Ren Yachao, Zhang Jun, Wang Guiying, Liu Xiaojie, Li Li, Wang Jinmao, Yang Minsheng
Forest Department, Forestry College, Hebei Agricultural University, Baoding, China.
Hebei Key Laboratory for Tree Genetic Resources and Forest Protection, Baoding, China.
Front Plant Sci. 2018 Jan 26;9:53. doi: 10.3389/fpls.2018.00053. eCollection 2018.
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 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 and 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 and among different transgenic lines. The average corrected mortality rates of and 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 larval mortality rates to exceed 80%, whereas only 26.3% of the 6-year-old transgenic lines. The mortality rates of exhibited the same trend: 89.5% of 1-year-old transgenic lines and 84.2% of 2-year-old transgenic lines caused 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 in different developmental stages also showed the larvae that fed on the leaves of 1-year-old trees were killed mostly during L and L stages, whereas the proportion of larvae that died in L and L 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.
为探究转基因抗虫树木发育过程中抗虫性的稳定性,本研究调查了抗虫性如何随转基因树木年龄的增长而变化。我们选择了19个转基因抗虫三倍体品系作为植物材料。使用聚合酶链反应(PCR)和酶联免疫吸附测定(ELISA)分析来验证外源基因的存在和Cry1Ac蛋白的表达。在树木种植于田间2年后和第6年后,通过向一龄幼虫投喂新鲜叶片来评估对[具体害虫1]和[具体害虫2]的毒性。PCR结果表明外源基因长期存在于杨树基因组中。ELISA分析表明6年生转基因品系存在显著差异。昆虫取食实验表明不同转基因品系对[具体害虫1]和[具体害虫2]的死亡率存在显著差异。[具体害虫1]和[具体害虫2]的平均校正死亡率分别为5.6 - 98.7%和35.4 - 7.2%。不同年龄品系的幼虫死亡率差异显著。1年生转基因品系中高达52.6%以及2年生转基因品系中42.1%导致[具体害虫1]幼虫死亡率超过80%,而6年生转基因品系中只有26.3%。[具体害虫2]的死亡率呈现相同趋势:1年生转基因品系中89.5%以及2年生转基因品系中84.2%导致[具体害虫2]幼虫死亡率超过80%;6年生植株这一比例降至63.2%。6年生树木幼虫死亡率超过80%的比例明显低于幼龄树木。[具体害虫1]在不同发育阶段的死亡分布还表明,取食1年生树木叶片的幼虫大多在L和L阶段死亡,而取食6年生树木叶片时,在L和L阶段死亡的幼虫比例显著增加。相关性分析结果表明,不同年龄树木的幼虫死亡率之间以及6年生树木的Cry1Ac蛋白含量与幼虫死亡率之间存在显著相关性。