摘要
4-香豆酸:辅酶A连接酶(4-coumarate:CoA ligase,4CL)基因是植物木质素、类黄酮和其他次生代谢产物合成的关键基因之一。为探究毛竹(Phyllostachys edulis)4CL基因的分子特征和表达模式,利用生物信息学方法对毛竹4CL同源基因进行鉴定,并对其基因结构、保守结构域和系统进化等进行全面分析,应用实时荧光定量PCR(qPCR)技术对基因表达模式进行研究。结果表明,在毛竹中共鉴定出15个4CL同源基因(Pe4CL1~Pe4CL15),均含有内含子,数量为2~5个。Pe4CLs编码蛋白长度为520~671 aa,分子量为55~72 kDa。Pe4CLs均含有2个4CL家族高度保守的结构域,保守基序数量为6~10个,且均定位在过氧化酶体上。系统进化分析显示,毛竹、水稻、二穗短柄草、拟南芥、大豆和毛果杨的4CL家族成员分为2个亚家族(A和B),其中亚家族A又分为3个类型(TypeⅠ~TypeⅢ)。基于转录组数据的基因组织特异性表达分析发现,15个Pe4CLs在毛竹不同组织和不同生长时期中的表达量均存在明显差异。qPCR结果显示,除Pe4CL6的表达下调外,其他基因均随着竹笋高度增加呈上调趋势。组织化学染色结果表明,随着笋高度的增加,其木质化程度加深,与大多数Pe4CLs表达上调相一致。本研究为进一步探究毛竹中4CL的功能提供了参考依据。
In plants,4-coumanic acid:coenzyme A ligase(4 CL)gene is one of the key genes involved in the synthesis of lignin,flavonoids and other secondary metabolites.In order to understand the molecular characteristics and expression patterns of 4 CL genes in Phyllostachys edulis,the homologous genes of 4 CL family were identified in P.edulis and a comprehensive analysis of gene structure,conserved domain and phylogenetic relationship was conducted using bioinformatics methods,and the gene expression patterns were studied by quantitative real-time PCR(qRT-PCR).The results showed that there were fifteen homologous genes of 4 CL in P.edulis(Pe4CL1~Pe4CL15),which all have introns with the numbers varying from 2 to 5.The proteins encoded by Pe4CLs ranged from 520 to 671 amino acids,with the molecular weight of about 55~72 kDa.All Pe4 CLs contained two highly conserved domains unique to the 4 CL family,with 6 to 10 conserved motifs,and all located on peroxidases.Phylogenetic analysis showed that all the 4 CLs of P.edulis,Oryza sativa,Brachypodium distachyon,Arabidopsis thaliana,Glycine max and Populus trichocarpa were clustered into two subfamilies(A and B),and those in A subfamily were divided into three Types(TypeⅠ~Ⅲ).Tissue specific expression analysis based on transcriptome data indicated that Pe4CLs had significantly different expression patterns in different tissues,as well as bamboo shoots at different development stages.The results of qPCR demonstrated that all the Pe4CLs were up-regulated with the increasing height of shoots except Pe4CL6 with a down-regulated expression pattern.The result of histochemical staining showed that the degree of lignification continuously increased with the increase of shoot height,which was consistent with the up-regulation of most Pe4CLs.This study provided a reference for further functional analysis of Pe4CLs in moso bamboo.
作者
杨克彬
单雪萌
史晶晶
朱成磊
高志民
YANG Kebin;SHAN Xuemeng;SHI Jingjing;ZHU Chenglei;GAO Zhimin(Institute of Gene Science and Industrialization for Bamboo and Rattan Resources,International Center for Bamboo and Rattan,National Forestry and Grassland Administration/Key Laboratory of Beijing for Bamboo&Rattan Science and Technology,Beijing 100102)
出处
《核农学报》
CAS
CSCD
北大核心
2021年第1期72-82,共11页
Journal of Nuclear Agricultural Sciences
基金
国际竹藤中心基本科研业务费专项资金项目(1632020004)
国家自然科学基金项目(31930075)。
