基于cpDNA条形码鉴定铁皮石斛种质资源  

Identification of Germplasm Resources in Dendrobium officinale Based on cpDNA Barcoding

作　　者：付涛[1] 胡仲义[1] 何月秋[1] 周亚美[1] 毛阳正 

机构地区：[1]宁波城市职业技术学院,浙江宁波315502

出　　处：《核农学报》2017年第2期255-262,共8页Journal of Nuclear Agricultural Sciences

基　　金：浙江省教育厅科研项目(Y201533793);宁波城市职业技术学院校内重点项目(ZZX15064);宁波市自然科学基金(2015A610267)

摘　　要：为选择适合鉴定铁皮石斛的条形码,以5份铁皮石斛种质资源和1份串珠石斛(外类群)为试验材料,采用cpDNA条形码技术进行分子鉴定。结果表明,5个条形码序列(matK、rbcL、trnH-psbA、petApsbJ和rpl32-trnL)种间遗传距离均远高于种内遗传距离,符合作为条形码的基本要求,其中petA-psbJ变异位点最多(8.03%),说明其进化速率亦最快,petA-psbJ可作为石斛属候选条形码之一;单个条形码鉴定率均较低(<60%),核心条形码matK+rbcL鉴定率亦较低(68.75%),而在核心条形码基础上结合其它序列(≥3)却可以显著提高鉴定率(≥81.25%),matK+rbcL+petA-psbJ鉴定率达到100%,可作为铁皮石斛分子鉴定的最适条形码组合。综上,利用cpDNA条形码技术,可将不同种质资源的铁皮石斛区分开,基本上可将不同采集地的铁皮石斛聚类;同时,建议matK+rbcL+petA-psbJ作为铁皮石斛种质资源鉴定的DNA条形码。本研究结果为铁皮石斛种质资源的快速鉴定提供了一定的参考。In order to select suitable barcoding for the identification of Dendrobium officinales. Fifteen Dendrobium officinales and one Dendrobium falconeri were selected to study the molecular identification through DNA barcode technology. The results showed that the interspecific genetic distance were significantly higher than the intraspecific genetic distance with the 5 barcode sequences （ matK, rbcL, trnH - psbA, petA - psbJ and rpl32 - trnL ） , that reached the basic requirements of the barcode, among them, petA - psbJ owned highest variable site （8.03%） , indicating its evolutionary rate was the fastest, it could be used as one candidate barcode of Dendrobium. The results also showed that the identification rate was lower （ 〈 60% ） with single barcode sequence, and the identification rate of core barcode matK ＋ rbcL was also not high （68.75%）, combination the core barcode with other sequences （ more than 3） could significantly improve the identification rate （ I〉 81. 25% ）, the matK ＋ rbcL ＋ petA - psbJ was the optimum barcode combination in the molecular identification of Dendrobium officinale whose identification rate was 100%. In this study, , different germplasm resources of Dendrobium officinale could be distinguished and clustered roughly according different areas with cpDNA barcoding technology, moreover, the combined sequence of matK ＋ rbcL ＋ petA - psbJ could be suggested as the authentication code for the identification of medicinal plant species in the genus Dendrobium officinale. The study provided an important reference for the rapid identification of Dendrobium officinale germplasm resource.

关 键 词：铁皮石斛 CPDNA 条形码 种质鉴定 

分 类 号：S567.239[农业科学—中草药栽培]