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为提高马铃薯品种(系)地上部分糖苷生物碱(SGAs)的含量,增强其抗逆性并改良块茎的品质。克隆了马铃薯茄啶鼠李糖基转移酶基因(sgt3)cDNA和1,5–二磷酸核酮糖羧化酶小亚基基因启动子(rbcS P);对sgt3亚细胞定位预测显示,该蛋白不具有叶绿体转运肽、线粒体导肽和分泌信号肽序列,推测可能位于细胞质;将克隆的sgt3 cDNA片段及rbcS重组到pCEPSP载体上,构建了具有草甘膦抗性标记的绿色组织特异表达sgt3基因植物表达载体。通过农杆菌介导法对马铃薯品种‘陇薯3号’和‘夏波蒂’进行转化,共获得了12株抗草甘膦的阳性转基因植株。对转基因植株的目的基因表达水平和SGAs含量分析发现,地上部sgt3基因相对表达量较未转化植株提高1.3~3.0倍,SGAs含量增加20%~37%,而转基因植株的块茎中SGAs含量变化不显著。本研究的结果为进一步培育枝、叶中高SGAs而块茎中低SGAs的马铃薯抗性品种提供了理论依据。
In order to improve the content of glycoside alkaloids (SGAs) in potato aboveground, enhance their resistance and improve tuber quality. The srs3 cDNA and the ribulose-1, 5-bisphosphate carboxylase small subunit gene promoter (rbcS P) were cloned. The prediction of sgt3 subcellular localization showed that the protein It is speculated that it may be located in the cytoplasm. The cloned sgt3 cDNA fragment and rbcS were recombined into pCEPSP vector to construct green tissue-specific sgt3 gene with glyphosate resistance marker Plant expression vector. Agrobacterium-mediated transformation of potato varieties ’Longshu 3’ and ’Xia Boti’ were transformed, a total of 12 glyphosate-resistant transgenic plants were obtained. Analysis of target gene expression level and SGAs content in transgenic plants showed that the relative expression level of sgt3 in shoots increased by 1.3-3.0 times and that of SGAs increased by 20% -37% compared with that of untransformed plants. However, the content of SGAs in tubers of transgenic plants did not change Significant. The results of this study provide a theoretical basis for further cultivating potato varieties with low SGAs in tuber and leaves with high SGAs.