中文摘要：本文研发了具有多个抗虫和耐盐基因的新型转基因杨树。作者将两个抗虫Bt基因Cry1Ac、Cry3A和耐盐基因BADH导入载体p209-Cry1Ac-Cry3A-BADH中，且通过农杆菌介导方法来转化该载体对欧美杨的克隆，并采用遗传检测和抗性表达分析对三种转基因株系进行了评估。PCR结果显示，在三种转基因株系种都存在外源基因Cry1Ac、Cry3A、BADH和选择标记基因NPTII。实时定量PCR显示，不同的转基因株系间三个外源基因的转录丰度有差异显著。Bt毒蛋白的检测结果表明，每一种转基因株系中Cry1Ac和Cry3A毒蛋白的含量分别为12.83-26.32和2108.91-2724.79 ng/g；转基因株系间Cry1Ac毒蛋白的含量明显不同，Cry3A毒蛋白含量约比Cry1Ac毒蛋白含量高100倍。昆虫抗药性的试验显示，转基因株系美国白蛾幼虫的死亡率明显高于非转基因株系，为42.2-66.7%；柳蓝叶甲L1和L2幼虫的死亡率为100%。转基因株系对柳蓝叶甲幼虫的杀虫效果明显高于美国白蛾幼虫。另外，在3-6%的NaCl浓度下三种转基因株系NaCl忍耐胁迫显著高于非转基因株系。
外文摘要：We characterized new transgenic varieties of poplar with multiple insect-resistant and salt stress tolerant genes. Two insect-resistant Bacillus thuringiensis (Bt) genes, Cry1Ac and Cry3A, and a salt-tolerant gene, Betaine aldehyde dehydrogenase (BADH) were inserted into a vector, p209-Cry1Ac-Cry3A-BADH. The clone of Populus x euramericana was transformed by the vector using the Agrobacterium-mediated method. Three transgenic lines were assessed using genetic detection and resistance expression analysis. PCR revealed that exogenous genes Cry1Ac, Cry3A, BADH and selective marker gene NPTII were present in three transgenic lines. Quantitative realtime PCR (qPCR) showed significant differences in the transcriptional abundance of three exogenous genes in different lines. Results of assays for Bt toxic proteins showed that the Cry1Ac and Cry3A toxic protein content of each line was 12.83-26.32 and 2108.91-2724.79 ng/g, respectively. The Cry1Ac toxic protein content of different lines was significantly different; the Cry3A toxic protein content was about 100 times higher than that of the Cry1Ac toxic protein. The insect-resistance test revealed the mortality rate of transgenic lines to Hyphantria cunea L1 larvae varied by 42.2-66.7%, which was significantly higher than non-transgenic lines. The mortality rate of L1 and L2 Plagiodera versicolora larvae was 100%. The insecticidal effect of transgenic lines to P. versicolora larvae was higher than that to H. cunea larvae. NaCl stress tolerance of three transgenic lines under 3-6% NaCl concentration was significantly higher than that of non-transgenic lines.
外文关键词：Transgenic poplar；Multiple transgenes；Populus x euramericana；Insect-resistance；Salt tolerance
作者：Yang, R. L；Wang, A. X；Zhang, J；等
作者单位：河北农业大学
期刊名称：GENETICS AND MOLECULAR RESEARCH
期刊影响因子：0.764
出版年份：2016
出版刊次：15
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