The Analysis Of TaCAD12 And TaPK-R1 Overexpress Transgenic Wheat Lines With Characterization And Function

Wheat is the main food crops in China. Wheat sharp eyespot is a soil-borne disease causing mainly by Rhizoctonia cerealis. Freezing is the one of the main natural disasters to affect the growth of wheat. Wheat cultivars resistant to sharp eyespot and freezing were very scarce, therefore it is important significance to that isolation and cloned resistance to anti-R. cerealis freezing injury genes, studies its function, providing theoretical basis for breeding of tolerance to both abiotic and biotic stress. In this study, we construction of the expression vector and embryo calli of wild-type were bombarded by the gold particle containing P20-Rssp2897:6MYC-TaCAD12 and pAHC25-MYC-TaPK-R1 vector DNA, achieved the TaCAD12 and TaPK-R1 transgenic wheat lines.The main results of TaCAD12 transgenic wheat lines are as follows:(1) Transgenic wheat plants in T0-T2 generations were subjected to PCR, and qRT-PCR analyses. Five transgenic wheat lines were obtained. (2)Western blot analysis showed that five transgenic wheat lines c-MYC-TaCAD12 fusion proteins were expressed. (3)Sharp eyespot severity assessments analysis showed that the transgenic wheat plants expressing TaCAD12 displayed significantly enhanced resistance to sharp eyespot compared with non-transgenic wheat WT.The main results of TaPK-R1 transgenic wheat lines are as follows:(1) Analyses of the transgenic wheat plants in T0-T4 generations with PCR, RT-PCR and qRT-PCR, and three transgenic wheat lines were obtained. (2)Western blot analysis showed that five transgenic wheat lines c-MYC-TaPK-R1 fusion proteins were expressed. (3) We were statistical its survival rate, observational of proline content and electrolyte leakage rate after freezing the transgenic wheat plants in T4 generations, the results showed that TaPK-R1 overexpression transgenic wheat lines adaptability to cold environment. (4) After inoculation with R. cerealis virulent-isolations R0301 and WK207, the sharp eyespot resistance tests showed that the transgenic wheat plants expressing TaPK-R1 displayed significantly enhanced resistance to sharp eyespot compared with non-transgenic wheat WT. (5) After inoculation with R. cerealis and freezing stress, three TaPK-R1 transgenic wheat lines defense sharp eyespot and freezing tolerance were obtained.