关键词: 抗虫转基因欧洲黑杨, Barnase基因, 雄性不育, 苏云金杆菌的晶体毒蛋白(Bt)基因, 花粉污染

Abstract:

Chimaeric TA29 Barnase gene was introduced into anti insect transgenic Populus nigra of good quality and high yield by Agrobacterium tumefaciens(A.t)and biolistics transformation from tender leaves of 15-n-192 tube plantlets, which growing at the medium 1/2MS+NAA0.2mg/L for 4～6 weeks, the leaves was cultured with MS+BA0.2mg/L+NAA0.02mg/L about 16 hours. The transformation frequency of Barnase gene by biolistics and A.t was 16 1 percent and 23 percent respectively, the number of repropagation plantlets were 175/12 and 71/10 respectively. The experiment showed that, the suitable developing leaves were selected, which was good for plantlets regeneration after Barnase gene transformation. Further more, we extracted the total DNA from transgenic plants by SDS method. The PCR analysis showed that, the DNA fragment from trangenic genome was amplified were completely identical with the Barnase gene; Southern Blot analysis showed that the transgenic plants genome carry about with Barnase gene fragment; Dot Blot analysis showed that the transgenic plants passes a cross spot clearly, and it was positive transformed plants further. The trans Barnasegenic plants of anti insect Populus nigra were obtained by the tissue culture and the methods of normal forest tree breeding.

Key words: The anti-insect transgenic Populus nigra, Barnase gene, Male sterility, Bacillus thuringiensis insecticidal crystal protein(B.t)gene, Pollen pollution