紫甘蓝COP1基因的克隆及表达模式分析

光照是植物生长发育的一个重要的环境因素,COP1是光调控植物发育的一个分子开关。本实验以"早红"紫甘蓝为实验材料,分两组置于光照(光照16h,28℃;黑暗8h,18℃)及完全遮光培养箱培养,至幼苗时提取RNA,进行COP1的cDNA克隆,并通过半定量RT-PCR方法分析COP1在不同处理下的表达情况。结果表明:COP1 cDNA全长为1863bp,编码620aa,分子量为70.325kD;在无光条件下生长的紫甘蓝幼苗虽然仍有少量花青素合成,但是其花青素含量明显低于光照条件下生长的紫甘蓝幼苗;COP1基因在无光条件下的表达量明显强于有光条件,表明COP1对紫甘蓝中花青素的合成起负调控作用。本文旨在探索在不同光照条件下紫甘蓝中COP1基因与其花青素合成的关系,为后期揭示花青素表达调控机制奠定基础。

Cloning and Expression Pattern Analysis of Purple Cabbage COP1 Gene

Light is the most influential environmental factor,affecting the plant growth and development,otherwise COP1 is a molecular switch of light regulation for the plant development.In this experiment, ＂Zaohong＂ purple cabbage as experimental material,is divided into two groups placed in the light（light 16 h,28℃;dark 8h,18℃）and completely shading incubator.RNA was extracted when it grew to seedlings,the cDNA of COP1 gene was cloned.We analyzed COP1 expression pattern under different processing by semi-quantitative RT-PCR method.The full-length cDNA sequence of COP1 was 1863 bp,encoding 620 amino acids,with a molecular weight of 70.325 kD.Specially,the purple cabbage seedling still produce anthocyanins in dark conditions,and the content of anthocyanins of the purple cabbage seeding grown in dark conditions significant lower than the seeding grown in light conditions.The expression level of COP1 gene in dark conditions is clearly higher than that in light conditions,it indicated that COP1 gene played a negative regulatory role to the synthesis of anthocyanins in purple cabbage.The purpose of this paper is to explore the relationship between the synthesis of anthocyanin and COP1 gene under the different illumination condition in the purple cabbage,and to lay the foundation for revealing anthocyanins expression regulation mechanism.