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转双抗虫基因741毛白杨的选择及抗虫性

SELECTION AND INSECT RESISTANCE OF TRANSGENIC HYBRID POPLAR 741 CARRYING TWO INSECT\|RESISTANT GENES

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【作者】 郑均宝梁海永高宝嘉王永芳田颖川

【Author】 Zheng Junbao 1) \ Liang Haiyong 2) \ Gao Baojia 4) \ Wang Yongfang 5) (Hebei Agricultural University Baoding 071000) Tian Yingchuan 3) (Institute of Microbiology, Academia sinica Beijing100080)

【机构】 河北农业大学林学院!保定071000中国科学院微生物研究所!北京100080

【摘要】 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 %

【Abstract】 Explants of hybrid poplar 741\%\[Populus alba\% L.×(\%P.davidiana \%Dode+\%P.simonii\% Carr.)×\%P.tomentosa\% Carr.\] were transformed with Agrobacterium tumefaciens LBA4404 harboring the expression vector pBtiA containing two insect\|resistant genes. Conditions for selection of poplar transforments, especially the critical concentrations of Kanamycin(Km’) were determined. 80% of the transforments selected on rooting medium containing Km’ gave out positive results by PCR detection. The gene\|transformed plants preliminaries selected were given to insect feeding tests with \%Clostera anachoreta \%Fabricius and \%Lymantria dispar\% Linnacus.And the tested insects were divided into four groups, high, middle, low and control according to mortality. In the meantime, the insect mortality in fixed period and the effects of delaying the growth and development of insects were comparatively studied to provide the basis for selecting plants or clones, which significantly resist to insect. The results of detailed insect\|bioassay and molecular biology analysis demonstrate that the two foreign genes have been integrated into the genomes of insect resistant plants. Transforments with normal phenotype, growth rate as that of the non\|transformed control and high insect\|resistant were selected and accounted for 38.1% of the transformed tested.

【基金】 河北省自然科学基金;河北省教委基金;中国科学院院长基金;联合国环境规划署基金
  • 【文献出处】 林业科学 ,SCIENTIA SILVAE SINICAE , 编辑部邮箱 ,2000年02期
  • 【分类号】S792.117
  • 【被引频次】223
  • 【下载频次】285
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