Molecular Cloning And Functional Analysis Of Flavonoids Synthase Genes In Oil Peony

Paeonia ostii and Paeonia rockii are perennial shrub belonging to Paeonia.As they contain a higher and quality oil,so they are used as oil tree peony.The oil contains much linoleic linolenic acid oleic acid and ect, which have the functions such as anti-cancer,anti-aging and preventing diabetes and others. The peony oil contains a good quality and possesses a higher developed potential, but conventional production has a shelled difficult problem. That is the limitation that it could not be used largely. As to the molecular points, the mediation of controlling seed coat color and astringency of genes is expected to become the fundamental solution to the peony seed color and astringency problems. It was reported that,in the oil of peony, anthocyanin is the main component of the control testa black and astringency is the main component of tannins. Thus, cloning and identification of synthase gene is good to learn oil peony seed coat anthocyanin and tannin biosynthesis, and adjust the color and astringency as a mean for further finding of the knockout.In this study, the seeds of Paeonia ostii and Paeonia rockii were used as materials.1、We tested the component of the seed skin and coloed their flavonoid pigment synthesis enzyme genes. The results showed that the pigment composition of oil peony seed skin is mainly cyanidin- anthocyanin.2、The full-length of Paeonia ostii and Paeonia rockii anthocyanidin synthase(ANS) gene is 1065 bp, which encodes 354 amino acids. Paeonia ostii ANS relative molecular weight protein is 40.4755 kDa. Its isoelectric point reached 5.42. The relative molecular weight of Paeonia rockii ANS protein is 40.4496 kDa. Both the peony contained a PLN03178 domain. Secondary structure contained the α helix, random coil, β and rotation extending chains. Cytoplasm is the maximum possibility of subcellular localization.3 、Referring to the camellia plant varieties, sorbitol, soybean, etc, the conserved sequences of Leucoanthocyanidin reductase( LAR)and Anthocyanidin reductase(ANR)genes were cloed.We compared the homologous and the result showed that both the genes may belong tooil peony genes; 4、Real-time fluorescence quantitative methods were used to test the expression of ANS in the Paeonia ostii including five flowering times. The results showed:The expression of ANS during the flowering process in the Paeonia ostii is falling,rising and falling.What’s more, in flowering time it showed that the highest expression, and lowest in theending of flowering.So we can see that ANS gene played an important role in petal color 5、T-mANS was successfully constructed.