Cloning And Functional Identification Of DgWRKY5 And DgWRKY4 In Chrysanthemum

As one of the ten traditional famous flowers in China, 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.In this experiment, chrysanthemum as experimental material to obtain a database based on the new high-throughput sequencing technology Illumina HiSeq 2000 chrysanthemum transcriptome sequencing, screened out a complete open reading frame of the gene, the use of in silico cloning and RT-PCR cloning to obtain new the WRKY transcription factor, named DgWRKY5 and DgWRKY4,successfully obtained DgWRKY5 transgenic chrysanthemum plants and DgWRKY4 transgenic tobacco plants. Sequence analysis showed that:DgWRKY5 open reading frame of 1603bp, encoding 540 amino acid residues, the gene encoding the protein molecular 60.0KDa, isoelectric point of 7.07; For gene transfer DgWRKY5 be daisy salt stress test:found the blade than the wild-type transgenic chrysanthemum daisy yellow leaves and wilting, and chlorophyll content decreased.Sequence analysis showed that:DgWRKY4 open reading frame of 1534bp, encoding 482 amino acid residues, the gene encoding protein molecular weight 53.6KDa, an isoelectric point of 7.35;On genetically modified tobacco DgWRKY4 phenotype observed:Under normal circumstances, the wild-type gene and four transfer DgWRKY4 lines throughout the life cycle of no significant difference in phenotype; on genetically modified tobacco DgWRKY4 salt stress test carried out in 150mM NaClo under salt stress, transgenic tobacco germination rate (58-60%) was significantly higher than that of wild-type tobacco (25%).