| 转cry2Ah-vp基因玉米的抗虫性鉴定 |
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| 引用本文：李梦桃,李圣彦,汪海,张杰,苍晶,郎志宏.转cry2Ah-vp基因玉米的抗虫性鉴定.植物保护学报,2020,47(1):74-83 |
| DOI：10.13802/j.cnki.zwbhxb.2020.2019043 |
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| 中文摘要:为获得新型抗虫转基因玉米，将通过群体筛选获得的具有抗虫性的转cry2Ah-vp基因玉米VP1-5采用PCR、Southern blot、实时荧光定量PCR（qPCR）、酶联免疫吸附测定（ELISA）等方法进行阳性植株鉴定、拷贝数分析、转录水平和翻译水平分析，同时通过室内和田间生物活性测定鉴定转基因玉米VP1-5对东方黏虫Mythimna separata和亚洲玉米螟Ostrinia furnacalis的抗性。结果表明，在转基因玉米VP1-5中cry2Ah-vp基因已整合到玉米基因组，以单拷贝的形式插入；cry2Ah-vp基因在转基因玉米VP1-5不同部位组织中均可以正常转录，在灌浆期叶片中的mRNA表达量最高，相对表达量为32.67，在灌浆期穗轴中的mRNA表达量最低，相对表达量为3.74；Cry2Ah-vp蛋白在转基因玉米VP1-5的6叶期各组织中表达量均较高，其中在叶片中的表达量达到2 155.18 ng/g FW，在抽雄期花丝中的表达量最高，达到2 165.86 ng/g FW；且转基因玉米VP1-5对东方黏虫有很高的杀虫活性，接虫3 d后幼虫死亡率达到100.00%；对亚洲玉米螟幼虫也有明显的生长抑制作用。表明转基因玉米VP1-5可作为玉米抗虫育种和害虫防治的种质资源。 |
| 中文关键词:害虫防治 苏云金芽胞杆菌 cry2Ah-vp基因 转基因玉米 东方黏虫 亚洲玉米螟 |
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| Identification of insect resistance in the transgenic maize harboring cry2Ah-vp gene |
| Author Name | Affiliation | E-mail | | LI Mengtao | College of Life Sciences, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China
Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China | | | LI Shengyan | Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China | | | WANG Hai | Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China | | | ZHANG Jie | State Key Laboratory for Biology of Plant Diseases and Insects Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China | | | CANG Jing | College of Life Sciences, Northeast Agricultural University, Harbin 150030, Heilongjiang Province, China | cangjing2003@163.com | | LANG Zhihong | Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China | langzhihong@caas.cn |
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| Abstract:To obtain the novel insect-resistant maize, the insect-resistant transgenic event VP1-5 harboring cry2Ah-vp gene was detected for the foreign gene insertion, copy number, transcriptional and translational expression of cry2Ah-vp gene by using PCR, Southern blot, real-time fluorescence quantitative PCR (qPCR) and enzyme-linked immunosorbent assay (ELISA), and the bioassay in the laboratory and field was conducted to identify the resistance of VP1-5 transgenic maize to Mythimna separata and Ostrinia furnacalis. The results showed that cry2Ah-vp gene was integrated into maize genome with a single copy in transgenic maize VP1-5. The cry2Ah-vp gene was correctly transcribed in different tissues showing the highest expression in leaf and the lowest expression in cob during grain-filling stage, and the relative mRNA level were 32.67 and 3.74, respectively. The Cry2Ah-vp protein was expressed relatively high in all tissues at six-leaf stage, especially in leaf where the expression was 2 155.18 ng/g FW. Overall, the highest expression of Cry2Ah-vp protein was 2 165.86 ng/g FW in silk at tassel stage. VP1-5 transgenic maize was demonstrated to have high insecticidal toxicity towards armyworm by the fact that the mortality of armyworm larvae reached 100.00% 3 d after inoculation; VP1-5 obviously inhibited the weight gain of the corn borer larvae. This study indicated that VP1-5 transgenic maize could be a good germplasm resource for insect resistance and pest control. |
| keywords:pest control Bacillus thuringiensis cry2Ah-vp gene transgenic maize Mythimna separata Ostrinia furnacalis |
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