新疆医科大学学报
新疆醫科大學學報
신강의과대학학보
JOURNAL OF XINJIANG MEDICAL UNIVERSITY
2014年
12期
1614-1618
,共5页
周倩%高福春%马红梅%张帆
週倩%高福春%馬紅梅%張帆
주천%고복춘%마홍매%장범
毛茛科%乌头%ISSR%遗传多样性%鉴别
毛茛科%烏頭%ISSR%遺傳多樣性%鑒彆
모간과%오두%ISSR%유전다양성%감별
Ranunculaceae%Aconitum L%Inter-Simple Sequence Repeat%Genetic diversity%Identification
目的:探讨新疆特产准噶尔乌头、多根乌头、白喉乌头、那拉提乌头、空茎乌头、林地乌头、拟黄花乌头共7个种共41个居群间的遗传多样性和变异情况,为药材鉴定提供 DNA 特征。方法利用植物基因组提取试剂盒提取 DNA,采用紫外分光光度法检测 DNA 的浓度和纯度,使用60种哥伦比亚引物分别对7个种共41个居群的乌头基因进行 ISSR 分析,应用 NTSYS-PC2.10和 POPGEN32对所得数据进行处理,计算不同种乌头的遗传相似系数和遗传距离,采用非加权平均法(UPGMA)进行聚类,构建亲缘关系系统图,用主坐标分析法绘制二维、三维散点图。结果从60个引物中筛选出13个条带清晰、多态性明显且重复性好的引物用于扩增,共扩增得到163个条带,其中多样性条带151个。根据 ISSR 聚类分析,41个居群的乌头可聚为7大类群,与种的分类相当符合;而且乌头类药材在多个引物下具有特征的 DNA 标记。结论乌头种质资源间具有很高的多态性,遗传变异较大;ISSR 分子标记法可用于7种药材的鉴别,提供了分子水平依据,也为构建其 DNA 指纹图谱提供可能。
目的:探討新疆特產準噶爾烏頭、多根烏頭、白喉烏頭、那拉提烏頭、空莖烏頭、林地烏頭、擬黃花烏頭共7箇種共41箇居群間的遺傳多樣性和變異情況,為藥材鑒定提供 DNA 特徵。方法利用植物基因組提取試劑盒提取 DNA,採用紫外分光光度法檢測 DNA 的濃度和純度,使用60種哥倫比亞引物分彆對7箇種共41箇居群的烏頭基因進行 ISSR 分析,應用 NTSYS-PC2.10和 POPGEN32對所得數據進行處理,計算不同種烏頭的遺傳相似繫數和遺傳距離,採用非加權平均法(UPGMA)進行聚類,構建親緣關繫繫統圖,用主坐標分析法繪製二維、三維散點圖。結果從60箇引物中篩選齣13箇條帶清晰、多態性明顯且重複性好的引物用于擴增,共擴增得到163箇條帶,其中多樣性條帶151箇。根據 ISSR 聚類分析,41箇居群的烏頭可聚為7大類群,與種的分類相噹符閤;而且烏頭類藥材在多箇引物下具有特徵的 DNA 標記。結論烏頭種質資源間具有很高的多態性,遺傳變異較大;ISSR 分子標記法可用于7種藥材的鑒彆,提供瞭分子水平依據,也為構建其 DNA 指紋圖譜提供可能。
목적:탐토신강특산준갈이오두、다근오두、백후오두、나랍제오두、공경오두、임지오두、의황화오두공7개충공41개거군간적유전다양성화변이정황,위약재감정제공 DNA 특정。방법이용식물기인조제취시제합제취 DNA,채용자외분광광도법검측 DNA 적농도화순도,사용60충가륜비아인물분별대7개충공41개거군적오두기인진행 ISSR 분석,응용 NTSYS-PC2.10화 POPGEN32대소득수거진행처리,계산불동충오두적유전상사계수화유전거리,채용비가권평균법(UPGMA)진행취류,구건친연관계계통도,용주좌표분석법회제이유、삼유산점도。결과종60개인물중사선출13개조대청석、다태성명현차중복성호적인물용우확증,공확증득도163개조대,기중다양성조대151개。근거 ISSR 취류분석,41개거군적오두가취위7대류군,여충적분류상당부합;이차오두류약재재다개인물하구유특정적 DNA 표기。결론오두충질자원간구유흔고적다태성,유전변이교대;ISSR 분자표기법가용우7충약재적감별,제공료분자수평의거,야위구건기 DNA 지문도보제공가능。
Objective To investigate the genetic diversities and variations of sevenAconitum L.species,in-cluding Aconitum soongaricum Stapf ., A.karakolicum Rapaics ., A.leucostomum Worosch ., A. leucostomum var .nalatiensis F.Zhang, A.nemorum M.Pop., A.apetalum B.Fedtsch .and A. anthoroideum DC.,and to supply DNA characteristics for accurate identification of crude drugs of Aconi-tum L.Methods Plant genome extraction kit was applied to extract DNA,and ultraviolet spectrophotom-eter was used to detect the concentrations and purity of DNA.Sixty ISSR primers were screened to analyze the DNA of total forty-one provenances of seven Aconitum L.species for ISSR amplification.Biosofteares including NTSYS-PC2.10 and POPGEN32 were used to analyze the polymorphic bands obtained.UPGMA method was used to make up the systematic diagram of genetic relationship,with two-dimension scatter-graph and three-dimension scatter-graph constructed by using the principal cordinate analysis.Results Thirteen primers selected from 60 primers were used for amplification and a total of 163 DNA bands were obtained,including 151 polymorphic bands.According to cluster analysis result of ISSR,the forty-one provenances of Aconitum L.were classified into seven groups,which accorded pretty with the classification of species;and those medicinal materials had the characteristics of DNA markers when using multiple primers.Conclusion Germplasm resources of Aconitum L.showed abundant polymorphism and higher ge-netic variation.ISSR molecular maker technology is useful for identifying species of Aconitum L.plants, supplying molecular level basis and making DNA fingerprints building possible.