中国水仙NtSOC1基因克隆及亚细胞定位  

Cloning and Sub-cellular Localization of NtSOC1 Gene from Narcissus tazetta var. chinensis

作　　者：武丹[1] 吴菁华[1] 张志忠[1] Wu Dan;Wu Jinghua(School of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002,China)

机构地区：[1]福建农林大学园艺学院,福州350002

出　　处：《西北植物学报》2017年第10期1889-1895,共7页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金(31601783);福建省自然科学基金(2014J01083)

摘　　要：以中国水仙‘金盏银台’为实验材料,采用RACE和RT-PCR技术获得1个与开花相关的转录因子(SOC1)的同源基因NtSOC1。NtSOC1的cDNA全长1 603bp,含有1个687bp开放阅读框,编码228个氨基酸。生物信息学分析表明,NtSOC1与单子叶植物的SOC1同源基因的氨基酸序列较为相似,且在C末端同样含有一个保守性很高的SOC1motif序列,说明NtSOC1是属于SOC1/TM3亚家族基因。荧光定量PCR分析显示,NtSOC1在花芽分化阶段的表达量随着花芽的分化而升高,花芽分化结束时减少,表明NtSOC1基因可能参与中国水仙的花芽分化。成功构建了NtSOC1基因表达载体pCAMBIA1302-NtSOC1,通过农杆菌转化洋葱表皮对编码蛋白进行亚细胞定位结果显示,NtSOC1基因编码蛋白定位于细胞核,符合转录因子的亚细胞定位特征。该实验结果为进一步研究NtSOC1基因的生物学功能奠定了基础。A flowering regulated transcription factor SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1(SOC1)homologous gene,NtSOC1,was cloned from the flower bud of Narcissus tazetta var.chinensis using RT PCR and rapid amplification of cDNA ends.NtSOC1is1603bp in length with an open reading frame687bp which is capable of encoding228amino acids.Phylogenetic analysis indicated that the amino acid sequence of NtSOC1is similar with the SOC1s in other species,especially the SOC1in monocots.NtSOC1contains a highly conservative SOC1motif,which indicates that NtSOC1belongs to the SOC1/TM3subfamily gene.Fluorescence quantitative PCR analysis showed that the expression level of NtSOC1in flower bud differentiation stage increased with the differentiation of flower bud and decreased at the end of flower bud differentiation,which indicated that NtSOC1might be involved in flower bud differentiation of narcissus.The pCAMBIA1302NtSOC1expression vector was successfully constructed.Recombinant plasmid was introduced into onion epidermal cells by Agrobacterium tumefaciens method.Sub cellular localization result showed that the NtSOC1was localized in the nucleus,which was consistent with the general character of sub cellular localization of transcription factors.The results laid a foundation for further study of the function of NtSOC1gene.

关 键 词：中国水仙 NtSOC1基因 花发育 亚细胞定位 

分 类 号：Q785[生物学—分子生物学] Q786