稻曲病菌菌株GZ-14-07中携带的全病毒基因组特征研究

doi:10.13560/j.cnki.biotech.bull.1985.2020-0479

摘要/Abstract

摘要：

稻曲病菌是引起真菌类病害水稻稻曲病的病原。从稻曲病菌菌株GZ-14-07中分离到一株真菌病毒,命名为Ustilaginoidea virens RNA virus 6（UvRV6）,对其进行了序列分析。通过CF-11纤维素粉法提取菌株GZ-14-07中的真菌病毒UvRV6,采用随机引物扩增法获得UvRV6的全长基因组序列,使用DNAMAN、BlastP和MEGA等软件对UvRV6的全长序列进行拼接、比对和系统进化树分析。研究发现UvRV6全基因组长度为5 043个碱基,GC为48.8%,UvRV6基因组序列中存在两个开放阅读框（ORF1和ORF2）,ORF1长2 280个碱基,编码760个氨基酸（79.6 kD）,ORF2长2 385个碱基,编码830个氨基酸（91.5 kD）。ORF1和ORF2编码的蛋白,分别与全病毒科维多利亚属真菌病毒成员的衣壳蛋白和依赖于RNA的RNA聚合酶显著出较高的相似性。另外,UvRV6的ORF1和ORF2之间有一个AUGA序列,在AUGA序列上游存在一段能形成一个H型的假节序列,这两个序列元件的存在使UvRV6两个开放阅读框（ORF1和ORF2）能通过翻译终止后又重新启动翻译的机制,融合成一个大的开放阅读框。最后,通过UvRV6和其他全病毒科成员的依赖于RNA的RNA聚合酶蛋白序列构建的系统进化树分析得出,UvRV6与属于全病毒科维多利亚属的成员聚集成一簇。明确了UvRV6属于全病毒科维多利亚属的新成员。

关键词: 稻曲病菌, 真菌病毒, dsRNA, 全病毒科, 维多利亚属

Abstract:

Ustilaginoidea virens is a pathogen that causes fungi-like disease Ustilaginoidea oryzae. A mycovirus isolated from strain GZ-14-07 of U. virens was designated as Ustilaginoidea virens RNA virus 6（UvRV6）and sequenced. Mycovirus UvRV6 was extracted from the strain GZ-14-07 using CF-11 cellulose powder method and its full-length genomic sequence was obtained by random-primer amplification method. Sequence analysis,alignment and phylogenetic tree of the UvRV6 were performed using DNAMAN,BlastP and MEGA software. The complete genome sequence of the UvRV6 was determined to be 5 043 bp in length,with a GC content of 48.8%,and contained two open reading frames（ORF1 and ORF2）. ORF1 had a length of 2 280 bp and encoded a 760-amino-acid protein（79.6 kD）,and ORF2 was 2 385 bp long and encoded a 830-amino-acid protein（91.5 kD）. ORF1- and ORF2-encoded protein had a high percentage of sequence similarity to the capsid protein and the RNA-dependent RNA polymerases（RdRps）of genus Victorivirus in the family Totiviridae,respectively. Moreover,there was an AUGA between ORF1 and ORF2,and a sequence forming H-type pseudo-knot existed in the upstream of the AUGA. These 2 sequence elements allowed ORF1 and ORF2 to be merged as a bigger ORF via a mechanism of re-initiating translation after translation ended. A phylogenetic analysis based on the alignment of RdRps of UvRV6 and other totiviruses showed the UvRV6 was clustered with the members of Victorivirus in the family Totiviridae. Therefore,UvRV6 is confirmed as a new member of Victorivirus in the family Totiviridae.

Key words: Ustilaginoidea virens, mycovirus, dsRNA, Totiviridae, Victorivirus

张婷婷, 滕丽, 蔡小姚, 龙慧, 李升愉, 刘红美. 稻曲病菌菌株GZ-14-07中携带的全病毒基因组特征研究[J]. 生物技术通报, 2020, 36(10): 80-87.

ZHANG Ting-ting, TENG Li, CAI Xiao-yao, LONG Hui, LI Sheng-yu, LIU Hong-mei. Genomic Characterization of a Totiviridae from the Ustilaginoidea virens Strain GZ-14-07[J]. Biotechnology Bulletin, 2020, 36(10): 80-87.

图/表 5

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Virus names	Complete genome	Overlapping motif	5'UTR motif	3'UTR motif	Identity（CP）	Identity（RdRp）	H- type pseudoknot or stem-loop structure	Citation
U. virens RNA virus 6	5043	AUGA	GGGCU	CAUCG	/	/	GAUGCggccAcagcCACUgcugggccUGCGC- CGCAAGUAUGA（stem-loop structure）	This study
U. virens RNA virus 5	5221	UAAUG	UGAAA	UCAAA	38%	38%	GAAGGggcgACcggccGCCGAGAGCCUggc- cgcgccGAAUAAUG（stem-loop structure）	[30]
U. virens RNA virus L	5654	UAAUG	CGAAA	UUUUU	40%	32%	CCCCAagcgCUgccccgcuGAGCgggcCGACU- CAAUCGUAAUG（H- type pseudoknot）	[19]
U. virens RNA virus 3	5075	UAAUG	CGAAA	GCCAA	73%	75%	ggacACggcCACUGCCGguccUGCgccCAGG- AUAUAAUG（H- type pseudoknot）	[31]
U. virens RNA virus 1 strain Uv0901	5220	UAAUG	UGAAA	GGCAA	39%	31%	UAGCGcugccccgcagAGUgggCCGACCCC- CGAGUAAUG（H- type pseudoknot）	[31]
U. virens RNA virus 1 strain JYH-ZT	5567	UAAUG	UGAAA	GGCAA	39%	31%	ACCUAgcgCUGccccgcAGAGUgggCCGAC- CCCCGAGUAAUG（H- type pseudoknot）	[17]
U. virens RNA virus 1 strain HNHS-1	5142	UAAUG	AAGAU	ACAAA	39%	31%	UCAGUcgcugcUGCUCCgcaGgcgGGUGCU- GCUCAGUAAUG（stem-loop structure）	[24]

Virus names	Complete genome	Overlapping motif	5'UTR motif	3'UTR motif	Identity（CP）	Identity（RdRp）	H- type pseudoknot or stem-loop structure	Citation
U. virens RNA virus 6	5043	AUGA	GGGCU	CAUCG	/	/	GAUGCggccAcagcCACUgcugggccUGCGC- CGCAAGUAUGA（stem-loop structure）	This study
U. virens RNA virus 5	5221	UAAUG	UGAAA	UCAAA	38%	38%	GAAGGggcgACcggccGCCGAGAGCCUggc- cgcgccGAAUAAUG（stem-loop structure）	[30]
U. virens RNA virus L	5654	UAAUG	CGAAA	UUUUU	40%	32%	CCCCAagcgCUgccccgcuGAGCgggcCGACU- CAAUCGUAAUG（H- type pseudoknot）	[19]
U. virens RNA virus 3	5075	UAAUG	CGAAA	GCCAA	73%	75%	ggacACggcCACUGCCGguccUGCgccCAGG- AUAUAAUG（H- type pseudoknot）	[31]
U. virens RNA virus 1 strain Uv0901	5220	UAAUG	UGAAA	GGCAA	39%	31%	UAGCGcugccccgcagAGUgggCCGACCCC- CGAGUAAUG（H- type pseudoknot）	[31]
U. virens RNA virus 1 strain JYH-ZT	5567	UAAUG	UGAAA	GGCAA	39%	31%	ACCUAgcgCUGccccgcAGAGUgggCCGAC- CCCCGAGUAAUG（H- type pseudoknot）	[17]
U. virens RNA virus 1 strain HNHS-1	5142	UAAUG	AAGAU	ACAAA	39%	31%	UCAGUcgcugcUGCUCCgcaGgcgGGUGCU- GCUCAGUAAUG（stem-loop structure）	[24]