甘蔗花青素转录因子基因ScRS克隆与基因枪转化研究

摘要

花青素不仅是天然色素,而且具有保健功能.花青素转录因子基因对于调控花青素的合成具有关键作用.本研究根据玉米花青素转录因子基因RS序列设计引物,利用同源克隆技术,得到甘蔗花青素转录因子基因ScRS的全长序列;在ScRS序列两端分别添加KpnⅠ、SalⅠ酶切位点,构建植物表达载体pCAMBIA1301-35SN-ScRS,包被微粒载体通过基因枪对甘蔗愈伤组织进行轰击,转化的愈伤组织明显呈紫红色;将紫红色愈伤组织分化培养获得再生甘蔗植株,经PCR检测,获得转ScRS基因阳性植株,初步证实利用克隆的基因转化甘蔗可以使愈伤组织显色.研究结果对于建立新型的遗传转化可视化示踪体系和培育高花青素甘蔗品种具有重要意义.%Anthocyanins are not only natural pigments, but also health care ingredients. The anthocyanin transcriptional factor genes play import roles in anthocyanin biosynthesis. In this study, the specific primer was designed according to the sequence of maize anthocyanin transcriptional factor gene RS. The full sequence of putative sugarcane anthocyanin transcription factor gene ScRS was cloned by homology cloning technology. The plant expression vector of pCAMBIA1301-35SN-ScRS was constructed by adding endonuclease sites of KpnⅠ and SalⅠ at the ends of ScRS sequence, which was transformed into sugarcane calli via gene gun bombardment, and the calli bombarded turned purple-red. The calli with purple-red color were selected and regenerated into plants. The PCR results were positive in the transgenic detection of the regenerated sugarcane plants with specific primers of ScRS. The results preliminarily confirmed that the transformation of ScRS gene would make sugarcane calli colorated. Cloning and characterization of anthocyanin transcription factor genes would contribute to develop anthocyanin-rich varieties and establish visual tracing system in genetic transformation.