摘要
油茶(Camellia oleifera Abe.)是我国重要木本油料树种,具有重要的经济价值和社会效益.隐花色素(Cryptochrome)是植物蓝光受体之一,参与植物开花调控、光形态建成等生长发育过程.本研究从油茶华硕中克隆到CoCRY1基因,通过生物信息学分析发现,其CDS序列长度为2262 bp,编码的蛋白质序列包含684个氨基酸,蛋白分子式为C3454H5281N971O1027S18,分子量77.42 kDa,是一种不稳定的疏水蛋白,进一步分析表明其蛋白序列包含3个结构域,分别为DNA_photolyase、FAD_binding_7和Cryptochrome_C,可证明CoCRY1蛋白属于隐花色素家族.同源序列及系统发育树分析发现CoCRY1蛋白与茶树(Camellia sinensis)CsCRY1序列相似性最高.组织表达分析显示,CoCRY1的表达量在茎中最高,在花中最低.通过农杆菌转化得到CoCRY1基因异源表达的拟南芥(Arabidopsis thaliana)植株,并且从DNA水平及RNA水平鉴定到阳性植株,对CoCRY1异源表达的拟南芥进行表型分析发现,CoCRY1基因的过量表达导致拟南芥提前开花,并对下胚轴伸长具有光特异性的抑制作用.本研究通过生物信息学、定量分析及异源表达发现,油茶CoCRY1基因在油茶开花等生物学过程中具有重要作用.
Abstract
Camellia oleifera Abe.is an important woody oil tree in China,which has important economic and social benefits.Cryptochrome is one of the blue light receptors in plants,which is involved in the growth and development of plants such as flowering regulation and photomorphogenesis.In this study,CoCRY1 gene was cloned from C.oleifera'Huashuo'.Bioinformatics analysis showed that CDS sequence length of CoCRY1 was 2262 bp encoding for 684 amino acids,the molecular formula of the CoCRY1 protein was C3454H5281N971O1027S18,and the molecular weight was 77.42 kDa,further analysis showed that this protein sequence contained three domains,namely DNA_photolyase,FAD_binding_7 and Cryptochrome_C,which proved that CoCRYl protein belongs to cryptochrome family.Homologous sequence and phylogenetic tree analysis showed that CoCRYl protein had the highest similarity with the homologous sequence of tea tree(Camellia sinensis).Tissue expression analysis showed that the transcripts of CoCRY1 were the highest in stem and the lowest in flower.Arabidopsis thaliana plants with heterogenous expression of CoCRY1 gene were obtained by Agrobacterium transformation,and transgenic plants were identified from DNA and RNA level.Phenotypic analysis of transgenic Arabidopsis showed that overexpression of CoCRY1 gene induced early flowering under long day londition and had photospectific inhibitory effect on hypocotyl elongation.Through bioinformatics,quantitative analysis and heterologous expression,it was found that CoCRY1 gene plays an important role in the biological process of C.oleifera flowering.
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