研究者詳細 - 近藤　秀樹

論文 - 近藤　秀樹

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Mitochondrial alterations related to programmed cell death in tobacco cells under aluminium stress 査読

Sanjib Kumar Panda, Yoko Yamamoto, Hideki Kondo, Hideaki Matsumoto

COMPTES RENDUS BIOLOGIES 331 ( 8 ) 597 - 610 2008年8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：ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

The present investigation was undertaken to verify whether mitochondria play a significant role in aluminium(Al) toxicity, using the mitochondria isolated from tobacco cells (Nicotiana tabacum, non-chlorophyllic cell line SL) under Al stress. An inhibition of respiration was observed in terms of state-III, state-IV, succinate-dependent, alternative oxidase (AOX)-pathway capacity and cytochrome (CYT)-pathway capacity, respectively, in the mitochondria isolated from tobacco cells subjected to Al stress for 18 h. In accordance with the respiratory inhibition, the mitochondrial ATP content showed a significant decrease under Al treatment. An enhancement of reactive oxygen species (ROS) production under state-III respiration was observed in the mitochondria isolated from Al-treated cells, which would create an oxidative stress situation. The opening of mitochondrial permeability transition pore (MPTP) was seen more extensively in mitochondria isolated from Al-treated cells than in those isolated from control cells. This was Ca2+ dependent and well modulated by dithioerythritol (DTE) and Pi, but insensitive to cyclosporine A (CsA). The collapse of inner mitochondrial membrane potential (Delta Psi(m)) was also observed with a release of cytochrome c from mitochondria. A great decrease in the ATP content was also seen under Al stress. Transmission electron microscopy analysis of Al-treated cells also corroborated our biochemical data with distortion in membrane architecture in mitochondria. TUNEL-positive nuclei in Al-treated cells strongly indicated the occurrence of nuclear fragmentation. From the above study, it was concluded that Al toxicity affects severely the mitochondrial respiratory functions and alters the redox status studied in vitro and also the internal structure, which seems to cause finally cell death in tobacco cells.

DOI： 10.1016/j.crvi.2008.04.008

Identification of amino acids of the beet necrotic yellow vein virus p25 protein required for induction of the resistance response in leaves of Beta vulgaris plants 査読

Soutaro Chiba, Masaki Miyanishi, Ida Bagus Andika, Hideki Kondo, Tetsuo Tamada

JOURNAL OF GENERAL VIROLOGY 89 ( Pt 5 ) 1314 - 1323 2008年5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：MICROBIOLOGY SOC

The RNA3-encoded p25 protein of beet necrotic yellow vein virus (BNYVV) is responsible for the production of rhizomania symptoms of sugar beet roots (Beta vulgaris subsp. vulgaris). Here, it was found that the presence of the p25 protein is also associated with the resistance response in rub-inoculated leaves of sugar beet and wild beet (Beta vulgaris subsp. maritima) plants. The resistance phenotype displayed a range of symptoms from no visible lesions to necrotic or greyish lesions at the inoculation site, and only very low levels of virus and viral RNA accumulated. The susceptible phenotype showed large, bright yellow lesions and developed high levels of virus accumulation. In roots after Polymyxa betae vector inoculation, however, no drastic differences in virus and viral RNA accumulation levels were found between plants with susceptible and resistant phenotypes, except at an early stage of infection. There was a genotype-specific interaction between BNYVV strains and two selected wild beet lines (MR1 and MR2) and sugar beet cultivars. Sequence analysis of natural BNYVV isolates and site-directed mutagenesis of the p25 protein revealed that 3 aa residues at positions 68, 70 and 179 are important in determining the resistance phenotype, and that host-genotype specificity is controlled by single amino acid changes at position 68. The mechanism of the occurrence of resistance-breaking BNYVV strains is discussed.

DOI： 10.1099/vir.0.83624-0

RNA4-encoded p31 of beet necrotic yellow vein virus is involved in efficient vector transmission, symptom severity and silencing suppression in roots 査読

Muhammad Danial Rahim, Ida Bagus Andika, Chenggui Han, Hideki Kondo, Tetsuo Tamada

JOURNAL OF GENERAL VIROLOGY 88 ( Pt 5 ) 1611 - 1619 2007年5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：SOC GENERAL MICROBIOLOGY

RNA3 and RNA4 of beet necrotic yellow vein virus (BNYVV) are not essential for virus multiplication, but are associated with vector-mediated infection and disease development in sugar beet roots. Here, a unique role for RNA4 in virus transmission, virulence and RNA silencing suppression was demonstrated. Mutagenic analysis revealed that the RNA4-encoded p31 open reading frame (ORF) was involved in efficient vector transmission and slight enhancement of symptom expression in some Beta species. No effects of RNA4 on virus accumulation in infected tissue were observed. Furthermore, the p31 ORF was involved in the induction of severe symptoms by BNYVV in Nicotiana benthamiana plants without affecting viral RNA accumulation. In contrast, RNA3-encoded p25, previously identified as a major contributor to symptom induction in sugar beet, had no such effect on N. benthamiana. In two different silencing suppression assays, neither p31 nor p25 was able to suppress RNA silencing in leaves, but the presence of p31 enhanced a silencing suppressor activity in roots without alteration in viral RNA accumulation. Thus, BNYVV p31 plays a multifunctional role in efficient vector transmission, enhanced symptom expression and root-specific silencing suppression.

DOI： 10.1099/vir.0.82720-0

Orchid fleck virus is a rhabdovirus with an unusual bipartite genome 査読

Hideki Kondo, Takanori Maeda, Yukio Shirako, Tetsuo Tamada

JOURNAL OF GENERAL VIROLOGY 87 ( Pt 8 ) 2413 - 2421 2006年8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：SOC GENERAL MICROBIOLOGY

Orchid fleck virus (OFV) has an unusual bipartite negative-sense RNA genome with clear sequence similarities to those of nucleorhabdoviruses. The OFV genome consists of two single-stranded RNA molecules, RNA1 and RNA2 that are 6413 and 6001 nt long, respectively, with open reading frame (ORF) information in the complementary sense. RNA1 encodes 49 (ORF1), 26 (ORF2), 38 (ORF3), 20 (ORF4) and 61 kDa (ORF5) proteins, and RNA2 encodes a single protein of 212 kDa (ORF6). ORF1, ORF5 and ORF6 proteins had significant similarities (21-38 % identity) to the nucleocapsid protein (N), glycoprotein (G) and polymerase (L) gene products, respectively, of other rhabdoviruses, especially nucleorhabdoviruses, whereas ORF2, ORF3 and ORF4 proteins had no significant similarities to other proteins in the international databases. Similarities between OFV and rhabdoviruses were also found in the sequence complementarity at both termini of each RNA segment (the common terminal sequences are 3'-UGUGUC---GACACA-5'), the conserved intergenic sequences and in being negative sense. It was proposed that a new genus Dichorhabdovirus in the family Rhabdoviridae of the order Mononegavirales should be established with OFV as its prototype member and type species.

DOI： 10.1099/vir.0.81811-0

Lower levels of transgene silencing in roots is associated with reduced DNA methylation levels at non-symmetrical sites but not at symmetrical sites 査読

IB Andika, H Kondo, MD Rahim, T Tamada

PLANT MOLECULAR BIOLOGY 60 ( 3 ) 423 - 435 2006年2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

Transgene transcripts were recently shown to accumulate at higher levels in roots, relative to leaves, of silenced-transgenic Nicotiana benthamiana plants and to be inversely related with the accumulation of small interfering RNAs (siRNAs), suggesting that RNA silencing is less active in roots than in leaves (Andika et al., 2005. Mol. Plant-Microbe Interact. 18: 194). Here we show that the lower transgene RNA silencing activity in roots was associated with lower transgene methylation levels at non-symmetrical CpNpN context but not at symmetrical CpG or CpNpG context in three sets of transformant plants with different exogenous genes. In contrast, such a difference between roots and leaves was not observed for the Tnt1 retrotransposon: no Tnt1 transcript was detected in roots or in leaves of N. benthamiana, while equal levels of Tnt1-derived siRNA accumulation and Tnt1 methylation were found. From our data and previously reported information, we suggest that roots have less of an activity that acts at the step of generation of siRNAs.

DOI： 10.1007/s11103-005-4429-7

Evidence that RNA silencing-mediated resistance to Beet necrotic yellow vein virus is less effective in roots than in leaves 査読

IB Andika, H Kondo, T Tamada

MOLECULAR PLANT-MICROBE INTERACTIONS 18 ( 3 ) 194 - 204 2005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：AMER PHYTOPATHOLOGICAL SOC

In plants, RNA silencing is part of a defense mechanism against virus infection but there is little information as to whether RNA silencing-mediated resistance functions similarly in roots and leaves. We have obtained transgenic Nicotiana benthamiana plants encoding the coat protein readthrough domain open reading frame (54 kDa) of Beet necrotic yellow vein virus (BNYVV), which either showed a highly resistant or a recovery phenotype following foliar rub-inoculation with BNYVV. These phenotypes were associated with an RNA silencing mechanism. Roots of the resistant plants that were immune to foliar rub-inoculation with BNYVV could be infected by viruliferous zoo-spores of the vector fungus Polymyxa betae, although virus multiplication was greatly limited. In addition, virus titer was reduced in symptomless leaves of the plants showing the recovery phenotype, but it was high in roots of the same plants. Compared with leaves of silenced plants, higher levels of transgene mRNAs and lower levels of transgene-derived small interfering RNAs (siRNAs) accumulated in roots. Similarly, in nontransgenic plants inoculated with BNYVV, accumulation level of viral RNA-derived siRNAs in roots was lower than in leaves. These results indicate that the RNA silencing-mediated resistance to BNYVV is less effective in roots than in leaves.

DOI： 10.1094/MPMI-18-0194

A reovirus of the fungus Cryphonectria parasitica that is infectious as particles and related to the Coltivirus genus of animal pathogens 査読

BI Hillman, S Supyani, H Kondo, N Suzuki

JOURNAL OF VIROLOGY 78 ( 2 ) 892 - 898 2004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：AMER SOC MICROBIOLOGY

RNA viruses of filamentous fungi fall into two broad categories, those that contain double-stranded RNA (dsRNA) genomes in rigid particles and those that are more closely related to positive-sense, single-stranded RNA viruses with dsRNA replicative intermediates found within lipid vesicles. Effective infectivity systems have been described for the latter, using RNA transcripts, but not for the former. We report the characterization of a reovirus from Cryphonectria parasitica, the filamentous fungus that causes chestnut blight disease. The virus substantially reduces the virulence of the fungus and results in dramatically altered colony morphology, as well as changes in other associated fungal traits, relative to the virus-free isogenic strain. Virus particles from infected mycelium contained 11 segments of dsRNA and showed characteristics typical of the family Reoviridae. Sequences of the largest three segments revealed that the virus is closely related to the Coltivirus genus of animal pathogens, which includes the human pathogen Colorado tick fever virus. The introduction of purified virus particles into protoplasts from virus-free isolates of the fungus resulted in a newly infected mycelium with the same morphology and virus composition as the original virus-infected isolate. This represents the completion of Koch's postulates for a true dsRNA virus from a filamentous fungus and the description of a definitive fungal member of the family Reoviridae.

DOI： 10.1128/JVI.78.2.892-898.2004

Orchid fleck virus: Brevipalpus californicus mite transmission, biological properties and genome structure 査読国際誌

H Kondo, T Maeda, T Tamada

EXPERIMENTAL AND APPLIED ACAROLOGY 30 ( 1-3 ) 215 - 223 2003年

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

Orchid fleck virus (OFV) causes necrotic or chlorotic ring spots and fleck symptoms in many orchid species world-wide. The virus has non-enveloped, bacilliform particles of about 40 nm x 100-150 nm and is sap-transmissible to several plant species. OFV is transmitted by the mite Brevipalpus californicus (Banks) in a persistent manner and efficiently transmitted by both adults and nymphs, but not by larvae. Viruliferous mites retain their infectivity for 3 weeks on a virus-immune host. The genome of OFV consists of two molecules of 6431 (RNA1) and 6001 nucleotides (RNA2). The RNAs have conserved and complementary terminal sequences. RNA1 contains five open reading frames (ORF), and RNA2 encodes a single ORF. Although some of the encoded proteins of OFV have sequences similar to those of proteins of plant rhabdoviruses, OFV differs from viruses in the family Rhabdoviridae in having a bipartite genome.

DOI： 10.1023/B:APPA.0000006550.88615.10

Identification of Orchid fleck virus by reverse transcriptase-polymerase chain reaction and analysis of isolate relationships

AL Blanchfield, AM Mackenzie, A Gibbs, H Kondo, T Tamada, CR Wilson

JOURNAL OF PHYTOPATHOLOGY-PHYTOPATHOLOGISCHE ZEITSCHRIFT 149 ( 11-12 ) 713 - 718 2001年12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：BLACKWELL VERLAG GMBH

Using reverse transcription-polymerase chain reaction (RT-PCR) tests with a primer complementary to a region of its nucleoprotein gene together with a polydT/ SP6 primer, Orchid fleck virus (OFV) was detected in 34 samples of infected orchids of several different genera from world-wide sources. The resulting DNA fragments were approximately 800 bp in length and their sequences were determined directly. Analysis of sequences of the major open reading frame (ORF) within the DNA fragments obtained showed the presence of two virus strains. The first group was of two isolates, the original Japanese isolate from which the primers were derived and one from Germany, and the second group contained 33 isolates from four continents. The sequences of different groups differed from one another by at least 15.6% (nucleotide) and 1.8% (amino-acid), but the within-group differences were much less (< 1.7% difference). A search of the international nucleotide database with the OFV sequences showed them to be related, but distantly, to nucleoprotein regions of the genomes of four plant-infecting rhabdoviruses.

DOI： 10.1046/j.1439-0434.2001.00702.x

Comparative cytopathology and immunocytochemistry of Japan, Australian and Brazilian isolates of Orchid fleck virus.(共著)

Kitajima et al.

Journal of General Plant Pathology 67 ( 3 ) 231 - 237 2001年

記述言語：英語出版者・発行元：Phytopathological Society of Japan

Cytopathic effects in orchid leaf tissues infected with Australian, Japanese and Brazilian isolates of Orchid fleck virus(OFV) were indistinguishable and like those previously described in the literature. Cells had an electron-lucent viroplasm with unenveloped rod-shaped virions in the nucleus and cytoplasm, often associated with the inner membrane of the nuclear envelope and the endoplasmic reticulum. Antiserum raised against a Japanese isolate of OFV reacted with Brazilian and Australian isolates in ELISA, and when used for immuno-gold labelling, also reacted in situ with the rod-shaped virions and the intranuclear viroplasm of all three isolates. These results suggest that the viroplasm is where structural proteins accumulate and virions are formed.

DOI： 10.1007/PL00013018

その他リンク： http://id.nii.ac.jp/1141/00138246/

Calanthe mild mosaic virus, a new potyvirus causing a mild mosaic disease of Clanth orchid in Japan(共著)

Garae t al

Journal of Phytopathology 146 ( 7 ) 357 - 363 1998年8月

Protective roles of two aluminum (Al)-induced genes, HSP150 and SED1 of Saccharomyces cerevisiae, in Al and oxidative stresses

B Ezaki, RC Gardner, Y Ezaki, H Kondo, H Matsumoto

FEMS MICROBIOLOGY LETTERS 159 ( 1 ) 99 - 105 1998年2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：ELSEVIER SCIENCE BV

We isolated two yeast cDNA clones whose transcripts are induced by aluminum (Al) metal stress. Partial nucleotide sequencing showed that one is the HSP150 gene encoding a secreted heat shock protein, and the other corresponds to the SED1 gene encoding a putative membrane protein. To clarify the biological functions of these genes, we analyzed the sensitivity of gene-disrupted mutants to Al stress and to oxidative stresses. The Al tests indicated that the HSP150 protein served a basal protective role in Al stress, but SED1 did nor, both of the genes had protective roles for oxidative stresses. The results for the HSP150 gene suggest that there is an overlap between Al ion stress, oxidative stress and heat shock stress in yeast. (C) 1998 Published by Elsevier Science B.V. All rights reserved.

DOI： 10.1016/S0378-1097(97)00554-5

ラン科植物に発生するシンビジウムモザイクウイルスの血清学的検出

Gara I Wayan, 近藤秀樹, 前田孚憲

岡山大学資源生物科学研究所報告 5 ( 1 ) 39 - 46 1997年3月

日本においてCymbidium属植物から分離された生物学的性質の異なるOdontoglossum Ringspot Virus分離株のペプチドマッピングによる比較

近藤秀樹, 前田孚憲, 井上成信

岡山大学資源生物科学研究所報告 5 ( 1 ) 31 - 38 1997年3月

Stunt disease of Habenaria radiata caused by a strain of watermelon mosaic virus 2(共著)

Gara et al.

Annals of the phytopathological society of Japan 63 ( 2 ) 113 - 117 1997年

記述言語：英語出版者・発行元：The Phytopathological Society of Japan

1995年,岡山県倉敷市において採集した,激しい萎縮,葉の奇形とモザイクを示したサギソウから分離されたpotyvirusは,ウイルス粒子の形態,感染細胞内所見,アブラムシ伝搬性,宿主範囲,血清学的類縁関係からカボチャモザイクウイルス(WMV 2)と同定された。本ウイルスの宿主範囲および病徴は,数種の植物において,既報の分離株のそれらと異なっていた。外被タンパク質(CP)遺伝子を含むウイルスゲノムの3'末端領域の塩基配列を決定し,既報の5分離株のそれと比較した結果, CPのアミノ酸配列で87-97%, 3'非翻訳領域で94-97%の相同性が認められた。WMV 2のサギソウにおける発生の報告は初めてであり,ウイルス病の和名をサギソウ萎縮病としたい。

DOI： 10.3186/jjphytopath.63.113

Altered mitochondrial gene expression in a maternal distorted leaf mutant of Arabidopsis induced by chloroplast mutator

W Sakamoto, H Kondo, M Murata, F Motoyoshi

PLANT CELL 8 ( 8 ) 1377 - 1390 1996年8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：AMER SOC PLANT PHYSIOLOGISTS

chloroplast mutator (chm) of Arabidopsis is a recessive nuclear mutation that causes green and white variegation in leaves and is inherited in a non-Mendelian fashion. In this study, we have identified and characterized a mutant observed in F-1 and backcrossed BC1 populations from a cross between chm1-3 and ecotype Columbia. This mutant, maternal distorted leaf (MDL), grows very poorly and is distinguished by distorted rough leaves and aborted flowering organs. Electron microscopic observation showed that in MDL plants, a significant portion of mitochondria are abnormal and appear to be nonfunctional. DNA gel blot end sequence analysis of the MDL mitochondrial DNA (mtDNA) revealed rearrangements in two mtDNA fragments associated with rps3-rpl16 genes (encoding ribosomal proteins S3 and L16, respectively). One rearrangement resulted in the insertion of the rps3-rpl16 operon downstream of atp9. An independent deletion in this region had eliminated the majority of rps3. In contrast, another rearrangement deleted part of rpl16, whereas rps3 remained intact. RNA gel blot analysis indicated that expression of these genes is also altered as a consequence of the mtDNA rearrangements. Thus, a mutation at the CHM locus affects mitochondrial gene expression, and impaired mitochondrial function may result in the distorted phenotype.

DOI： 10.1105/tpc.8.8.1377

Ixiaから分離されたbean yellow mosaic virus

辻俊也, 前田孚憲, 近藤秀樹

岡山大学資源生物科学研究所報告 4 ( 2 ) 201 - 213 1996年3月

エビネ類に発生する黄色斑紋モザイク病の病原，Orchid Fleck Virusについて

井上成信, 松本純一, 前田孚憲, 光畑興二, 近藤秀樹, 田原望武

岡山大学資源生物科学研究所報告 4 ( 2 ) 119 - 135 1996年

記述言語：日本語出版者・発行元：岡山大学資源生物科学研究所

Orchid fleck virus(OFV) was isolated from Calanthe spp.(Cal. discolor,Cal. Bicolor,Cal. Hizen,Cal. triplicata,Cal longicalcarata,Cal Satusma) showing light-green and/or yellowish fleck mosaic on the leaves, which different from previously known viruses of Calanthe. OFV caused systemic infection in Calanthe, Chenopodium quinoa and Beta vulgasis var. cicla, and local infection in C.amaranticolor, C. murale, Spinacia oleracea, Tetragonia expansa, Nicotiana tabacum, N. clevelandii, N. glutinasa, N. rustica, Vigna unguiculata. C quinoa and T expansa are useful as indecator hosts and as a source of virus for inoculation, diagnosis and purification. Sap from C. quinoa was infective after dilution to 10-3 but not 10-4, after 10 min at 45 but not 50℃, and after 1 hr at 20℃ but not 2 hrs. For sap inoculation, it is best to use the homogenate of OFV-onfected leaves within about 7-8 min after homogenization in summer and within about 15 min in winter. The virus particles were bullet-shape or bacilliform, approximately 45-50×105-125 nm in a negatively stained praparations. In ultrathin sections, the viroplasms were observed in the nuclei, and the virus particles and the chracteristic spokewheel structures were found both in the nuclei and the cytoplasm. Antiserum (precipitin tiner:1/512) against the present virus reacted strongly with the isolates of OFV-Cy-50, similar to that of homologous virus. In agar gel diffusion tests, no spur formation occurred among Cal. 94-16 and OFV-Cy-50. In SDS-polyacrylamide gel electrophoresis, one major band of Mr 55,000, probably viral nucleocapsid-protein, and three minor proteins were detected, similar to those of OFV･So from Cymbidium.1988年以来、山口・宮崎・鹿児島県で発生が認められたエビネ類（エビネ、タカネ、ヒゼン、サツマ、ツルラン、オナガエビネ）の葉に淡緑色～黄色斑紋のモザイク病の病原ウイルスを調べたところ、orchid fleck virusと同定された。本ウイルスは11科46種（54品種）の植物に汁液接種を行ったところ、エビネに感染して原株と同様の病班を生じたほか、C.quinoa,フダンソウに全身感染し、C.murale,ホウレンソウ、ツルナ、ササゲ、タバコ、N.clevelandii,N.glutinosa,N.rusticaの5科12種の植物に局部感染が認められた。本ウイルスの粒子はDN法試料で被膜のない弾丸型であり、またときに短桿菌型も存在したが、その大きさは長さが約105～125nm、幅約45～50nmであった。病葉の粗汁液を用いた希釈限度は10-3～10-4、不活性化温度は45～50℃、保存限度は1～2時間であった。ウイルスの汁液接種には、接種源植物としてC.quinoa またはツルナがよく、病葉磨砕後夏期では7分以内、冬期では15分以内の病汁液を供試すると接種検定によい結果が得られることが認められた。純化ウイルスを家兎に注射して、微滴法で力値512倍の抗血清が得られた。本抗血清はCymbidium から分離されたOFV・Cy-50とよく反応し、寒天ゲル内二重拡散法では Cal.94-16およびOFV・Cy-50の沈降帯が完全に融合した。ウイルスの構造蛋白質の分子量は約55Kであった。病細胞の超薄切片の電顕観察像には、閣内にviroplasmが認められ、その内部や周辺に層状に集塊あるいは散在した粒子が認められた。さらに核内や細胞質内に膜に包まれた車輪状の粒子集塊も見られた。OFVによる本病をエビネ類黄色斑紋モザイク病とした。

バンダから分離されたCymubidium Mosaic Virusの諸性質

Gara I Wayan, Kondo Hideki, Maeda Takanori, Mitsuhata Koji, Inouye Narinobu

岡山大学資源生物科学研究所報告 4 ( 2 ) 164 - 174 1996年

記述言語：英語出版者・発行元：岡山大学資源生物科学研究所

A virus causing necrotic spots and necrotic flecks on the leaves of Vanda orchids in Japan was identified as cymbidium mosaic virus(Cymv) on the basis of host range,stabilly in crude sap, particle morphology, serological test and physico-chemical properties. The virus was transmitted by sap inoculation to 12 of 57 species in 6 of 12 families tested, but not by aphid Mizus persicae or through seeds. Systemic infection occurred in all Orchidaceae plants tested and only one in non-orchidaceae (Sesamum indicum). In Tetragonia expansa sap, the infective at a dilution of 10-5 but not at 10-6, after heating at 65℃ for 10 min, and was still active after 1 month aging in vitro. Flexuous rod particles, c. 475×13nm,were observed.In ultrahtin sections of leaf tissues from diseased plants, virus particles were found to aggregate in the cytoplasm. The molecular weight of the protein submit and RNA determined by gel electrophoresis, was 27.8×103 and 2.2×106, respectively. Double-stranded RNAs with estimated molecular weight of 5.4×106, 4.0×106, 3.6×106 and 3.0×106 were isolated from infected plants.日本において葉にえそ斑やえそ輪紋を示すバンダから分離されたウイルスは、宿主範囲、病葉汁液中での安定性、粒子長、物理化学的性質、外被タンパク質ならびにウイルスRNAの分子量等からCymbidiummosaic virus(CyMV) と同定された。本ウイルスを12科57種の植物に汁液接種したところ、数種のラン科植物とゴマに全身感染した。また、本ウイルスは汁液接種で容易に伝搬されず、種子伝染も認められなかった。ツルナの病葉汁液中での安定性は不活化温度が65～70℃（10分）、希釈限度が10-5～10-6、保存限度が20℃で１～2ヶ月であった。DN法による電顕観察では屈曲性のある長さ約475×13nmのひも状粒子が多数認められ、感染葉（Cymbidium) の超薄切片にはウイルス粒子がfingerprint状として細胞質内に存在することが確かめられた。ゲル電気泳動により外被タンパク質とウイルスRNAの分子量の解析を行ったところ、それぞれ27.8×103、2.2×106であった。また、本ウイルスに罹病したツルナ葉からは、主に4種の感染に特異的な二本鎖RNA（分子量約5.4×106、4.0×106、3.6×106 3.0×106）が検出された。

Ixia から分離された bean yellow mosaic virus

Tsuji Toshiya, Maeda Takanori, Kondo Hideki, Inouye Narinobu

岡山大学資源生物科学研究所報告 4 ( 2 ) 201 - 213 1996年

記述言語：英語出版者・発行元：岡山大学資源生物科学研究所

A strain (Ixia-B) of bean yellow mosaic virus (BYMV) isolated from Ixia hybrida was characterized and compared with other isolates of BYMV and clover yellow vein virus (CYVV). Ixia-B was transmitted by aphids,Myzus presicae in a non-presistent manner and by sap-inoculation to 11 of 46 species in 5 of 10 families tested, and had a similar host range to that of some BYMV isolates, althrough some defferences were detected. Sap from diseased C. quinoa was infective after 10 min heating at 55℃ but not 60℃, after a dilution to 10-3 but not 10-4, and after 2 days but not 4 days at 20℃.The Virus particles were filamentous rods of about 13×820 nm. Ixia-B contaied a single protein species with a molecular weight of 34,000 and a single viral RNA with approximately 9,000 bases. In ultrahtin sections of leaf tissues from infected plants, the virus particles, cylindrical cytoplasmic inclusions and dense bodies were obsserved in the cytoplasm. The antiserum to Ixia-B produced by immunizing a rabbit had a titer of 1/512. A close serological relationship was revealed between Ixia-B and two strains of BYMV from crocus and gladiolus, but no relationship to clover yellow vein virus was found in agar gel diffusion tests. However,Ixia-B could be distinguished from two strains of BYMV by the formation of spurs among them in agar gel and by the differences in the patterns of peptide mapping of coat proteins. From these findings, Ixia-B was identified as a strain of BYMV.1992年に岡山県倉敷市玉島で、葉に斑入りを生じた球根類花卉植物Ixia hybridaからpotyvirus(Ixia-B)が分離され、その諸性質から bean yellow masaic virus(BYMV)と同定された。本ウイルスを11科47種の植物に接種したとこと、フリージャ、Nicotiana clevelandii、Chenopodium amaranticolar、ソラマメ、クリムソンクローバー、インゲンマメ、ホウレンソウに全身感染し、またC.quinoa、フダンソウ、ツルナ、センニチコウなどに局部感染したが、エンドウ、ササゲ、ダイズなどには感染しなかった。本ウイルスをはモモアカアブラムシにより非永続的伝搬され、C.quinoa の病葉粗汁液中での安定性は耐熱性が55℃～60℃（10分）、耐希釈性10-3～10-4、耐保存性2～4日であった。DN法試料の電顕観察で多くのpotyvirusよりやや長い約820nmのひも状粒子と管状封入体の破片が見られた。感染葉の超薄切片では風車状、層板状の封入体、dence body、細胞質に散在するウイルス粒子が観察された。本ウイルスはfreesia mosaic virusおよびclover yellow vein virusの抗血清と反応せず、また本ウイルス抗血清を用いた寒天ゲル内二重拡散法ではBYMV分離株（Cro-4, BYMV-G）と反応したが、本ウイルスとBYMVのCro-4およびBYMV-G間にspurが形成され、異種抗原の存在が認められた。外被タンパク質の分子量は約34Kで、ssRNAのサイズは約9Kbであった。Papain,chymotrypsin,pepsinを用いた外被タンパク質のぺプタイドマッピングでは、本ウイルス、Cro-4、BYMV-G、Cal-35の部分分解パターンがそれぞれ異なり、外被タンパク質がアミノ酸配列レベルで異なっていることが示唆された。

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