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中国中医科学院 中药研究所,中药鉴定与安全性评估北京市重点实验室,北京 100700
Received:03 March 2023,
Published Online:11 May 2023,
Published:05 February 2024
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陈竹荫,俞越童,郝晨阳等.莲NnOMT和NnNMT基因家族的鉴定与分析[J].中国实验方剂学杂志,2024,30(03):152-160.
CHEN Zhuyin,YU Yuetong,HAO Chenyang,et al.Identification and Analysis of NnOMT and NnNMT Gene Families in Nelumbo nucifera[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(03):152-160.
陈竹荫,俞越童,郝晨阳等.莲NnOMT和NnNMT基因家族的鉴定与分析[J].中国实验方剂学杂志,2024,30(03):152-160. DOI: 10.13422/j.cnki.syfjx.20230965.
CHEN Zhuyin,YU Yuetong,HAO Chenyang,et al.Identification and Analysis of NnOMT and NnNMT Gene Families in Nelumbo nucifera[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(03):152-160. DOI: 10.13422/j.cnki.syfjx.20230965.
目的
2
莲苄基异喹啉生物碱(BIAs)的生物合成途径的解析具有重要的理论和经济价值,为了更好地促进莲BIAs生物合成途径的解析,本研究对莲氧甲基转移酶(NnOMT)、莲氮甲基转移酶(NnNMT)基因家族进行鉴定和生物信息学分析。
方法
2
在莲全基因组的基础上,利用UniPort、美国国家生物技术信息中心(NCBI)数据库对莲NnOMT、NnNMT基因家族进行鉴定,分析其理化性质及亚细胞定位。利用TBtools、MEME、MEGA 11.0、FigTree 1.4.4等工具对前期鉴定出的
NnOMT
、
NnNMT
基因的系统发育、序列特征、基因结构、功能注释、顺式作用元件等进行分析。
结果
2
共鉴定出61个莲
NnOMT
、
NnNMT
基因,编码的氨基酸数目介于168~580 aa,等电点范围为4.76~9.16,相对分子质量为18 699.52~64 934.53 Da,大部分表现为酸性且多为亲水蛋白,含有10个保守基序;京都基因与基因组百科全书(KEGG)分析共富集到12个通路,包括新陈代谢、异喹啉生物碱生物合成、苯丙烷的生物合成等;基因本体(GO)富集结果可视化显示,61个莲
NnOMT
、
NnNMT
基因共注释到32个类别,其中包括16项分子功能,如谷氨酸合成酶还原型烟酰胺腺嘌呤二核苷酸(NADH)活性、外肽酶活性,以及16项生物过程,如四氯化碳代谢过程、厌氧四氯化碳代谢过程、对外源生物刺激的反应。在莲
NnOMT
、
NnNMT
基因的启动子区存在多种顺式作用元件,主要为启动子和增强子区响应元件、光响应和茉莉酸甲酯响应元件。
结论
2
该研究基于莲的基因组数据对61个莲
NnOMT
、
NnNMT
基因进行了全面的生物信息学分析,为莲NnOMT、NnNMT基因家族的基因结构和功能研究奠定了基础,并为莲BIAs的生物合成途径解析提供了参考。
Objective
2
The biosynthetic pathways of benzylisoquinoline alkaloids(BIAs) in
Nelumbo nucifera
are of great theoretical and economic value. In this paper,
N. nucifera
O
-methyltransferase(NnOMT) and
N. nucifera N
-methyltransferase(NnNMT) gene families were identified and analyzed by bioinformatics in order to facilitate the biosynthetic pathway of BIAs in
N. nucifera
.
Method
2
Based on the whole genome of
N. nucifera
, UniPort and National Center for Biotechnology Information(NCBI) databases were used to identify the NnOMT and NnNMT gene families of
N. nucifera
, and analyze their physicochemical properties and subcellular localization, then TBtools, MEME, MEGA 11.0, FigTree 1.4.4 and other tools were used to analyze the phylogeny, sequence characteristics, gene structure, functional annotation and
cis
-acting elements of
NnOMT
and
NnNMT
genes identified in the previous stage.
Result
2
A total of 61
NnOMT
and
NnNMT
genes were identified in this paper, the number of amino acids encoded by these genes ranged from 168 aa to 580 aa, the isoelectric point ranged from 4.76 to 9.16, and the relative molecular weight ranged from 18 699.52 Da to 64 934.53 Da, most of which showed acidic and mostly hydrophilic proteins. There were 10 conserved motifs, Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis enriched a total of 12 pathways, including metabolism, biosynthesis of phenylpropane and isoquinoline alkaloids, etc. And Visualization of Gene Ontology(GO) enrichment results showed that 61
NnOMT
and
NnNMT
genes were annotated to 32 items, which included 16 molecular functions[such as reduced nicotinamide adenine dinucleotide(NADH) activity and exopeptidase activity] and 16 biological processes(such as metabolic process of carbon tetrachloride, anaerobic carbon tetrachloride metabolic process and responses to exogenous biological stimuli). There were a variety of
cis
-acting elements in the promoter regions of
NnOMT
and
NnNMT
genes, mainly promoter and enhancer regions element, light responsive element and methyl jasmonate responsive element.
Conclusion
2
In this study, a comprehensive bioinformatics analysis of 61
NnOMT
and
NnNMT
genes is carried out based on the genome data of
N. nucifera
, which lays a foundation for research on the gene structure and function of NnOMT and NnNMT gene families, and provides a reference for biosynthetic pathway elucidation of BIAs in
N. nucifera
.
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