【摘要】：以芍藥屬牡丹組全部9個(gè)野生種、5個(gè)紫斑牡丹栽培品種及3個(gè)中原牡丹品種為試材,進(jìn)行核糖體內(nèi)轉(zhuǎn)錄間隔區(qū)(ITS)和葉綠體成熟酶K(matK)基因片段測(cè)序分析,探討ITS和matK序列為牡丹組所有野生種種間關(guān)系提供分子證據(jù)。從GeneBank中選取了1個(gè)牡丹及3個(gè)外類群芍藥、川赤芍和草芍藥的ITS及matK序列。對(duì)試驗(yàn)樣品進(jìn)行DNA提取、PCR擴(kuò)增并雙向測(cè)序得到44條序列,人工校正后將所得44條序列進(jìn)行比對(duì);計(jì)算堿基組成頻率、變異位點(diǎn)、簡(jiǎn)約信息位點(diǎn)數(shù)、轉(zhuǎn)換/顛換比率、種內(nèi)及種間遺傳距離,以鄰接法進(jìn)行系統(tǒng)發(fā)育分析。結(jié)果表明,牡丹組所有個(gè)體ITS序列長(zhǎng)度在750~800 bp,含有86個(gè)多態(tài)位點(diǎn),74個(gè)簡(jiǎn)約信息位點(diǎn),轉(zhuǎn)換/顛換比率(R)為1.2;而matK序列含有20個(gè)簡(jiǎn)約信息位點(diǎn),轉(zhuǎn)換/顛換比率(R)為1.7。ITS序列分析將牡丹組野生種分為兩大枝,稷山牡丹、紫斑牡丹、卵葉牡丹和楊山牡丹聚為一枝,狹葉牡丹、滇牡丹、黃牡丹和大花黃牡丹聚為另一枝,這兩枝分別與革質(zhì)花盤亞組和肉質(zhì)花盤亞組相對(duì)應(yīng),而四川牡丹位于革質(zhì)花盤亞組最底端,支持前人研究將四川牡丹歸為革質(zhì)花盤亞組。matK序列分析的牡丹組野生種間遺傳距離結(jié)果不理想,未能清晰的表明野生種之間的親緣關(guān)系。由此說明,ITS序列更適合牡丹組野生種間親緣關(guān)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.
【作者單位】： 甘肅農(nóng)業(yè)大學(xué)林學(xué)院;
【基金】：以核基因ITS序列為分子標(biāo)記建立甘肅省紫斑牡丹品種鑒定標(biāo)準(zhǔn)的研究(GNSW-2014-7);省農(nóng)牧廳生物技術(shù)專項(xiàng)~~
【分類號(hào)】：S685.11

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