The Study On The Technique Of Pollen Tube Path Way Used To Introduce Insect-resistant And Disease-resistant Gene Into Soybeans

In order to study the optimum introducing period, survival rate and inversion rate, thetechnique of pollen tube path way was used to introduce insect-resistant and disease-resistant gene into soybeans. PCR was also done to test the Dland D2 generations of transgenic plants. All these have proved that the technique of pollen tube path is feasible to study soybean.The results showed that:1.The way of drop of liquid was used to introduce chitinase and Bt gene into 27 soybean varieties. The totaliniroduced flower counts was 1667 and the pods obtained was 712. The mean value of survival rate was 42.7%.2.ln the following year, 15 transgenic varieties among 27 of Dl generation were planted. The total number of seeds was 1060 and the number of maturity plants was 676. The mean rate of maturity plants was 63.8%.3 .The suviral rate and matarity rate of different genes soybeans and different transmiting genes have no obvious differnce. The main effect factor exist in the operation technique.4.X-Gluc was used to test the plants of Dl generation of Bt-transgenic plants. 132 were done and no positive reaction was found.5.The DNA of the young leaves of 132 Bt transgenic plants was extracted to do PCR Five positive inversion plants were obtained and the inversion rate was 3.8%.6.The DNA of fire positive inversion plants obtained in Dl generation was extracted and tested. Two stable positive inversion plants were obtained in D2 generations.7 The way of flourescent mirror flat was used to observe the pollen tube extension circumstance in the flower post after self-pollination. 1) According to flower bud, we can choose the optimum ones to transfer gene by pollen tube path way 2) When the tube is used in genetic lrunsformation, the reason of the low inversion rate is that soybeans seed capsule is smell and the pouen is slim,comparing with other plants (such as cotton,cucumber and tomato etc),so the DNAis difficult to permeate into.The experiment shows that pollen-tube pathway method is fully feasible in soybeans plants,and it provides a practical and feasible pathway of genetic transformation to create varieties of new soybeans insect-resistantand disease resistan.Postgraduate:Cui Yan Supervisor: Yang QingkaiMajor:Crop genetic and breeding...