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利用RT-PCR技术从牡丹组7个野生种和1个栽培种中分别克隆得到泛素延伸蛋白基因(ubiquitin,UBI)、素环蛋白基因(cyclophilin,CYP)、肌动蛋白基因(Actin)、葡萄糖六磷酸脱氢酶基因(glucose-6-phosphate 1-dehydrogenase,G6PD)和β–微管蛋白基因(beta-tubulin,TUB)等5个管家基因的编码序列(coding sequence,CDS)。序列比对分析发现,5个管家基因序列均相对保守,其在牡丹7个野生种和1个栽培种中的保守性在99.04%~99.69%。利用5个管家基因分别构建了其在牡丹组8份材料中的系统进化树。5个系统进化树均将8份材料分成两组,即革质花盘亚组和肉质花盘亚组;由于5个管家基因核苷酸序列各自的保守性不同,在区分亚组成员间略有差异。综合分析5个管家基因构建的系统进化树可知,杨山牡丹(Paeonia ostii)和栽培牡丹(P.suffruticosa‘Luoyanghong’)常聚为一支,二者亲缘关系较近,因而推测杨山牡丹可能参与了栽培牡丹的形成;紫斑牡丹(P.rockii)和四川牡丹(P.decomposita)一直聚为一支,推测二者亲缘关系较近;黄牡丹(P.lutea)和紫牡丹(P.delavayi)一直聚为一支,推测这二者亲缘关系较近。狭叶牡丹(P.potaninii)与黄牡丹和紫牡丹分化较早,结合前人研究结果建议将狭叶牡丹作为一个独立的种。
The ubiquitin (UBI), cyclophilin (CYP), actin (Actin) genes were cloned from 7 wild and 1 cultivated species of Peony by RT-PCR. Five coding genes of housekeeping genes such as glucose-6-phosphate 1-dehydrogenase (G6PD) and beta-tubulin (TUB). Sequence alignment analysis showed that the five housekeeping genes were relatively conserved, and their conservatism was 99.04% -99.69% in seven wild and one cultivated species of Peony. Five housekeeping genes were used to construct phylogenetic tree among 8 materials of Peony. Five phylogenetic trees divided the eight materials into two subgroups: the subfascia and the subfascal group. Because of the different conservativeness of the nucleotide sequences of the five housekeeping genes, there was a slight difference among the subgroups . Based on the phylogenetic tree analysis of five housekeeping genes, we can see that Paeonia ostii and P.suffruticosa’Luoyanghong ’often clustered together and the genetic relationship between the two was close Participation in the formation of cultivated peony; P.rockii and P.decomposita have been grouped together, suggesting that the two are closely related; P.lutea and P.delavayi ) Has been a polygamy, speculated that the two are relatively close relationship. P.potaninii and yellow peony and purple peony differentiation earlier, combined with the results of previous studies suggest that the thin-leaf peony as an independent species.