岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996ホウレンソウ（Spinacia oleracea L.）種子のαグルコシダーゼの精製とその性質239252ENManabuSugimotoSatoshiFuruiYukioSuzukiFour molecular forms of α-glucosidase were isolated from spinach seeds by several kinds of chromatography. The molecular masses of α-glucosidases T,U,V,and W were 78,78,82 and 82kDa by SDS-PAGE, and 62,62,190,and 70kDa by gel filtration, respectively. α-Glucosidases Tand U showed similar enzymatic properties. The Km for soluble starch was about 10 times lower than that for maltose, and they had higher activity not only towards malto-oligosaccharides but also towards α-glucans. The optimum pH was 4.5-5.5 and about 50% of the activity remained after incubation at 71℃ for 20 min. On the other hand, α-glucosidases V and W showed similar enzymatic propreties. The Km for maltose was 3-4 times lower than for solble starch, and they had high activity toward malto-oligosaccharides but faint activity towards α-glucnas. The optimum pH was 4.5-5.0 and no activity was found after incubation at 70℃ for 20 min. However, anti-α-glucosidase V serum precipitated specifically with α-glucosidase V.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996コムギうどんこ病菌株間の種内交雑におけるオオムギおよびカモジグサに対する病原性菌系の作出229237ENHideoHetaNarinobuInouyeTwo isolates of Erysiphe graminis f. sp. tritici, T3 and T4, were crossed and selected for pathogenicity. The corssing and selection process were repeated five times with the progenies. Isolates of each generation were inoculated on the non-hosts, Hordeum vulgare and Agropyron tsukushiense var. transiens OHWI, and parasitism and pathogenicity were investigated with respect to host specificity.As shown in Tables 2 to 5, the process of crossing and selection produced isolates which were pathogenic on barley cultivars, 'Kagoshimahadaka' or 'Russian 9' or Agropyron tsukushiense ecotypes 'Agropyron Early ecotype' or 'Agropyron Yezoense 3'.Factors affecting the degree of pathogenicity appear to be under polygenic control. The cumulative effect of minor genes may play a role in the differentiation of formae speciales suggesting a new concept of pathogenicity in the powdery mildew of grasses.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996茨城県でダイズから分離されたアズキモザイクウイルスの諸性質215227ENKokichiTakahashiNarinobuInouyeTakanoriMaedaKojiMitsuhataShigekiChidaYoshiyukiTakahashiAzuki bean mosaic virus (ABMV) was isolated from Glycine max showing mild mosaic and stunt on the leaves, collected in Ibaraki-Prefecture, in July, 1989. ABMV was transmitted by sap-inoculation to 8 species systemically and 14 species locally out of 33 species in 8 families and by aphids, Aulacorthum solani and Myzus persicae in a non-persistent manner.Seed transmission was recognized in 0.8%
of the seeds of soybean cv."Iwate-wase-kurome" inoculated at seeding stage. The soybean plant infected with ABMV produced mottled seeds, which were radical- or saddle-like. The virus particles were flexuous rods, bout 750 nm in length. In ultrathin virus-infected tissues, cytoplasmic inclusions containing pinwheels and scrolls were observed in the cytoplasms. In DAS-ELISA, the virus reacted strongly with antiserum to ABMV. The weight of plants and seed of two soybean cv."Okuhara 1" and "Iwate-wase-kurome" inoculated with ABMV at the seedling stage was about 40〜50% less than that of healthy plants.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996Characterization of Bean Yellow Mosaic Virus from Ixia hybrida201213ENToshiyaTsujiTakanoriMaedaHidekiKondoNarinobuInouyeA 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.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996エビネ（Calanthe spp.）から分離されたCymbidium mosaic virus187199ENJun-ichiMatsumotoShinjiUrabeTakanoriMaedaKojiMitsuhataHidekiKondoMochimuTaharaNarinobuInouyeCymbidium mosaic virus(CyMV) was isolated from Calanthe spp. showing mosaic on the leaves, collected in Yamaguchi and Kyoto Prefectures in 1986〜1993. CyMV, Cal. 90-1 isolate was transmitted by sapinoculation to 12 out of 37 species in 7 out of 9 families. Sap from diseaded Tetragonia expansa was infective to Chenopodium amaranticolor after dilution to 10-5 but not 10-6, after heating at 65℃ for 10 min but not 70℃, and after 1 month at 20℃ but not 2 months. The virus particles were flexuous rod, about 475 nm long. The virus was purified from diseased T. expansa leaves and contained a single protein species of Mr27,800. The Mr of the capsid proteins(Cal. 90-1) was similar to those of two ohter CyMV isolates(Cal. 90-4, Cal. 93-14).Cal. 90-1 and Cal. 93-14 reacted with antiserum to the Cymbidium isolate (Cy-16), suggesting that Cal. 90-1 was serologically very similar to the other two CyMV isolates. Two species of dsRNA were isolated from plants infected with Cal-1 and they were similar to those of two other CyMv isolates.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996キュウリモザイクウイルスの血清型と病原性との関連性175185ENTakanoriMaedaKojiMitsuhataNarinobuInouyeSixty-eight isolates of cucumber mosaic virus (CMV) belonging to the Y serotype (serogroup) or P serotype (serotype U) in Japan were inoculated to Nicotiana tabacum cv. White Burley and the symptoms were observed up to three months after inoculation. These isolates were divided into four goups accoding to symptomatology on tabacco plant. None of the isoletes obtained from lilies infected tobacco plants systemically. The plants inoculated with isolates of the P serotype showed systemic mosaic, but only occasional mild symptoms were observed on newly developed leaves. On the other hand, all isolates of Y serotype showed symptoms consisting of mosaic, distortion and/or necrosis throughout the experimental period. In another experiment, reactions of some selected test plants to 30 isolates belonging to the Y or P serotype were examined. Lily isolates may be unique strain of CMV as they did not cause systemic infections on Lycopersicon esculentum, Solanum melongena or Cucumis sativus. Host range tests using 16 isolates originally obtained from plants other than lilies demonstrated a close relationship between serotype and pathogenicity to some test plants including N.tabacum, N. clevelandii, L.esculentum, Pisum sativum, phaseolus angularis and Zea mays.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996Further Characterization of Cymbidium Mosaic Virus from Vanda Orchid164174ENI WayanGaraHidekiKondoTakanoriMaedaKojiMitsuhataNarinobuInouyeA 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.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996東洋ランに発生するウイルスの検索・同定149162ENHidekiKondoTakanoriMaedaKojiMitsuhataNarinobuInouyeA survey of virus diseases occurring in Oriental Cymbidium collected from a commerical nursery and home garden in Japan was conducted in 1991-1994. Identification of the vurus was based on partcle morphology, symptomatology in indicator plants, ultrastructure of infected cells and serology. Four viruses, odontoglossum ringspot tabamovirus(ORSV), cymbidium mosaic potexvirus(CyMV), orchid fleck virus (ORV) and a previously underscribed spherical virus, were found in 27 out of 37 Cymbidium plants tested. ORSV was detected from 11 plants belinging to Cym. ensifolium, Cym. forrestii, Cym. goeringii, Cym. kanran, Cym. sinense and Cymbidium spp. showing chlorotic streaks and/or mild mosaic. CyMV was isolated from only one plant of Cymbidium sp. showing mosaic and necrotic spots on leaves. In negatibvely stained dip preparations from plants infected with ORSV and CyMV, rod shaped particles of ca. 310 nm and flexuous rod-shaped ca. 475 nm in length were observed, respectively. The viruses were reacted strongly with respective antiserum to each virus in immunosorbent electron microcopy and inderect ELISA. OFV was isolated from four plants of Cym. formosanum, Cym. kanran, Cym. sinense and Cymbidium sp. showing mosaic and necrotic flecks. The virus had non-enveloped, bullet-shaped particles about 40×120〜150 nm in dip preparation. The undescribed spherical virus, ca. 28 nm diameter, was isolated from 11 plants of Cym. forrestii, Cym. goeringii and Cymbidium spp. showing stunting and chlorotic streaks on newly developed leaves. The virus was mechanically transmitted only to Cymbidium orchids. Previously, we designated it as cymbidium chlorotic mosaic sobemovirus(CyCMV)(Kondo et al,1994),as the virus was considered to be a new member of the genus Sobemovirsu.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996Genomic Organization of Odontoglossum Ringspot Virus (Cy-1 Strain) RNA and Comparison with That of Korean Strain137147ENMasatoIkegamiNarinobuInouyeThe complete nucleotide sequince of the genomic RNA of odontoglossum ringspot virus Cy-1 strain(ORSV Cy-1) was determined using cloned cDNA. This sequence is 6611 nucleotides long containing four open reading frames, which correspond to 126 K,183 K,31 K and 18 K proteins. The 5' non-coding region of ORSV Cy-1 is 62 nucleotides. The ORFs encoded a 126 K polypeptide and a 183 K read-through product in which helicase-sequence and polymerase-sequence motifs are found. The5' non-coding region,which extends from bases 1 to 62 has 2G residues and the ribosome binding site (AUU). The3' non-coding region of ORSV Cy-1 composes 414 nucleotides in length. The genomic organization of ORSV Cy-1 is nearly identical to that of ORSV Korean strain(ORSV-K). However, the ORF encoding 183 K protein overlapes the ORF encoding 31 K protein in ORSV Cy-1, but not in ORSV-K. The 183 K read-through product of ORSV Cy-1 is 16 amino acids longer than that of ORSV-K. The homology of the nucleotide sequences of ORSV Cy-1 and ORSV-K is 96%.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996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119135ENNarinobuInouyeJun-ichiMatsumotoTakanoriMaedaKojiMitsuhataHidekiKondoMochimuTaharaOrchid 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.No potential conflict of interest relevant to this article was reported.岡山大学資源生物科学研究所Acta Medica Okayama0916-930X421996Detection and Identification of Viruses of Orchids in Indonesia109118ENNarinobuInouyeI WayanGaraThree viruses, Cymbidium mosaic virsu(CyMV), odontoglossum ringspot virus (ORSV) and an unidentified potyvirus were found in the orchids in Indonesia. CyMV was detected from orchids in 8 genera, namely Aranthera, Calanthe, Cattleya, Cymbidium, Gromatophyllum, Phalaenopsis,Oncidium and Vanda. The virus was widespread in many orchids in Indonesia and was common in Aranthera and Calanthe, thus being an economically important virus in Indonesia.ORSV was also detected in orchids of 5 genera, namely Bulvophyllum,Calanthe, Cattleya,Oncidium and Phalaenopsis. The unidentified potyvirus was found in Aranthera.No potential conflict of interest relevant to this article was reported.