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The Relationship between Insect Resistance and Tree Age of Transgenic Triploid Populus tomentosa Plants

机译:转三倍体毛白杨植物的抗虫性与树龄的关系

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摘要

To explore the stability of insect resistance during the development of transgenic insect-resistant trees, this study investigated how insect resistance changes as transgenic trees age. We selected 19 transgenic insect-resistant triploid Populus tomentosa lines as plant material. The presence of exogenous genes and Cry1Ac protein expression were verified using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) analyses. The toxicity for Clostera anachoreta and Lymantria dispar was evaluated by feeding fresh leaves to first instar larvae after the trees were planted in the field for 2 years and after the sixth year. Results of PCR showed that the exogenous genes had a long-term presence in the poplar genome. ELISA analyses showed significant differences existed on the 6-year-old transgenic lines. The insect-feeding experiment demonstrated significant differences in the mortality rates of C. anachoreta and L. dispar among different transgenic lines. The average corrected mortality rates of C. anachoreta and L. dispar ranged from 5.6–98.7% to 35.4–7.2% respectively. The larval mortality rates differed significantly between the lines at different ages. Up to 52.6% of 1-year-old transgenic lines and 42.1% of 2-year-old transgenic lines caused C. anachoreta larval mortality rates to exceed 80%, whereas only 26.3% of the 6-year-old transgenic lines. The mortality rates of L. dispar exhibited the same trend: 89.5% of 1-year-old transgenic lines and 84.2% of 2-year-old transgenic lines caused L. dispar larval mortality rates to exceed 80%; this number decreased to 63.2% for the 6-year-old plants. The proportion of 6-year-old trees with over 80% larval mortality rates was clearly lower than that of the younger trees. The death distribution of C. anachoreta in different developmental stages also showed the larvae that fed on the leaves of 1-year-old trees were killed mostly during L1 and L2 stages, whereas the proportion of larvae that died in L3 and L4 stages was significantly increased when fed on leaves of 6-year-old trees. Results of correlation analysis showed there was a significant correlation between the larvae mortality rates of trees at different ages, as well as between Cry1Ac protein contents and larvae mortality rates of 6-year-old trees.
机译:为了探索在转基因抗虫树发育过程中昆虫抗性的稳定性,本研究调查了昆虫抗性如何随着转基因树龄的变化而变化。我们选择了19种转基因抗昆虫三倍体毛白杨(Populus tomentosa)系作为植物材料。使用聚合酶链反应(PCR)和酶联免疫吸附测定(ELISA)分析验证了外源基因和Cry1Ac蛋白表达的存在。在田间种植树木两年后和第六年后,通过将新鲜叶子喂入第一龄幼虫来评估对无梭梭菌和狼尾草的毒性。 PCR结果表明,外源基因在杨树基因组中具有长期存在。 ELISA分析显示6岁的转基因品系存在显着差异。昆虫饲养实验表明,在不同的转基因品系中,C。anachoreta和L. dispar的死亡率存在显着差异。 C. anachoreta和L. dispar的平均校正死亡率分别为5.6–98.7%至35.4–7.2%。不同年龄的品系之间的幼虫死亡率有显着差异。高达52.6%的1年龄转基因品系和42.1%的2年龄转基因品系导致无翅念珠菌幼虫死亡率超过80%,而只有66.3年转基因品系的死亡率高达26.3%。 Dispar的死亡率呈现出相同的趋势:1年龄转基因品系占89.5%,2年龄转基因品系占84.2%,导致Dispar幼虫死亡率超过80%。对于使用6年的植物,这一数字下降到63.2%。幼虫死亡率超过80%的6岁树的比例明显低于年轻树。 C. anachoreta在不同发育阶段的死亡分布也表明,以1年生树木的叶片为食的幼虫大部分在L1和L2阶段被杀死,而在L3和L4阶段死亡的幼虫比例显着以6岁树的叶子为食时,其增加。相关分析结果表明,不同年龄树木的幼虫死亡率之间,以及6岁龄树木的Cry1Ac蛋白含量与幼虫死亡率之间存在显着相关性。

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