biology of endophytism in plants with particular reference to woody perennials In Fokkema N.J. and van den Heuvel J. Eds Microbiology of the Phyllosphere. Cambridge:Cambridge University Press.1986.205-222.
[73]Guo L D,Hyde K D,Liew E C Y. Identification of endophytic fungi from Livistona chinensis based on morphology and rDNA sequences [J].New Phytology, 2000,147(3):617-630.
[74]Guo B. Dai,JR, Ng S, et al. Cytonic:acids A and B:novel tridepside inhibitors of hCMV protease from the endophytic:fungus Cytonaema species[J].J Nat Prod, 2000, May,63(5):602-604.
[75]HUANG Yaojian,WANG Jianfeng,LIGuiling,et al.Antitumor and antifungal activities in endophytic fungi isolated from pharmaceutical plants Taxus mairei,Cephalataxus fortunei and Torreya grandis[J].FEMSI mmunology and Medical Microbiology,2001,31 (2):163-167.
[76]Hao XY, Tan NH, Zhou J. Anewdi-(p-hydroxybenzyl)hydroxyhmine from Gastrodiaelata B1[J]. Chinese Chemical Letters(中国化学快报),1999,10(6):
467.
[77]Hsieh MT, Wu CR, Chen CF. Gastrodin and p-hydroxybenzyl alchol facilitate moemory consolidation and retrieval, but not acquisition, on the passive avoidance task in rata[J]. J of Ethnopharmacology,1997,56(1):45—54.
[78]Jia-yao Li,et al. Etal. Endophytic taxol-producing fungi from bald cypress,Taxodium distichum. Microbiology.1996,142:2223-2226
[79]Judy F. Shearer.2002. The Potential Role of an Endophytic Fungus in the Decline of Stressed Eurasian Watermilfoil. J. Aquat. Plant Manage,40:76-78.
[80]Kim HJ, Moon KD, Lee DS, et al. Ethyl ether fraction of Gastrodia elata Blume protects amyloid betapeptideinduced cell death[J]. J Ethnopharmacol,2003,84 (1):95-98.
[81]L i J Y, Strobel G A. Jesterone and hydroxyl-jesterone antioomycete cyclohexenone epoxides from the endophytic fungus Pestalotiop sis jesteri[J]. Phytochemistry,2001,57:261～265.
[82]Li J Y,Strobel G A, Harper J,et al. Cryptocin,a potent tetramic acid antimycotic from the endophytic fungus Cryptosporiopsis cf.quercina[J].Org Lett,2000,2:767～770.
[83]Lu H, Zou W X, Meng J C,et al. New bioactive metabolites produced by Colletotrichum sp.,an endophytic fungus in Artemisia annua[J].Plant Sci,2000,151:67～73
[84]Petrini O. Fungal endophytes of tree leaves [M] New York, Springer-Verlag, Andrews J H, Hirano S S. eds,1991,179-197.
[85]Schutz B.2001.Endophytic fungi:a source of novel biologically active secondary metabolites. British Mycological Society, International Symposium Proceedings, Bioactive Fungal Metabolites-Impact and Exploitation, University of Wales, Swansea.
[86]Siegel MR,Johnson MC,Varney DR,et al.1984.A fungal endo-phyte in tall fescue: incidence and dissemination[J].Phytopatho-logy,74:932～937.
[87]Southcott K A,Johnson J A. Isolation of endophytes from two Species of palm,from Bermuda [J].Canadian Journal of microbiology,1997,43(4):789-792.
[88]Strobel G,DaisyB,Castillo U,et al.Naturalproductsfrom endophytic microorganisms.[J].JNat prod.2004,67(2):257～68
[89]Strobel G A.2003. Endophytes as sources of bioactive products. Microbes and Infection,5:535-544.
[90]Strobel G A, Miller R V, Martinez-Miller C,et al.Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina [J]. Mcrobiology,1999,145:1919～1926.
[91]Strobel Qet al. Taxomyces andreanae,A proposed new Taxon for aBubilliferous hyphomyceteassociated with pa-cific yew(Taxus brevifolia).Mycotaxon,1993,X Ⅳ Ⅱ:71～80
[92]Stierle A,et al.Taxol and taxane production by Taxomyces and reanal,an endophytic fangus of pacific yew comments. Science,1993,260:214～216
[93]Stierle A, Strobel G, Stierle D.Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew [J].Science,1993,260:214-216.
[94]Sutton B C. The Coelomycetes[M]. Kew:Commonwealth Mycological Institute,1980.
[95]Taxon for a Bubilliferous hyphomyceteassociated with pa-cific yew(Taxus brevifolia).Mycotaxon,1993, ⅩⅣⅡ:71～80
[96]Zou W X, Meng J C, Lu H,et al. Metabolites of Colletotrichum gloeosporioides, an endophytic fungus in Artemisia mongolica [J]. J Nat Prod,2000,63:1529 ～1530.
[97]Wang Yu,Guo liang-Dong,Morphological and Molecular indentification of an endophytic fungus Epicoccum Nigrum,Mucosystema,2004,23(4):2004.
[98]Wagenaar MM, Clardy J.Dicerandrols, new antibiotic and cytotoxic:dimmers produce by the fungus Phomcpsis Longicolla isolated from an endangered mint[J].J Nat Prod.2001, Aug,64(8):1006-1009.
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