中国水仙FOMT2基因及启动子的克隆与分析  

Cloning and Ananysis of Flavonoid O-methyltransferase Gene and Promoter in Narcissus tazetta.var chinensis

作　　者：张蒙[1] 潘腾飞[1,2,3] 余磊 杨华丽[1] 潘东明[1,2,3] ZHANG Meng;PAN Tengfei;YU Lei;YANG Huali;PAN Dongming(College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China;Institute of Storage Science and Technology of Horticultural Products, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China;Fujian Prorincial Narcissus Breeding Engineering Technology Research Center, Fuzhou, Fujan 350002, China)

机构地区：[1]福建农林大学园艺学院,福建福州350002 [2]福建农林大学园艺产品贮运保鲜研究所,福建福州350002 [3]福建省水仙花品种选育工程技术研究中心,福建福州350002

出　　处：《热带作物学报》2018年第4期726-732,共7页Chinese Journal of Tropical Crops

基　　金：闽林种站[2013]42号;闽财指[2013]891号;福建省林业科学研究项目(No.H2014020)

摘　　要：为探究花色基因FOMT2在中国水仙(Narcissus tazetta var.chinensis)类黄酮代谢途径中的功能作用,以期明确小分子化合物甲基化修饰在水仙类黄酮代谢中的作用,本研究以多花水仙"金盏银台"(Narcissus tazetta var.chinensis‘jinzhanyintai’)不同时期的花器官为材料,提取总RNA,采用RACE与RT-PCR结合的方法,克隆得到NtFOMT2基因的cDNA全长。结果表明:NtFOMT2基因的cDNA全长为1 404 bp,其中5′UTR长度为33 bp,3′UTR长度为279 bp,开放阅读框为1 092 bp,共编码363个氨基酸。定量PCR分析结果表明,NtFOMT2基因表达水平在水仙花器官的不同时期差异很大,其表达模式表现为Ⅱ>Ⅰ>Ⅲ>Ⅳ。此外,为分析水仙花色素基因的调节因素,以基因组DNA为模板,采用染色体步移法,成功克隆并分析NtFOMT2基因的5’端调控序列。结果表明,NtFOMT2基因启动子序列除包含起始转录位点(transcription start site,TSS)TATA-box、CAAT增强子外,还包含多个顺式作用元件,如ACE、ATCT-motif、G-box、Sp1等多个光响应元件。此外,该启动子序列还含有MYB参与抗旱诱导的结合位点MBS元件、MYB转录因子结合位点等。In order to explore the function of FOMT2 gene in the flavonoid metabolic pathway in Narcissus tazetta. Var chinensis, and to clarify the function of the methylation modification of small molecule compounds in the flavonoid metabolism in Narcissus, the research used Narcissus tazetta. Var chinensis ‘Jinzhanyintai＇ as the meterial to extract total RNA, and the full-length cDNA of NtFOMT2 was cloned with the method of RACE and RT. Results showed that the full length cDNA of NtFOMT2 was 1 404 bp, including a 33 bp 5′ UTR, a 279 bp 3′ UTR and a 1 092 bp ORF encoding 363 amino acids. The results of quantitative PCR showed that the expressions of NtFOMT2 in flower had big difference at different periods. The expression pattern was expressed as： Ⅱ〉Ⅰ〉Ⅲ〉Ⅳ. Besides, to analyze the regulating factors of pigment genes in N. tazetta, the regulation sequence of the 5＇ end of NtFOMT2 gene was cloned with the method of chromosome walking. Results showed that the promoter sequence of NtFOMT2 gene also included cis-acting elements such as ACE, ATCT-motif, G-box, Sp1 and many light responsive elements, besides necessary transcription starting site TATA-box and CAAT enhancer. Furthermore, this promoter sequence included a binding site MBS element participating in drought inducement of MYB, and a MBS binding site of MYB transcription factor and so on.

关 键 词：中国水仙 基因 克隆 FOMT 启动子 类黄酮 

分 类 号：S682.21[农业科学—观赏园艺]