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用部分改造BtCrylAc基因与慈菇蛋白酶抑制剂 (API)基因构建的双抗虫基因表达载体 ,通过农杆菌介导法转化了 741毛白杨 [PopulusalbaL .× (P .davidianaDode +P .simoniiCarr .)×P .tomentosaCarr .]。本文报道了选择转化植株的条件。在含卡那霉素 (Km’)的生根培养基中初步选择到转化植株 ,PCR检测表明 80 %的植株呈现阳性反应。对初步选择的转基因植株用杨扇舟蛾 (ClosteraanachoretaFabricius)和舞毒蛾 (LymantriadisparLinnacus)幼虫进行饲虫试验。其结果按照死亡率划分了高、中、低和对照 4类昆虫死亡率的植株 ,并对这 4类的定期死亡率、生长发育的延迟性后效应等进行了比较研究 ,为选择具有显著抗虫性的植株 (无性系 )提供依据。分子生物学检测结果 ,证明被选择植株 (无性系 )的基因已转入 ,且生长发育正常 ,占参试饲虫试验植株的 38 1 %
The Populus alba L. [Populus alba L. × (P. davidiana Dode + P. Simiiiiiiiiiiiiiiiiiiii) × Agrobacterium tumefaciens was transformed with a double insect gene expression vector partially modified by BtCrylAc gene and arrowhead protease inhibitor (API) P. TomentosaCarr.]. This article reports the conditions for selecting transformed plants. Transformed plants were initially selected in rooting media containing kanamycin (Km ’), and PCR assays showed that 80% of the plants showed a positive reaction. Insect tests were conducted on the initially selected transgenic plants with larvae of the genus Clostera anachoreta Fabrius and Lymantriadis par Linnus. The results were divided into high, medium, low and control four kinds of insect mortality according to the mortality rate of plants, and these four types of regular mortality, delayed development and development after a comparative study were carried out to select for a significant anti Insect-resistant plants (clones) provide the basis. Molecular biology test results showed that the selected plants (clones) genes have been transferred into, and the growth and development of normal, accounted for 38.1% of test insects test plants,