作 者：
李伟;其其格;黄旭;岳阳;尹靖琨;王瑞俭
关键词：
越桔属;蓝莓;栽培品种;DNA条形码;遗传分化
摘 要：
【目的】筛选出适合鉴别蓝莓品种的DNA条形码,用于蓝莓不同品种的区分及遗传分化研究。【方法】以40个蓝莓品种108份样品总DNA为模板,对8个条形码片段(rpo B、psb A-trnH、ycf5、rbc L、rpo C、mat K、ITS、ITS2)进行扩增与测序,分析扩增效率及测序成功率、遗传距离、barcoding gap,并对遗传距离计算结果进行Wilcoxon检验,利用NJ树法对各蓝莓品种进行聚类分析。【结果】在8个DNA条形码中,ITS与ycf5在所有蓝莓样本中均无扩增产物。除mat K序列扩增成功率(96. 30%)、测序成功率(99. 04%)相对较低外,其他5个条形码扩增与测序成功率均为100%。蓝莓的rpo B序列完全一致,为高度保守序列。各条形码的变异位点数依次为:ITS2(11个)>mat K(4个)>rbc L(3个)>psb A-trnH(2个)>rpo C(1个)>rpo B(0个)。各蓝莓品种的品种内、品种间遗传距离较小,介于0. 000 16～0. 002 58之间,且品种间遗传距离大于品种内。Barcoding gap分析结果显示,各条形码均未形成明显的间隔区,但从分布情况看,ITS2、psb A-trnH、mat K 3个条形码有偏向两端分布的趋势,尤其是ITS2。Wilcoxon检验显示,ITS2、psb A-trnH品种间变异较大,psb A-trnH品种内的变异较大。聚类分析结果表明,psb A-trnH和rpo C可将蓝莓品种分为2个类群,rbc L和mat K将蓝莓划分为3个类群,ITS2将蓝莓分为4个类群。利用条形码组合可提高蓝莓品种鉴定率,其中ITS2+mat K+rpo C+rbc L鉴定成功率最高,为20%。【结论】ITS2对蓝莓品种的鉴定结果优于其他条形码,条形码组合ITS2+mat K+rpo C+rbc L将40个蓝莓品种分为14组,能够将山东省主要的蓝莓栽培品种如伯克利、阳光蓝、北陆等品种区分开来,较适于蓝莓的品种鉴定。
译 名：
DNA Barcodes of Blueberry Cultivars
作 者：
Li Wei;Qi Qige;Huang Xu;Yue Yang;Yin Jingkun;Wang Ruijian;Forestry College of Beihua University;
关键词：
Vaccinium;;blueberry;;cultivar;;DNA barcode;;genetic divergence
摘 要：
【Objective】 The purpose of this study was screening the suitable DNA barcodes for identifying cultivars,which were used for studying the discrimination and the heredity differentiation of blueberry(Vaccinium) cultivars.【Method】Eight DNA barcodes(rpo B,psb A-trnH,ycf5,rbc L,rpo C,matK,ITS and ITS2) of 108 individuals from 40 blueberry cultivars were amplified and sequenced. The amplification efficiency,sequencing success rate,genetic distance and barcoding gap were analyzed,and the calculations of genetic distance were verified using the Wilcoxon test. Finally,cluster analysis of blueberry cultivars was performed by N-J tree method.【Result】 Among the eight DNA barcodes,ITS and ycf5 could not be amplified in all samples. The amplification and sequencing rate of all the five barcodes(rpo B,psb AtrnH,rbc L,rpo C and ITS2) were 100%,except matK was 96. 3% and 99. 04% respectively. The rpo B sequences of all blueberry cultivars were completely consistent,so this fragment is highly conserved in blueberry. The mutation points of six barcodes were as follows: ITS2(11) > matK(4) > rbc L(3) > psb A-trnH(2) > rpo C(1) > rpo B(0). The genetic distances among or within blueberry cultivars were smaller,range from 0. 000 16-0. 002 58. The genetic distances were greater among cultivars than within cultivars. Barcoding gap analysis showed that,there was no obvious interval area formed in all barcodes,but according to distribution,barcodes ITS2,psb A-trnH and matK tended to be distributed at both ends,especially ITS2. Wilcoxon test showed that,the interspecific variation of barcodes ITS2 and psb A-trnH were more obvious than others,and greater intraspecific variation was found in psb A-trnH. The cluster analysis indicated that,blueberry cultivars could be divided into two groups by psb A-trnH or rpo C,three groups by rbc L or matK,and four groups by ITS2.Furthermore,the identification rate of blueberry cultivars could be improved by the combination of barcodes and with the highest success rate of 20% by ITS2+matK+rpo C+rbc L.【Conclusion】The identification of blueberry cultivars by ITS2 was superior to other barcodes. A total of 40 blueberry cultivars were divided into 14 groups by the barcode combination of ITS2+matK+rpo C+rbc L,which can distinguish the major blueberry cultivars in Shandong province,such as ‘Berkeley',‘Sunshineblue'and ‘Northland'respectively. The barcode combination is suitable for the identification of blueberry cultivars.
