马铃薯地上部绿色组织中糖苷生物碱合成调控的研究  

Regulation of SGAs Biosynthesis of Green Tissues in Aerial Part of the Potato

作　　者：郭海霞[1] 张晶晶[2] 安然[1] 乔岩[1] 石文慧[1] 石菁[1] 张金文[1] 

机构地区：[1]甘肃农业大学农学院,甘肃省作物遗传改良与种质创新重点实验室,甘肃省干旱生境作物学重点实验室,兰州730070 [2]甘肃农业大学资源与环境学院,兰州730070

出　　处：《园艺学报》2017年第6期1105-1115,共11页Acta Horticulturae Sinica

基　　金：国家自然科学基金项目(31260343);甘肃省生物技术专项(GNSW-2015-11)

摘　　要：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 accumulation of steroidal glycoalkaloids（SGAs） in the aerial part of potato cultivar and thus to enhance its resistance and tuber quality improvement,the solanidine rhamnosyltransferase（sgt3）gene and ribulose-1,5-bisphoshate carboxylase（Ruisco）promoter were cloned in this research. Subcellular localization prediction showed that SGT3 protein did not contain chloroplast transit peptide,mitochondrial peptide and signal peptide sequence. Therefore,we speculated that SGT3 protein was located in the cytoplasm. Furthermore,cDNA fragment of sgt3 and rbcS promoter was recombinated into plasmid pCEPSP. And we got a plant vector with glyphosate resistance marker that has sgt3 gene driven by a green-tissue promoter rbc S（rbc S：sgt3）. In addition,Longshu 3 and Shepody were transformed by Agrobacterium-mediated protocol,and we obtained 12 glyphosate-tolerant transgenic plantlets. The analysis of gene expression levels indicated that the relative expression of sgt3 transcript in transformation plantlets were higher（1.3–3.0 times）than non-transformation plantlets. Also,the content of SGAs has increased by 20% to 37% for all transformation plantlets. Whereas,SGAs content did not display a significant difference between transgenic tuber and non-transgenic tuber in potato. Our result of this study will provide a theoretical basis for breeding varieties with SGAs content enriched in the foliage but diminished in the tuber in the further.

关 键 词：马铃薯 糖苷生物碱 绿色组织特异性启动子 茄啶鼠李糖基转移酶 遗传转化 

分 类 号：S532[农业科学—作物学]