建兰花叶病毒的遗传变异及其寄主适应性

doi:10.16420/j.issn.0513-353x.2023-0170

摘要/Abstract

摘要：

建兰花叶病毒（Cymbidium mosaic virus，CymMV）是危害兰花并造成重要经济损失的病毒之一。为揭示CymMV的遗传变异及其适应性进化特征，新测定了16个CymMV石斛兰（Dendrobium nobile）分离物的外壳蛋白基因（coat protein，CP）序列，并结合已报道的52个CymMV中国分离物CP基因序列进行遗传变异及进化分析。遗传多样性分析结果显示，68个CymMV分离物CP基因的核苷酸多样性（P_i）和单倍型多样性（H_d）平均值分别为0.039和0.992，表明该病毒具有较高的遗传多样性。分子变异分析（analysis of molecular variance，AMOVA）显示，CymMV变异源主要来自于病毒个体，约占总变异的71.72%。群体遗传结构分析显示，该病毒可以分为两个大簇（Cluster），CymMV建兰（Cymbidium ensifolium）分离物独立为一簇（Cluster 1），其他寄主分离物聚为另一大簇（Cluster 2），Cluster 2内相同寄主来源的病毒分离物倾向于相聚成簇。进一步的系统发育与性状关联分析结果显示CymMV与寄主来源存在显著的关联性。上述结果表明，CymMV的遗传变异具有寄主特异性，其适应性进化主要受寄主因素所驱动。

关键词: 建兰花叶病毒, 外壳蛋白基因, 遗传多样性, 主成分判别分析, 适应性进化

Abstract:

Cymbidium mosaic virus（CymMV）is a highly destructive plant pathogen that inflicts significant losses on orchid production. This study investigates the genetic variability and adaptive evolution of CymMV. The coat protein（CP）gene of 16 CymMV isolates from Dendrobium nobile was sequenced and analyzed alongside published CP sequences of 52 CymMV isolates from various host plants in China. Analysis reveals a substantial genetic diversity within the CP gene，with an overall nucleotide diversity of 0.039 and a haplotype diversity of 0.992. AMOVA tests demonstrate that the molecular variation among individuals accounts for 71.72% of the total variation of CymMV. Population structure analysis classifies CymMV into two distinct clusters. Cluster 1 exclusively consists of CymMV isolates from Cymbidium ensifolium，while Cluster 2 encompasses CymMV isolates from multiple host species，yet isolates from the same host species tend to form subclusters. Furthermore，a robust association between the phylogeny and host species of CymMV is revealed through phylogeny-trait association analysis. Overall，the findings of this study indicate that the genetic variation of CymMV is specific to its host species，suggesting that its evolution is primarily driven by selection pressures imposed by its hosts.

Key words: Cymbidium mosaic virus, coat protein gene, genetic diversity, discriminant analysis of principal components, adaptive evolution

关夏玉, 陈细红, 郭菁, 吕浩阳, 梁晨媛, 高芳銮. 建兰花叶病毒的遗传变异及其寄主适应性[J]. 园艺学报, 2024, 51(1): 203-212.

GUAN Xiayu, CHEN Xihong, GUO Jing, LÜ Haoyang, LIANG Chenyuan, GAO Fangluan. Genetic Variability and Host Adaption of Cymbidium Mosaic Virus[J]. Acta Horticulturae Sinica, 2024, 51(1): 203-212.

图/表 10

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寄主 Host	采样省份 Location	分离物 Isolate	GenBank 登录号 Accession No.	寄主 Host	采样省份 Location	分离物 Isolate	GenBank 登录号 Accession No.
国兰 Cymbidium sinense	福建Fujian	CHN-FJZZ12	DQ067878	蝴蝶兰 Phalaenopsis aphrodite	河南Henan	CHN-HNZZ02	KU873001
		CHN-FJZZ13	DQ067879	蕙兰C. faberi	西藏Tibet	C-LZ02	KP137368
	湖北Hubei	CHN-HBWH21	DQ067880			C-LZ06	KP137369
		CHN-HBWH22	DQ067881			C-LZ14	KP137370
	浙江Zhejiang	CHN-ZJHZ52	DQ067882			C-LZ18	KP137371
		CHN-ZJHZ61	DQ067883			C-LZ21	KP137372
		CHN-ZJHZ62	DQ067884	石斛兰 Dendrobium nobile	中国台湾 Taiwan，China	CyMV-1 ^*	OQ615774
		CHN-ZJHZ63	DQ067885			CyMV-2 ^*	OQ615782
		CHN-ZJHZ67	DQ067886			CyMV-3 ^*	OQ615783
		CHN-ZJHZ69	DQ067887			CyMV-4 ^*	OQ615784
		CHN-ZJHZ612	DQ067888			CyMV-5 ^*	OQ615785
蝴蝶兰 Phalaenopsis aphrodite	浙江Zhejiang	CyMV-hangzhou	AM398153			CyMV-6 ^*	OQ615786
		CrMV-HZ	DQ915439			CyMV-7 ^*	OQ615787
	福建Fujian	xm	GQ507023			CyMV-8 ^*	OQ615788
	江苏Jiangsu	CrMV-NJ-1	JQ860108			CyMV-9 ^*	OQ615789
	广东Guangdong	FS2	KF853247			CyMV-10 ^*	OQ615775
		FS3	KF853248			CyMV-11 ^*	OQ615776
		FS4	KF853249			CyMV-12 ^*	OQ615777
		FS5	KF853250			CyMV-13^*	OQ615778
		GZ1	KF853251			CyMV-14 ^*	OQ615779
		CH	KF853252			CyMV-15 ^*	OQ615780
		GZ2	KF853253			CyMV-16 ^*	OQ615781
		GZ3	KF853254		广东Guangdong	ZHD1	EU672821
		FS6	KF853255	建兰 C. ensifolium	浙江Zhejiang	ZJXTHC1	GU295167
		ZH1	KF853256		四川Sichuan	SCJJC7	GU295168
		ZH2	KF853257			SCDXZC4	GU295172
		FS7	KF853258		广东 Guangdong	GZKTLC6	GU295170
		FS8	KF853259			GDDHLC5	GU295173
		FS9	KF853260		北京 Beijing	BJKTLC3	GU295169
		FS10	KF853261		云南 Yunnan	KMMLC2	GU295171
		SZ1	KF853262	墨兰 C. haematodes	广东 Guangdong	gd2	AY360409
		SZ2	KF853263			gd3	AY360410
		SZ3	KF853264		中国台湾 Taiwan，China	CrMV-CS	AY429021
		SZ4	KF853265			Taiwan，China	AY571289

寄主 Host	采样省份 Location	分离物 Isolate	GenBank 登录号 Accession No.	寄主 Host	采样省份 Location	分离物 Isolate	GenBank 登录号 Accession No.
国兰 Cymbidium sinense	福建Fujian	CHN-FJZZ12	DQ067878	蝴蝶兰 Phalaenopsis aphrodite	河南Henan	CHN-HNZZ02	KU873001
		CHN-FJZZ13	DQ067879	蕙兰C. faberi	西藏Tibet	C-LZ02	KP137368
	湖北Hubei	CHN-HBWH21	DQ067880			C-LZ06	KP137369
		CHN-HBWH22	DQ067881			C-LZ14	KP137370
	浙江Zhejiang	CHN-ZJHZ52	DQ067882			C-LZ18	KP137371
		CHN-ZJHZ61	DQ067883			C-LZ21	KP137372
		CHN-ZJHZ62	DQ067884	石斛兰 Dendrobium nobile	中国台湾 Taiwan，China	CyMV-1 ^*	OQ615774
		CHN-ZJHZ63	DQ067885			CyMV-2 ^*	OQ615782
		CHN-ZJHZ67	DQ067886			CyMV-3 ^*	OQ615783
		CHN-ZJHZ69	DQ067887			CyMV-4 ^*	OQ615784
		CHN-ZJHZ612	DQ067888			CyMV-5 ^*	OQ615785
蝴蝶兰 Phalaenopsis aphrodite	浙江Zhejiang	CyMV-hangzhou	AM398153			CyMV-6 ^*	OQ615786
		CrMV-HZ	DQ915439			CyMV-7 ^*	OQ615787
	福建Fujian	xm	GQ507023			CyMV-8 ^*	OQ615788
	江苏Jiangsu	CrMV-NJ-1	JQ860108			CyMV-9 ^*	OQ615789
	广东Guangdong	FS2	KF853247			CyMV-10 ^*	OQ615775
		FS3	KF853248			CyMV-11 ^*	OQ615776
		FS4	KF853249			CyMV-12 ^*	OQ615777
		FS5	KF853250			CyMV-13^*	OQ615778
		GZ1	KF853251			CyMV-14 ^*	OQ615779
		CH	KF853252			CyMV-15 ^*	OQ615780
		GZ2	KF853253			CyMV-16 ^*	OQ615781
		GZ3	KF853254		广东Guangdong	ZHD1	EU672821
		FS6	KF853255	建兰 C. ensifolium	浙江Zhejiang	ZJXTHC1	GU295167
		ZH1	KF853256		四川Sichuan	SCJJC7	GU295168
		ZH2	KF853257			SCDXZC4	GU295172
		FS7	KF853258		广东 Guangdong	GZKTLC6	GU295170
		FS8	KF853259			GDDHLC5	GU295173
		FS9	KF853260		北京 Beijing	BJKTLC3	GU295169
		FS10	KF853261		云南 Yunnan	KMMLC2	GU295171
		SZ1	KF853262	墨兰 C. haematodes	广东 Guangdong	gd2	AY360409
		SZ2	KF853263			gd3	AY360410
		SZ3	KF853264		中国台湾 Taiwan，China	CrMV-CS	AY429021
		SZ4	KF853265			Taiwan，China	AY571289

簇 Cluster	亚簇 Subcluster	样本量大小 Sample size	单倍型 Haplotypes	单倍型多样性H_d Haplotype diversity	核苷酸多样性P_i Nucleotide diversity
1		7	3	0.524 ± 0.209	0.002 ± 0.009
2		61	53	0.995 ± 0.004	0.040 ± 0.004
2	1	17	14	0.971 ± 0.032	0.041 ± 0.008
	2	11	9	0.961 ± 0.051	0.041 ± 0.012
	3	33	30	0.994 ± 0.009	0.036 ± 0.007
总计Total		68	56	0.992 ± 0.005	0.039 ± 0.004

簇 Cluster	亚簇 Subcluster	样本量大小 Sample size	单倍型 Haplotypes	单倍型多样性H_d Haplotype diversity	核苷酸多样性P_i Nucleotide diversity
1		7	3	0.524 ± 0.209	0.002 ± 0.009
2		61	53	0.995 ± 0.004	0.040 ± 0.004
2	1	17	14	0.971 ± 0.032	0.041 ± 0.008
	2	11	9	0.961 ± 0.051	0.041 ± 0.012
	3	33	30	0.994 ± 0.009	0.036 ± 0.007
总计Total		68	56	0.992 ± 0.005	0.039 ± 0.004

变异源 Source of variation	自由度 df	平方和 Sum of squares	变量组分 Variance components	变异比例 Percentage of variation	固定系数 Fixation index
簇间Among clusters	1	65.903	3.580	21.880	0.220 ^ns
簇内亚簇间Among sub-clusters within clusters	2	61.563	1.046	6.390	0.082^*
亚簇内Within sub-clusters	64	750.931	11.733	71.720	0.282^***
总计Total	67	878.397	16.359