锦香草及其近缘种的亲缘关系ITS序列分析

doi:10.3969/j.issn.1004-1524.2021.08.11

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (8): 1436-1444.DOI: 10.3969/j.issn.1004-1524.2021.08.11

锦香草及其近缘种的亲缘关系ITS序列分析

陆彭城¹(), 张文春¹, 陈建芳², 黄学敏³, 叶菁⁴, 彭东辉¹^,^*()

1.福建农林大学,福建福州 350000
2.龙岩新罗区林业局,福建龙岩 364000
3.泉州永春县林业局,福建泉州 362000
4.福州植物园,福建福州 350000

收稿日期:2020-11-06 出版日期:2021-08-25 发布日期:2021-08-27
通讯作者: 彭东辉
作者简介:*彭东辉,E-mail: fjpdh@126.com
陆彭城(1996—),男,福建宁德人,硕士研究生,研究方向为园林植物及其应用。E-mail: 471317886@qq.com
基金资助:
福建省林业推广项目闽林推〔2017〕(SJ15)

Analysis of inter-species relationship of Phyllagathis cavaleriei and its related species by use of ITS sequences

LU Pengcheng¹(), ZHANG Wenchun¹, CHEN Jianfang², HUANG Xuemin³, YE Jing⁴, PENG Donghui¹^,^*()

1. Fujian Agriculture and Forestry University, Fuzhou 350000, China
2. Longyan Xinluo District Forestry Bureau, Longyan 364000, China
3. Quanzhou Yongchun County Forestry Bureaus, Quanzhou 362000, China
4. Fuzhou Botanical Garden, Fuzhou 350000, China

Received:2020-11-06 Online:2021-08-25 Published:2021-08-27
Contact: PENG Donghui

摘要/Abstract

摘要：

为厘清锦香草及其近缘种的亲缘关系,为其杂交育种提供数据基础,收集我国锦香草及其近缘种短毛熊巴掌与长柄熊巴掌等样品46份,对供试样品核糖体基因(nrDNA)的内转录间隔区(internal transcribed spacer,ITS)序列进行分析,并构建系统进化树,对其系统发育关系进行研究。结果表明:供试材料的ITS1、ITS2和5.8S的序列长度分别为246、230、160~161 bp;ITS1与ITS2序列存在变异位点426个,信息位点232个;5.8S序列存在202个变异位点,信息位点84个。根据ITS序列构建进化树:供试材料可分为2支,锦香草与短毛熊巴掌在进化树中聚为一支,分支内部互相嵌入;长柄熊巴掌单独聚成一支。以上结果显示,锦香草、短毛熊巴掌亲缘关系较近,它们与长柄熊巴掌亲缘关系较远,该结果支持《Flora of China》将短毛熊巴掌处理为锦香草的异名,长柄熊巴掌为独立种。

关键词: 锦香草, 短毛熊巴掌, 长柄熊巴掌, ITS, 亲缘关系

Abstract:

In oder to clarify the phylogenetic relationship between Phyllagathis cavaleriei and its related species and provide data basis for its cross breeding,46 samples were collected from China, including P. cavaleriei and its related species P. cavaleriei var. tankahkeei and P. cavaleriei var. wilsoniana in China. Sequence of the internal transcribed spacer (ITS) of the ribosomal gene (nrDNA) of the tested samples was analyzed, and phylogenetic tree was constructed to study the phylogenetic relationship. The results showed that length of ITS1, ITS2, and 5.8S sequences of 46 materials were 246, 230, and 160-161 bp, respectively. Total 426 variable sites were found in ITS1 and ITS2 sequences with 232 informative sites. There were 202 mutation sites and 84 informative sites in 5.8S sequence. Based on ITS sequence, an evolutionary tree was constructed, materials could be divided into two branches. P. cavaleriei and P. cavaleriei var. tankahkeei were clustered into one branch in the cluster tree, and the branches were embedded into each other, while P. cavaleriei var. wilsoniana was clustered into a single branch. The above results showed that P. cavaleriei was closely related to P. cavaleriei var. tankahkeei, while P. cavaleriei var. wilsoniana was far related. The results supported Flora of China to deal with P. cavaleriei var. tankahkeei as a different name of P. cavaleriei, P. cavaleriei var. wilsoniana was a separate species.

Key words: Phyllagathis cavaleriei, Phyllagathis cavaleriei var. tankahkeei, Phyllagathis cavaleriei var. wilsoniana, internal transcribed spacer(ITS), phylogenetic relationship

中图分类号:

陆彭城, 张文春, 陈建芳, 黄学敏, 叶菁, 彭东辉. 锦香草及其近缘种的亲缘关系ITS序列分析[J]. 浙江农业学报, 2021, 33(8): 1436-1444.

LU Pengcheng, ZHANG Wenchun, CHEN Jianfang, HUANG Xuemin, YE Jing, PENG Donghui. Analysis of inter-species relationship of Phyllagathis cavaleriei and its related species by use of ITS sequences[J]. Acta Agriculturae Zhejiangensis, 2021, 33(8): 1436-1444.

图/表 5

参考文献 18

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编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
1	锦香草 P.cavaleriei	HNSHS011	湖南新宁县舜皇山	111°0'	26°24'	712
2	锦香草 P.cavaleriei	HNSHS016	Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	706
3	锦香草 P.cavaleriei	HNSHS024	湖南新宁县舜皇山	111°01'	26°24'	686
			Shunhuang Mountain, Xinning, Hunan
4	锦香草 P.cavaleriei	GDDXS028	广东仁化县丹霞山	113°71'	25°02'	317
			Danxia Mountain, Dehua, Guangzhou
5	锦香草 P.cavaleriei	GXJXLHS030	广西金秀县莲花山	110°11'	24°16'	861
			Lianhua Mountain, Jinxiu, Guangxi
6	锦香草 P.cavaleriei	GDSMT042	广东英德石门台	113°31'	24°41'	289
			Yingdeshimentai, Guangzhou
7	锦香草 P.cavaleriei	GXYL045	广西玉林市Yulin, Guangxi	110°18'	22°66'	673
8	锦香草 P.cavaleriei	GXGPSBH046	广西桂平市十八河	110°02'	23°61'	356
			Shibahe, Guiping, Guangxi
9	锦香草 P.cavaleriei	GDNL049	广东乳源县Ruyuan, Guangdong	113°04'	24°94'	300
10	锦香草 P.cavaleriei	GZXDJ054	贵州榕江县小丹江村	108°34'	26°34'	578
			Xiaodanjiang Village, Rongjiang, Guizhou
11	锦香草 P.cavaleriei	GZXDJ056	贵州榕江县小丹江村	108°34'	26°34'	728
			Xiaodanjiang Village, Rongjiang, Guizhou
12	锦香草 P.cavaleriei	GZXDJ058	贵州榕江县小丹江村	108°34'	26°34'	640
			Xiaodanjiang Village, Rongjiang, Guizhou
编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
13	锦香草 P.cavaleriei	GZXDJ060	贵州榕江县小丹江村	108°34'	26°34'	710
			Xiaodanjiang Village, Rongjiang, Guizhou
14	锦香草 P.cavaleriei	GZLGS062	贵州雷山县雷公山	108°07'	26°38'	1 345
			Leigong Mountain, Leishan, Guizhou
15	锦香草 P.cavaleriei	GZLGS067	贵州雷山县雷公山	108°07'	26°38'	1 456
			Leigong Mountain, Leishan, Guizhou
16	锦香草 P.cavaleriei	GZLGS069	贵州雷山县雷公山	108°07'	26°38'	1 277
			Leigong Mountain, Leishan, Guizhou
17	锦香草 P.cavaleriei	GXJX076	广西金秀县下古呈	110°06'	23°89'	780
			Xiagucheng, Jinxiu, Guangxi
18	锦香草 P.cavaleriei	GXJX081	广西金秀县下古呈	110°06'	23°89'	690
			Xiagucheng, Jinxiu, Guangxi
19	锦香草 P.cavaleriei	GXJX084	广西金秀县下古呈	110°06'	23°89'	805
			Xiagucheng, Jinxiu, Guangxi
20	锦香草 P.cavaleriei	GXJX086	广西金秀县下古呈	110°06'	23°89'	710
			Xiagucheng, Jinxiu, Guangxi
21	锦香草 P.cavaleriei	GXJXLHS093	广西金秀莲花山	23°89'	24°16'	850
			Lianhua Mountain, Jinxiu, Guangxi
22	锦香草 P.cavaleriei	GZLGS096	贵州雷山县雷公山	108°07'	26°38'	1 259
			Leigong Mountain, Leishan, Guizhou
23	锦香草 P.cavaleriei	HNYZ098	湖南永州祁阳县	112°09'	26°28'	323
			Qiyang, Yunzhou, Hunan
24	锦香草 P.cavaleriei	JXQYS100	江西崇义县齐云山	113°59'	25°54'	633
			Qiyun Mountain, Chongyi, Jiangxi
25	短毛熊巴掌 P.cavaleriei var. tankahkeei	HNSHS018	湖南新宁县舜皇山 Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	893
26	短毛熊巴掌 P. cavaleriei var. tankahkeei	HNSHS025	湖南新宁县舜皇山 Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	720
27	短毛熊巴掌 P. cavaleriei var. tankahkeei	FJFZQS031	福建闽侯县旗山 Qi Mountain, Minhou, Fujian	119°13'	25°97'	694
28	短毛熊巴掌 P.cavaleriei var. tankahkee	GXJXSTS035	广西金秀县圣堂山 Shengtang Mountain, Jinxiu, Guangxi	110°11'	23°97'	835
29	短毛熊巴掌 P.cavaleriei var. tankahkeei	GXJXYS036	广西金秀县银杉公园 Yinshan Park, Jinxiu, Guangxi	110°24'	24°21'	1 140
30	短毛熊巴掌 P.cavaleriei var. tankahkeei	FJGYS039	福建华安县贡鸭山 Gongya Mountain, Hua'an, Fujian	117°42'	24°91'	735
31	短毛熊巴掌 P.cavaleriei var. tankahkeei	GZLGS066	贵州雷山县雷公山 Leigong Mountain, Leishan, Guizhou	108°07'	26°38'	1 340
32	短毛熊巴掌 P.cavaleriei var. tankahkeei	FJJDS097	福建上杭县吉当山 Jidang Mountain, Shanghang, Fujian	116°82'	25°18'	849
33	短毛熊巴掌 P.cavaleriei var. tankahkeei	ZJWYL099	浙江泰顺县乌岩岭自然保护区 Wuyanling Nature Reserve, Taishun, Zhejiang	119°44'	27°48'	510
34	短毛熊巴掌 P. cavaleriei var. tankahkeei	FJNPWML101	福建南平万木林保护区 Wanmulin Nature Reserve, Nanping, Fujian	118°14'	27°05'	316
35	长柄熊巴掌 P. cavaleriei var. wilsoniana	SCELS102	四川天全县二郎山 Erlang Mountain, Tianquan Country, Sichuan	102°35'	29°87'	1 807
36	长柄熊巴掌 P. cavaleriei var. wilsoniana	SCWWS104	四川瓦屋山 Wawu Mountain, Sichuan	102°53'	29°38'	2 670
编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
37	长柄熊巴掌 P. cavaleriei var. wilsoniana	MG993330.1	四川瓦屋山 Wawu Mountain, Sichuan	102°53'	29°38'	#
38	异药花 Fordiophyton faberi	J344	四川峨眉市峨眉山 Mount Emei, Emei, Sichuan	#	#	760
39	药囊花 Cyphotheca montana	J361	云南红河州金平县 Jinping, Honghezhou, Yunnan	#	#	1 150
40	光萼八蕊花 Sporoxeia latifolia var. fengii	J376	广西那坡县 Napo, Guangxi	#	#	1 220
41	偏瓣花 Plagiopetalum esquirolii	J377	广西那坡县 Napo, Guangxi	#	#	700
42	柏拉木 Blastus cochinchinensis	J440	香港 Hongkong	#	#	560
43	野海棠 Bredia hirsute	J448	砂拉越州西里京镇 Serikin, Sarawak	#	#	#
44	长穗花 Styrophyton caudatum	J569	云南麻栗坡 Malipo, Yunnan	#	#	1 459
45	红敷地发 P. elattandra	J679	广东江门台山 Taishan, Jiangmen, Guangdong	#	#	220
46	圆叶锦香草 P. rotundifolia	J699	金斯堡社区学院 KBCC	#	#	#

编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
1	锦香草 P.cavaleriei	HNSHS011	湖南新宁县舜皇山	111°0'	26°24'	712
2	锦香草 P.cavaleriei	HNSHS016	Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	706
3	锦香草 P.cavaleriei	HNSHS024	湖南新宁县舜皇山	111°01'	26°24'	686
			Shunhuang Mountain, Xinning, Hunan
4	锦香草 P.cavaleriei	GDDXS028	广东仁化县丹霞山	113°71'	25°02'	317
			Danxia Mountain, Dehua, Guangzhou
5	锦香草 P.cavaleriei	GXJXLHS030	广西金秀县莲花山	110°11'	24°16'	861
			Lianhua Mountain, Jinxiu, Guangxi
6	锦香草 P.cavaleriei	GDSMT042	广东英德石门台	113°31'	24°41'	289
			Yingdeshimentai, Guangzhou
7	锦香草 P.cavaleriei	GXYL045	广西玉林市Yulin, Guangxi	110°18'	22°66'	673
8	锦香草 P.cavaleriei	GXGPSBH046	广西桂平市十八河	110°02'	23°61'	356
			Shibahe, Guiping, Guangxi
9	锦香草 P.cavaleriei	GDNL049	广东乳源县Ruyuan, Guangdong	113°04'	24°94'	300
10	锦香草 P.cavaleriei	GZXDJ054	贵州榕江县小丹江村	108°34'	26°34'	578
			Xiaodanjiang Village, Rongjiang, Guizhou
11	锦香草 P.cavaleriei	GZXDJ056	贵州榕江县小丹江村	108°34'	26°34'	728
			Xiaodanjiang Village, Rongjiang, Guizhou
12	锦香草 P.cavaleriei	GZXDJ058	贵州榕江县小丹江村	108°34'	26°34'	640
			Xiaodanjiang Village, Rongjiang, Guizhou
编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
13	锦香草 P.cavaleriei	GZXDJ060	贵州榕江县小丹江村	108°34'	26°34'	710
			Xiaodanjiang Village, Rongjiang, Guizhou
14	锦香草 P.cavaleriei	GZLGS062	贵州雷山县雷公山	108°07'	26°38'	1 345
			Leigong Mountain, Leishan, Guizhou
15	锦香草 P.cavaleriei	GZLGS067	贵州雷山县雷公山	108°07'	26°38'	1 456
			Leigong Mountain, Leishan, Guizhou
16	锦香草 P.cavaleriei	GZLGS069	贵州雷山县雷公山	108°07'	26°38'	1 277
			Leigong Mountain, Leishan, Guizhou
17	锦香草 P.cavaleriei	GXJX076	广西金秀县下古呈	110°06'	23°89'	780
			Xiagucheng, Jinxiu, Guangxi
18	锦香草 P.cavaleriei	GXJX081	广西金秀县下古呈	110°06'	23°89'	690
			Xiagucheng, Jinxiu, Guangxi
19	锦香草 P.cavaleriei	GXJX084	广西金秀县下古呈	110°06'	23°89'	805
			Xiagucheng, Jinxiu, Guangxi
20	锦香草 P.cavaleriei	GXJX086	广西金秀县下古呈	110°06'	23°89'	710
			Xiagucheng, Jinxiu, Guangxi
21	锦香草 P.cavaleriei	GXJXLHS093	广西金秀莲花山	23°89'	24°16'	850
			Lianhua Mountain, Jinxiu, Guangxi
22	锦香草 P.cavaleriei	GZLGS096	贵州雷山县雷公山	108°07'	26°38'	1 259
			Leigong Mountain, Leishan, Guizhou
23	锦香草 P.cavaleriei	HNYZ098	湖南永州祁阳县	112°09'	26°28'	323
			Qiyang, Yunzhou, Hunan
24	锦香草 P.cavaleriei	JXQYS100	江西崇义县齐云山	113°59'	25°54'	633
			Qiyun Mountain, Chongyi, Jiangxi
25	短毛熊巴掌 P.cavaleriei var. tankahkeei	HNSHS018	湖南新宁县舜皇山 Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	893
26	短毛熊巴掌 P. cavaleriei var. tankahkeei	HNSHS025	湖南新宁县舜皇山 Shunhuang Mountain, Xinning, Hunan	111°0'	26°24'	720
27	短毛熊巴掌 P. cavaleriei var. tankahkeei	FJFZQS031	福建闽侯县旗山 Qi Mountain, Minhou, Fujian	119°13'	25°97'	694
28	短毛熊巴掌 P.cavaleriei var. tankahkee	GXJXSTS035	广西金秀县圣堂山 Shengtang Mountain, Jinxiu, Guangxi	110°11'	23°97'	835
29	短毛熊巴掌 P.cavaleriei var. tankahkeei	GXJXYS036	广西金秀县银杉公园 Yinshan Park, Jinxiu, Guangxi	110°24'	24°21'	1 140
30	短毛熊巴掌 P.cavaleriei var. tankahkeei	FJGYS039	福建华安县贡鸭山 Gongya Mountain, Hua'an, Fujian	117°42'	24°91'	735
31	短毛熊巴掌 P.cavaleriei var. tankahkeei	GZLGS066	贵州雷山县雷公山 Leigong Mountain, Leishan, Guizhou	108°07'	26°38'	1 340
32	短毛熊巴掌 P.cavaleriei var. tankahkeei	FJJDS097	福建上杭县吉当山 Jidang Mountain, Shanghang, Fujian	116°82'	25°18'	849
33	短毛熊巴掌 P.cavaleriei var. tankahkeei	ZJWYL099	浙江泰顺县乌岩岭自然保护区 Wuyanling Nature Reserve, Taishun, Zhejiang	119°44'	27°48'	510
34	短毛熊巴掌 P. cavaleriei var. tankahkeei	FJNPWML101	福建南平万木林保护区 Wanmulin Nature Reserve, Nanping, Fujian	118°14'	27°05'	316
35	长柄熊巴掌 P. cavaleriei var. wilsoniana	SCELS102	四川天全县二郎山 Erlang Mountain, Tianquan Country, Sichuan	102°35'	29°87'	1 807
36	长柄熊巴掌 P. cavaleriei var. wilsoniana	SCWWS104	四川瓦屋山 Wawu Mountain, Sichuan	102°53'	29°38'	2 670
编号 No.	名称 Name	凭证标本或GenBank 登录号 Voucher specimen or GenBank accession number	来源 Origin	经度 Longitude	纬度 Latitude	海拔 Altitude/m
37	长柄熊巴掌 P. cavaleriei var. wilsoniana	MG993330.1	四川瓦屋山 Wawu Mountain, Sichuan	102°53'	29°38'	#
38	异药花 Fordiophyton faberi	J344	四川峨眉市峨眉山 Mount Emei, Emei, Sichuan	#	#	760
39	药囊花 Cyphotheca montana	J361	云南红河州金平县 Jinping, Honghezhou, Yunnan	#	#	1 150
40	光萼八蕊花 Sporoxeia latifolia var. fengii	J376	广西那坡县 Napo, Guangxi	#	#	1 220
41	偏瓣花 Plagiopetalum esquirolii	J377	广西那坡县 Napo, Guangxi	#	#	700
42	柏拉木 Blastus cochinchinensis	J440	香港 Hongkong	#	#	560
43	野海棠 Bredia hirsute	J448	砂拉越州西里京镇 Serikin, Sarawak	#	#	#
44	长穗花 Styrophyton caudatum	J569	云南麻栗坡 Malipo, Yunnan	#	#	1 459
45	红敷地发 P. elattandra	J679	广东江门台山 Taishan, Jiangmen, Guangdong	#	#	220
46	圆叶锦香草 P. rotundifolia	J699	金斯堡社区学院 KBCC	#	#	#

种名 Species name	序列长度 Sequence length/bp	5.8S		ITS1		ITS2
种名 Species name	序列长度 Sequence length/bp	长度 Length/bp	GC含量 GC content/%	长度 Length/bp	GC含量 GC content/%	长度 Length/bp	GC含量 GC content/%
锦香草P. cavaleriei	733~777	160~161	53.1~53.4	246	63.4~65.8	230	64.8~66.5
短毛熊巴掌	717~763	160	53.1	246	63.4~65.0	230	63.9~65.7
P. cavaleriei var. tankahkeei
长柄熊巴掌	778~783	160	53.1	246	63.4~63.8	230	62.6
P. cavaleriei var. wilsoniana

种名 Species name	序列长度 Sequence length/bp	5.8S		ITS1		ITS2
种名 Species name	序列长度 Sequence length/bp	长度 Length/bp	GC含量 GC content/%	长度 Length/bp	GC含量 GC content/%	长度 Length/bp	GC含量 GC content/%
锦香草P. cavaleriei	733~777	160~161	53.1~53.4	246	63.4~65.8	230	64.8~66.5
短毛熊巴掌	717~763	160	53.1	246	63.4~65.0	230	63.9~65.7
P. cavaleriei var. tankahkeei
长柄熊巴掌	778~783	160	53.1	246	63.4~63.8	230	62.6
P. cavaleriei var. wilsoniana

种名(来源) Species name and origin	锦香草 (湖南) P. cavaleriei	锦香草 (广东) P. cavaleriei (Guangdong)	锦香草 (贵州) P. cavaleriei (Guizhou)	锦香草 (广西) P. cavaleriei (Guangxi)	锦香草 (江西) P. cavaleriei (Jiangxi)	短毛熊巴掌 (湖南) P. cavaleriei var. tankahkeei (Hunan)	短毛熊巴掌 (广西) P. cavaleriei var. tankahkeei (Guangxi)	短毛熊巴掌 (福建) P. cavaleriei var. tankahkeei (Fujian)	短毛熊巴掌 (贵州) P. cavaleriei var. tankahkeei (Guizhou)	短毛熊巴掌 (浙江) P. cavaleriei var. tankahkeei
锦香草(广东)	0.001
P. cavaleriei (Guangdong)
锦香草(贵州)	0.011	0.012
P. cavaleriei (Guizhou)
锦香草(广西)	0.008	0.009	0.003
P. cavaleriei (Guangxi)
锦香草(江西)	0.011	0.012	0.005	0.003
P. cavaleriei (Jiangxi)
短毛熊巴掌(湖南)	0.000	0.001	0.011	0.008	0.011
P. cavaleriei var. tankahkeei(Hunan)
短毛熊巴掌(广西)	0.009	0.011	0.001	0.001	0.004	0.009
P. cavaleriei var. tankahkeei(Guangxi)
短毛熊巴掌(福建)	0.012	0.013	0.007	0.004	0.007	0.012	0.005
P. cavaleriei var. tankahkeei(Fujian)
短毛熊巴掌(贵州)	0.014	0.016	0.009	0.006	0.009	0.015	0.008	0.003
P. cavaleriei var. tankahkeei(Guizhou)
短毛熊巴掌(浙江)	0.016	0.017	0.011	0.008	0.011	0.016	0.009	0.004	0.001
P. cavaleriei var. tankahkeei(Zhejiang)
长柄熊巴掌(四川)	0.034	0.036	0.041	0.037	0.041	0.035	0.040	0.041	0.044	0.036
P. cavaleriei var. wilsoniana(Sichuan)

锦香草及其近缘种的亲缘关系ITS序列分析

Analysis of inter-species relationship of Phyllagathis cavaleriei and its related species by use of ITS sequences

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