摘要
MOC1属于植物特有的GRAS家族蛋白基因,是调控植物腋芽形成发育的关键基因。启动子对基因转录效率起直接调控作用,其功能分析可以精确定位基因的表达部位、发育阶段和调控机制,克隆甘蔗腋芽形成发育关键基因ScMOC1的启动子序列,研究其功能对该基因表达调控机制具有重要意义。本研究以我国主栽甘蔗品种新台糖22号(ROC22)的基因组DNA为模板,通过基因组步移和巢式PCR技术克隆到ScMOC1起始密码子ATG上游1874 bp的启动子序列。PlantCARE在线分析预测表明,该序列包含多个真核生物启动子必需的核心元件TATA-box、CAAT-box以及与光响应、激素响应和分生组织表达等相关的顺式作用元件,推测ScMOC1启动子可通过激素诱导调控ScMOC1表达,且该启动子可能通过分生组织表达顺式调控元件CAT-box参与ScMOC1对甘蔗分蘖的调控。将获得的启动子序列替换pBI121质粒中的CaMV35S启动子驱动下游GUS基因表达进行活性分析,结果表明:本研究克隆的启动子片段能驱动GUS基因在甘蔗嫩叶中瞬时表达。5′缺失分析表明该启动子的基础启动子序列在起始密码子ATG上游350～500 bp之间。该结果为后续ScMOC1的调控机制研究奠定了良好的基础。
MOC1 encodes for a plant-specific GRAS family protein,and this gene plays key role in the formation and development of axillary bud.Since the gene promoter is directly involved into transcriptional regulation,functional analysis can accurately locate the expression site,development stage and regulation mechanism of gene.Cloning the sequence and function analysis of ScMOC1 promoter will be of great signi?cance in illustrating the regulation mechanism of ScMOC1.In this study,the 1874 bp promoter sequence upstream of ScMOC1 was isolated from the genomic DNA of sugarcane main cultivar ROC22 by using nested PCR and genomic walking methods.This isolated fragment was verified to be the promoter of ScMOC1 via sequence structure analysis.The results indicated that ScMOC1 promoter contained a few of core elements of the eukaryotic promoter such as TATA-box,CAAT-box,several light,hormone responsive elements and a cis-acting regulatory element related to meristem expression(CAT-box).We speculated that the ScMOC1 promoter could regulate the expression of ScMOC1 in a manner of hormone treatment,required for the sugarcane tillering regulation by the CAT-box cis-acting element.By constructing the target promoter into pBI121 plasmid that contains a GUS reporter,the results showed that this promoter could result in transient expression of GUS gene in sugarcane young leaves,and the deletion analysis indicated that the basic promoter sequence of the promoter was between350 bp and 500 bp upstream of the starting codon ATG of ScMOC1.Thus,these results obtained above will provide the foundation information for ScMOC1 on transcriptional regulation.
作者
李旭娟
林秀琴
字秋艳
李纯佳
徐超华
吴转娣
朱建荣
刘洪博
方志存
刘新龙
LI Xu-juan;LIN Xiu-qin;ZI Qiu-yan;LI Chun-jia;XU Chao-hua;WU Zhuan-di;ZHU Jian-rong;LIU Hong-bo;FANG Zhi-cun;LIU Xin-long(Sugarcane Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Key L aboratory of SugarcaneGenetic Improvement, Kaiyuan 661699;Bureau of Agriculture and Science and Technology of Pingbian Miao Autonomous County, Pingbian Yunnan 661200)
出处
《植物遗传资源学报》
CAS
CSCD
北大核心
2019年第3期709-717,共9页
Journal of Plant Genetic Resources
基金
国家自然科学基金青年项目(31601362)
云南省应用基础研究计划青年项目(2015FD063)
云南省中青年学术技术带头人后备人才(2014HB038)~~
