基于ITS和matK基因对牡丹组序列分析及其亲缘关系的研究

发布时间：2018-11-21 17:05

【摘要】：以芍药属牡丹组全部9个野生种、5个紫斑牡丹栽培品种及3个中原牡丹品种为试材,进行核糖体内转录间隔区(ITS)和叶绿体成熟酶K(matK)基因片段测序分析,探讨ITS和matK序列为牡丹组所有野生种种间关系提供分子证据。从GeneBank中选取了1个牡丹及3个外类群芍药、川赤芍和草芍药的ITS及matK序列。对试验样品进行DNA提取、PCR扩增并双向测序得到44条序列,人工校正后将所得44条序列进行比对;计算碱基组成频率、变异位点、简约信息位点数、转换/颠换比率、种内及种间遗传距离,以邻接法进行系统发育分析。结果表明,牡丹组所有个体ITS序列长度在750~800 bp,含有86个多态位点,74个简约信息位点,转换/颠换比率(R)为1.2;而matK序列含有20个简约信息位点,转换/颠换比率(R)为1.7。ITS序列分析将牡丹组野生种分为两大枝,稷山牡丹、紫斑牡丹、卵叶牡丹和杨山牡丹聚为一枝,狭叶牡丹、滇牡丹、黄牡丹和大花黄牡丹聚为另一枝,这两枝分别与革质花盘亚组和肉质花盘亚组相对应,而四川牡丹位于革质花盘亚组最底端,支持前人研究将四川牡丹归为革质花盘亚组。matK序列分析的牡丹组野生种间遗传距离结果不理想,未能清晰的表明野生种之间的亲缘关系。由此说明,ITS序列更适合牡丹组野生种间亲缘关系的研究分析。
[Abstract]:All 9 wild species of Paeonia (Paeonia) group, 5 cultivars of Violet peony and 3 cultivars of Central Plain Peony were used to sequence the (ITS) and K (matK) gene fragments of ribosomal intertranscription spacer and chloroplast mature enzyme. To explore ITS and matK sequences to provide molecular evidence for all wild relationships in the peony group. The ITS and matK sequences of one peony and three other groups of Paeonia lactiflora, Radix Paeoniae rubra and Paeonia lactiflora were selected from GeneBank. DNA was extracted from the sample and 44 sequences were obtained by PCR amplification and bidirectional sequencing. Base composition frequency, mutation site, reduced information site number, conversion / transversion ratio, genetic distance between species and species were calculated, and phylogenetic analysis was carried out by adjacency method. The results showed that all individuals in peony group had 86 polymorphic loci and 74 parsimonic information sites in 750,800 bp, and the conversion / transversion ratio (R) was 1.2; The matK sequence contained 20 parsimonious information sites, and the conversion / transversion ratio (R) was used as 1.7.ITS sequence analysis. The wild species of peony group were divided into two branches: Jishan peony, purple peony, egg leaf peony and Yangshan peony, and narrow leaf peony. Dian peony, yellow peony and large flower yellow peony gathered as another branch, which corresponded to leathery disk subgroup and fleshy disk subgroup, respectively, while Sichuan peony was located at the bottom of leathery disk subgroup. In support of previous studies, Sichuan peony was classified as leathery disk subgroup. The result of genetic distance between wild species in matK sequence analysis was not ideal, and the genetic relationship between wild species could not be clearly indicated. Therefore, ITS sequence is more suitable for the study of wild interspecific genetic relationship of peony group.
【作者单位】：甘肃农业大学林学院;
【基金】：以核基因ITS序列为分子标记建立甘肃省紫斑牡丹品种鉴定标准的研究(GNSW-2014-7);省农牧厅生物技术专项~~
【分类号】：S685.11

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