厚朴Dirigent基因家族的挖掘与生物信息学分析  

Mining and Bioinformatics Analysis of Houpo(Houpoea officinalis)Dirigent Gene Families

作　　者：张柏年 赵明海 田徽[1] 张元元 王迁 王涛 冯大仁 席祯鹏 ZHANG Bainian;ZHAO Minghai;TIAN Hui;ZHANG Yuanyuan;WANG Qian;WANG Tao;FENG Daren;XI Zhenpeng(Mianyang Teachers’College,Mianyang 621000,Sichuan,China;Mianyang Hospital of Traditional Chinese Medicine,Mianyang 621000,Sichuan,China;Mianyang Institute for Food and Drug Control,Mianyang 621000,Sichuan,China;Beichuan Shennong Agriculture Technology Development Co.,Ltd,Mianyang 621000,Sichuan,China)

机构地区：[1]绵阳师范学院,绵阳市中药质量控制与评价重点实验室,四川绵阳621000 [2]绵阳市中医医院,中医药研究所,四川绵阳621000 [3]绵阳市食品药品检验所,四川绵阳621000 [4]北川神侬农业科技发展有限公司,四川绵阳621000

出　　处：《中华中医药学刊》2024年第1期189-193,I0017-I0019,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81202933);四川省自然科学基金项目(2022NSFSC0592);绵阳市科技计划项目(2018YFZJ025)。

摘　　要：目的木脂素是药用植物厚朴中一类主要的活性物质,其合成与Dirigent(DIR)基因家族密切相关,然而厚朴中的DIR基因尚未被研究。研究旨在鉴定厚朴中的DIR基因,同时筛选出与木脂素合成相关的DIR基因。方法在获得厚朴全基因序列的基础上,利用生物信息学分析,采用本地BLAST将厚朴全基因序列与已知的模式植物拟南芥的25个DIR基因进行比对后,共筛选最终得到20个含有dirigent保守结构域的HoDIR基因,然后对这些基因进行包括蛋白质理化性质、亚细胞定位、蛋白二级结构、蛋白三级结构、保守结构域等在内的一系列生物信息学分析,并与拟南芥的25个DIR同源基因构建发育树并进行相关的分析。结果20个HoDIR基因只含有1个完整或非完整dirigent保守结构域,以及普遍存在于各HoDIR中的5种保守基序。相对分子量为18736.48 Da左右,理论等电点为5.05~10.27,碱性蛋白居多;除HoDIR4外均表现为稳定性蛋白;除HoDIR9外,均为疏水性蛋白。亚细胞定位主要位于叶绿体和细胞壁,少数位于细胞膜和细胞质。蛋白质二级结构中主要结构元件为延伸链和无规则卷曲;蛋白质三级结构大致相似,其中HoDIR6和HoDIR9与拟南芥中DIR-a亚族的AtDIR6结构极为相似。结论系统进化分析结果表明厚朴中的DIR基因分为3个亚族:DIR-a、DIR-b/d、DIR-e。其中HoDIR6和HoDIR9属于DIR-a,可能参与调控厚朴木脂素合成。对厚朴DIR基因家族的挖掘与鉴定,为厚朴木脂素的生物合成及其药效、品质的提高提供了一定的理论依据。Objective Lignans are a major class of active compounds in the medicinal plant Houpo(Houpoea officinalis),and their synthesis is closely linked to the Dirigent(DIR)gene family.However,DIR genes in Houpo(Houpoea officinalis)have not been previously studied.This study aims to identify DIR genes in Houpo(Houpoea officinalis)and screen for those associated with lignan synthesis.Methods Using the complete genomic sequence of Houpo(Houpoea officinalis),a bioinformatics analysis was conducted by comparing it with the sequences of 25 known DIR genes from the model plant Ninanjie(Arabidopsis thaliana)through local BLAST.This led to the selection of 20 HoDIR genes containing the conserved dirigent domain.Subsequently,a series of bioinformatics analyses was performed,encompassing physicochemical properties,subcellular localization,protein secondary and tertiary structures,as well as conserved domains.A phylogenetic tree was constructed using 25 DIR orthologous genes from Ninanjie(Arabidopsis thaliana)for comparative analysis.Results The findings revealed that these 20 HoDIR genes each contained either a complete or incomplete dirigent conserved domain and five prevalent conserved motifs in each HoDIR.Their relative molecular weight was approximately 18736.48 Da,with theoretical isoelectric points ranging from 5.05 to 10.27,indicating a predominance of alkaline proteins.All of these proteins,except HoDIR4,demonstrate stability,and with the exception of HoDIR9,they were all hydrophobic.Subcellular localization was primarily observed in chloroplasts and the cell wall,with a few instances in the cell membrane and cytoplasm.The main structural elements in the protein's secondary structure were extended chains and random coils.Protein tertiary structures were generally similar,with HoDIR6 and HoDIR9 displaying remarkable structural similarity to AtDIR6 of the DIR-a subfamily in Ninanjie(Arabidopsis thaliana).Conclusion Phylogenetic analysis results indicate that DIR genes in Houpo(Houpoea officinalis)can be classified into three

关 键 词：厚朴 木脂素 DIR蛋白 生物信息学 

分 类 号：R282.7[医药卫生—中药学]