| Sexual hybridization was studied in this article,choosing the White Poplar clones transformed two insect-resistant genes (Bt and API) which had different resistant as female parent and 84K Poplar which had choiceness character as male parent. In the end, we had got crossbreed strip.The inheretance and expression of foreign genes(marker gene NPt II and BtCry I Ac) and the relationship of two foreign genes were studied with materials of different crossbreed strip. Theory basis and original material to the utilization of transgenic polar and hibridization were provided according to exploring the regularity , stability and the relationship of foreign genes . The progress was as followed:l.With five years observation to the nature White Polar Hybrid 741 (Polar 741),the results showed that Polar 741 was lower than Chinese White Polar in fringe length, fringe thick, fringe number and capsule number per meter. The observation indicated that the fringe of Chinese White Polar fell off 5-10% before they crazed when they were ripe .And Polar 741 fell off 70-90% .So the Polar 741 did not fly garrulous basically, and reduced the feasibility of genetic drift of foreign genes greatly.The two insect-resistance genes transformed Polar 741 got seed hardly. The main reason was that the embryo of crossbreed stopped growthing and disappeared after three weeks growth. It lead to lower seed rate, at the same time most of the fringe fell off before they crazed when they were ripe. So the embryo rescue techniques was an effective method to conquer the steril of outbred cross . The optimal medium for culture immature embryo was MS+6-BA0.1mg/L.2. The experiment proved that the fastness bud rooted in the culture medium with 50mg·L-1Kan was a better course to filtrate .We could draw a conclusion that the cross offspring whether or not to contain Neomycin phosphotransferase according to the radication condition of tender stem in culture medium with 50mg·L-1Kan.We selected 41 offspring clony of 5 cross combination which growth better and used it Kan radication filtrate. The result showed that did not fit the Mendel's Law(1:1);And in the offspring of cross combination PB1× 84K that fit the Mendel's Law(1:1);And in the offspring of cross combination PB6x84K that did not fit the Mendel's Law(1:1).The veracity of experiment's should be validate because of little clony.3. In this experiment, sexual hybridization was studied,choosing 741 Poplar with twoinsect-resistant genes which had different resistant as female parent and 84K Poplar which had choiceness character as male parent. And it got 73 clony.Polymerase Chain Reaction (PCR) technique was used to investigate the inheritance and expression of the foreign genes BtCrylAc and Npt II of the 73 clony . The results showed that the segregation ratio of positive to negative plant in F| colony was 1.70 I 1 of the foreign gene BtCrylAc , which did not fit the Mendel's Law (1-1) by X2 test.Meanwhile the segregation ratio of positive to negative plant in the F| of PBl (who crossed with 84K Poplar) colony wasl : 1 of the foreign gene BtCrylAc , which fit the Mendel's Law (1 : 1) by X2 test,and the segregation ratio of positive to negative plant in Ft of PB6 colony was 2.43 I 1 of the foreign gene BtCrylAc, which did not fit the Mendel's Law (1 : 1) by X2 test.Polymerase Chain Reaction (PCR) technique was used to investigate the inheritance and expression of the foreign gene Npt II of the 73 clony. The results showed that the segregation ratio of positive to negative plant in Fi colony was 1.92 .' 1 of the foreign gene NptW , which did not fit the Mendel's Law (1 '. 1) by X2test.Meanwhile the segregation ratio of positive to negative plant in the F| of PBl (who crossed with 84K Poplar) colony wasl .' 1 of the foreign gene Npt II , which fit the Mendel's Law (1 .' 1) by X2 test,and the segregation ratio of positive to negative plant in F| of PB6 colony was 3.8 .' 1 of the foreign gene Npt II , which did not fit the Mendel's Law (1 .' 1) by X2 test.The two foreign genes BtCrylAc and Npt II 's the segregation ratio of positive to negative plant in the Fi of PB6 (who crossed with 84K) colony was not 1 ' 1 and approached 3 ." 1 ,which fit the Mendel's Law (3 ." 1) by Attest.T-DNA tabled gene of Neomycin phosphotransferase (Nptll) in Bt transgenic polar, but the cross separateing was occurred in offspring and the change rate was 2.7%.The study provided some basis for the application of transgenic technique in genetic melioration and some theoretical basis for the application and popularization of transgenic Poplar. |
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