FLS基因调控玉兰与紫玉兰花色形成的机制研究  

Study on the Mechanism of the FLS Gene Regulating Flower Color Formation in Yulania denudata and Y.liliiflora

作　　者：吴超 戴梦怡 张超[1] 石从广[2] 任明杰 马晶晶 申亚梅[1] WU Chao;DAI Mengyi;ZHANG Chao;SHI Congguang;REN Mingjie;MA Jingjing;SHEN Yamei(Zhejiang Provincial Key Laboratory of Germplasm Innovation and Utilization for Garden Plants,School of Landscape Architecture,Zhejiang A&F University,Hangzhou,Zhejiang 311300;Zhejiang Academy of Forestry,Hangzhou,Zhejiang 310023)

机构地区：[1]浙江农林大学风景园林与建筑学院,浙江省园林植物种质创新与利用重点实验室,浙江杭州311300 [2]浙江省林业科学研究院,浙江杭州310023

出　　处：《核农学报》2023年第10期1947-1956,共10页Journal of Nuclear Agricultural Sciences

基　　金：浙江省“十四五”育种专项花卉协作组项目(2021C02071-3);浙江农林大学科研发展基金人才启动项目(2021LFR051)。

摘　　要：黄酮醇合成酶(FLS)是植物黄酮醇合成途径中的关键酶。为探究FLS对玉兰属植物花色形成的影响,以玉兰(Yulania denudata)和紫玉兰(Yulania liliiflora)为研究对象,利用超高效液相色谱技术(UPLC)测定其花开放五个关键时期的花被片黄酮醇含量;基于转录组数据对玉兰和紫玉兰的FLSs基因进行克隆;利用实时荧光定量PCR(qRT-PCR)分析玉兰与紫玉兰不同组织和开花五个关键时期的FLSs基因表达量;通过洋葱表皮细胞瞬时转化对FLSs蛋白进行亚细胞定位;通过异源转化烟草进行FLSs基因功能验证,并利用qRT-PCR和UPLC分析转基因烟草花冠中黄酮醇含量差异。结果表明,玉兰花被片黄酮醇含量随花开放呈现先升高后降低的趋势,紫玉兰花被片黄酮醇含量逐渐下降。YdFLS和YlFLS具有序列保守性,在不同组织中均有表达,YdFLS在花中表达量最高,YlFLS在嫩叶中表达量最高;在花开放进程中,YdFLS表达量逐渐升高,至盛开期最高,而YlFLS则表现相反的表达模式。YdFLS与YlFLS都定位于细胞质;YdFLS和YlFLS过表达烟草植株花冠的颜色变浅,黄酮醇含量增加且花青素苷积累被抑制。综上,本研究验证了FLS基因在玉兰属植物花色形成过程中功能保守,发挥促进黄酮醇合成的作用,其转录水平的差异是各色花色形成的主要原因。本研究结果揭示了FLS基因在玉兰属植物花色形成中发挥着重要功能。Flavonol synthase(FLS)is a key enzyme in the flavonol biosynthesis pathway in plants.To investigate the mechanism of flower color formation in different species of Yulania,the flavonol contents of Y.denudata and Y.liliiflora were measured by the ultra-performance liquid chromatography(UPLC)at five key stages of flowering.The FLSs genes were cloned by RNA-seq in Y.denudata and Y.liliiflora.The expression patterns of FLSs genes in different tissues and tepals at five flowering stages were detected using quantitative real-time PCR(qRT-PCR).Subcellular localization of FLSs proteins were performed by transient transformation technique in onion epidermis.The functions of FLSs genes were verified by heterologous transformed in tobacco,and the difference in flavonol contents in the transgenic tobacco corolla was analyzed by UPLC.The results showed the flavonol content of Y.denudata tepals was first increased and then decreased during flowering,while the flavonol content of Y.liliiflora tepals gradually decreased.YdFLS and YlFLS were conserved in sequence.The highest expression level of the YdFLS gene was found in flowers,and YlFLS had the highest expression level in young leaves.The expression of YdFLS was gradually increased and peaked at the bloom stage,while YlFLS showed a declining expression pattern.Both YdFLS and YlFLS were located in the cytoplasm.Overexpression of YdFLS and YlFLS genes in tobacco caused a light corolla phenotype and flavonol accumulation.In conclusion,this study verified that FLSs genes conservatively functioned on promoting flavonol synthesis in Yulania species.This study revealed the important function of FLS gene in flower color formation of Magnolia.

关 键 词：玉兰 花色 黄酮醇合成酶基因 基因功能验证 

分 类 号：S685.15[农业科学—观赏园艺]