The Application And Research Of Herbicides Screening On Wheat Transgenic Plants

In the process of genetic transformation of wheat, bar gene is one of the most commonly used and effective selection marker gene. Using the herbicide resistance characteristics of bar gene, which can eliminate a lot of cells of non-transformant and improve the filtering efficiency of resistant regeneration plant in the process of vitro regeneration of immature embryo callus. In the process of homozygous and stable transformation of offspring of transgenetic plants, its needs a generational screening appraisal, eliminates non-transformant and it does not express and obtains stable transgenic plants. This experiment aimed at three excellent transformation receptor of wheat genotypes, selecting a appropriate concentration of herbicide sensitive and handling way in the process of regeneration of young embryo and plant. To establish a simple and intuitive conversion method of plant screening, and provide a reference for wheat genetic transformation and regeneration of plant.(1) Using the lunxuan987, zhengmai7698 and jimai19 as materials, and the herbicides of different concentrations have been used to screen immature embryo callus in the stage of differentiation and planting. The results indicated that: the critical concentration in these two stages are 1mg/L, and 5mg/L in rooting stage, the rate of inhibion are 85%, 84% and 82.5% respectively. The effects among these concentrations showed significcant difference. Between each type, zhengmai7698 and jimai19 showed no sianificant difference, but there are significant diffferences when compared to lunxuan987.(2) To the above three wheat varieties of indoor and field seedling, different concentrateons of herbicides have been used to daub and spray leaf and plant respectively. The results showed that the critical concentrations used for spray and seeds soaking of indoor seedling are 200mg/L and 120mg/L, the rate of inhibition of seedling germination was reaching out of 90%, while the rate of yellowing or necrosis of plant was reaching out of 83.3%; while the critical concentrations used for daub and spray of field plants are both of 80mg/l, the rate of yellowing or necrosis was reaching out of 80%.(3) PCR test has been used to indentify the positive plant of T1,T2 and T3 generations, And the herbicides as the screening agent to validate the intensity of resistance. The results indicated that 9 of 44 positive plant showed strong resistance, others showed yellowing or necrosis in different degrees.