Cloning And Analysis Of Sliaa9 And Sliaa14 Gene Promoters On Tomato

The eukaryotic promoters have been the focus of modern molecular biology. Expression of different genes in plant is mainly regulated by different transcription factors at the transcriptional level. Therefore, it is necessary and valuable to research the structure and regulation mode of promoters.Tomato(Solanum lycopersicum)is a world wide planting vegetable. Meanwhile, it is also a model plant for plants genetic engineering researchs. A lot of basic research has been done on tomato genetics and the genetic transformation system. This can faciliate our research on auxin regulations.In this study, the upstream regulatory elements of the SlIAA9 and the SlIAA14 gene which is the Aux/IAA Family Members was cloned by us using PCR-based homologous sequence and genomic DNA walking method. The YFP-GUS fusion expression vectors and the overexpressing vector SlIAA9::SlIAA9were constructed and transfered into tomato.The analysis of SlIAA9 and SlIAA14gene. promoter Structural Characteristics and the Expression pattern had been made.The main results were as follows:1.1891 bp of PSlIAA9 promoter DNA fragment and 2197 bp promoter DNA fragment PSlIAA14 were got by PCR.Results of sequencing analysis showed that the region contains several putative cis-elements such as elements which respond to ABA, GA and CTK, as well as several elements responding to different environmental stresses, such as dehydration and wounding.2. An expression vector containing the PSlIAA14-YFP-GUS,PSlIAA9-YFP-GUS fusion gene and the overexpressing vector SlIAA9::SlIAA9 were constructed3.Transgenic plants were generated with the SlIAA9::GUS+YFP, SlIAA14::GUS+YFP and SlIAA9::SlIAA9 vectors mediated by Agrobacterium transformation into tomato genome with cotyledon explants.4. Tissue specific expression of the promoter driving YFP and GUS were observed in roots of transgenic plants which confirmed the promoter activity of PSlIAA9 and PSlIAA14-The Expression of SlIAA9::GUS+YFP is mainly in the leaf base and the lateral root primordia while the SlIAA14::GUS+YFP is mainly in the leaf veins and root tip meristem.5. Leaf morphology of SlIAA9::SlIAA9 transgenic plants were shown that the SlIAA9:: SlL4A9can not recovery the leaf morphology of entire mutant lines. Further Research should be made to find out the probable mechanism.