草莓白化相关病毒中国分离物全基因组分析

doi:10.16420/j.issn.0513-353x.2019-0989

摘要/Abstract

摘要：

草莓白化相关病毒（strawberry pallidosis-associated virus,SPaV）属于长线形病毒科（Closteroviridae）毛形病毒属（Crinivirus）,可引起草莓病害,2017年在中国首次报道。采用高通量测序、RACE和RT-PCR技术获得了SPaV中国分离物（FJ）的基因组全长。该病毒含有两条正单链基因组RNA1和RNA2。RNA1全长8 048 nt,5′和3′非编码区序列分别为264和197 nt,含有3个开放阅读框（ORF）,分别编码ORF 1a/1b融合蛋白和p9蛋白。RNA2全长7 977 nt,5′和3′非编码区序列分别为248和186 nt,含有8个开放阅读框（ORF）,分别编码HSP70h、CPh、CP、CPm、p7、p6、p9和p28等8个蛋白。RNA1和RNA2与美国M1分离物分别具有98.5%和99.0%的核苷酸一致性;系统发育分析结果表明,SPaV中国分离物（FJ）单独处在一个分支。对SPaV来源的小RNA的分析表明,来源于SPaV的小RAN长度以21和22 nt为主。

Abstract:

Strawberry pallidosis-associated virus（SpaV）belongs to the genus Crinivirus of the family Closteroviridae,which causes strawberry diseases. It was first reported in China in 2017. In this study,high-throughput sequencing,RACE and RT-PCR techniques were used to obtain the full length of SPaV China isolate（FJ）. The virus contains two positive-sense single,stranded genome RNA1 and RNA2,RNA1 is composed of 8 048 nt and contains three open reading frames（ORFs）,encoding ORF 1a/1b fusion protein and p9 protein,flanked by 5′ and 3′ untranslated regions of 264 nt and 197 nt,respectively. RNA2 is composed of 7 977 nt and contains eight ORFs,encoding HSP70h,CPh,CP,CPm and other four unknown function proteins（p7,p6,p9 and p28）,flanked by 5′ and 3′ untranslated regions of 248 and 186 nt,respectively. RNA1 and RNA2 share 98.5% and 99.0% nucleotide sequence identity with that of M1 isolate（USA）,respectively. Phylogenetic analysis showed that SPaV China isolate （FJ）was clustered in a separate branch. The analysis of sRNA derived from SPaV showed that the dominant sizes were 21 nt and 22 nt.

Key words: Fragaria × ananassa, strawberry pallidosis-associated virus, NGS, RT-PCR, viral genome sequence

中图分类号:

S668.4

陈道, 张洁, 吴祖建, 丁新伦. 草莓白化相关病毒中国分离物全基因组分析[J]. 园艺学报, 2021, 48(1): 146-152.

CHEN Dao, ZHANG Jie, WU Zujian, DING Xinlun. Genome Sequence of Strawberry Pallidosis-associated Virus Isolated in China[J]. Acta Horticulturae Sinica, 2021, 48(1): 146-152.

图/表 5

参考文献 11

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引物名称 Primer name	引物序列（5′-3′） Primer pair
RNA1-1	F:GTAATGAAGTCGTCGAGACCA;R:GAATAGACTACTTCGCCATGTCTG	47
RNA1-2	F:GAGACGATAACGATGTGCGAC;R:TCGACAGTTGAGAGTCGTGAC	52
RNA1-3	F:TCTAAATTTAAGAGTTTGCTGTTTAA;R:ATAGCTCTAGAGGCTCTAGCATT	57
RNA1-4	F:TGCGTATCAGTATGTCACCAC;R:CATGGCCAAGGTCACCAGAGT	56
RNA1-5	F:TCTTATAATATTCATACCCTTGAAC;R:CGGGAATTTATTGTAAATCATAC	58
race1-3	ACACTGGAGTTAAAGTCTGAAACTCTAAATTTAAGAGTTTAG	57
race1-5	TTTAAAAGTAACTGATACATCTTCAGGCAGACATAATCTCT	61
RNA2-1	F:CCAAAGATAAATAACAAACAATCAA;R:ATATCTTCCCGCATACGAATTTATCAA	57
RNA2-2	F:GGGCAGATCAAAGAACAACTTGTT;R:CATTGATCGTTCCACTCAATTTGAAA	60
RNA2-3	F:ATGACGTCGCAAATGATAGTGCTTTC;R:CTTCCAACTCTCTGCTCGGTAACTT	55
RNA2-4	F:CTTGATAAACGTGACAAATCAAACCT;R:GGGGTCAGCTCTTTCAAAGCTTGA	57
RNA2-5	F:CTTTTTATTAGCGTCAATCTTAGCTTT;R:TTTCTCGGTCAATCGATCGAACTCTGA	58
RNA2-6	F:CAAGAATAATGGATTGGAGGATATGCCTT;R:TCACCAGGGGATTGCTTTCCAACTT	56
race2-3	GATTACGCCAAGCTTTGCTACCTCCTATACCAATGCGGA	67
race2-5	ACCTCCCAAACGAATTTATCAATCTCATAAGGTAAAGTGTT	65

引物名称 Primer name	引物序列（5′-3′） Primer pair
RNA1-1	F:GTAATGAAGTCGTCGAGACCA;R:GAATAGACTACTTCGCCATGTCTG	47
RNA1-2	F:GAGACGATAACGATGTGCGAC;R:TCGACAGTTGAGAGTCGTGAC	52
RNA1-3	F:TCTAAATTTAAGAGTTTGCTGTTTAA;R:ATAGCTCTAGAGGCTCTAGCATT	57
RNA1-4	F:TGCGTATCAGTATGTCACCAC;R:CATGGCCAAGGTCACCAGAGT	56
RNA1-5	F:TCTTATAATATTCATACCCTTGAAC;R:CGGGAATTTATTGTAAATCATAC	58
race1-3	ACACTGGAGTTAAAGTCTGAAACTCTAAATTTAAGAGTTTAG	57
race1-5	TTTAAAAGTAACTGATACATCTTCAGGCAGACATAATCTCT	61
RNA2-1	F:CCAAAGATAAATAACAAACAATCAA;R:ATATCTTCCCGCATACGAATTTATCAA	57
RNA2-2	F:GGGCAGATCAAAGAACAACTTGTT;R:CATTGATCGTTCCACTCAATTTGAAA	60
RNA2-3	F:ATGACGTCGCAAATGATAGTGCTTTC;R:CTTCCAACTCTCTGCTCGGTAACTT	55
RNA2-4	F:CTTGATAAACGTGACAAATCAAACCT;R:GGGGTCAGCTCTTTCAAAGCTTGA	57
RNA2-5	F:CTTTTTATTAGCGTCAATCTTAGCTTT;R:TTTCTCGGTCAATCGATCGAACTCTGA	58
RNA2-6	F:CAAGAATAATGGATTGGAGGATATGCCTT;R:TCACCAGGGGATTGCTTTCCAACTT	56
race2-3	GATTACGCCAAGCTTTGCTACCTCCTATACCAATGCGGA	67
race2-5	ACCTCCCAAACGAATTTATCAATCTCATAAGGTAAAGTGTT	65

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