摘要
本研究以转Bt基因‘南林895’杨12个株系扦插苗为试验材料,分析其对美国白蛾和杨小舟蛾的虫害抗性。利用PCR、ELISA等技术对转抗虫基因杨树进行分子检测,同时在室内控制条件下对美国白蛾和杨小舟蛾进行抗虫性分析。PCR分析结果显示在12个转基因株系中均检测到了Bt目的基因片段。ELISA分析结果显示Bt毒蛋白浓度范围为371.9~10 698.2 ng/g,且不同株系间存在一定差异。转Bt基因杨树饲虫分析结果表明,12个转基因株系对美国白蛾和杨小舟蛾均有一定的杀虫活性,其中转基因株系A-4-6、A-5-0、A-3-4、A-5-23、Z-1-3的抗虫性较显著。株系A-5-0对杨小舟蛾1龄幼虫12 d校正死亡率达87.2%,对美国白蛾的18 d校正死亡率为65.6%。且转Bt基因‘南林895’杨对美国白蛾和杨小舟蛾幼虫的取食和生长有明显的抑制作用。在对12个转Bt基因株系的分子检测和饲虫试验中,Bt毒蛋白的表达水平存在差异且抗虫效果与Bt毒蛋白浓度的高低有关。
In this study, 12 cuttings of Bt-transgenic poplar lines(NL-895) were used as experimental materials to analyze their resistance to Hyphantria cunea and Micromelalopha troglodyta. Polymerase chain reaction(PCR) and ELISA were used for molecular detection of the transgenic poplars. The insect resistance of Hyphantria cunea and Micromelalopha troglodyta were analyzed under indoor control conditions. The results of PCR analysis showed that Bt target gene fragments were detected in 12 transgenic lines. ELISA analysis showed that the concentration of Bt toxin protein ranged from 371.9 to 10 698.2 ng/g, and there were some differences among different strains. The results of insecticidal analysis of Bt-transgenic poplar feeding insects showed that 12 transgenic lines had certain insecticidal activity against Hyphantria cunea and Micromelalopha troglodyta, among which the transgenic lines A-4-6, A-5-0, A-3-4, A-5-23 and Z-1-3 had significant insecticidal resistance. The 12 day corrected mortality of strain A-5-0 to the first instar larvae of Micromelalopha troglodyta reached 87.2%, and 65.6% to Hyphantria cunea at the 18 day corrected mortality. Transgenic poplars significantly inhibited the feeding and growth of the larvae of Micromelalopha troglodyta and Hyphantria cunea, and the feeding and weight of the larvae of Micromelalopha troglodyta and Hyphantria cunea were significantly lower than those of the control. In the molecular detection and feeding test of 12 Bt transgenic strains, the expression level of Bt toxin protein was different and the effect of insect resistance was related to the concentration of Bt toxin protein.
作者
王璞
郭同斌
魏辉
孙伟博
诸葛强
Wang Pu;Guo Tongbin;Wei Hui;Sun Weibo;Zhuge Qiang(Key Laboratory of Forest Genetics and Biotechnology of Ministry of Education,Co-Innovation Center for Sustainable Forestry in Southern China,College of Biology and the Environment,Nanjing Forestry University,Nanjing,210037;Xuzhou Forestry Technology Extension Service Center,Xuzhou,221004)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第14期4645-4656,共12页
Molecular Plant Breeding
基金
转基因生物新品种培育重大专项(2018ZX08020002)
江苏高校优势学科建设工程项目(PAPD)共同资助。
