基于DNA条形码序列的天目竹节人参及其近缘种系统进化分析

doi:10.16178/j.issn.0528-9017.20240913

浙江农业科学 ›› 2025, Vol. 66 ›› Issue (3): 623-626.DOI: 10.16178/j.issn.0528-9017.20240913

基于DNA条形码序列的天目竹节人参及其近缘种系统进化分析

肖文斐(), 严建立, 忻雅, 柴伟国()

杭州市农业科学研究院,浙江杭州 310024

收稿日期:2024-11-29 出版日期:2025-03-11 发布日期:2025-04-02
通讯作者: 柴伟国
作者简介:柴伟国(1970—),男,浙江杭州人,研究员,从事蔬菜、中药材新品种引选、品质鉴定,E-mail:kuni@21cn.com。
肖文斐(1979—),女,湖南祁东人,高级农艺师,从事蔬菜、中药材种质资源保护与评价利用,E-mail:xiao_wenfei@126.com。
基金资助:
浙江省公益性技术应用研究项目(LGN21C020003)

Phylogenetic analysis of Panax japonicus (TianMu) and its relatives based on DNA barcode sequences

XIAO Wenfei(), YAN Jianli, XIN Ya, CHAI Weiguo()

Hangzhou Academy of Agricultural Sciences, Hangzhou 310024, Zhejiang

Received:2024-11-29 Online:2025-03-11 Published:2025-04-02
Contact: CHAI Weiguo

摘要/Abstract

摘要： 为探讨天目竹节人参的进化地位,以天目竹节人参为试材,利用PCR法扩增ITS1/ITS2、ITS2、psbA-trnH、rbcL等4种DNA条形码标记片段序列,同时从NCBI下载8种人参属植物及3种外属植物的同源序列,利用MEGA 11.0软件构建系统发育树,从分子水平上对天目竹节人参及其近缘种植物进行比较与系统进化分析。结果表明,ITS1/ITS2序列在种间差异较大,种内较为保守;ITS2序列多态性最丰富,种内也存在分离;psbA-trnH及rbcL序列对某些近缘种无法有效区分。基于ITS1/ITS2序列采用邻接法(neighbor joining)构建进化树,结果显示,天目竹节人参与人参、西洋参亲缘关系较近,归为同一类群;与三七和恩施竹节参差异大于种间最小遗传差异。上述人参属植物种间的遗传距离在0.003~0.033,表明这些物种亲缘关系较近。该研究对人参属的遗传资源保护和探索具有重要意义。

关键词: 天目竹节人参, 人参属, DNA条形码, 系统进化

Abstract:

To explore the evolutionary status of Panax japonicus (TianMu), DNA barcode markers including ITS1/ITS2, ITS2, psbA-trnH, and rbcL were amplified using PCR from Panax japonicus (TianMu) samples. Additionally, the homologous sequences of 8 Panax species and 3 exogenous plants were downloaded from NCBI. A phylogenetic tree was constructed using MEGA 11.0 software to analyze the phylogenetic relationships of P. japonicus (TianMu) and its closely related species at the molecular level. The results indicated that the ITS1/ITS2 sequences showed significant differences between species but were conserved within species; ITS2 sequences had the richest polymorphism and also exhibited intraspecific variation. The psbA-trnH and rbcL sequences were not effective in distinguishing some closely related species. A phylogenetic tree was constructed using the ITS1/ITS2 sequences with the neighbor joining method, which showed that Panax. japonicus (TianMu) was closely related to Panax ginseng and Panax quinquefolius, and they were classified into the same group; the differences between Panax. japonicus (TianMu) and Panax notoginseng and Panax japonicus (Enshi) were greater than the minimum genetic differences within species. The genetic distances between the above-mentioned Panax species ranged from 0.003 to 0.033, indicating that these Panax species are closely related. This study is significant for the protection and exploration of genetic resources of Panax japonicus (TianMu).

Key words: Panax japonicus (TianMu), Panax, DNA barcode, phylogenetics

中图分类号:

S567.51

肖文斐, 严建立, 忻雅, 柴伟国. 基于DNA条形码序列的天目竹节人参及其近缘种系统进化分析[J]. 浙江农业科学, 2025, 66(3): 623-626.

XIAO Wenfei, YAN Jianli, XIN Ya, CHAI Weiguo. Phylogenetic analysis of Panax japonicus (TianMu) and its relatives based on DNA barcode sequences[J]. Journal of Zhejiang Agricultural Sciences, 2025, 66(3): 623-626.

图/表 4

参考文献 12

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DNA区段	正/反向引物	引物序列(5'-3')
ITS1/ITS2	ITS4	TCCTCCGCTTATTGATATGC
	ITS5	GGAAGTAAAAGTCGTAACAAGG
ITS2	ITS2F	ATGCGATACTTGGTGTGAAT
	ITS3R	GACGCTTCTCCAGACTACAAT
rbcL	rbcLF	ATGTCACCACAAACAGARACTAAAGCAAG
	1024R	CCCTAAGGGTGTCCTAAAGTTTCTCCACC
psbA-trnH	psbA3F	GTTATGCATGAACGTAATGCTC
	trnH3R	CGCGCATGGTGGATTCACAATC

DNA区段	正/反向引物	引物序列(5'-3')
ITS1/ITS2	ITS4	TCCTCCGCTTATTGATATGC
	ITS5	GGAAGTAAAAGTCGTAACAAGG
ITS2	ITS2F	ATGCGATACTTGGTGTGAAT
	ITS3R	GACGCTTCTCCAGACTACAAT
rbcL	rbcLF	ATGTCACCACAAACAGARACTAAAGCAAG
	1024R	CCCTAAGGGTGTCCTAAAGTTTCTCCACC
psbA-trnH	psbA3F	GTTATGCATGAACGTAATGCTC
	trnH3R	CGCGCATGGTGGATTCACAATC

对应DNA区段	片段大小/bp	人参属内同源性/%	与楤木的同源性/%	与刺五加的同源性/%	与胡萝卜的同源性/%
ITS1/ITS2	681	96.74~99.70	95.25~95.85	92.59~93.48	70.25~70.31
ITS2	465	95.63~99.56	95.64~97.39	93.91~94.61	72.37~73.68
rbcL	938	97.73~100.00	98.27~99.36	97.73~99.04	95.57~96.81
psbA-trnH	505	96.56~100.00	95.13~96.79	94.61~96.72	88.72~89.47

对应DNA区段	片段大小/bp	人参属内同源性/%	与楤木的同源性/%	与刺五加的同源性/%	与胡萝卜的同源性/%
ITS1/ITS2	681	96.74~99.70	95.25~95.85	92.59~93.48	70.25~70.31
ITS2	465	95.63~99.56	95.64~97.39	93.91~94.61	72.37~73.68
rbcL	938	97.73~100.00	98.27~99.36	97.73~99.04	95.57~96.81
psbA-trnH	505	96.56~100.00	95.13~96.79	94.61~96.72	88.72~89.47

DNA区段	人参属内遗传距离	与楤木遗传距离	与刺五加遗传距离	与胡萝卜遗传距离
ITS2	0.004~0.031	0.030	0.054	0.251
ITS1/ITS2	0.004~0.033	0.045	0.070	0.268
rbcL	0~0.007	0.006	0.012	0.035
psbA-trnH	0~0.021	0.012	0.034	0.088

基于DNA条形码序列的天目竹节人参及其近缘种系统进化分析

Phylogenetic analysis of Panax japonicus (TianMu) and its relatives based on DNA barcode sequences

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