Isolation And Characterization Of Three WRKY Transcription Factors From Hevea Brasiliensis

WRKY trancription factors constitute a gene superfamily in higher plants, which are not only widely involved in plant responsed to biotic and abiotic stresses, but also involved in the growth, development and regulation of various metabolic pathways. In this paper, three WRKY trancription factors were cloned from Hevea brasiliensis(PR107), which were named HbWRKY041, HbWRKY043and HbWRKY045, based on this, we had analysised their sequence features, subcellular localization and expression regulation characteristics.1. The full-length sequence of HbWRKY041, HbWRKY043and HbWRKY045were cloned by RT-PCR from Heveabrasiliensis which translated three WRKY transcription factors that have full sequence of471bp、606bp and605bp, the sequence contain438bp、567bp and588bp open read frame(ORF) which encode145、188and195amino acids;2. The bioinformatics were analysised for the3trancription factors that contained a highly conserved sequence(WRKYGQK) and a zinc finger(C2H2), belonging to the second class in WRKY transcription factor family, which was conserved during evolution;3. The green fluorescent protein fused expression vectors of HbWRKY041, HbWRKY043and HbWRKY045were constructed and transformed into tobacco plants using agrobacterium-mediated transient expression. The results indicated that three transcription factors were all localized in the nucleus;4. The gene expression patterns of HbWRKY041, HbWRKY043and HbWRKY045were studied by real time RT-PCR technology during different tissues and developmental stages leaves, different phytohormone and stress treatments.All of the three genes showed different expression patterns, both HbWRKY041and HbWRKY043with predominance expression in leaves, HbWRKY045had predominance expression in roots followed by leaves; During different developmental stages leaves, they all had predominance expression in senescence leaf;The expressions of HbWRKY043and HbWRKY045were up-regulated by ETH、SA and MeJA treatment, but decreased by ABA, The expressions of HbWRKY041were up-regulated by ABA and SA, decreased by ETH and MeJA;Under oxidative stress conditions, the expressions of three genes were all increased; HbWRKY043was up-regulated by salt stress, whereas HbWRKY041and HbWRKY045were not affected; HbWRKY043and HbWRKY045were affected by drought, the expressions of was decreased firstly then went down; HbWRKY041and HbWRKY045were increased by wounding, but HbWRKY043was not affected.