广藿香青枯病菌rep-PCR基因指纹图谱分析  

DNA Fingerprint Analysis of Ralstonia solanacearum Strains Isolated from Pogostemon cablin Benth. by Rep-PCR

作　　者：李颛[1] 邓志成[1] 贺红[1] 金华[1] 张宇瑶[1] 

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《中药新药与临床药理》2016年第6期871-876,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金(81373901);高等学校博士学科点专项科研基金(20134425110012)

摘　　要：目的对广藿香青枯病病原菌进行鉴定,并利用rep-PCR基因指纹图谱技术,对广藿香青枯病菌进行遗传多样性分析,以了解青枯病菌的遗传结构及菌株间的遗传分化情况。方法利用青枯菌特异性引物OLI1/Y2对广藿香青枯菌疑似菌株进行PCR鉴定;利用rep-PCR技术,分别用3组引物(BOX A1R;ERIC 1R,ERIC2;REP 1R,REP 2-1)对广藿香青枯病菌进行PCR扩增及基因指纹图谱分析。结果共有12株广藿香青枯菌疑似菌株扩增出288 bp大小的目标条带;BOX-PCR、ERIC-PCR及REP-PCR指纹图谱分析结果表明,供试菌株在相似系数为0.5处,分别分成6,7,8个簇群;在相似系数为0.8处,均分为8个簇群,其中簇群Ⅰ均包括5个亲缘关系相近的菌株。结论 rep-PCR基因指纹图谱技术能有效地区分供试菌株间的遗传差异;广藿香青枯病菌具有较丰富的遗传多样性,部分菌株间遗传相似度较高,组成优势菌群。Objective To identify Ralstonia solanacearum strains isolated from Pogostemon cablin Benth.,and then to study the genetic diversity of the Ralstonia solanacearum strains by repetitive sequence-based polymerase chain reaction（rep-PCR）.Methods The identification of the Ralstonia solanacearum strains was conducted via PCR amplification,using specific primer OLI1/Y2 designed based on 16 S r RNA gene sequences of R.solanacearum strain.The genetic diversity among R.solanacearum strains was analyzed by Rep-PCR fingerprinting with three groups of primers which were BOX and A1 R,ERIC 1R and ERIC 2,and REP 1R and REP 2-1.Results The objective band with 288 bp was obtained from 12 strains which were identified as R.solanacearum.The results of BOX-PCR,ERIC-PCR and REP-PCR genetic fingerprinting showed that R.solanacearum strains were divided into 6,7,8 clusters at 0.5similarity level,and were similarly divided into 8 clusters at 0.8 similarity level,respectively.At 0.8 similarity level,clusterⅠwas predominant,including 5 strains which possessed high genetic similarity.Conclusion Rep-PCR is an effective technique in discriminating genetically related R.solanacearum strains isolated from Pogostemon cablin Benth..Significantly genetic diversity exists among the tested strains.Some dominant strains were also found in this study.

关 键 词：广藿香 青枯病病原菌 鉴定 重复序列PCR 遗传多样性 

分 类 号：R284.2[医药卫生—中药学]