[1]朱伟康,贾小明*,李周岐.转Bt基因欧洲黑杨转化TA29-Barnase基因的研究[J].西北林学院学报,2017,32(1):121-125.[doi:10.3969/j.issn.1001-7461.2017.01.19]
 ZHU Wei-kang,JIA Xiao-ming*,LI Zhou-qi. Transformation of T29-Barnase Gene of Bt Transgenic Populus nigra[J].JOURNAL OF NORTHWEST FORESTRY UNIVERSITY,2017,32(1):121-125.[doi:10.3969/j.issn.1001-7461.2017.01.19]
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Bt基因欧洲黑杨转化TA29-Barnase基因的研究()
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《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]

卷:
第32卷
期数:
2017年第1期
页码:
121-125
栏目:
出版日期:
2017-01-31

文章信息/Info

Title:
 Transformation of T29-Barnase Gene of Bt Transgenic Populus nigra
文章编号:
1001-7461(2017)01-0121-05
作者:
 朱伟康贾小明*李周岐
 (西北农林科技大学 林学院,陕西 杨陵 712100)
Author(s):
 ZHU Wei-kangJIA Xiao-ming*LI Zhou-qi
 (College of Forestry,Northwest A&F University,Yangling,Shaanxi 712100,China)
关键词:
Bt基因欧洲黑杨Barnase基因转化
Keywords:
 Bt transnenic Populus nigra Barnase gene transform
分类号:
S792.119
DOI:
10.3969/j.issn.1001-7461.2017.01.19
文献标志码:
A
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+0.05 mg·L-1NAA+0.01 mg·L-1 TDZ+30 g·L-1蔗糖+6 g·L-1琼脂;不定芽生根最适培养基为:1/2MS+0.05 mg·L-1 NAA +0.2 mg·L-1 IBA+20 g·L-1蔗糖+6 g·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。
Abstract:
 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.

备注/Memo

备注/Memo:
 收稿日期:2016-03-22修回日期:2016-06-12
基金项目: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)。
作者简介:朱伟康,男,硕士研究生,研究方向:林木生物技术。E-mail:578094061@qq.com
*通信作者:贾小明,男,讲师,研究方向:林木生物技术。E-mail: jxm0601@163.com
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