Research of the high temperature response core element and upstream-downstream regulators of tomato LeDREBA4 transcription factor

Tomato is a main kind of vegetable whose growth and development is affected by increasingly serious high temperature. DREB transcription factors play a significant role under high temperature and other abiotic stress. This study used the model system Micro-Tom tomato as experimental material and the previously discovered high-temperature response LeDREBA4 transcription factor as research object to firstly make clear its basic properties, including the characteristic of subcellular localization. Overexpression and VIGS (virus induced gene silencing) method was used to transform the Micro-Tom to analyze the function of LeDREBA4 under high temperature. Furthermore, the downstream target genes of LeDREBA4 were identified through chromatin immunoprecipitation combining high-throughput sequencing (ChIP-Seq). The high-temperature response core element in LeDREBA4 gene’s promoter was identified by using the promoter deletion and site-directed mutation approach. Based on the high-temperature response core element, the upstream regulatory factors of LeDREBA4 gene were obtained though the construction of yeast one-hybrid cDNA library from Micro-Tom tomato induced by high temperature, electrophoresis mobility shift assay (EMSA) and ChIP-PCR approach. The research results will demonstrate the function of LeDREBA4 under high temperature and its molecular regulation mechanism preliminarily. In addition, theoretical foundation and gene resources will be furnished to verify the underlying molecular mechanism by which tomato accommodate high temperature and breeders carry out gene engineering breeding under high temperature stress.