Preliminary Research On One Tomato Gene Involved In Genetic Transformation And Disease Resistance

Tomato resistance gene contains coiled-coil structure (CC), nucleotide-binding site (NBS), and Leucine-rich repeat domain (LRR) is a maximum resistance gene family in plants, the majority of R genes have nucleotide binding site (NBS) and Leucine-rich repeat domain (LRR). CC-NBS-LRR domains are important structural of R genes, disease-resistant plants to play an important role. Bacterial wilt, early blight, late blight, fusarium wilt, leaf mold and other diseases of tomato production has brought devastating impact, but adverse external environment and a variety of pathogenic bacteria will usually cause some damage to plant growth and development. Plant has formed a complete self-protection mechanisms in the long process of evolution to resist pathogen invasion and damage of adverse external environment. Studies have shown that plant disease resistance R gene can identify a pathogen-induced anaphylaxis (Hypersensitive response, HR), thereby improving plant disease resistance.In this study, obtained by yeast two-hybrid screening approach a DDB1IP9 gene interacte with important DDB1 gene of tomato, and by Sol Genomics Network and NCBI (National Center for Biotechnology Information Search database) information compare, containing CC-NBS-LRR domain, predicted to have R gene resistance, so by overexpression transgenic in tomato to further explore. Molecular cloning has a good effect of expression through a strong promoter CaMV 35S, and DDB1IP9 build on pBI121 carrier, and get pBI121::DDB1IP9 recombinant vector constructed overexpression vector of DDB1IP9 expression by phylloxera Agrobacterium-mediated transformation into wild-type tomato, which were screened DDB1IP9 high expression of positive overexpression lines. And by constructing tobacco overexpression vector PBTEX::35S::DDB1IP9-Flag. Simultaneous resistance Experiment, T1 generation of transgenic positive DDB1IP9 transgenic and wild-type tomato AC inoculated Pst.DC3000, tomato plants were observed and compared the apparent disease situation and the relatively flat dash calculate the number of colonies 1cm2 difference with position of the blade. By pathogen inoculation experiments, the preliminary analysis can be found in tomato DDB1IP9 antiviral function.The results of this study show that, DDB1IP9 transgenic plants was significantly lower than the incidence of wild-type AC tomato, the incidence of more serious wild-type plants; drilling sampling 1cm2 colonies found of wild-type AC is 2-4 times DDB1IP9 transgene, indicating DDB1IP9 genetically modified to have a significantly higher Prt.DC3000 resistant wild tomato, with obvious resistance. Therefore, over-expression DDB1IP9 gene can increase tomato Pst.DC3000 resistance, improve disease resistance in tomato, tomato planting with a good economic value, make a new attempt to use genetic engineering methods modified tomato disease resistance, but also with tomato DDB1 have interaction, functional pathways DDB1 research can provide a new direction.