TvNHX1基因对矮牵牛的遗传转化  

Transformation of Petunia hybrida with TvNHX1 Gene

作　　者：崔继哲[1] 乔磊[1] 弭晓菊[1] 付寅生[1] 李想[1] 杨慧[1] 

机构地区：[1]哈尔滨师范大学生命科学与技术学院,黑龙江哈尔滨150025

出　　处：《黑龙江农业科学》2012年第12期14-18,共5页Heilongjiang Agricultural Sciences

基　　金：黑龙江省教育厅科技研究资助项目(11551144)

摘　　要：为获得耐盐碱特性的矮牵牛新种质,提高其在广大盐碱地区的适应性,以矮牵牛品种梦幻兰和利宝紫为试材,简要介绍了TvNHX1基因的植物表达载体构建及用农杆菌介导法将TvNHX1基因转化矮牵牛的初步结果,并分析了6-BA和NAA组合对矮牵牛叶片分化的影响。结果表明:对PCR筛选得到的阳性菌落摇菌、提取质粒后进行BamHI/SacI双酶切鉴定,在1 640bp左右得到了单一条带,与预期一致。证明pBI121-TvNHX1构建成功;在不同浓度激素组合的培养基中,梦幻兰的分化率都高于利宝紫,梦幻兰品种在6-BA为2.0mg.L-1、NAA为0.2mg.L-1时分化率达90.69%;以梦幻兰品种的叶片为外植体诱导生根,得到12株在筛选培养基上正常生根的矮牵牛苗,以TvNHX1基因特异引物WFF和JWR1对获得的Kan抗性植株和非转基因对照进行PCR检测,12个抗性植株中7株扩增出特异的条带,说明TvNHX1基因已转入矮牵牛。In order to obtain new germplasm of saline-alkaline tolerance Petunia hybrida ,improve its adaptabili- ty in the saline area,taking varieties of Petunia hybrida Menghuanlan and Libaozi as test materials,the TvN- HX1 gene construction of plant expression vector and transformation results of Petunia hybrida using agrobacterium-mediated method with TvNHX1 gene were briefly introduced, and the influence of the 6-BA and NAA combinations on P. hybrid leaf differentiation were analyzed. The result of BamHI/SacI double enzyme i- dentification showed that there was a single band in 1 640 bp or so,which was consistent with the expected. It proved that pBI121-TvNHX1 building success. In different concentration hormone combinations of medium, the differentiation rate of Menghuanlan was higher than that of Libaozi. The differentiation rate of Menghuan- lan was 90.69% in 6-BA 2.0 mg-L-1 ,NAA 0.2 mg.L-1. Taking leaves of Menghuanlan as explants to induced root, got 12 seedlings of normal rooted P, hybrid in screening medium. Kan resistant plant and the non-trans- genic control were PCR detected by TvNHX1 gene specific primer WFF and JWR1,7 plants amplified specific bands of 12 resistance plants,which displaying that TvNHX1 gene had turned into P. hybrid.

关 键 词：矮牵牛 TvNHX1基因 遗传转化 

分 类 号：S681.9[农业科学—观赏园艺]