The Construction And Genetic Transformation Of Peas Gene PBR-F3H Whose Vector Is Expressed Through RNAi

Flavonoids are a class of plant secondary metabolites formed during growth, including isoflavones, flavonoids, chalcones, anthocyanins and other biologically active substances, flavone synthase gene in pea flavonoid biosynthetic pathway(FNS), flavanone 3-hydroxylase gene(F3H) and isoflavone synthase(IFS) is an important function of genes.Isoflavones have weak estrogenic activity, antioxidant activity, anti-hemolytic activity and antifungal activity, can effectively prevent and suppress the occurrence of leukemia, osteoporosis, cancer and other diseases, but their distribution limited in nature Papilionoideae and a few other plants in the legume. Not only time consuming and less potential to improve by conventional breeding methods.Isoflavones metabolic pathway is a very active area, but the reported use of secondary metabolites of RNAi technology research peas isoflavones little.In this study, the use of RNAi technology to build interference flavanone 3-hydroxylase gene(F3H) vector pBR-F3 H, assisted by Agrobacterium-mediated vacuum pea shoot tips transformation and pollen tube pathway method vector pBR-F3 H integrated into the genome of the pea, so that more of the precursor(flavone synthase, flavanone 3-hydroxylase and isoflavone synthase have a common precursor 4,5,7-trihydroxy flavanone) to the iso flavonoid synthesis branch. To improve the role of pea isoflavone content.After the experiment, we get interference flavanone- 3- hydroxylase gene(pBR F3H)- 4 in F3 H carrier and positive transgenic plants(two kinds of transformation methods respectively 2 tree), and the positive transgenic plant isoflavones content determination, found that the auxiliary vacuum stem tip conversion system for 2 transgenic positive pea plant isoflavones content compared with the non-transgenic control pea plant by 1076.01(including including g/g and 968.86 g/g, isoflavones content increased by 53.01% and 47.74% respectively; However, pollen tube channel transformation system for 2 transgenic positive pea plant isoflavones content compared with the non-transgenic control pea plant by 1069.04(including including g/g and 1204.15 g/g, isoflavones content increased by 38.37% and 43.23% respectively. In future experiments and Southern blot, northern blot, western blot molecular detection, such as to improve the detection system.Is obtained by means of genetically modified(gm) of the pea varieties, not only the isoflavone content increased significantly, but also greatly save the cycle of germplasm improvement needed, for the further study of class flavonoids play a role.