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糖苷生物碱(steroidal glycoalkaloids,SGAs)是一类存在于茄科和某些百合科植物的重要次生代谢物,与植物的抗逆性和产品品质有密切关系.茄啶半乳糖基转移酶(solanidine galactosyltransferases,SGT1)是SGAs合成代谢途径的末端关键酶之一,研究其编码基因的启动子序列对于SGAs生物合成代谢调控有重要的作用和意义.研究采用染色体步移技术(Genome walking),首次克隆到马铃薯Sgt1基因起始密码子上游2 183 bp的启动子序列,已注册到GenBank(注册号:KC759163).构建该启动子驱动融合报告基因gfp::gus的植物双元表达载体p1304Sgt1p,转化野生型烟草获得Sgt1p::gfp::gus转基因植株,通过GUS组织化学染色分析Sgt1p::gfp::gus转基因植株中Sgt1p启动子的活性.结果表明,gus基因在转基因烟草的根、茎和叶中均表达,在叶中Sgt1p启动子的活性低于CaMV35S启动子,而在根和茎中二者基本相同;光诱导结果显示,光照处理明显增强了Sgt1p::gfp::gus转基因烟草叶片中Sgt1p启动子的活性,表明庄薯3号马铃薯Sgt1p启动子是一种光诱导型启动子.
Steroidal glycoalkaloids (SGAs) are important secondary metabolites that exist in Solanaceae and certain Liliaceae plants and are closely related to plant stress resistance and product quality. solanidine galactosyltransferases, SGT1) is one of the key end-enzymes involved in the metabolic pathway of SGAs, and it is important to study the promoter sequence of its coding genes for the biosynthesis and metabolism regulation of SGAs.Methods Genome walking, The promoter sequence of 2 183 bp upstream of the start codon of Sgt1 gene of potato was cloned into GenBank (Accession No .: KC759163). The promoter was used to construct the plant binary expression vector p1304Sgt1p with the fusion reporter gfp :: gus, Wild type tobacco Sgt1p :: gfp :: gus transgenic plants were obtained and the activity of Sgt1p promoter in Sgt1p :: gfp :: gus transgenic plants was analyzed by GUS histochemical staining.The results showed that the gus gene was expressed in roots, stems and leaves of transgenic tobacco , And the activity of Sgt1p promoter in leaves was lower than that in CaMV35S promoter, while the content of Sgt1p promoter was the same in roots and stems. The light-induced results showed that the light treatment significantly Enhances the activity of the Sgt1p promoter in Sgt1p :: gfp :: gus transgenic tobacco leaves, indicating that the potato Sgt1p promoter of Zhuzhu No.3 is a light-inducible promoter.