转Bt基因‘南林895’杨时空表达及生物安全分析  

Spatio-temporal Expression and Biosafety Analysis of Transgenic'Nanlin-895'Poplar with Bt

作　　者：周培军 李玲玲 李红岩 魏辉 王璞 孙伟博 诸葛强 Zhou Peijun;Li Lingling;Li Hongyan;Wei Hui;Wang Pu;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)

机构地区：[1]南京林业大学生物与环境学院,南方现代林业协同创新中心,林木遗传与生物技术教育部重点实验室,南京210037

出　　处：《分子植物育种》2022年第5期1568-1580,共13页Molecular Plant Breeding

基　　金：转基因生物新品种培育重大专项(2018ZX08020002);国家林业局生物安全与遗传资源管理项目(KJZXSA-2018004);江苏高校优势学科建设工程项目(PAPD)共同资助。

摘　　要：为探究田间种植的转Bt基因‘南林895’杨对周围生态系统的影响以及Bt基因的时空表达特征。本研究利用PCR技术检测了外源基因的稳定性,结果显示外源基因Bt依然存在于转基因杨树中,且没有发生基因水平转移现象;利用RT-qPCR和ELISA技术分别检测了Bt基因在转录和翻译水平的时空表达量,结果显示Bt基因的转录和翻译水平均在时间上呈现出明显的季节性变化,而在空间上表达变化规律不显著;利用田间测量、稀释平板培养和Sandwich等方法分别检测了转Bt基因杨树的生长、对根系周围可培养土壤微生物数量的影响及其化感活性,结果显示转基因杨树与非转基因杨树之间无显著性差异。初步检测结果表明转Bt基因‘南林895’杨没有对周围生态系统造成显著影响。本研究为今后该转Bt基因抗虫杨树的环境释放提供了坚实基础,为建立和完善转基因杨树的安全性评价体系提供了理论依据。The purpose of this study was to investigate the effects of the transgenic'Nanlin895'poplar with Bt planted in the field on the surrounding ecosystem and the temporal and spatial expression characteristics of Bt gene.In this study,PCR was used to detect the stability of exogenous genes.The results showed that exogenous genes Bt were still existed in the transgenic poplar without horizontal gene transfer.The temporal and spatial expression quantity of Bt gene in transcription and translation levels were detected by RT-qPCR and ELISA respectively.The results showed that the transcription and translation of Bt gene showed significant seasonal changes in time,but not significant changes in space.Field measurement,dilution plate culture and Sandwich methods were used to detect the growth of Bt transgenic poplar,the influence on the number of soil microorganisms culturable around the roots and the allelopathy activities,respectively.The results showed that there was no significant difference between transgenic poplar and non-transgenic poplar.Preliminary test results indicated that the transgenic'Nanlin895'poplar with Bt had no significant impact on the surrounding ecosystem.This study will provide a solid foundation for the environmental release of the transgenic insect-resistant poplar in the future and a theoretical basis for the establishment and improvement of the safety evaluation system of transgenic poplar.

关 键 词：杨树 时空表达 生物安全性 BT 

分 类 号：S792.11[农业科学—林木遗传育种]