Molecular Cloning Of BcWRKY1 Transcriptional Factor Gene From Boea Crassifolia Hems1 And Its Preliminary Functional Analysis

The WRKY proteins, mainly present in plants, constitute a super family of transcription factors. They are extensively involved in some developmental and metabolic processes, as well as in biotic and abiotic stress responses in plants. In this paper, The BcWRKY1 gene partial cDNA sequence has been cloned by RT-PCR from Boea crassifolia Hemsl that have the trait of resurrection plant. The BcWRKY1 gene full sequence have been cloned by RACE (Rapid Amplification of cDNA Ends), the sequence multiple alignment analysis, gene revolution analysis and the primary expression research have been done; the functions of BcWRKY1 gene in the plant have been discussed.The cDNA sequences that code a protein belong to WRKY1 subgroup which contains two conserved domains--WRKYGQK and the C2H2 motif is about 444 amino acids. The full long of BcWRKY1 cDNA is about 1803bp. It shares low identity with other WRKY proteins, indicating a new member of WRKY family.The multiple alignment of nucleotide between BcWRKY1 and members of WRKY1 subgroup of WRKY transcriptional factor super family, the result of multiple alignment between BcWRKY1 and members of WRKY1 subgroup showed that the comparability of cDNA full long between BcWRKY1 and WRKY3 is highest, the rate is 43.7%.and the rates of 5'and 3' conserved domains are 80% and 83%. But the BcWRKY1 get the highest score of the comparability when aligned with SPF1 at the 3' conserved domain. According to the conclusion that the 3' WRKY conserved domain play a key role in the process of the WRKY transcriptional factor binding with the W-box, so we conferred BcWRKY1 and SPF 1 belonged to the same evolution generation.The result of Southern blot indicated there may have two copies of BcWRKY1 gene in the Boea crassifolia Hemsl genomic DNA. The BcWRKY1 can be induced by abiotic stress such as drought,low temperature,salty and signal molecular as SA,JA,MeJA and ABA. The expression behaviors of BcWRKY1 gene are different by different inducing conditions. The BcWRKY1 can be induced intensively and especially by150mmol/L NaCL.The endogen and dicotyledonous plant expression vectors p3300-35S-Tnos and p3300-ubi-tnos had been constructed for further research. The BcWRKY1 gene will be transformed into different plant for the research of the gene expression behaviors in different target plant.