Cloning And Functional Identification Of DgZFP5 And BZIP1 In Chrysanthemum

As one of the four most important cut flowers, chrysanthemum’s ornamental, medicinal and edible value have been known to the word. In addition, it contains a very high economic value. In the actual production of chrysanthemum, however, it is often adversely affected by environmental factors such as drought, high salt, etc. Nowadays, cultivating new varieties of chrysanthemum with strong stress tolerance is an urgent goals in chrysanthemum breeding.Either in plant growth, development or in the process of stress appropriate, C2H2 zinc finger protein and bZIP transcription factor plays an extremely important role. In this study, the chrysanthemum as experimental material, access the database based on the new high-throughput sequencing technology Illumina HiSeq 2000 Chrysanthemum transcriptome sequencing, screened out with a complete open reading frame of the gene, by the use of silico cloning and RT-PCR, we cloned a new C2H2 zinc finger protein and a bZIP transcription factor, named DgZFP5 and bZIP1. Sequence analysis showed that:The open reading frame of DgZFP5 is 450bp, encoding a protein of 149 amino acids residues, protein molecular weight of 16.7KDa and isoelectric point of 9.11. And the protein encoded by this gene contains two C2H2 zinc finger domains and an EAR-box (DLNL). bZIP1’s open rending frame is 417bp amino acids residues, encoding a protein of 138 amino acids residues, protein molecular weight of 16.363 KDa and isoelectric point of 9.50.We use DgZFP5 plasmid and plant expression vector pBI 121 to construct expression vector pBI121-DgZFP5, at the same time, bZIP1 plasmid and plant expression vector pCAMBIA 2300 was constructed to plant expression vector pCAMBIA 2300-bZIP 1 By means of Agrobacterium mediated transformation, we got DgZFP5 and bZIPl transgenic tobacco plants. Through the phenotype, under normal situation,we observed that there is no significant difference in DgZFP5-overexpression transgenic tobacco and wide type in phenotypic throughout the life cycle; Counter rotating DgZFP5 gene tobacco salt stress tests are carried out, In the 150mM NaCl salt stress conditions, transgenic tobacco germination rate (62-73%) was significantly higher than that of wild-type tobacco (28%). Under normal growth conditions, the wild-type gene and 6 turn bZIP1 lines throughout the life cycle without significant phenotypic differences; on genetically modified tobacco bZIPl salt stress tests conducted at 150mM NaCl salt stress, compared with wild-type tobacco, the germination rate of bZIPl gene transfer tobacco turn out 67-100%.