Abstract
Apple rubbery wood virus 2 (ARWV-2) and citrus virus A (CiVA) belong to a recently approved family Phenuiviridae in the order Bunyavirales and possess negative-sense single-stranded RNA genomes. In this study, the genome sequence of three ARWV-2 isolates (S17E2, LYC2, and LYXS) and a CiVA isolate (CiVA-P) infecting pear trees grown in China were characterized using high-throughput sequencing combined with conventional reverse-transcription PCR (RT-PCR) assays. The genome-wide nt sequence identities were above 93.6% among the ARWV-2 isolates and above 93% among CiVA isolates. Sequence comparisons showed that sequence diversity occurred in the 5′ untranslated region of the ARWV-2 genome and the intergenic region of the CiVA genome. For the first time, this study revealed that ARWV-2 proteins Ma and Mb displayed a plasmodesma subcellular localization, and the MP of CiVA locates in cell periphery and can interact with the viral NP in bimolecular fluorescence complementation assays. RT-PCR tests disclosed that ARWV-2 widely occurs, while CiVA has a low incidence in pear trees grown in China. This study presents the first complete genome sequences and incidences of ARWV-2 and CiVA from pear trees and the obtained results extend our knowledge of the viral pathogens of pear grown in China.
Highlights
ARWV-1 and Apple rubbery wood virus 2 (ARWV-2) have been identified in apple grown in China [3,4], and ARWV-2 was reported to be able to infect pear in China [5] and apple in Italy [6]
We report the complete genome sequences of ARWV-2 and citrus virus A (CiVA) infecting pear by using high throughput sequencing combined with conventional Sanger sequencing and their infection status in pear trees grown in China
high-throughput sequencing (HTS) was used to identify viruses in four diseased pear samples and revealed the presence of two recently described -ssRNA viruses, ARWV-2 and CiVA, and revealed the presence of two recently described -ssRNA viruses, ARWV-2 and CiVA, and three common pear-infecting viruses in the samples. This is the first report of the three common pear-infecting viruses in the samples. This is the first report of the complete genome sequences of the two viruses from pear trees and the first detection of complete genome sequences of the two viruses from pear trees and the first detection of
Summary
(ARWV-2) were discovered in apple trees showing rubbery wood disease symptoms in Germany and USA [1] Both ARWV-1 and ARWV-2 possess a tripartite negative-sense single-stranded RNA (-ssRNA) genome and are associated with apple rubbery wood disease [1]. ARWV-1 and ARWV-2 have been identified in apple grown in China [3,4], and ARWV-2 was reported to be able to infect pear in China [5] and apple in Italy [6]. In addition to PCLSaV, two other -ssRNA viruses ARWV-2 and CiVA were identified. This is the first finding of these two viruses infecting pear grown in China. We report the complete genome sequences of ARWV-2 and CiVA infecting pear by using high throughput sequencing combined with conventional Sanger sequencing and their infection status in pear trees grown in China. For the first time the subcellular localization of the predicated movement proteins encoded by the two viruses was determined
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