转Bt基因欧洲黑杨转化TA29-Barnase基因的研究  

Transformation of T29-Barnase Gene of Bt Transgenic Populus nigra

作　　者：朱伟康[1] 贾小明[1] 李周岐[1] 

机构地区：[1]西北农林科技大学林学院,陕西杨陵712100

出　　处：《西北林学院学报》2017年第1期121-125,149,共6页Journal of Northwest Forestry University

基　　金：国家林业局948项目"转基因林木生物安全控制及早期评价技术引进"(2013-4-38)

摘　　要：为了获得雄性不育转Bt基因欧洲黑杨,在以叶片为外植体建立转Bt基因欧洲黑杨叶片组培再生体系的基础上,利用农杆菌介导叶盘法将TA29-Barnase基因转化到转Bt基因欧洲黑杨中。结果表明,转Bt基因欧洲黑杨叶片不定芽分化最适培养基为:MS+0.5 mg·L^(-1) 6-BA+0.05mg·L^(-1) NAA+0.01mg·L^(-1) TDZ+30g·L^(-1)蔗糖+6g·L^(-1)琼脂;不定芽生根最适培养基为:1/2MS+0.05mg·L^(-1) NAA+0.2mg·L^(-1) IBA+20g·L^(-1)蔗糖+6g·L^(-1)琼脂;经过在叶片不定芽诱导及生根诱导培养基添加除草剂PPT连续筛选,共获得9株抗性植株。对抗性植株进行基因特异性PCR检测,其中有5株呈阳性,转化阳性率达55.6%。初步表明获得了转Barnase基因的转Bt基因欧洲黑杨植株。In order to obtain male sterility Bt transgenic Populus nigra ,we established the leaf regeneration system,and transformed TA29-Barnase gene into Bt transgenic P. nigra with Agrobacterium-mediated leaf disc. The results showed that the best differentiation medium of Bt transgenic P. nigra was MS＋0.5 mg·L^-1 6-BA＋0.05 mg·L^-1 NAA＋0.01 mg·L^-1 TDZ＋30 g ·L^-1 sucrose＋6 g·L^-1 agar,and the optimal rooting medium was 1/2MS＋0.05 mg·L^-1 NAA ＋0.2 mg·L^-1 IBA＋20 g·L^-1 sucrose＋6 g·L^-1 agar. Through successive selection in the stages of shooting and root induction with the addition of PPT （phosphinothricin）, 9 phosphinothricin-resistant plants were obtained. According to the TA29-Barnase sequence designing specific primers,5 positive plants were detected through PCR analysis with a conversion rate of 55.6 %. Preliminary result showed that the Barnase gene had been Successfully transformed into Bt transgenic P. nigra.

关 键 词：转Bt基因欧洲黑杨 Barnase基因 转化 

分 类 号：S792.119[农业科学—林木遗传育种]