花粉管通道法转EPSPS基因创制耐草甘膦玉米种质  

Glyphosate-tolerant Maize Plants Generated by Pollen Tube Pathway Method

作　　者：李向龙[1] 郑登俞 张春[1] 张中保[1] 吴忠义[1] LI Xianglong;ZHENG Dengyu;ZHANG Chun;ZHANG Zhongbao;WU Zhongyi(Institute of Biotechnology,Beijing Academy of Agriculture and Forestry Sciences/Beijing Key Laboratory of Agricultural Genetic Resources and Biotechnology,Beijing 100097,China)

机构地区：[1]北京市农林科学院生物技术研究所/农业基因资源与生物技术北京市重点实验室,北京100097

出　　处：《福建农业学报》2023年第5期524-529,共6页Fujian Journal of Agricultural Sciences

基　　金：北京市农林科学院科技创新能力建设专项(KJCX20230603)。

摘　　要：【目的】对T_(0)-T_(3)代转基因植株进行抗草甘膦EPSPS基因的筛选检测,以期得到遗传稳定的转基因株系。【方法】利用花粉管通道法将EPSPS基因转入优良自交系京92中,通过田间草甘膦筛选和分子检测鉴定外源基因在世代间的遗传表达。【结果】在田间用200 mg·L^(-1)的草甘膦除草剂筛选,T_(0)代15个抗性植株经PCR鉴定,共获得10个阳性植株,转化率为1.03%;对各株系从T_(1)代到T_(3)代逐代筛选淘汰,株系内阳性株数逐渐增多,在T_(3)代5个株系中分离出K3和K8两个整合了EPSPS外源基因的稳定遗传株系;进一步对K3和K8两个株系的T_(3)代采用试纸条进行功能表达分析,发现目的基因均能正常表达。【结论】K3和K8两个转基因玉米株系后期可作为培育耐草甘膦的基础材料。【Objective】Stable transgenic glyphosate-tolerant strains of maize were obtained by screening T_(0)-T_(3) generations of the genetically modified plants for the presence of EPSPS after an artificial exposure to glyphosate in the field.【Method】The gene,EPSPS,responsible for a maize plant to resist the herbicide was transferred into the excellent inbred line Jing 92 by means of the pollen tube pathway method.Expression of the introduced gene in maize was verified by molecular detection after exposing to glyphosate in the field.【Result】Fifteen resistant plants from T_(0) generation of the transgenic maize plants were identified after a treatment of 200 mg·L^(-1) glyphosate,and 10 by PCR.The genetic transformation rate was 1.03%.The identified plants were further scrutinized to eliminate non-resistant-gene-carriers.The proportion of confirmed carriers among the plants increased from the T_(1) to the T_(3) generation.Out of 5 strains in T_(3) generation,K3 and K8 were confirmed to have positively integrated EPSPS and be hereditarily stable.A test strip functional analysis on the two strains further verified the presence of the target gene.【Conclusion】The K3 and K8 transgenic lines identified by this study could plausibly be used to breed glyphosate-tolerant maize plants for cultivation.

关 键 词：玉米 花粉管通道法 EPSPS基因 草甘膦耐性 

分 类 号：S513[农业科学—作物学]