Cloning Of PeZFP And PeMYB Genes And Functional Analysis Of PeZFP Gene In Phyllostachys Edulis

Cold, drought and high salinity are the main abiotic stresses which affect plant growth. So cultivating the new kinds is critical for the crop. Expression of many inducible genes is regulated on the transcriptional level by transcriptional factors. So far, hundreds of genes related with drought, high salinity, cold, hormone, pathogen response, growth and development were isolated from high plants. Phyllostachys edulis was one of the most extensive distribution and highest economy value bamboo. Drought and high salinity not only affect the yield of Phyllostachys edulis, but also restrict the distribution of it. Many areas were not suitable for the growth of Phyllostachys edulis.PeZFP and PeMYB related with stress tolerance were isolated through homological cloning with RT-PCR and RACE from Phyllostachys edulis. The sequences were analyzed by the biotical tools. The expression patterns of two genes with semi-quantitative RT-PCR were studied under treatments of high salinity and drought stress conditions. Expression vector constructed was transferred into Arabidopsis thaliana to research the function.Sequence analysis shows PeZFP gene is 495bp and codes 164 amino acids. The A20 domain is sited between 16 and 50 amino acids, while AN1 domain is sited between 102 and 145 amino acid. The homological percentage among the znic finger proteins of Oryza sativa, Sorghum bicolor and Zea mays is above 90%; PeMYB gene is 1766bp and codes 427 amino acids. The sequence consisted two MYB domains which was in 13-63 amino acid and 66-114 amino acid. PeMYB gene was a classic R2R3 MYB transcriptional factor;The results of semi-quantitative RT-PCR indicated PeZFP and PeMYB showed different inducible expression under 200 m mol/L NaCl and drought stresses in different time, while the expression of them were lasting;In order to research better the function of PeZFP gene, the pCAMBIA1301 harboring sense and antisense- PeZFP gene was transformed into Arabidopsis thaliana by Agrobacterium-mediated method. We obtained the lines transformed with PeZFP gene selected through Hygromycin, DNA and RNA. The results of semi-quantitative RT-PCR showed overexpression of gene was in lines with sense- PeZFP gene, improving the transgenic plants tolerance to high salinity and drought. Expression of gene was in lines with antisense- PeZFP gene was restricted.The function and regulation of znic finger protein was analyzed with transgenic plants, on the one hand, it can further identify molecular regulation and mechanism of znic finger protein in Phyllostachys edulis, on another hand it also selects the resources could tolerance stress and spread them.