Zuokun Yang

Actinidia chlorotic ringspot‐associated virus: a novel emaravirus infecting kiwifruit plants

By integrating next-generation sequencing (NGS), bioinformatics, electron microscopy and conventional molecular biology tools, a new virus infecting kiwifruit vines has been identified and characterized. Being associated with double-membrane-bound bodies in infected tissues and having a genome composed of RNA segments, each one containing a single open reading frame in negative polarity, this virus shows the typical features of members of the genus Emaravirus. Five genomic RNA segments were identified. Additional molecular signatures in the viral RNAs and in the proteins they encode, together with data from phylogenetic analyses, support the proposal of creating a new species in the genus Emaravirus to classify the novel virus, which is tentatively named Actinidia chlorotic ringspot-associated virus (AcCRaV). Bioassays showed that AcCRaV is mechanically transmissible to Nicotiana benthamiana plants which, in turn, may develop chlorotic spots and ringspots. Field surveys disclosed the presence of AcCRaV in four different species of kiwifruit vines in five different provinces of central and western China, and support the association of the novel virus with symptoms of leaf chlorotic ringspots in Actinidia. Data on the molecular features of small RNAs of 21-24 nucleotides, derived from AcCRaV RNAs targeted by host RNA silencing mechanisms, are also reported, and possible molecular pathways involved in their biogenesis are discussed.

Characterization of a novel double-stranded RNA mycovirus conferring hypovirulence from the phytopathogenic fungus Botryosphaeria dothidea

A novel double-stranded RNA (dsRNA) virus, designated as Botryosphaeria dothidea RNA virus 1 (BdRV1), isolated from a hypovirulent strain YZN115 of Botryosphaeria dothidea was biologically and molecularly characterized. The genome of BdRV1 comprises of five dsRNAs. Each dsRNA contains a single open reading frame. The proteins encoded by dsRNA1-4 shared significant amino acid identities of 55%, 47%, 43% and 53% with the corresponding proteins of Aspergillus fumigatus tetramycovirus-1. DsRNA1, 3, and 4 of BdRV1 encoded an RNA-dependent RNA polymerase, a viral methyltransferase, and a P-A-S-rich protein, respectively. Function of proteins encoded by the dsRNA2 and dsRNA5 were unknown. BdRV1 conferred hypovirulence for its host and could be transmitted through conidia and hyphae contact.

Deep sequencing reveals the first fabavirus infecting peach

A disease causing smaller and cracked fruit affects peach [Prunus persica (L.) Batsch], resulting in significant decreases in yield and quality. In this study, peach tree leaves showing typical symptoms were subjected to deep sequencing of small RNAs for a complete survey of presumed causal viral pathogens. The results revealed two known viroids (Hop stunt viroid and Peach latent mosaic viroid), two known viruses (Apple chlorotic leaf spot trichovirus and Plum bark necrosis stem pitting-associated virus) and a novel virus provisionally named Peach leaf pitting-associated virus (PLPaV). Phylogenetic analysis based on RNA-dependent RNA polymerase placed PLPaV into a separate cluster under the genus Fabavirus in the family Secoviridae. The genome consists of two positive-sense single-stranded RNAs, i.e., RNA1 [6,357 nt, with a 48-nt poly(A) tail] and RNA2 [3,862 nt, with a 25-nt poly(A) containing two cytosines]. Biological tests of GF305 peach indicator seedlings indicated a leaf-pitting symptom rather than the smaller and cracked fruit symptoms related to virus and viroid infection. To our knowledge, this is the first report of a fabavirus infecting peach. PLPaV presents several new molecular and biological features that are absent in other fabaviruses, contributing to an overall better understanding of fabaviruses.

Identification of a divergent variant of grapevine berry inner necrosis virus in grapevines showing chlorotic mottling and ring spot symptoms

A new variant of grapevine berry inner necrosis virus (GINV) was identified by sequencing of small RNA extracted from ‘Beta’ and Thompson seedless grapevines showing leaf mottle and ring spot symptoms. However, GINV was not found in symptomless samples used as a control. The complete genome sequences of two GINV isolates (KU234316-17) were determined, and these showed 75.76–89.74% sequence identity to the genome of a previously reported Japanese GINV isolate. The new variants appear to be evolutionarily distinct from the original GINV isolate. This is the first report of GINV outside of Japan.

Genetic diversity and evolution of two capsid protein genes of citrus tristeza virus isolates from China

The genetic diversity and population structure of citrus tristeza virus (CTV) isolates from China were investigated based on partial sequences spanning the C-terminal end of p61 and the complete sequences of the CPm and CP genes. Phylogenetic analysis revealed five known groups (RB, T30, T36, HA and VT) and one new group (VI) consisting of only Chinese CTV isolates. Incongruent phylogenetic trees coupled with recombination analysis suggested several recombination events in the CPm gene. Positive selection was detected at codon 9 of CPm and codons 31, 41 and 68 of CP. The widespread CTV subpopulation AT-1 found in China has a unique amino acid insertion at the C-terminus of p61, which could increase CTV population complexity with implications for the evolutionary history of the virus. Our results suggest relevant roles for gene flow, purifying selection and recombination in shaping the CTV population in China.

Molecular characterization of an Apple stem grooving virus isolate from kiwifruit (Actinidia chinensis) in China

Abstract Apple stem grooving virus (ASGV) is the type species of the genus Capillovirus in the family Betaflexiviridae. The virus naturally infects several important horticultural crops. Although many ASGV isolates from apple (Malus spp.), pear (Pyrus spp.), citrus (Citrus spp.) and lily (Lilium spp.) have been characterized, the occurrence status and molecular characteristics of the virus from kiwifruit trees are still largely unknown. In this study, by employing the strategy of small RNA deep sequencing coupled with conventional RT-PCR, we provide the first complete genome sequence of an ASGV isolate (named ASGV-Ac) infecting a kiwifruit plant (Actinidia chinensis) grown in China. The complete genome of ASGV-Ac consisted of 6496 nucleotides, excluding poly (A) tail at its 3ʹ end. ASGV-Ac shared high nucleotide sequence identities of 96.9% and 93.5% with a kiwifruit ASGV isolate reported from New Zealand (accession no. AF522459) for their CP and MP coding regions. However, the isolate was phylogenetically distal to ASGV isolates from all other hosts by sharing 79.5–82.4% genome sequence identity, and likely represented a novel variant. RT-PCR detection revealed the presence of ASGV-infected kiwifruit in two provinces in China. The partial CP gene of nine ASGV isolates determined in this study shared 87.8–99.6% nucleotide sequence identity with the corresponding sequences of other reported ASGV isolates from kiwifruit, indicating molecular diversity of ASGV isolates from kiwifruit plants. This study provides important evidence for monitoring the viral disease and novel molecular information for developing efficient detection techniques to prevent virus spread.