The complete genome of a Watermelon silver mottle virus (WSMoV) (genus Tospovirus, family Bunyaviridae) isolate (WSMoV-GZ) from Guangdong province, China was sequenced. The genomes of WSMoV-GZ contained 3,603, 4,909, and 8,914 nt of small (S), medium (M), and large (L) RNA segments, respectively, and had a genomic organization characteristic of members of the genus Tospovirus. The amino acid sequence of the nucleocapsid (N) protein, S RNA-encoded nonstructural (NSs) protein, M RNA-encoded nonstructural (NSm) protein, Gn/Gc glycoprotein precursor, and RNA-dependent RNA polymerase (RdRp) protein showed 94.3-97.5 % identity with those of other WSMoV isolates. Phylogenetic analysis showed that the N protein of WSMoV-GZ was clustered together with those of the WSMoV isolates. The full sequence of WSMoV-GZ provides a reference genome for comparison with other tospoviruses.

<正>兰花是花卉产业最重要的观赏植物之一[1],一旦感染上病毒,其观赏价值和经济价值将受到严重影响。近年来,随着兰花产业的迅速发展,兰花国际国内贸易和种质交流十分频繁,建兰花叶病毒(Cymbidium mosaic virus,CyMV)和齿兰环斑病毒(Odontoglossum ringspot virus,ORSV)是严重影响兰花生产的2种主要病毒[2-3],有必要建立这两种病毒的高效检测方法。一些研究者对这2种病毒建立了双重RT-PCR检测方法[4-6],但对其检测体系的影响因素研究不多[5]。本试验对此进行了研究,旨在为兰花进出口贸易和安全生产奠定基础。

A watermelon sample possibly infected by tospovirus in Guangdong was detected by ELISA and RT-PCR.Crude extracts from yellow spot watermelon leaves were not react with antisera of Tomato spotted wilt virus(TSWV) and Watermelon silver mottle virus(WSMoV) by ELISA.When they were tested by RT-PCR using primer pair J13/UHP,about 1400bp fragment was amplified,which contained 840bp ORF of nucleocapsid protein(NP).Sequence analysis showed that the NP shared 99% identity at the amino acid level with those reported NP of Melon yellow spot virus(MYSV),phylogenetic analysis confirmed that the virus infected watermelon in Guangdong(designated as MYSV-GZ) was belonged to MYSV serogroup of Tospovirus.

According to the conserved sequences of Banana bunchy top virus(BBTV),Cucumber mosaic virus(CMV) and Banana streak virus(BSV),three specific primer pairs were designed,through optimizing the system,a Multiplex-PCR were established to detect the RNA(CMV) and DNA(BBTV,BSV) simultaneously

Cymbidium mosaic virus(CyMV) and Odontoglossum ringspot virus(ORSV) were evaluated by DAS-ELISA respectively in 44 diseased orchid samples collected from Guangdong province,the results showed that CyMV and ORSV were detected in 59.1% and 36.4% respectively,mixed infections with CyMV and ORSV were detected in 22.7% of the 44 diseased orchid samples.CyMV and ORSV were evaluated by one step RT-PCR respectively in 24 diseased orchid samples collected from Guangdong province,the results showed that CyMV and ORSV were detected in 83.3% and 75% respectively,mixed infections with CyMV and ORSV were detected in 66.7% of the 24 diseased orchid samples.These results suggested that the sensitivity of RT-PCR method is higher than that of DAS-ELISA method.

Some tomato samples possibly infected by tospovirus in Guangdong were detected with indirect ELISA and RT-PCR.The results showed that the virus infected tomato did not react with the antiserum of Tomato spotted wilt virus(TSWV),but about 500 bp fragment of RT-PCR shared 83%-84% nucleotide identities with N gene of those reported tospoviruses.The phylogenetic tree of the N gene fragment compared with those of other tospoviruses indicated that the virus infected tomato was belonged to Tospovirus.

辣椒是一类重要的经济作物,在世界各地广泛种植.辣椒感染病毒后,对其产量和品质都有较大的影响.目前可侵染辣椒的病毒至少有45种,其中包括番茄斑萎病毒属Tospovirus的番茄斑萎病毒Tomato spotted wilt virus(TSWV)、凤仙花坏死病毒Impatiens necrotic spot virus(INSV)、番茄褪绿斑病毒Tomato chlorosis spot virus(TCSV)以及辣椒褪绿病毒Capsicum chlorosis virus(CaCV).