Flower Pigment Analysis Of Phalaenopsis And Selection Of Transformation Vatieties With The Blue Genes

The study was based on the results of the our team's research on the blue flowers of monocotyledons.18 kinds of Phalaenopsis species were used as materials and identified by epigenetic and anatomical observation.The pigment elements of Phalaenopsis were qualitatively and quantitatively analyzed by high performance liquid chromatography,and then genotypes of flavonoids were analyzed and transiently expressed.In this study,The reveal of the mechanism of flavonoid metabolic pathway in Phalaenopsis could be used for the systematic regulation of Phalaenopsis spp.The results are as follows:1.The method for the analysis of flavonoid compounds was established by high performance liquid chromatography(HPLC).The results showed that the mobile phase A was 0.1% formic acid water,B was 80% acetonitrile,elution program 0 min,12 % Min,25% B;32min,38% B;40min,38% B;45min,12% B;50min,12% B,which can achieve good separation effect.2.The metabolic analysis showed that different patterns of Phalaenopsis and Grape hyacinth were completely different.The main anthocyanin types were Cy and Pg type pigment derivatives,flavonoids and flavonols were mainly apigenin derivatives.The grape hyacinth was isolated from the same wavelength,the main anthocyanin type was Dp and its methylated pigments Pt,Mv derivatives.The main substances of flavonoids and flavonols were kaempferol derivatives.3.The expression of CHS,F3'H and F3'5'H genes in different flavonoids of Phalaenopsis were different.The results showed that the expression patterns of different genotypes were different,but the three genes Expression consistent.In terms of the gene expression level,the purple variety was higher than the white variety.4.The transient overexpression of Phalaenopsis was performed usingAgrobacterium tumefaciens with three RNA interference vectors constructed with PhCHS?PhDFR?PhF3'H and six overexpression vectors with IhDFR?IhF3'5'H?MaDFR?MaF3'5'H?ThDFR?CcF3'5'H.The RNA interference of the Three genes had the phenomenon that the petals were lighter in color,pKANNIBAL-PhDFR and pKANNIBAL-PhCHS interference results were similar,but the former performs better with a 70% anthocyanin in petals;PhF3'H-RNAi reached 60% and its Antisense fragment vector only got 47% decrease.We didn't get the desired result.The effect of strain GV3101 was better than that of EHA105.5.In summary,we selected low Cy and Pg-type pigment,high Dp-type pigment high as well as low flavonol content of varieties such as P.‘Red angle',and also simple pigment type,small and easy to transform varieties such as P.‘Big chili'as a suitable variety transformed with blue gene.