Cold Related Genes Colning And Transformation In Petunia Hybrida

Petunia hybrida is known as the "king of the world flower and flower bed" and widely used in the landscape.But it is sensitive to frost especially in the seedling stage,which made it impossible to live through the winter in most regions of the country.The quality and quantity of the flower will seriously affected even it can overwinter successfully.Therefore,It has important value that breeding new varieties.In the early stage of the team,Gene chip technology was applied to select cold responsive genes of the selfing line H.Based on the microarray results,four genes was chosen for further study(The names are 8-cn9604,10-C16,ZPT2-9 and ZPT2-13).Their stress induced expression profile were analyzed.The main results are as follows:1.Cloning,sequence analysis and induced expression analysis of 8-cn9604,10-C16,ZPT2-9 and ZPT2-13:Analyzing the CDS sequences and then designing the primers of the clone.Using the software to analysis the target gene sequence,The results show that 8-cn9604 and 10-C16 are the ERF transcription factor genes;ZPT2-9 and ZPT2-13 are the C2H2 zinc-finger protenins.The induced expression profile analysis of cold,salt,hypertonicity,ABA and JA through quantitative technology,the results showed that the four genes were up regulated in different stress under.Suggesting that they are associated with the responses of cold and the other abiotic stresses.2.Vector construction and genetic transformation of 8-cn9604,10-C16,ZPT2-9 and ZPT2-13:The overexpression vectors and RNAi vectors were constructed,and then made genetic transform on the petunia W115.The positive transform plants are as follows:8-cn9604,10-C16,ZPT2-9 and ZPT2-13 overexpression plants are 3,2,2 and 2 respectively;and 8-cn9604-R2,10-C16-R1,ZPT2-9-R1 and ZPT2-13-R1 RNAi transgenic plants are 2,1,1 and 2.