Study On The Blue Functional Genes Of Phalaenopsis

Phalaenopsis orchid is the most widely cultivated in orchid,one of the most popular type,its flower-like ovrflows,rich design and color is gorgeous,leaf shape is beautiful,can do cut flowers and potted flower can do,is of great ornamental value and commercial value,in the tropical orchid known as“orchid queen”,always regard them as the treasures of both at home and abroad,is deep consumers.However because of the limitation of genetic resources by conventional breeding methods cannot obtain the difference of blue color,but the use of transgenic technology can break species limits,can import new gene to the moth orchid gene pool,so that the blue flower cultivation possible.The results are as follows:1.In this study,the transient transformation system of phalaenopsis was established.The results showed that the OD₆₀₀ of agrobacterium bacilli was 0.6,and the infection time was 60 s.In the heavy suspension,150 of the mol/L acetylacetone was added,and 3 d was cultivated,and GUS instantaneous expression rate was the highest,reaching 85.01%.The RNAi interference vector of chalcone synthase?CHS?was transformed into the petals of phalaenopsis,and the changes of the patterns and pigments in the transformed materials were observed after 3 d.The color of the transgenic butterfly orchid was significantly reduced,the pigment content was decreased,and the transcription activity of the CHS gene was significantly reduced compared with that of the control group.This experiment has successfully established a rapid gene function verification method in the phalaenopsis orchid organ,which provides technical support for the research and breeding of phalaenopsis.2.A semi-quantitative RT-PCR method was used to analyze the expression patterns of the flavonoid pathway structural genes CHS,DFR and F3'H in different development stages of the orchid red variety.The results showed that there was a difference in the expression of petals and sepals in different periods,but the overall trend was consistent in the red variety.3.By using agrobacter mediated method,the interference vectors of phDFR,phCHS,phF3'H,and IhF3'5'H,Ma3'5'H,IhDFR,ThDFR,MaDFR and other genes were used to transform the phalaenopsis.Phalaenopsis orchid endogenous CHS,DFR,F3'H interference carrier phalaenopsis orchid petal,the results showed that the interference of phCHS and phDFR gene carrier into petals color significantly lighter than the control group,the total contents cyanine results show that the control group respectively was 5.75 times pKANNIBAL-phCHS group,is pKANNIBAL-phDFR group 2.56 times,and pKANNIBAL-phF3'H and water control group there is almost no difference.4.The DFR gene that interferes with the internal source of phalaenopsis and overexpresses the DFR and F3'H gene of the foreign grape hyacinth,can make the color darker.The total anthocyanin content in the group was 1.52 times higher than that in the control group.Further HPLC analysis of anthocyanin content and composition in the control group and the experimental group showed that anthocyanin content increased significantly,but no new pigment components were detected.