不同品种蝴蝶兰黄酮代谢途径分析

Analysis of flavonoid metabolic pathways of different varieties of Phalaenopsis and their molecular mechanism

采用高效液相色谱(HPLC)研究18种蝴蝶兰的黄酮代谢途径。建立HPLC技术分析黄酮代谢途径(调控颜色)。使用荧光定量PCR(Real-Time PCR)进一步分析蝴蝶兰黄酮代谢途径关键基因表达水平;农杆菌介导瞬时转化蝴蝶兰,获得转基因株系。蝴蝶兰HPLC分离技术可成功分离29种组分,与其它物种葡萄、风信子差别较大;使用Real-Time PCR分析不同品种蝴蝶兰黄酮代谢途径关键基因,发现不同品种的查尔酮合酶(Chalcone synthase,CHS),黄烷酮-3-羟化酶(Flavonol-3-hydroxylase,F3H)以及查尔酮异构酶(Chalcone isomerase,CHI)时空表达模式基本一致;农杆菌介导法构建关键酶基因RNAi干扰载体瞬时转化蝴蝶兰,黄酮代谢途径中关键酶基因干扰表达会淡化蝴蝶兰颜色。建立了蝴蝶兰黄酮代谢途径的HPLC研究方法,并对其分子机制进行了初步研究,对蝴蝶兰花色调控机理的揭示提供了理论和实践基础。

High performance liquid chromatography(HPLC)was used to study the flavonoid metabolism pathways of 18 Phalaenopsis.A HPLC technology was established to analyze the pathway of flavonoid metabolism(regulate color).Fluorescence quantitative PCR(Real-Time PCR)was used to further analyze the expression level of key genes in Phalaenopsis flavonoid synthesis pathway,and an Agrobacteriummediated transient transformation of Phalaenopsis was used to obtain transgenic strains.Phalaenopsis HPLC separation technology can successfully separate 29 components,which is quite different from other species of grape and hyacinth.Real-Time PCR analysis of the key genes in the flavonoid synthesis pathway of different species of Phalaenopsis finds that the temporal and spatial expression patterns of Chalcone synthase(CHS),Flavonol-3-hydroxylase(F3H)and Chalcone isomerase(CHI)of different species are basically the same.The key enzyme gene RNAi interference vector is constructed by Agrobacterium-mediated method to transform Phalaenopsis transiently.The interference expression of key enzyme genes in the flavonoid synthesis pathway will reduce the color of Phalaenopsis.In conclusion,the HPLC research method of Phalaenopsis flavonoid metabolism pathway is established,and its molecular mechanism is preliminary studied,which provids a theoretical and practical basis for revealing the color regulation mechanism of Phalaenopsis orchids.