The Insect-resistant Genes Transformation And Its Environmental Risk Assay Of The Key Shelter Forest Tree (Polulus Tomentasa) In The North Of China

P.tomentosa is the key shelter forest tree with nice biological characteristics.The sterile male P. tomentosa(NO.D) was used as materials of the insect-resistant genes transformation,its sterility and the environmental risk of transgenic poplar were studied.The purpose is to get new clone of sterile male P. tomentosa with the insect-resistance and environmental safety nature The results were shown as below.1.The anther morphology and histology of the sterile male P.tomentosa was observed for 4 years, from 2004 to 2007.The results is that there is no pollination step during the process of anthers growth. The mature anther was not able to split.The tapetum cell delayed disintegration.The sporule couldn't grow normall.2.Both shoots regeneration system and leaf regeneration systme were established by using its buds and leaf as explants seperately.3.50mg/L of kanamycin was the selection limite concentration for pBtiA transformation of the sterile male P.tomentosa screened by agrobacterium-medium transformation methos.4.21 Kan-resistant clones were obtained by agrobacterium-medium transformation method,they were identified by checking Bt gene and NPTâ…¡gene using PCR method,and 90%of kan-resistant clones were positive5.Feeding Clostera anachoreta(Fabricius) and Lymantria dispar(Linnaeus) with the young tender leaves of transgenic sterile male P.tomentosa were carried out for checking the biological effects. Toxicity of transgenic sterile male P.tomentosa was significantly higher than that of normal sterile male P.tomentosa.According to the insect total death rate,four high insect-resistance clones(D--84,D-87,D-94,D-95) and three middling insect-resistance clones(D-74,D-85,D-88 ) were sieved.Both high and middle insect-resistant clones had the negative effects on the insects growth and their development. The southern blot results of 3 clones,D-84,D-88 and D-95,were positive.6.The environmental risk of transgenic 741 poplar was studied.The species of the bacterium which were got from the rootage soil of the transgenic 741 poplus was much more than that of normal 741 poplar.The species of the fungi of the transgenic 741 poplus was less than that of normal 741 poplar. 7.Both of Bt gene and NPTâ…¡gene were not detected in 860 colonies of bacteria and 120 colonies of fungi isolated in the rhizosphere of transgenic 741 poplar by PCR method.