研究者詳細 - 近藤　秀樹

論文 - 近藤　秀樹

分割表示 144 件中 1 - 20 件目／全件表示 >>

Evidence for the replication of a plant rhabdovirus in its arthropod mite vector. 国際誌

Hideki Kondo, Miki Fujita, Paul Telengech, Kazuyuki Maruyam, Kiwamu Hyodo, Aline Daniele Tassi, Ronald Ochoa, Ida Bagus Andika, Nobuhiro Suzuki

Virus research 351 199522 - 199522 2025年1月

記述言語：英語掲載種別：研究論文（学術雑誌）

Transmission of plant viruses that replicate in the insect vector is known as persistent-propagative manner. However, it remains unclear whether such virus-vector relationships also occur between plant viruses and other biological vectors such as arthropod mites. In this study, we investigated the possible replication of orchid fleck virus (OFV), a segmented plant rhabdovirus, within its mite vector (Brevipalpus californicus s.l.) using quantitative RT-qPCR, western blotting and next-generation sequencing. Time-course RT-qPCR and western blot analyses showed an increasing OFV accumulation pattern in mites after virus acquisition. Since OFV genome expression requires the transcription of polyadenylated mRNAs, polyadenylated RNA fractions extracted from the viruliferous mite samples and OFV-infected plant leaves were used for RNA-seq analysis. In the mite and plant datasets, a large number of sequence reads were aligned to genomic regions of OFV RNA1 and RNA2 corresponding to transcribed viral gene mRNAs. This includes the short polyadenylated transcripts originating from the leader and trailer regions at the ends of the viral genome, which are believed to play a crucial role in viral transcription/replication. In contrast, a low number of reads were mapped to the non-transcribed regions (gene junctions). These results strongly suggested that OFV gene expression occurs both in mites and plants. Additionally, deep sequencing revealed the accumulation of OFV-derived small RNAs in mites, although their size profiles differ from those found in plants. Taken together, our results indicated that OFV replicates within a mite vector and is targeted by the RNA-silencing mechanism.

DOI： 10.1016/j.virusres.2024.199522

Metatranscriptomic Sequencing of Sheath Blight-Associated Isolates of Rhizoctonia solani Revealed Multi-Infection by Diverse Groups of RNA Viruses. 査読国際誌

Michael Louie R Urzo, Timothy D Guinto, Ana Eusebio-Cope, Bernard O Budot, Mary Jeanie T Yanoria, Gilda B Jonson, Masao Arakawa, Hideki Kondo, Nobuhiro Suzuki

Viruses 16 ( 7 ) 2024年7月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Rice sheath blight, caused by the soil-borne fungus Rhizoctonia solani (teleomorph: Thanatephorus cucumeris, Basidiomycota), is one of the most devastating phytopathogenic fungal diseases and causes yield loss. Here, we report on a very high prevalence (100%) of potential virus-associated double-stranded RNA (dsRNA) elements for a collection of 39 fungal strains of R. solani from the rice sheath blight samples from at least four major rice-growing areas in the Philippines and a reference isolate from the International Rice Research Institute, showing different colony phenotypes. Their dsRNA profiles suggested the presence of multiple viral infections among these Philippine R. solani populations. Using next-generation sequencing, the viral sequences of the three representative R. solani strains (Ilo-Rs-6, Tar-Rs-3, and Tar-Rs-5) from different rice-growing areas revealed the presence of at least 36 viruses or virus-like agents, with the Tar-Rs-3 strain harboring the largest number of viruses (at least 20 in total). These mycoviruses or their candidates are believed to have single-stranded RNA or dsRNA genomes and they belong to or are associated with the orders Martellivirales, Hepelivirales, Durnavirales, Cryppavirales, Ourlivirales, and Ghabrivirales based on their coding-complete RNA-dependent RNA polymerase sequences. The complete genome sequences of two novel RNA viruses belonging to the proposed family Phlegiviridae and family Mitoviridae were determined.

DOI： 10.3390/v16071152

The intriguing phenomenon of cross-kingdom infections of plant and insect viruses to fungi: Can other animal viruses also cross-infect fungi? 査読国際誌

Ida Bagus Andika, Xinran Cao, Hideki Kondo, Liying Sun

PLoS pathogens 19 ( 10 ) e1011726 2023年10月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Fungi are highly widespread and commonly colonize multicellular organisms that live in natural environments. Notably, studies on viruses infecting plant-associated fungi have revealed the interesting phenomenon of the cross-kingdom transmission of viruses and viroids from plants to fungi. This implies that fungi, in addition to absorbing water, nutrients, and other molecules from the host, can acquire intracellular parasites that reside in the host. These findings further suggest that fungi can serve as suitable alternative hosts for certain plant viruses and viroids. Given the frequent coinfection of fungi and viruses in humans/animals, the question of whether fungi can also acquire animal viruses and serve as their hosts is very intriguing. In fact, the transmission of viruses from insects to fungi has been observed. Furthermore, the common release of animal viruses into the extracellular space (viral shedding) could potentially facilitate their acquisition by fungi. Investigations of the cross-infection of animal viruses in fungi may provide new insights into the epidemiology of viral diseases in humans and animals.

DOI： 10.1371/journal.ppat.1011726

Cross-Kingdom Interactions Between Plant and Fungal Viruses. 査読国際誌

Ida Bagus Andika, Mengyuan Tian, Ruiling Bian, Xinran Cao, Ming Luo, Hideki Kondo, Liying Sun

Annual review of virology 2023年7月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

The large genetic and structural divergences between plants and fungi may hinder the transmission of viruses between these two kingdoms to some extent. However, recent accumulating evidence from virus phylogenetic analyses and the discovery of naturally occurring virus cross-infection suggest the occurrence of past and current transmissions of viruses between plants and plant-associated fungi. Moreover, artificial virus inoculation experiments showed that diverse plant viruses can multiply in fungi and vice versa. Thus, virus cross-infection between plants and fungi may play an important role in the spread, emergence, and evolution of both plant and fungal viruses and facilitate the interaction between them. In this review, we summarize current knowledge related to cross-kingdom virus infection in plants and fungi and further discuss the relevance of this new virological topic in the context of understanding virus spread and transmission in nature as well as developing control strategies for crop plant diseases. Expected final online publication date for the Annual Review of Virology, Volume 10 is September 2023. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

DOI： 10.1146/annurev-virology-111821-122539

Discovery and Genome Characterization of a Closterovirus from Wheat Plants with Yellowing Leaf Symptoms in Japan 査読

Hideki Kondo, Hitomi Sugahara, Miki Fujita, Kiwamu Hyodo, Ida Bagus Andika, Hiroshi Hisano, Nobuhiro Suzuki

Pathogens 12 ( 3 ) 358 - 358 2023年2月

担当区分：筆頭著者,　責任著者掲載種別：研究論文（学術雑誌）出版者・発行元：MDPI AG

Many aphid-borne viruses are important pathogens that affect wheat crops worldwide. An aphid-transmitted closterovirus named wheat yellow leaf virus (WYLV) was found to have infected wheat plants in Japan in the 1970s; however, since then, its viral genome sequence and occurrence in the field have not been investigated. We observed yellowing leaves in the 2018/2019 winter wheat-growing season in an experimental field in Japan where WYLV was detected five decades ago. A virome analysis of those yellow leaf samples lead to the discovery of a closterovirus together with a luteovirus (barley yellow dwarf virus PAV variant IIIa). The complete genomic sequence of this closterovirus, named wheat closterovirus 1 isolate WL19a (WhCV1-WL19a), consisted of 15,452 nucleotides harboring nine open reading frames. Additionally, we identified another WhCV1 isolate, WL20, in a wheat sample from the winter wheat-growing season of 2019/2020. A transmission test indicated that WhCV1-WL20 was able to form typical filamentous particles and transmissible by oat bird-cherry aphid (Rhopalosiphum pad). Sequence and phylogenetic analyses showed that WhCV1 was distantly related to members of the genus Closterovirus (family Closteroviridae), suggesting that the virus represents a novel species in the genus. Furthermore, the characterization of WhCV1-WL19a-derived small RNAs using high-throughput sequencing revealed highly abundant 22-nt-class small RNAs potentially derived from the 3′-terminal end of the WhCV1 negative-strand genomic RNA, indicating that this terminal end of the WhCV1 genome is likely particularly targeted for the synthesis of viral small RNAs in wheat plants. Our results provide further knowledge on closterovirus diversity and pathogenicity and suggest that the impact of WhCV1 on wheat production warrants further investigations.

DOI： 10.3390/pathogens12030358

Mycovirus Diversity and Evolution Revealed/Inferred from Recent Studies. 査読国際誌

Hideki Kondo, Leticia Botella, Nobuhiro Suzuki

Annual review of phytopathology 2022年5月

担当区分：筆頭著者記述言語：英語掲載種別：研究論文（学術雑誌）

High-throughput virome analyses with various fungi, from cultured or uncultured sources, have led to the discovery of diverse viruses with unique genome structures and even neo-lifestyles. Examples in the former category include splipalmiviruses and ambiviruses. Splipalmiviruses, related to yeast narnaviruses, have multiple positive-sense (+) single-stranded (ss) RNA genomic segments that separately encode the RNA-dependent RNA polymerase motifs, the hallmark of RNA viruses (members of the kingdom Orthornavirae). Ambiviruses appear to have an undivided ssRNA genome of 3∼5 kb with two large open reading frames (ORFs) separated by intergenic regions. Another narna-like virus group has two fully overlapping ORFs on both strands of a genomic segment that span more than 90% of the genome size. New virus lifestyles exhibited by mycoviruses include the yado-kari/yado-nushi nature characterized by the partnership between the (+)ssRNA yadokarivirus and an unrelated dsRNA virus (donor of the capsid for the former) and the hadaka nature of capsidless 10-11 segmented (+)ssRNA accessible by RNase in infected mycelial homogenates. Furthermore, dsRNA polymycoviruses with phylogenetic affinity to (+)ssRNA animal caliciviruses have been shown to be infectious as dsRNA-protein complexes or deproteinized naked dsRNA. Many previous phylogenetic gaps have been filled by recently discovered fungal and other viruses, which have provided interesting evolutionary insights. Phylogenetic analyses and the discovery of natural and experimental cross-kingdom infections suggest that horizontal virus transfer may have occurred and continue to occur between fungi and other kingdoms. Expected final online publication date for the Annual Review of Phytopathology, Volume 60 is August 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

DOI： 10.1146/annurev-phyto-021621-122122

First Report of Chinese wheat mosaic virus Infecting Barley in Japan. 査読国際誌

Hideki Kondo, Hidekazu Masejima, Kazuyuki Maruyama, Miki Fujita, Takehiro Ohki

Plant disease 2022年1月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Chinese wheat mosaic virus (CWMV), a member of the genus Furovirus in the family Virgaviridae (Adams et al. 2017), has a positive-sense RNA genome and is transmitted by Polymyxa graminis. CWMV is a causal agent of yellow mosaic disease in winter wheat in China (Guo et al. 2019). CWMV has also been detected in wheat plants in limited areas of the northern Japan (Nagano and Iwate Prefectures) (Fuji et al. 2022; Maeshima et al. 2010; Shirako and Maejima 2008). In preliminary tests using Western blotting with an antiserum raised against CWMV capsid protein (by Dr Y. Shirako, Tokyo University), we detected a furovirus in a breeding line of barley, "Tozan Kawa 111" (Hordeum vulgare L.) (collected in April 2012), grown in an experimental field infested with CWMV and wheat yellow mosaic virus (genus Bymovirus) in Nagano Prefecture. To investigate the infection of barley plants with cereal plant-associated soil-borne viruses, we collected leaf samples of "Tozan Kawa 111" plants (ten plants) showing yellow mosaic symptoms, but with not apparent wilting or stunting (Fig. S1A) in April 2016 (2015/16 growing season). Total RNA was extracted from symptomatic leave samples with TaKaRa RNAiso reagent (TaKaRa Bio) and subjected to reverse transcription polymerase chain reaction (RT-PCR) to detect virus agents. After cDNA synthesis using Moloney murine leukemia virus reverse transcriptase (Thermo Fisher Scientific) with random hexamers, PCR amplification with QuickTaq HS Dye Mix (Toyobo Co.) was conducted using primer sets specific to two furoviruses and three bymoviruses (Fig. S1B) known to infect wheat and/or barley plants in Japan (Tamada and Kondo 2013). RT-PCR analysis detected infection with CWMV in the leaf samples of "Tozan Kawa 111" plants (Fig. S1B), but not the other soil-borne viruses tested. The amplified PCR products (752 and 718 bp for CWMV RNA1 and RNA2, respectively) were purified by Wizard® SV Gel and PCR Clean-Up System (Promega) and subjected to Sanger sequencing to confirm their nucleotide sequences. The virus sequences from PCR amplicons were deposited in GenBank/DDBJ/ENA with accession numbers LC657081 (RNA1) and LC657082 (RNA2). Nucleotide Basic Local Alignment Search Tool (BLASTn) analysis showed that the sequences have 98.7% and 98.8% nucleotide sequence identity with RNA1 of CWMV Japanese northern isolate (accession No, AB299271) and RNA2 of CWMV Nagano-A isolate (AB935554), respectively. Identical sequences were also found in symptomatic wheat leaf samples ("Fukuho Komugi" cultivar) obtained from the same field (Fig. S1B). Rod-shaped particles were observed by transmission electron microscopy (Hitachi H-7650) in symptomatic "Tozan Kawa 111" leaf samples (Fig. S1C). Using reverse transcription loop-mediated isothermal amplification (Fukuta et al. 2013), CWMV was detected in "Tozan Kawa 111" plants in the same field in the 2015/16-2017/18 growing seasons, but not in the 2018/19-2020/21 growing seasons (Fig. S1D). In the 2015/16-2017/18 growing seasons, CWMV was detected in the barley plants (pooled five plant samples) cultivar "Kashima-mugi", which showed similar yellow mosaic symptoms (Fig. S1D), but not in most of the other barley variants planted in the field. To our knowledge, this is the first report of CWMV field infection in plants other than wheat (Kuhne 2009). Further extensive virus screening in the fields and virus inoculation experiments are necessary to understand the pathology of CWMV in barley and possibly in other cereal crops.

DOI： 10.1094/PDIS-12-21-2803-PDN

Identification of a Novel Quinvirus in the Family Betaflexiviridae That Infects Winter Wheat. 査読国際誌

Hideki Kondo, Naoto Yoshida, Miki Fujita, Kazuyuki Maruyama, Kiwamu Hyodo, Hiroshi Hisano, Tetsuo Tamada, Ida Bagus Andika, Nobuhiro Suzuki

Frontiers in microbiology 12 715545 - 715545 2021年

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Yellow mosaic disease in winter wheat is usually attributed to the infection by bymoviruses or furoviruses; however, there is still limited information on whether other viral agents are also associated with this disease. To investigate the wheat viromes associated with yellow mosaic disease, we carried out de novo RNA sequencing (RNA-seq) analyses of symptomatic and asymptomatic wheat-leaf samples obtained from a field in Hokkaido, Japan, in 2018 and 2019. The analyses revealed the infection by a novel betaflexivirus, which tentatively named wheat virus Q (WVQ), together with wheat yellow mosaic virus (WYMV, a bymovirus) and northern cereal mosaic virus (a cytorhabdovirus). Basic local alignment search tool (BLAST) analyses showed that the WVQ strains (of which there are at least three) were related to the members of the genus Foveavirus in the subfamily Quinvirinae (family Betaflexiviridae). In the phylogenetic tree, they form a clade distant from that of the foveaviruses, suggesting that WVQ is a member of a novel genus in the Quinvirinae. Laboratory tests confirmed that WVQ, like WYMV, is potentially transmitted through the soil to wheat plants. WVQ was also found to infect rye plants grown in the same field. Moreover, WVQ-derived small interfering RNAs accumulated in the infected wheat plants, indicating that WVQ infection induces antiviral RNA silencing responses. Given its common coexistence with WYMV, the impact of WVQ infection on yellow mosaic disease in the field warrants detailed investigation.

DOI： 10.3389/fmicb.2021.715545

Virome Analysis of Aphid Populations That Infest the Barley Field: The Discovery of Two Novel Groups of Nege/Kita-Like Viruses and Other Novel RNA Viruses. 査読国際誌

Hideki Kondo, Miki Fujita, Hiroshi Hisano, Kiwamu Hyodo, Ida Bagus Andika, Nobuhiro Suzuki

Frontiers in microbiology 11 509 - 509 2020年

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）

Aphids (order Hemiptera) are important insect pests of crops and are also vectors of many plant viruses. However, little is known about aphid-infecting viruses, particularly their diversity and relationship to plant viruses. To investigate the aphid viromes, we performed deep sequencing analyses of the aphid transcriptomes from infested barley plants in a field in Japan. We discovered virus-like sequences related to nege/kita-, flavi-, tombus-, phenui-, mononega-, narna-, chryso-, partiti-, and luteoviruses. Using RT-PCR and sequence analyses, we determined almost complete sequences of seven nege/kitavirus-like virus genomes; one of which was a variant of the Wuhan house centipede virus (WHCV-1). The other six seem to belong to four novel viruses distantly related to Wuhan insect virus 9 (WhIV-9) or Hubei nege-like virus 4 (HVLV-4). We designated the four viruses as barley aphid RNA virus 1 to 4 (BARV-1 to -4). Moreover, some nege/kitavirus-like sequences were found by searches on the transcriptome shotgun assembly (TSA) libraries of arthropods and plants. Phylogenetic analyses showed that BARV-1 forms a clade with WHCV-1 and HVLV-4, whereas BARV-2 to -4 clustered with WhIV-9 and an aphid virus, Aphis glycines virus 3. Both virus groups (tentatively designated as Centivirus and Aphiglyvirus, respectively), together with arthropod virus-like TSAs, fill the phylogenetic gaps between the negeviruses and kitaviruses lineages. We also characterized the flavi/jingmen-like and tombus-like virus sequences as well as other RNA viruses, including six putative novel viruses, designated as barley aphid RNA viruses 5 to 10. Interestingly, we also discovered that some aphid-associated viruses, including nege/kita-like viruses, were present in different aphid species, raising a speculation that these viruses might be distributed across different aphid species with plants being the reservoirs. This study provides novel information on the diversity and spread of nege/kitavirus-related viruses and other RNA viruses that are associated with aphids.

DOI： 10.3389/fmicb.2020.00509

Two novel fungal negative-strand RNA viruses related to mymonaviruses and phenuiviruses in the shiitake mushroom (Lentinula edodes). 査読

Lin YH, Fujita M, Chiba S, Hyodo K, Andika IB, Suzuki N, Kondo H

Virology 533 125 - 136 2019年7月

A novel insect-infecting virga/nege-like virus group and its pervasive endogenization into insect genomes. 査読

Kondo H, Chiba S, Maruyama K, Andika IB, Suzuki N

Virus research 262 37 - 47 2019年3月

Evidence for a novel negative-stranded RNA mycovirus isolated from the plant pathogenic fungus Fusarium graminearum 査読

Luan Wang, Hao He, Shuangchao Wang, Xiaoguang Chen, Dewen Qiu, Hideki Kondo, Lihua Guo

Virology 518 232 - 240 2018年5月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Academic Press Inc.

Here we describe a novel (−)ssRNA mycovirus, Fusarium graminearum negative-stranded RNA virus 1 (FgNSRV-1), isolated from Fusarium graminearum strain HN1. The genome of FgNSRV-1 is 9072 nucleotides in length, with five discontinuous but linear ORFs (ORF I-V). Phylogenetic analysis based on entire L polymerase sequences indicated that FgNSRV-1 is related to the (−)ssRNA mycovirus Sclerotinia sclerotiorum negative-stranded RNA virus 1 (SsNSRV-1), and other mycoviruses. Our data suggest that FgNSRV-1 can be classified into the family Mymonaviridae, order Mononegavirales. Putative enveloped virion-like structures with filamentous morphology similar to SsNSRV-1 were observed in virion preparation samples. The L proteins of FgNSRV-1, and other fungal mononegaviruses, were found to be related to L protein-like sequences in some fungal genome, supporting the hypothesis that there is coevolution occurring between mycoviruses and fungi. Besides, clearing the virus from the infected host fungus resulted in no discernable phenotypic change.

DOI： 10.1016/j.virol.2018.03.008

A possible occurrence of genome reassortment among bipartite rhabdoviruses 査読

Hideki Kondo, Keisuke Hirota, Kazuyuki Maruyama, Ida Bagus An'dika, Nobuhiro Suzuki

VIROLOGY 508 18 - 25 2017年8月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE

Orchid fleck virus (OFV) represents a rhabdovirus with a unique bipartite genome. OFV genetic diversity at the whole genome level has not been described. Using the partial genome sequence of RNA1, we have determined that several OFV isolates derived from orchids in Japan belong to two genetically distant subgroups: subgroup I, the members of which are distributed worldwide but previously not known in Asia, and subgroup II, which is commonly distributed in Japan. However, complete genome sequence analysis of a novel Japanese subgroup I isolate revealed that although its RNA1 sequence differs considerably from those of subgroup II isolates, its RNA2 sequence is almost identical to them. Based on phylogenetic and recombination analyses, the genome reassortment events were predicted to occur between OFV subgroups including other unseen strains. Our data show that genome reassortment contributes to the genetic diversities of the bipartite rhabdoviruses and its occurrence may be geographically constrained.

DOI： 10.1016/j.virol.2017.04.027

Sequence and phylogenetic analyses of novel totivirus-like double-stranded RNAs from field-collected powdery mildew fungi 査読

Hideki Kondo, Sakae Hisano, Sotaro Chiba, Kazuyuki Maruyama, Ida Bagus Andika, Kazuhiro Toyoda, Fumihiro Fujimori, Nobuhiro Suzuki

Virus Research 213 353 - 364 2016年2月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：Elsevier B.V.

The identification of mycoviruses contributes greatly to understanding of the diversity and evolutionary aspects of viruses. Powdery mildew fungi are important and widely studied obligate phytopathogenic agents, but there has been no report on mycoviruses infecting these fungi. In this study, we used a deep sequencing approach to analyze the double-stranded RNA (dsRNA) segments isolated from field-collected samples of powdery mildew fungus-infected red clover plants in Japan. Database searches identified the presence of at least ten totivirus (genus Totivirus)-like sequences, termed red clover powdery mildew-associated totiviruses (RPaTVs). The majority of these sequences shared moderate amino acid sequence identity with each other (&lt
44%) and with other known totiviruses (&lt
59%). Nine of these identified sequences (RPaTV1a, 1b and 2-8) resembled the genome of the prototype totivirus, Saccharomyces cerevisiae virus-L-A (ScV-L-A) in that they contained two overlapping open reading frames (ORFs) encoding a putative coat protein (CP) and an RNA dependent RNA polymerase (RdRp), while one sequence (RPaTV9) showed similarity to another totivirus, Ustilago maydis virus H1 (UmV-H1) that encodes a single polyprotein (CP-RdRp fusion). Similar to yeast totiviruses, each ScV-L-A-like RPaTV contains a -1 ribosomal frameshift site downstream of a predicted pseudoknot structure in the overlapping region of these ORFs, suggesting that the RdRp is translated as a CP-RdRp fusion. Moreover, several ScV-L-A-like sequences were also found by searches of the transcriptome shotgun assembly (TSA) libraries from rust fungi, plants and insects. Phylogenetic analyses show that nine ScV-L-A-like RPaTVs along with ScV-L-A-like sequences derived from TSA libraries are clustered with most established members of the genus Totivirus, while one RPaTV forms a new distinct clade with UmV-H1, possibly establishing an additional genus in the family. Taken together, our results indicate the presence of diverse, novel totiviruses in the powdery mildew fungus populations infecting red clover plants in the field.

DOI： 10.1016/j.virusres.2015.11.015

Cymbidium chlorotic mosaic virus, a new sobemovirus isolated from a spring orchid (Cymbidium goeringii) in Japan 査読

Hideki Kondo, Shogo Takemoto, Kazuyuki Maruyama, Sotaro Chiba, Ida Bagus Andika, Nobuhiro Suzuki

ARCHIVES OF VIROLOGY 160 ( 8 ) 2099 - 2104 2015年8月

担当区分：筆頭著者,　責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SPRINGER WIEN

Cymbidium chlorotic mosaic virus (CyCMV), isolated from a spring orchid (Cymbidium goeringii), was characterized molecularly. CyCMV isometric virions comprise a single, positive-strand RNA genome of 4,083 nucleotides and 30-kDa coat protein. The virus genome contains five overlapping open reading frames with a genomic organization similar to that of sobemoviruses. BLAST searches and phylogenetic analysis revealed that CyCMV is most closely related to papaya lethal yellowing virus, a proposed dicot-infecting sobemovirus (58.8 % nucleotide sequence identity), but has a relatively distant relationship to monocot-infecting sobemoviruses, with only modest sequence identities. This suggests that CyCMV is a new monocot-infecting member of the floating genus Sobemovirus.

DOI： 10.1007/s00705-015-2460-9

Differential contributions of plant Dicer-like proteins to antiviral defences against potato virus X in leaves and roots 査読

Ida Bagus Andika, Kazuyuki Maruyama, Liying Sun, Hideki Kondo, Tetsuo Tamada, Nobuhiro Suzuki

PLANT JOURNAL 81 ( 5 ) 781 - 793 2015年3月

担当区分：責任著者記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：WILEY-BLACKWELL

Members of the plant Dicer-like (DCL) protein family are the critical components of the RNA-silencing pathway that mediates innate antiviral defence. The distinct antiviral role of each individual DCL protein has been established with mostly based on observations of aerial parts of plants. Thus, although the roots are closely associated with the life cycle of many plant viruses, little is known about the antiviral activities of DCL proteins in roots. We observed that antiviral silencing strongly inhibits potato virus X (PVX) replication in roots of some susceptible Solanaceae species. Silencing of the DCL4 homolog in Nicotiana benthamiana partially elevated PVX replication levels in roots. In Arabidopsis thaliana, which was originally considered a non-host plant of PVX, high levels of PVX accumulation in inoculated leaves were achieved by inactivation of DCL4, while in the upper leaves and roots, it required the additional inactivation of DCL2. In transgenic A.thaliana carrying the PVX amplicon with a green fluorescent protein (GFP) gene insertion in the chromosome (AMP243 line), absence of DCL4 enabled high levels of PVX-GFP accumulation in various aerial organs but not in the roots, suggesting that DCL4 is critical for intracellular antiviral silencing in shoots but not in roots, where it can be functionally compensated by other DCL proteins. Together, the high level of functional redundancies among DCL proteins may contribute to the potent antiviral activities against PVX replication in roots.
Significance StatementThis study demonstrates the differential contributions of DCL proteins between leaves and roots.

DOI： 10.1111/tpj.12770

Transcriptional mapping of the messenger and leader RNAs of orchid fleck virus, a bisegmented negative-strand RNA virus 査読

Hideki Kondo, Kazuyuki Maruyama, Sotaro Chiba, Ida Bagus Andika, Nobuhiro Suzuki

VIROLOGY 452 166 - 174 2014年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：ACADEMIC PRESS INC ELSEVIER SCIENCE

The transcriptional strategy of orchid fleck virus (OFV), which has a two-segmented negative-strand RNA genome and resembles plant nucleorhabdoviruses, remains unexplored. In this study, the transcripts of six genes encoded by OFV RNA1 and RNA2 in the poly(A)-enriched RNA fraction from infected plants were molecularly characterized. All of the OFV mRNAs were initiated at a start sequence 3'-UU-5' with one to three non-viral adenine nucleotides which were added at the 5' end of each mRNA, whereas their 3' termini ended with a 5'-AUUUAAA(U/G)AAAA(A)n-3' sequence. We also identified the presence of polyadenylated short transcripts derived from the 3'-terminal leader regions of both genomic and antigenomic strands, providing the first example of plus- and minus-strand leader RNAs in a segmented minus-strand RNA virus. The similarity in the transcriptional strategy between this bipartite OFV and monopartite rhabdoviruses, especially nucleorhabdoviruses (family Rhabdoviridae) is additional support for their close relationship. (C) 2014 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.virol.2014.01.007

Complete genome sequence of Habenaria mosaic virus, a new potyvirus infecting a terrestrial orchid (Habenaria radiata) in Japan 査読

Hideki Kondo, Takanori Maeda, I. Wayan Gara, Sotaro Chiba, Kazuyuki Maruyama, Tetsuo Tamada, Nobuhiro Suzuki

ARCHIVES OF VIROLOGY 159 ( 1 ) 163 - 166 2014年1月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：SPRINGER WIEN

The complete genomic sequence of Habenaria mosaic virus (HaMV), which infects terrestrial orchids (Habenaria radiata), has been determined. The genome is composed of 9,499 nucleotides excluding the 3'-terminal poly(A) tail, encoding a large polyprotein of 3,054 amino acids with the genomic features typical of a potyvirus. Putative proteolytic cleavage sites were identified by sequence comparison to those of known potyviruses. The HaMV polyprotein showed 58 % amino acid sequence identity to that encoded by the most closely related potyvirus, tobacco vein banding mosaic virus. Phylogenetic analysis of the polyprotein amino acid sequence and its coding sequences confirmed that HaMV formed a cluster with the chilli veinal mottle virus group, most of which infect solanaceous plants. These results suggest that HaMV is a distinct member of the genus Potyvirus.

DOI： 10.1007/s00705-013-1784-6

Characterization of burdock mottle virus, a novel member of the genus Benyvirus, and the identification of benyvirus-related sequences in the plant and insect genomes 査読

Hideki Kondo, Shuichi Hirano, Sotaro Chiba, Ida Bagus Andika, Makoto Hirai, Takanori Maeda, Tetsuo Tamada

Virus Research 177 ( 1 ) 75 - 86 2013年10月

記述言語：英語掲載種別：研究論文（学術雑誌）

The complete nucleotide sequence of the burdock mottle virus (BdMoV) isolated from an edible burdock plant ( Arctium lappa) in Japan has been determined. BdMoV has a bipartite genome, whose organization is similar to RNA1 and RNA2 of benyviruses, beet necrotic yellow vein virus (BNYVV), beet soil-borne mosaic virus (BSBMV), and rice stripe necrosis virus (RSNV). BdMoV RNA1 (7038 nt) contains a single open reading frame (ORF) encoding a 249-kDa polypeptide that consists of methyl-transferase, helicase, papain-like protease, AlkB-like, and RNA-dependent RNA polymerase domains. The AlkB-like domain sequence is not present in the proteins encoded by other known benyviruses, but is found in replication-associated proteins of viruses mainly belonging to the families Alfaflexiviridae and Betaflexiviridae. BdMoV RNA2 (4315 nt) contains six ORFs that are similar to those of benyviruses: these are coat protein (CP), CP readthrough, triple gene block movement and cysteine-rich proteins. Phylogenetic analyses showed that BdMoV is more closely related to BNYVV and BSBMV than to RSNV. Database searches showed that benyvirus replicase-related sequences are present in the chromosomes of a chickpea plant ( Cicer arietinum) and a blood-sucking insect ( Rhodnius prolixus). Some other benyvirus-related sequences are found in the transcriptome shotgun libraries of a few species of plants and a bark beetle. Our results show that BdMoV is a distinct species of the genus Benyvirus and that ancestral and extant benyviruses may have infected or currently infect a wide range of hosts, including plants and insects. © 2013 Elsevier B.V.

DOI： 10.1016/j.virusres.2013.07.015

Orchid fleck virus structural proteins N and P form intranuclear viroplasm like structures in the absence of viral infection 査読

Hideki Kondo, Sotaro Chiba, Ida Bagus Andika, Kazuyuki Maruyama, Tetsuo Tamada, Nobuhiro Suzuki

Journal of Virology 87 ( 13 ) 7423 - 7434 2013年7月

記述言語：英語掲載種別：研究論文（学術雑誌）

Orchid fleck virus (OFV) has a unique two-segmented negative-sense RNA genome that resembles that of plant nucleorhabdoviruses. In infected plant cells, OFV and nucleorhabdoviruses induce an intranuclear electron-lucent viroplasm that is believed to be the site for virus replication. In this study, we investigated the molecular mechanism by which OFV viroplasms are produced in vivo. Among OFV-encoded proteins, the nucleocapsid protein (N) and the putative phosphoprotein (P) were present in nuclear fractions of OFV-infected Nicotiana benthamiana plants. Transient coexpression of N and P, in the absence of virus infection, was shown to be sufficient for formation of an intranuclear viroplasm-like structure in plant cells. When expressed independently as a fluorescent protein fusion product in uninfected plant cells, N protein accumulated throughout the cell, while P protein accumulated in the nucleus. However, the N protein, when coexpressed with P, was recruited to a subnuclear region to induce a large viroplasm-like focus. Deletion and substitution mutagenesis demonstrated that the P protein contains a nuclear localization signal (NLS). Artificial nuclear targeting of the N-protein mutant was insufficient for formation of viroplasm-like structures in the absence of P. A bimolecular fluorescence complementation assay confirmed interactions between the N and P proteins within subnuclear viroplasm-like foci and interactions of two of the N. benthamiana importin-α homologues with the P protein but not with the N protein. Taken together, our results suggest that viroplasm formation by OFV requires nuclear accumulation of both the N and P proteins, which is mediated by P-NLS, unlike nucleorhabdovirus viroplasm utilizing the NLS on protein N. © 2013, American Society for Microbiology.

DOI： 10.1128/JVI.00270-13

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