| Wheat is one of the world’s major food crops, wheat production often suffered the harm of high salt, drought and low temperature and other abiotic stresses.Biotechnology study and understanding of the genes, resulting in a bio-genetically modified technology.Plant transgenic technology uses recombinant DNA technology, excellent target genes into plant cells or tissues for expression, to obtain plants with new traits. Use of transgenic technology, to improve the quality of wheat breeding drought, salinity, frosting the new varieties of wheat, has become one of the means of wheat quality improvement.Stress of drought, high salt and low temperature, plants can sense and pass the stress signal, induced the expression of functional genes. Transcription factors play an important role in stress signal transmission, the regulatory function of gene expression. The transcription factor OsCBF1from rice, to belong AP2/EREBP family of transcription factors, mainly involved in plant drought, salt, low temperature stress response. Transformation of the resilience function of the genes is also another important aspect to improve plant resilience. Mangrin salt tolerance genes from mangrove plants sexangula.This gene encodes a protein of265amino acid residues.16-86of70amino acids from the protein composition similar to the LEA protein is rich in serine region. LEA proteins rich in serine region play a role to protect the cell membrane structure and protein in the plant stress.In this study,OsCBFl,Mangrin of a cDNA construct expression vectors YM7252, YM7262.genes gun-mediated method transcription factor OsCBFl and functional genes Mangrin conversion to wheat. Molecular detection and resistance identification of future generations of transgenic plants to filter out the good resistance of transgenic plants. To expect to improve the resilience of the wheat to get new varieties of drought, salt and cold. In this study, the following results:1.Strss resistance genes in wheat genetic transformation(1differences in wheat genotypes is one of the important factors affecting wheat tissue culture regeneration. Under the same experimental conditions, transformation of immature embryo callus number Ningmai13less than the conversion of nong33wheat callus. Ningmai13seedling number115. The nong33failed to differentiate into plants.(2) The same genotype Ningmai13immature embryo as the acceptor material. Conversion OsCBF1gene115To generation plants.PCR preliminary test positive for a total of nine plants. transformation of the gene of wheat immature embryo differentiation rate of6.65%, the positive conversion rate of0.52%.Conversion Mangrin genes a total of40To progeny were identified by PCR and preliminary inspection of the five positive plants, conversion of the gene of wheat immature embryo differentiation rate of8.23%, the positive conversion rate of1.02%.2.Transfer the gene expression of the gene T1plants and resistance detectionOsCBF1gene into the wheat immature embryo115To generation plants, which survived and the seeds of positive plants9, PCR detection of RNA expression levels, the conversion of7gene was expressed in9OsCBF1gene positive plants. Real-time quantitative results were expressed in the T1generation OsCBF1gene into the wheat, but different expression levels in different plants. Compared with the control group, transgenic plants N13-CBF38, N13-CBF89relative expression levels of more than290times, the transgenic plants55relative expression of about72.1times, N13-CBF105of the transgenic plants relative expression level was about16.5times, other transgenic plants express about three times. physiological and experimental results of T1transgenic plants:(1) the proline experimental results show that:N13-CBF36、N13-CBF86, N13-CBF87, N13-CBF89, N13-CBF105proline content of the transgenic plants was significantly higher than that in the control group instructions exogenous transformation the factor OsCBFl antiretroviral increase wheat’s resistance.(2) malondialdehyde experimental results show that:MDA content in transgenic plants N13-CBF38> N13-CBF55, N13-CBF87, N13-CBF89, N13-CBF105accumulated at different times for the two control groups, indicating that the conversion of exogenous gene OsCBFl of resistance genes to increase the resistance of wheat.(3) superoxide dismutase experimental results show that:the transgenic plants N13-CBF38、 N13-CBF55, N13-CBF87, N13-CBF89superoxide dismutase activity was significantly higher than the two control groups, indicating that the conversion of exogenous genes to increase plant resistance, these two groups transgenic experimental group measured the MDA content was also significantly lower than the control group, superoxide dismutase activity may be one of the factors affecting the content of MDA.(4) the peroxidase experimental results show that:N13-CBF38of the transgenic plants, the peroxidase activity is significantly higher than the two control groups.3Mangrin function gene transformation of immature wheat embryos and40To generation plants by PCR Initial examination identified a total of five positive plants, now planted in experimental fields, pending further identification positive and resistant physiology... |
PDF Full Text Request