CUI Ji-zhe,QIAO Lei,MI Xiao-ju,et al.Transformation of Petunia hybrida with TvNHX1 Gene[J].HEILONGJIANG AGRICULTURAL SCIENCES,2012,(12):14-18.
TvNHX1基因对矮牵牛的遗传转化
- Title:
- Transformation of Petunia hybrida with TvNHX1 Gene
- 文章编号:
- 1002-2767（2012）12-0014-05
- Keywords:
- Petunia hybrida; TvNHX1 gene; genetic transformation
- 分类号:
- S681.6
- 文献标志码:
- A
- 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1 640 bp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2.0 mg·L-1、NAA为0.2 mg·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。
- Abstract:
- In order to obtain new germplasm of saline-alkaline tolerance Petunia hybrida,improve its adaptability in the saline area,taking varieties of Petunia hybridaMenghuanlan and Libaozi as test materials,the TvNHX1 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 identification 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 Menghuanlan 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-transgenic control were PCR detected by TvNHX1gene specific primer WFF and JWR1,7 plants amplified specific bands of 12 resistance plants，which displaying that TvNHX1 gene had turned into P.hybrid.
参考文献/References:
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备注/Memo
基金项目：黑龙江省教育厅科技研究资助项目（11551144）