基于黄檗全基因组的苄基异喹啉生物碱生物合成相关O-甲基转移酶基因筛选及功能鉴定

doi:10.7525/j.issn.1673-5102.2026.02.007

摘要/Abstract

摘要：

黄檗（Phellodendron amurense）是一种传统的药用树种，其富含的苄基异喹啉生物碱（BIA）具有显著的药理活性。O-甲基转移酶（OMT）是植物次生代谢中催化酚羟基甲基化的关键酶类，对苄基异喹啉生物碱结构多样化与稳定性具有重要作用。为系统解析黄檗OMT基因家族特征并挖掘参与BIA合成的关键基因，该研究基于黄檗全基因组数据，系统鉴定82个OMT基因家族成员。系统发育分析表明：所有成员均属于ClassⅠ类群，进一步划分为ClassⅠa和ClassⅠb 2个亚族。保守基序分析显示，2个亚族在保留核心SAM结合域的同时，其N端与C端基序组成存在显著差异。染色体定位分析表明：PaOMT基因呈不均匀分布且存在明显串联复制现象。结合黄檗多组织转录组数据筛选及体外酶活验证发现，PaOMT30能够催化（S）-金黄紫堇碱C-2位羟基的O-甲基化，生成（S）-四氢巴马汀红碱。该研究首次系统解析黄檗OMT基因家族特征，并明确PaOMT30在黄檗BIA合成途径中的C-2位甲基化活性，为阐明黄檗药用成分生物合成网络及BIA药物开发奠定了基础。

关键词: 黄檗, 苄基异喹啉生物碱, O-甲基转移酶, 甲基化, 功能表征

Abstract:

Phellodendron amurense is a traditional medicinal plant recognized for its abundant benzylisoquinoline alkaloids（BIAs）， showing substantial pharmacological properties. O-methyltransferases（OMTs）， pivotal enzymes in plant secondary metabolism， facilitate the methylation of phenolic hydroxyl groups and are crucial for the structural diversity and stability of BIAs. To comprehensively characterize the OMT gene family in P.amurense and identify key genes involved in BIA biosynthesis， this study performed a genome-wide analysis， identifying 82 OMT gene family members. Phylogenetic analysis indicated that all members belonged to the ClassⅠgroup， which can be further subdivided into two subgroups， ClassⅠa and ClassⅠb. Analysis of conserved motifs revealed that， while both subgroups maintained the core S-adenosylmethionine （SAM）-binding domain， they displayed notable differences in the composition of N- and C-terminal motifs. Chromosomal localization analysis illustrated the dispersed distribution of PaOMT genes. By integrating transcriptomic data from multiple tissues with in vitro enzymatic assays， PaOMT30 was identified to catalyze the O-methylation of the C-2 hydroxyl group of（S）-scoulerine， yielding（S）-tetrahydropalmatrubine. This study represented the first comprehensive analysis of the OMT gene family in P. amurense and successfully identified the C-2 methylation activity of PaOMT30 in BIA biosynthesis， establishing a foundation for elucidating the biosynthetic pathways of medicinal components in P.amurense and advancing BIA-based drug development.

Key words: Phellodendron amurense, benzylisoquinoline alkaloids, O-methyltransferase, methylation, functional characterization

中图分类号:

Q945.1

曹畅, 田亚, 刘成伟. 基于黄檗全基因组的苄基异喹啉生物碱生物合成相关O-甲基转移酶基因筛选及功能鉴定[J]. 植物研究, 2026, 46(2): 270-279.

Chang CAO, Ya TIAN, Chengwei LIU. Genome-wide Screening and Functional Verification of O-methyltransferase Genes Involved in Benzylisoquinoline Alkaloid Biosynthesis in Phellodendron amurense[J]. Bulletin of Botanical Research, 2026, 46(2): 270-279.

图/表 8

表1

表2

引物序列

基因ID Gene ID	引物名称 Primer name	引物序列（5′→3′） Primer name（5′→3′）
Pamu15G000614	PaOMT6-F	gtcgcggatccgaattcATGGTCATGAAATCAGCGATC
Pamu28G000660	PaOMT20-F	gtcgcggatccgaattcATGGCTTCAATAGCAGATCAGAAA
Pamu14G000851	PaOMT22-F	gtcgcggatccgaattcATGGCTTCTTTAGCAGAGGAAGA
Pamu02G001197	PaOMT28-F	gtcgcggatccgaattcATGGGTTCTGTAACAGAAACCAAG
Pamu22G000567	PaOMT30-F	gtcgcggatccgaattcATGGCTTCTTCAGCAGAATATATTG
Pamu08G000114	PaOMT45-F	gtcgcggatccgaattcATGAGTTCATTCGAAGAGCAAGTG
Pamu30G000130	PaOMT47-F	gtcgcggatccgaattcATGGCTTGTTCAGAGGAGAAGA
Pamu24G000306	PaOMT50-F	gtcgcggatccgaattcATGGAAGAGCAAGATCAAGAGTTG
Pamu37G000004	PaOMT52-F	gtcgcggatccgaattcATGGCAGCTGAAGAAGAA
Pamu13G000164	PaOMT53-F	gtcgcggatccgaattcATGGGTTCAACAAGTTCAGAAAC
Pamu10G001367	PaOMT63-F	gtcgcggatccgaattcATGAGTTTGATTGAAAATGCTGAGG
Pamu19G000073	PaOMT70-F	gtcgcggatccgaattcATGAATGGTCAAGGGGGCA
Pamu16G000056	PaOMT71-F	gtcgcggatccgaattcATGTCTAATATGAAAAATG
Pamu21G000065	PaOMT72-F	gtcgcggatccgaattcATGTCGAATATGAAAGATG
Pamu08G001171	PaOMT79-F	gtcgcggatccgaattcATGGAATCAGTAGAAGCTGAAGCA
Pamu08G000723	PaOMT81-F	gtcgcggatccgaattcATGGGATCAATGGAAGTTCATG
Pamu15G000614	PaOMT6-R	cgacggagctcgaattcTCAAGAAGTTGAAACATATGC
Pamu28G000660	PaOMT20-R	cgacggagctcgaattcCTATTTTAAGAGTTCCATAACATAACAATT
Pamu14G000851	PaOMT22-R	cgacggagctcgaattcCTATTTGAAAAATTCCATAACATATAAAT
Pamu02G001197	PaOMT28-R	cgacggagctcgaattcCTATTTATAAAACTCCATGACGCAG
Pamu22G000567	PaOMT30-R	cgacggagctcgaattcTTACTTGTAGAACTCCATAACCCAGT
Pamu08G000114	PaOMT45-R	cgacggagctcgaattcTTAGATGGTTTTGTAGAATTC
Pamu30G000130	PaOMT47-R	cgacggagctcgaattcCTATTTGAAGAATTCAGTAGC
Pamu24G000306	PaOMT50-R	cgacggagctcgaattcCTACTTGCAGAACTCCATAACCCA
Pamu37G000004	PaOMT52-R	cgacggagctcgaattcCTATTTATAGAATTCCGT
Pamu13G000164	PaOMT53-R	cgacggagctcgaattcTCAAGGCTTTTTAAGAAATTCC
Pamu10G001367	PaOMT63-R	cgacggagctcgaattcCTATGGAAACACTTCAATAAAGGACC
Pamu19G000073	PaOMT70-R	cgacggagctcgaattcCTATGGATACACTTCAATGAAG
Pamu16G000056	PaOMT71-R	cgacggagctcgaattcTTAAGCATCTGCCCTCTTATAGAG
Pamu21G000065	PaOMT72-R	cgacggagctcgaattcTTAAGCATCTGCCCTCTTATAGAG
Pamu08G001171	PaOMT79-R	cgacggagctcgaattcTCACTCAGGATATGCCTCGATAAT
Pamu08G000723	PaOMT81-R	cgacggagctcgaattcTCACTGTGGATAGGCCTCAATA

表2

表3

图1

图2

图3

图4

图5

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基因分类 Gene classification	基因名称 Gene name	基因序列号 Accession number	基因分类 Gene classification	基因名称 Gene name	基因序列号 Accession number
ClassⅠ	CbCOMT	AAB71141	ClassⅠ	RgOMT	AAX82431
	Cj4'OMT	BAB08005		RhcOMT	AAM23004
	Cj6OMT	BAB08004		TaFOMT	Q84N28
	CjCoOMT	BAC22084		ZeCOMT	Q43239
	CjSOMT	BAA06192	Class Ⅱ	VvCCoAOMT	CAA90969
	CsTCS1	BAB12278		PtCCoAOMT	AAA80651
	Ct7OMT	AXC09387		VvAOMT	C7AE94
	EgCOMT	P46484	Class Ⅲ	BAMTa	EOX91020
	MpF4'OMT	AAR09602		CaXMT	BAB39215
	MtHI4'OMT	AAY18581		SAMTa	EOX91014
	Ps6OMT	AAP45315		IAMT	EOY12219
	PsSOMT1	AFK73709		JMTa	EOY14825
	PtCOMT	AAC49708		MT1b	EOY19969
	RcEOMT	BAC78826		MT4a	EOY01378
	RgANMT	ABI93949		MT5a	EOY21224

基因ID Gene ID	基因名称 Gene name	基因ID Gene ID	基因名称 Gene name	基因ID Gene ID	基因名称 Gene name
Pamu08G000115	PaOMT1	Pamu02G001216	PaOMT29	Pamu11G000549	PaOMT57
Pamu06G000283	PaOMT2	Pamu22G000567	PaOMT30	Pamu10G001364	PaOMT58
Pamu20G000479	PaOMT3	Pamu05G000726	PaOMT31	Pamu11G000550	PaOMT59
Pamu20G000485	PaOMT4	Pamu05G000752	PaOMT32	Pamu10G001366	PaOMT60
Pamu20G000482	PaOMT5	Pamu05G000750	PaOMT33	Pamu11G000547	PaOMT61
Pamu30G000130	PaOMT6	Pamu05G000751	PaOMT34	Pamu38G000436	PaOMT62
Pamu10G000639	PaOMT7	Pamu05G000753	PaOMT35	Pamu10G001367	PaOMT63
Pamu06G000143	PaOMT8	Pamu05G000746	PaOMT36	Pamu11G000548	PaOMT64
Pamu18G000972	PaOMT9	Pamu05G000747	PaOMT37	Pamu02G001590	PaOMT65
Pamu36G000631	PaOMT10	Pamu05G000745	PaOMT38	Pamu02G001589	PaOMT66
Pamu36G000632	PaOMT11	Pamu05G000749	PaOMT39	Pamu02G001588	PaOMT67
Pamu36G000634	PaOMT12	Pamu05G000748	PaOMT40	Pamu19G000823	PaOMT68
Pamu36G000641	PaOMT13	Pamu05G000725	PaOMT41	Pamu34G000129	PaOMT69
Pamu36G000645	PaOMT14	Pamu05G000720	PaOMT42	Pamu19G000073	PaOMT70
Pamu36G000640	PaOMT15	Pamu05G000727	PaOMT43	Pamu16G000056	PaOMT71
Pamu36G000651	PaOMT16	Pamu05G000714	PaOMT44	Pamu21G000065	PaOMT72
Pamu06G000144	PaOMT17	Pamu08G000114	PaOMT45	Pamu10G000363	PaOMT73
Pamu18G000976	PaOMT18	Pamu08G000104	PaOMT46	Pamu10G000364	PaOMT74
Pamu18G000971	PaOMT19	Pamu15G000614	PaOMT47	Pamu34G000498	PaOMT75
Pamu28G000660	PaOMT20	Pamu15G000613	PaOMT48	Pamu25G000711	PaOMT76
Pamu28G000775	PaOMT21	Pamu24G000307	PaOMT49	Pamu10G000273	PaOMT77
Pamu14G000851	PaOMT22	Pamu24G000306	PaOMT50	Pamu08G001172	PaOMT78
Pamu09G001243	PaOMT23	Pamu35G000447	PaOMT51	Pamu08G001171	PaOMT79
Pamu14G000813	PaOMT24	Pamu37G000004	PaOMT52	Pamu08G001174	PaOMT80
Pamu14G000512	PaOMT25	Pamu13G000164	PaOMT53	Pamu08G000723	PaOMT81
Pamu30G000533	PaOMT26	Pamu10G001363	PaOMT54	Pamu34G000553	PaOMT82
Pamu22G000566	PaOMT27	Pamu11G000546	PaOMT55
Pamu02G001197	PaOMT28	Pamu10G001365	PaOMT56