转基因杨树嫁接苗中外源Bt-cry3A基因的表达及其产物运输  

Expression and Products Transportation of Bt-cry3A Gene in the Grafted Seedlings of Transgenic Poplar

作　　者：王连荣[1] 薛拥志[2] 张德健[3] 甄志先 张军 Wang Lianrong;Xue Yongzhi;Zhang Dejian;Zhen Zhixian;Zhang Jun(College of Agriculture and Forestry Science and Technology,Hebei North University,Zhangjiakou,075131;College of Animal Science and Technology,Hebei North University,Zhangjiakou,075131;School of Life Sciences,Inner Mongolia University,Hohhot,010021;Hebei Key Laboratory for Tree Genetic Resources and Forest Protection,College of Forestry Science,Hebei Agricultural University,Baoding,071000)

机构地区：[1]河北北方学院农林科技学院,张家口075131 [2]河北北方学院动物科技学院,张家口075131 [3]内蒙古大学生命科学学院,呼和浩特010021 [4]河北农业大学林学院,河北省林木种质资源与森林保护重点实验室,保定071000

出　　处：《分子植物育种》2021年第9期2906-2911,共6页Molecular Plant Breeding

基　　金：国家自然基金项目(31660211);河北省基础研究计划项目(18966801D)共同资助。

摘　　要：为探索外源Bt-cry3A基因在转基因杨树嫁接苗中的表达情况以及其表达产物在嫁接砧木和接穗间的运输机制,本试验以转Bt-cry3A基因741杨高抗株系CC71与未转基因的741杨、毛白杨嫁接的1 a生嫁接苗为材料,利用RT-PCR技术研究Bt-cry3A基因的mRNA的表达与运输规律,结果表明同砧嫁接杨树苗中CC71部位外源Bt-cry3A基因均能表达出mRNA,非转基因741杨部位未检测到Bt-cry3A基因的mRNA;利用ELISA技术研究Bt-Cry3A蛋白的表达与运输规律,结果表明同砧嫁接、异砧嫁接和中间砧嫁接的杨树苗,CC71部位的韧皮部、木质部、髓以及叶片中均能表达出Bt-Cry3A蛋白,同时,未转基因的741杨、毛白杨部位的叶片、韧皮部、木质部和髓中也检测出Bt-Cry3A蛋白的存在,但检测到的量很低。本试验研究结果表明外源Bt-cry3A基因的mRNA不能在转基因嫁接杨树苗的砧木和接穗间进行长距离的运输,Bt-Cry3A蛋白可以在砧木和接穗间运输,但运输的量极低。此研究结果为合理安全利用转Bt-cry3A基因杨树提供了理论依据。In the study,expression of Bt-cry3A gene and the mode of migration of its expression products between rootstock and scion were investigated in the grafted seedlings of transgenic poplar.Experimental materials were 1 a-grafted poplar seedlings that were grafted using CC71(Transgenic poplar 741 with Bt-cry3A gene),non-transgenic poplar 741 and non-transgenic Populous tomentosa,either as rootstocks or as scions.Expression and migration of mRNA of Bt-cry3A gene were studied by using RT-PCR technology.The results showed that mRNA of Bt-cry3A gene of homoplastic grafted poplar seedling was detected in the CC71 positions and was not detected in non-transgenic poplar 741 positions.ELISA technology was used to study the expression and migration of Bt-Cry3A protein.The results showed that Bt-Cry3A protein were detected in the phloem,xylem,pith and leaves of the CC71 positions of the grafted poplar seedlings with homoplastic graft,heteroplastic graft and interstock graft,were also detected that of non-transgenic poplar 741 and non-transgenic Populous tomentosa at the same time,but the amount of Bt-Cry3A protein was very low.The results showed that the mRNA of Bt-cry3A gene could not be transported between the rootstock and scion of the grafted poplar seedlings,and Bt-cry3A protein could be transported between the rootstock and scion,but the amount of transportation was very low.The results provide a theoretical basis for the rational and safe use of transgenic poplar with Bt-cry3A gene.

关 键 词：转基因杨树 嫁接苗 Cry3A蛋白 MRNA 运输 

分 类 号：S792.11[农业科学—林木遗传育种]