Study Ansterring Ghd7Gene Into Wheat And The Effect Of Wheat Seedling With Agrobacterium Tumefaciens Soaking Root

Wheat is one of the major food crops in the world. However, wheat is the last successfulfood crops by means of genetic engineering.currently, the mainly way for wheattransformation is particle bombardment, Agrobacterium-mediated and pollen tube pathway..With the reduction in arable land area and population increase, the demand for wheatproduction is very greatly. The genes Ghd7cloning from rice, simultaneous control of riceplant height, grain number and growth period, this genes plays an important role to controlrice yield, therefore, this study intends to put Ghd7into wheat.and improve wheat productiontraits.Using Huanghuai Wheat planting varieties as material,by pollen tube pathway andparticle bombardment for genetic transformation of wheat and preliminary study aboutAgrobacterium dip genetic transformation system, obtained the following results:1, Using Wheat varieties Zhengmai9023, Xiaoyan22and Xinong1376as materials,immature embryo as explants, imported Ghd7into wheat genome by particle bombardment,and examined their offspring by PCR. The results showed that1140survived in176resistantseedlings after transplanting, and140resistant plants have nine positive plants, six of whichwere Zhengmai9023,3strains were Xinong1376, there were not positive plants in Xiaoyan22.the conversion efficiency was about0.225%.Using Mature embryo as receptortransformated of wheat by gene gun, compared domestic split film with imported split filmquality,we found they were stable after conversion of the wheat, there were some differencesbetween the wheat callus rate and survival rate, but the differences were not significant.2, Using wheat varieties Xinong979,Xinong889and Lunxuan9871as materials,imported Ghd7into wheat genome through the pollen tube pathway,harvested contemporaryseed about1650, extremed the positive plants by PCR.the T1generation have11positiveplants,Xinong979have seven positive plants,Lunxuan9871have three positive plants andXinong889have one positive plants.tested the T2generation again and found that only a partof plant contained the positive plants, a portion of the target gene was lost, which shows thepollen tube pathway method is feasible and instability. 3,The Effect of Physiological and Biochemical Characteristics of Wheat Seedling withAgrobacterium tumefaciens Soaking Root, It was found that the growth rate, root activity,POD activity and SOD activity of Xinong889were higher than Xinong979and Zhengmai9023; with the increasing of root length by Agrobacterium tumefaciens soaking, the rootgrowth rate, seedling root length, seedling fresh weight, root activity, and POD activityincreased as well, while the seedling height showed a downward trend, MDA contentincreased when root length increasing in the range of4-7cm, and in the range of7-10cm theMDA content did not change significantly; compared with the control, when Agrobacteriumtumefaciens concentration were at0.5OD, wheat root growth rate, seedling fresh weight, androot activity increased slightly, with the increasing of concentration of Agrobacteriumtumefaciens, wheat root growth rate, seedling fresh weight, and root activity significantlydecreased, while MDA levels had been increasing, POD activity and SOD activity increasedand then showed a downward trend. According to the effect of soaking root treatment withAgrobacterium tumefaciens to the main physiological and biochemical characteristics ofwheat seedling, the Agrobacterium tumefaciens bacterial concentration of0.5OD and rootlength of10cm, are appropriate for soaking root transformation, and appropriate genotype forsoaking root is Xinong889.