贵州天麻主产区蜜环菌的分离及rDNA-ITS序列分析  

On RDNA-ITS Analysis and Isolate of Armillarilla Collected from Main Producing Areas of Gastrodia Elatain Guizhou

作　　者：黄万兵 桂阳[2,3] 朱国胜[2,3] 刘朝贵[1] 李青风[1,2,3] 

机构地区：[1]西南大学园艺园林学院,重庆400715 [2]贵州省现代中药材研究所,贵阳550006 [3]贵州省农业生物技术重点实验室,贵阳550006

出　　处：《西南师范大学学报（自然科学版）》2014年第6期35-42,共8页Journal of Southwest China Normal University(Natural Science Edition)

基　　金：贵州省优秀青年科技人才培养对象专项资金项目[黔科合人字(2011)35];贵州省科技计划项目[黔科合院所创能(2010)4002];中央补助地方科技基础条件专项基金项目[黔科条中补地(2010)4002];贵州省中药现代化科技产业研究开发专项项目[黔科合中药字(2011)5035];贵州省农业科学院研究生创新基金项目[黔农科合(创新基金)2011016];贵州省农业科学院专项资金项目[黔农科院院专项(2012)020]

摘　　要：[目的]了解贵州天麻主产区蜜环菌的生物种及其地理分布.[方法]在代表性主产区采集蜜环菌样本,利用双抗培养基分离菌索或子实体;从液体培养的菌丝中提取总DNA,利用真菌通用引物ITS1和ITS4扩增rDNAITS片段,扩增产物进行TA克隆并送测序;利用SeqMan软件对序列进行拼接,并用MEGA软件构建进化树.[结果]从大方、德江等县分离到35株蜜环菌;得到DF1等17个菌株的rDNA-ITS完整序列,GenBank中蜜环菌rDNA-ITS序列比对鉴定与系统发育分析一致.[结论]17个rDNA-ITS序列分析的蜜环菌株中,4株属于生物种Armillaria gallica;8株属于Armillaria cepistipes;5株与Armillaria mellea亲缘关系较近,可能为其同源种.贵州省内A.mellea,A.cepistipes的分布最广,其次是A.gallica.To investigate biological species and geographic distribution of Armillarillaisolated from the main producing areas of Gastrodia elatain Guizhou.Rhizomorph or fruiting body of Armillarilla mellea have been collected from representative areas and isolated on medium with penicillin and gentamicin.Total DNA has been extracted fromArmillarilla mycelium cultured in fluid medium.General primer ITS1 and ITS4 have been used to amplificate rDNA-ITS sequences which are TA Cloned and Sequenced.SeqMan and MEGA software has been adopted to do sequence splicing and construct evolutionary tree.35 Armillarillastrains have been isolated from samples collected from Dafang County,Dejiang County etc.rDNAITS complete sequence of 17 strains have been obtained.The result of sequence blast and phylogenetic analysis shows that they are consistent.Among the 17 Armillaria strains,there are 4 strains that are known as Armillarilla gallica,8 strains belong to Armillaria cepistipes,5 strains may be Armillaria mellea subspecies.In guizhou province,A.melleaand A.cepistipesis are the most geographically widespread of Armillaria biological species,followed by A.gallica.

关 键 词：蜜环菌 分离 生物种 RDNA-ITS 

分 类 号：Q939[生物学—微生物学]