[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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转Bt基因欧洲黑杨转化TA29-Barnase基因的研究()
《西北林学院学报》[ISSN:1001-7461/CN:61-1202/S]
- 卷:
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第32卷
- 期数:
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2017年第1期
- 页码:
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121-125
- 栏目:
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- 出版日期:
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2017-01-31
文章信息/Info
- Title:
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Transformation of T29-Barnase Gene of Bt Transgenic Populus nigra
- 文章编号:
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1001-7461(2017)01-0121-05
- 作者:
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朱伟康; 贾小明*; 李周岐
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(西北农林科技大学 林学院,陕西 杨陵 712100)
- Author(s):
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ZHU Wei-kang; JIA Xiao-ming*; LI Zhou-qi
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(College of Forestry,Northwest A&F University,Yangling,Shaanxi 712100,China)
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- 关键词:
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转Bt基因欧洲黑杨; Barnase基因; 转化
- Keywords:
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Bt transnenic Populus nigra; Barnase gene; transform
- 分类号:
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S792.119
- DOI:
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10.3969/j.issn.1001-7461.2017.01.19
- 文献标志码:
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A
- 摘要:
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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:
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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:
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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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