新疆蟠桃中发现油桃茎痘相关病毒和亚洲李属病毒

doi:10.16420/j.issn.0513-353x.2020-0215

摘要/Abstract

摘要：

采用高通量测序（High-throughput sequencing,HTS）技术检测新疆蟠桃感染病毒的情况。采集具有穿孔、脉间褪绿等症状的蟠桃嫩叶,提取总RNA,用于高通量测序,共获得13.36 Gb的数据,包含44 563 187对reads,其中比对到油桃茎痘相关病毒（nectarine stem pitting-associated virus,NSPaV,KT273409）和亚洲李属病毒（asian prunus virus,APV2,KT893294）基因组的分别有9 943对和40 036对,经拼接各得到长4 978和9 393 nt的contig,基因组覆盖率均接近100%（5′端分别差10个和7个碱基）,核酸一致度分别为95.0%和92.9%,将此分离物分别命名为NSPaV-Tao和APV2-Tao4。用RT-PCR方法对23个蟠桃树样品进行了检测,结果NSPaV和APV2检出率分别为43.5%和69.6%。通过比较已知NSPaV病毒的核苷酸序列,发现NSPaV-Tao可能是其他分离物重组的结果。

关键词: NSPaV, APV2, 桃, 高通量测序, RT-PCR, 序列分析

Abstract:

In this study,the High-throughput sequencing technology（HTS）was used to detect the virus infection of Prunus persica var. compressa. The peach leaves showing shot hole and chlorosis between leaf veins were collected to extract total RNA for HTS. A total of 13.36 Gb sequence data,including 44 563 187 paired reads,were obtained. The sequence alignment analysis showed that there were 9 943 and 40 036 paired reads belong to the genome of nectarine stem pitting-associated virus（NSPaV）（KT273409）and asian prunus virus（APV2）（KT893294）. The contigs of 4 978 nt and 9 393 nt with nearly completed coverage of viruses genome（10 and 7 bases missing at 5′ end,respectively）were assembled and the sequence identity with the genome sequence of NSPaV（KT273409）and APV2（KT893294）were 95.0% and 92.9% respectively. We named them as isolates NSPaV-Tao and APV2-Tao4.To confirm the result of HTS,23 samples of P. persica var. compressa were detected by the RT-PCR method for the infection of NSPaV and APV2. The results showed that the infection rate of NSPaV and APV2 was 43.5% and 69.6%,respectively. Moreover,NSPaV-Tao may be a recombination of other NSPaV isolates based on the nucleotide sequence comparison.

Key words: NSPaV, APV2, Prunus persica var. compressa, high-throughput sequencing, RT-PCR, sequence analysis

中图分类号:

S662.1

卜方迪, 陈俊光, 刘贞, 向本春, 申冕, 崔百明, 郑银英. 新疆蟠桃中发现油桃茎痘相关病毒和亚洲李属病毒[J]. 园艺学报, 2021, 48(1): 49-59.

BU Fangdi, CHEN Junguang, LIU Zhen, XIANG Benchun, SHEN Mian, CUI Baiming, ZHENG Yinying. Nectarine Stem Pitting-associated Virus and Asian Prunus Virus Found in Prunus persica of Xinjiang[J]. Acta Horticulturae Sinica, 2021, 48(1): 49-59.

图/表 9

参考文献 29

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病毒 Virus	登录号 Accession No.	寄主 Host	分离物 Isolate	序列长度/nt Nucleic acid length	国家 Country
NSPaV	KT273409	油桃Prunus persica var. nectarina	SF04522E	4 991	美国USA
	KT273410	油桃Prunus persica var. nectarina	12P42	4 991	美国USA
	MN095353	油桃Prunus persica var. nectarina	T04	4 991	中国China
	MF326520	桃Prunus persica	SK	4 991	韩国Korea
	NC027211	油桃Prunus persica var. nectarina	nectarine	4 991	美国USA
APV2	KR998048	桃Prunus persica	Q65a	9 393	美国USA
	KR998049	桃Prunus persica	Q86	9 400	美国USA
	KT893294	梅Prunus mume	Bungo Q-1256-01	9 362	日本Japan
	KT893295	梅Prunus mume	Bonsai	9 375	日本Japan
	KT893297	桃Prunus persica	Tatao25 Q-375-02	9 007	中国China
	KY310581	桃Prunus persica	BC	9 402	韩国Korea
	KY310582	桃Prunus persica	JH	9 393	韩国Korea
	KY445748	梅Prunus mume	WY	9 389	韩国Korea

病毒 Virus	登录号 Accession No.	寄主 Host	分离物 Isolate	序列长度/nt Nucleic acid length	国家 Country
NSPaV	KT273409	油桃Prunus persica var. nectarina	SF04522E	4 991	美国USA
	KT273410	油桃Prunus persica var. nectarina	12P42	4 991	美国USA
	MN095353	油桃Prunus persica var. nectarina	T04	4 991	中国China
	MF326520	桃Prunus persica	SK	4 991	韩国Korea
	NC027211	油桃Prunus persica var. nectarina	nectarine	4 991	美国USA
APV2	KR998048	桃Prunus persica	Q65a	9 393	美国USA
	KR998049	桃Prunus persica	Q86	9 400	美国USA
	KT893294	梅Prunus mume	Bungo Q-1256-01	9 362	日本Japan
	KT893295	梅Prunus mume	Bonsai	9 375	日本Japan
	KT893297	桃Prunus persica	Tatao25 Q-375-02	9 007	中国China
	KY310581	桃Prunus persica	BC	9 402	韩国Korea
	KY310582	桃Prunus persica	JH	9 393	韩国Korea
	KY445748	梅Prunus mume	WY	9 389	韩国Korea

分离物 Isolate	核苷酸 Nucleotide	5′ UTR	P1		P2		CP		CP-RT		3′ UTR
分离物 Isolate	核苷酸 Nucleotide	5′ UTR	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	3′ UTR
T04	96.0	90.9	95.7	98.2	94.5	98.3	97.5	97.6	96.6	97.4	94.6
SK	94.4	89.4	90.3	90.9	96.1	98.3	95.5	97.1	95.6	96.2	94.9
12P42	96.8	91.6	96.4	8.5	96.8	96.8	97.5	98.6	96.2	96.8	95.4
nectarina	94.8	88.6	94.4	96.1	96.0	97.9	96.8	96.2	94.2	93.9	95.0
SF04522E	95.0	86.3	95.1	96.4	96.0	98.5	97.0	97.1	94.4	94.1	95.0

分离物 Isolate	核苷酸 Nucleotide	5′ UTR	P1		P2		CP		CP-RT		3′ UTR
分离物 Isolate	核苷酸 Nucleotide	5′ UTR	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleotide	氨基酸Amino acid	3′ UTR
T04	96.0	90.9	95.7	98.2	94.5	98.3	97.5	97.6	96.6	97.4	94.6
SK	94.4	89.4	90.3	90.9	96.1	98.3	95.5	97.1	95.6	96.2	94.9
12P42	96.8	91.6	96.4	8.5	96.8	96.8	97.5	98.6	96.2	96.8	95.4
nectarina	94.8	88.6	94.4	96.1	96.0	97.9	96.8	96.2	94.2	93.9	95.0
SF04522E	95.0	86.3	95.1	96.4	96.0	98.5	97.0	97.1	94.4	94.1	95.0

分离物 Isolate	核苷酸 Nucleotide	5′ UTR	Rep		基因盒 Triple gene block						CP		3′ UTR
			Rep		TGB1		TGB2		TGB3		CP
			核苷酸Nucleotide	氨基酸Amino acid	核苷酸Nucleo- tide	氨基酸Amino acid	核苷酸Nucleo- tide	氨基酸Amino acid	核苷酸Nucleo- tide	氨基酸Amino acid	核苷酸Nucleo- tide	氨基酸Amino acid
BC	92.6	86.9	92.5	97.3	93.1	98.4	91.9	97.4	86.9	90.8	91.9	94.1	95.6
JH	92.8	96.3	92.4	97.3	93.0	98.4	92.7	97.4	93.0	97.0	91.7	94.4	95.7
WY	90.9	90.8	90.0	95.9	92.1	97.5	94.2	98.3	94.5	98.5	90.3	93.8	93.6
Q86	93.1	85.0	92.4	97.2	93.4	95.9	94.8	97.4	96.0	98.5	93.5	96.8	94.8
Q65a	79.1	83.4	77.7	89.1	78.1	89.6	83.7	92.1	83.9	80.0	78.0	84.3	88.0
Bonsai	93.0	82.0	92.8	97.2	93.4	97.1	93.0	98.3	95.0	95.4	92.9	95.3	93.1
BungoQ	92.9	86.9	92.3	96.8	93.9	98.4	94.8	98.3	96.0	98.5	92.7	94.4	95.6
Tatao25^*	89.4	-	87.4	91.5	93.9	98.0	94.8	98.3	96.0	98.5	93.3	95.0	95.3