甘蔗花青素转录因子基因ScRS克隆与基因枪转化研究  

Cloning of Sugarcane Transcriptional Factor Gene ScRS and Transformation by Microprojectile Bombardment

作　　者：刘迪[1,2] 施桂姣[1,2] 肖乃衍 Khalil Farghama 许跃语 陈平华[1,2] 张卓[1,2] 王恒波[1,2] 高三基[1] 许莉萍[3] 张华[1,2] 

机构地区：[1]福建农林大学国家甘蔗工程技术研究中心,福建福州350002 [2]福建农林大学农业部甘蔗及制品质量监督检验测试中心转基因检测室,福建福州350002 [3]福建农林大学农业部福建甘蔗生物学与遗传育种重点实验室,福建福州350002

出　　处：《热带作物学报》2017年第7期1295-1302,共8页Chinese Journal of Tropical Crops

基　　金：福建省自然科学基金(No.2014J01092);国家自然科学基金项目(No.31070330);科技发展基金项目(No.KF2015080;KF2015118)

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 p CAMBIA1301-35SN-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.

关 键 词：甘蔗 花青素转录因子基因 ScRS 基因枪轰击 

分 类 号：S566.1[农业科学—作物学]