Differential Expression Analysis Of Stripe Rust-Resistance Related Genes And Gene Cloning

Stripe rust in wheat is one of the most important diseases. It could lead to grain output reduced in large-scale and cause a serious threat to Chinese food security. Using stripe rust-resistant varieties is the best way to control the disease. Currently, most of the stripe rust-resistant genes of disease-resistant varieties are unknown, So that it is difficult to achieve rational distribution of the rust-resistant varieties. Therefore, understanding disease-resistant gene expression, disease-resistant mechanisms and the search for new disease-resistant genes in molecular level are very important to access the development of stable and lasting disease-resistant varieties. cDNA-AFLP, Subtractive Stress Hybridization, Semi-QRT-PCR, In silico cloning and bio-informatics technologies were combined to research germplasm N95175 with stripe rust resistance and analysis of the Stripe rust resistance related gene expression. The main results are as follows :(1) cDNA-AFLP technology was adopted in analysis of stripe rust-resistance related genes expression in N95175. Using BLASTN software and BLASTX software separately to analyze EST sequences in the GenBank dbEST database and protein sequence databases in NCBI. Based on the sequences homologous comparison and function querying, the results showed: one of nine ESTs was high homologous (>86%) with pathogen infection EST; three ESTs were high homologous with zinc finger protein, transcriptional regulator and transposable element which were presumed to related to stripe rust resistance; three ESTs were high homologous with hypothetical protein and unnamed protein which functions were still need further study.(2) Suppression subtractive hybridization was adopted to analyze stripe rust-resistance related genes differential expression in N95175.EST sequences were analyzed by BLASTN software. The results indicated that the ESTs were high homologous with biotic and abiotic intimidate (contains pathogen-infection, salt-treated, hormone-treated, oxidatively-stressed, drought-stressed and trace elements...