Co-Transferring Of Transcription Factors Related To Disease Resistance And Replicase Gene Into Wheat And Cloning Of Gene Responsive To Adverse Abiotic Stresses

Transcription factors play vital roles in signal transduction, expression and modulation of related functional genes in plants. ERF-like transcription factors participate in the defense response to pathogens invasion and abiotic stresses by interacting with GCC-binding elements, and DREB transcription factors respond to abiotic stresses by binding to DRE/CRT elements. To create new wheat germplasms with comprehensive disease resistances, broad disease-resistant ERF-like transcription factors and wheat yellow mosaic virus (WYMV) replicase genes were transferred into wheat through microprojectile bombardment in this study. At the same time, several DREB-like transcription factors related to abiotic stresses were also isolated to provide potential gene sources for crop improvement on the resistance to abiotic stresses using engineering technique.1. By molecular detection and disease resistance test in field, T6 transgenic plants in which Yangmai 158 was used as receptor with WYMV replicase gene Nib8 were to be homozygous or stable in both genetics and disease resistance.2. Nib8 gene and broad disease-resistant ERF gene W17 were integrated into high-efficient and constitutive expression vector of monocots and then co-transferred into wheat varieties of Yangmai 12 and Yangmai 16 through microprojectile bombardment. As a result, 42 plants with Nib8, 48 with W17 and 6 with both were achieved.3. High-efficient and constitutive expression vector for monocots with W17 and selective marker gene Bar were constructed and then co-transferred into wheat varieties of Yangmai 12 and Yangmai 16 through microprojectile bombardment. In total, 17 individuals with W17 were obtained.4. Thirty-nine gene fragments encoding DREB-like transcription factors were spliced using electric cloning technique. The homologous analysis on amino acid revealed that the fragments can be grouped into 4 families. Among them, 10 DREB-like transcription factors were chose to analyze the expression character, and the result showed that their expression can be induced by high salinity, low temperature, dehydration, and ABA. Further analysis on tissue expression character manifested that the expression level was the highest in roots, the middle in leaves, and the lowest in stems. 5. Sub-celluar localization of two full-length DREB transcription factors, DRF72 and DRF100, displayed that DRF100 can reach the nucleus by the aid of its NLS, while DRF72 can be localized into cytoplasm and nucleus. So we concluded that DRF72 may direct into nucleus by integrating with other transcription factors.