Xu Chen, Wei Hui, Wang Like, Yin Tongming, Zhuge Qiang
Co-Innovation Center for Sustainable Forestry in Southern China, Key Laboratory of Forest Genetics and Biotechnology, Ministry of Education, College of Biology and the Environment, Nanjing Forestry University, Nanjing, China.
Jiangsu Provincial Key Construction Laboratory of Special Biomass Resource Utilization, Nanjing Xiaozhuang University, Nanjing, China.
Front Plant Sci. 2019 Mar 26;10:335. doi: 10.3389/fpls.2019.00335. eCollection 2019.
Increased expression of the insect control protein genes of in has been critical to the development of genetically improved plants with agronomically acceptable levels of insect resistance. () proteins with highly specific toxicity against were screened using an indoor bioactivity assay to obtain hyper-resistant transgenic poplars. Then, the sequence was optimized and transformed according to the optimal codon in poplar using software of our own design (http://120.79.60.226:8080/u/chen/w/codonpoplar). A vector was constructed to transform poplar NL895. The gene was transformed to poplar NL895 and six transgenic lines were obtained. The expression and insecticidal effect of the gene in transgenic poplar were evaluated by PCR and ELISA, and the specific indoor activity and field insecticidal activity against . were compared with a control. We concluded that the insecticidal activity of the transgenic NL895 was significantly better against lower instar larvae of than against higher instar larvae. The mortality and pupation rates clearly differed among the various instar larvae and between transgenic and non-transgenic poplar. We obtained poplar seedlings with hyper-resistance to . by screening Bt genes and optimizing their genetic sequence.
苏云金芽孢杆菌(Bt)杀虫蛋白基因的过量表达对于培育具有农艺学上可接受的抗虫水平的转基因植物至关重要。利用室内生物活性测定法筛选对杨树蛀干害虫具有高度特异性毒性的Bt蛋白,以获得高抗转基因杨树。然后,使用我们自己设计的软件(http://120.79.60.226:8080/u/chen/w/codonpoplar)根据杨树中的最优密码子对Bt序列进行优化并转化。构建载体转化杨树NL895。将Bt基因转化到杨树NL895中,获得了6个转基因株系。通过PCR和ELISA评估Bt基因在转基因杨树中的表达和杀虫效果,并将其对杨树蛀干害虫的室内特异性活性和田间杀虫活性与对照进行比较。我们得出结论,转基因NL895对杨树蛀干害虫低龄幼虫的杀虫活性明显优于高龄幼虫。不同龄期幼虫以及转基因杨树和非转基因杨树之间的死亡率和化蛹率明显不同。通过筛选Bt基因并优化其基因序列,我们获得了对杨树蛀干害虫具有高抗性的杨树幼苗。