Japanese Journal of Phytopathology
Online ISSN : 1882-0484
Print ISSN : 0031-9473
ISSN-L : 0031-9473
Volume 56 , Issue 5
Showing 1-21 articles out of 21 articles from the selected issue
  • Kazusato OHSHIMA, Jumanto HARJOSUDARMO, Yo ISHIKAWA, Eishiro SHIKATA
    1990 Volume 56 Issue 5 Pages 569-576
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    More than two hundred and fifty-five monoclonal antibody (MoAb)-secreting hybridomas against 3 plant luteoviruses, 2 plant reoviruses and a potyvirus were produced. The hybridomas for potato leafroll virus, beet western yellows virus, tobacco necrotic dwarf virus, rice dwarf virus, rice ragged stunt virus and potato virus Y-ordinary strain, were screened by four different procedures of enzyme-linked immunosorbent assay (ELISA); procedure 1, antigen adsorption indirect ELISA (AAI-ELISA) in which purified virus in phosphate buffered saline (PBS) at pH 7.4 was adsorbed onto the microplate wells, procedure 2, AAI-ELISA in which purified virus in sodium carbonate-bicarbonate buffer at pH 9.6 was adsorbed onto the microplate wells, procedure 3, indirect double antibody sandwich ELISA (IDAS-ELISA) in which polyclonal antibody was used as trapping antibody and purified virus preparations diluted in PBS-T (containing Tween-20) as antigens were used, procedure 4, IDAS-ELISA in which polyclonal antibody was used for trapping antibody and crude saps of virus infected plants extracted in PBS-T as antigens were used. Based on the MoAb reactivities against homologous viruses in four different ELISA procedures, MoAb-secreting hybridomas were divided into ten groups. Using purified MoAbs from ascitic fluids, direct double antibody sandwich ELISA (DAS-ELISA) was examined for the detection of virus antigens in infected plants. All the MoAbs reacted in DAS-ELISA belonged to group 1 in which the MoAbs were reactive in each four of the screening procedures 1-4, or group 2 in which MoAbs were reactive in three of screening procedures 1, 3 and 4. These results indicate that MoAbs being reactive in DAS-ELISA can be readily selected in hybridomas in group 1 or 2.
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  • Kazuo SUYAMA, Budi TJAHJONO, Hiroshi FUJII
    1990 Volume 56 Issue 5 Pages 577-583
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    A slimy rot symptom was found on the potato tubers transported from Hokkaido (Abashiri) to Tokyo. The symptom appeared only on the tubers which has been washed with tap water immediately after harvest and packed in plastic bags, but did not appear on unwashed ones. Same symptom was observed on the potato tubers piled up temporarily at the fields or on the seed-tubers stored in the air-conditioned store house (5-10C) for several months. Causal organisms were identified as pectolytic clostridia based on their bacteriological properties which will be reported in a succeeding publication. Pectolytic clostridia were detected on most of potato tubers collected from various potato fields in Japan. Not only on harvested matured tubers, they were also found on young and small tubers in their early growth stages. Slimy rot and gas production in rotten tubers are important criterions to distinguish this disease from soft rot which is caused by Erwinia spp., though other symptoms are almost similar. The name of “slimy rot” was proposed for this new post-harvest disease of potato tuber.
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  • Hideki YAMAMOTO, Kiroku KOBAYASHI, Akira OGOSHI
    1990 Volume 56 Issue 5 Pages 584-590
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Isozyme patterns for 15 enzymes from mycelial extract were compared with 10 isolates each of Phialophora gregata from adzuki bean and soybean and one isolate each of Acremonium sp. from adzuki bean and Cephalosporium gramineum from wheat. These three species showed considerable different patterns in all the enzymes tested with the coefficient of similarity of 0.05. The adzuki bean and soybean isolates of P. gregata exhibited the same patterns in 11 enzymes. Four other enzymes, i.e. lactate dehydrogenase, peroxidase, α-esterase and acid phosphatase, distinguished these two groups of isolates. The coefficient of similarity within each group of the adzuki bean and soybean isolates and between the groups were 0.83, 0.97 and 0.64, respectively. These results support the recognition of the two groups as separate formae speciales of P. gregata.
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  • Carlos Hidemi UESUGI, Kenichi TSUCHIYA, Kazunori TSUNO, Nobuaki MATSUY ...
    1990 Volume 56 Issue 5 Pages 591-596
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    SDS-polyacrylamide gel electrophoresis patterns of Erwinia carotovora subsp. carotovora strains were compared with those of other plant pathogenic bacteria such as E. c. subsp. atroseptica, E. herbicola pv. milletiae, E. chrysanthemi pv. zeae, Pseudomonas syringae pv. tabaci, P. gladioli pv. gladioli, Xanthomonas campestris pv. glycines, X. c. pv. phaseoli, and X. c. pv. vignicola. Since the 60.2kd peptide band was consistently detected from all E. c. subsp. carotovora strains while not from others, usefulness of this band as taxonomic determinant was suggested. The reaction of membrane proteins extracted from bacterial strains to an anti-E. c. subsp. carotovora N7129 serum was similar to that of whole cell antigen suggesting important roles of membrane proteins as antigen determinants of the bacteria.
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  • Carlos Hidemi UESUGI, Kenichi TSUCHIYA, Kazunori TSUNO, Nobuaki MATSUY ...
    1990 Volume 56 Issue 5 Pages 597-604
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Membrane proteins of Erwinia chrysanthemi strains isolated from various host plants were extracted by LiCl and analyzed by SDS-polyacrylamide gel electrophoresis. The strains showed various polypeptide patterns associating with the kinds of hosts from which they were isolated. Especially, polypeptide bands near 40.0kd were closely related to pathovars indicating usefulness of the membrane protein profiles as a taxonomic tool for grouping E. chrysanthemi strains at infraspecific level of pathovars. From these results it was surmised that polypeptides of membrane protein playing some important roles for determination of host specificity.
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  • Jin-zhong XIAO, Shin-ichi NAKATSUKA, Mitsuya TSUDA, Noriyuki DOKE, Syo ...
    1990 Volume 56 Issue 5 Pages 605-612
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Bipolaris zeicola race 3, which was originally reported as a pathogen causing a leaf spot disease of corn, was found to cause a leaf spot disease on rice plants when inoculated in greenhouse. A factor inducing susceptibility to a non-pathogenic isolate of Alternaria alternata only in rice plant leaves was isolated from the culture filtrates of race 3 by ethyl acetate extraction, TLC, and HPLC. The factor was characterized as a compound with the unique molecular formula of C30H44O8 from 1H-NMR, 13C-NMR and mass spectra analyses and was named as SIF 1. SIF 1 at concentrations higher than 10μg/ml induced the susceptibility, but neither inhibited root growth of rice seedlings nor caused necrosis or chlorosis on rice leaves even at 250μg/ml. However, SIF 1 may not be the primary determinant of B. zeicola race 3 responsible for the pathogenicity in rice plants, because SIF 1 was not detected in spore-germination fluids of race 3, even though the fluids strongly induced a susceptibility on rice leaves, and also because B. zeicola race 1, B. victoriae and B. bicolor, which are non-pathogenic to rice plants and known to be closely related to B. zeicola race 3, also produced SIF 1 during culture in nutrient medium. SIF 1 may be used as a suppressor probe for the analysis of resistance of rice plants.
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  • Naoto NITTA, Yoichi TAKANAMI
    1990 Volume 56 Issue 5 Pages 613-620
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Cucumber mosaic virus (CMV) RNA replication complex was prepared from a crude membrane fraction of infected tobacco leaves and was further purified by a zwitterionic detergent, Zwittergent, and Sepharose 4B column chromatography. The active fraction was found at void volume of the column and in the precipitate after following ultracentrifugation. In vitro products synthesized by the enzyme fraction as well as by the crude enzyme were full-length viral RNAs 1-4, suggesting that the characteristic of the enzyme essentially unchanged by the Zwittergent treatment. Analysis of proteins labeled with radioactivity in CMV-infected tobacco leaf disks showed that the enzyme fraction contained three infection-specific proteins, Mr 105, 000, Mr 32, 000 and Mr 25, 000, which comigrated with viral 1a, 3a and coat proteins, respectively, synthesized in vitro. However, a protein corresponding to 2a protein encoded by CMV RNA 2 was not identified in in vitro translation system nor in the enzyme fraction. Considering our previous results that inoculation of RNAs 1 and 2 is necessary to induce replicase activity, this result implies that at least 1a protein is closely related to the replicase activity, while the role of 2a protein in RNA replication is yet to be solved.
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  • Yoshiyuki TAKAHASHI, Kenichiro SHOHARA
    1990 Volume 56 Issue 5 Pages 621-627
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Nine monoclonal antibodies (MABs) were examined for their reactivities to wasabi strain of tobacco mosaic virus (TMV-W) in two indirect ELISA and competitive ELISA. Based on the results, seven epitopes were discriminated on the surface of a particle of TMV-W and one cryptotope was found inside of the virus particle. For practical detection of TMV-W, mixture of different types of MABs, i.e., cocktail MABs, were examined in double sandwich antibody form ELISA (DAS-ELISA). Five MABs with high titers, which recognized different epitopes were chosen to compound the cocktail MABs. Among the combinations of five MABs, that with the highest sensitivity was selected for coating γ-globulin. To obtain the sensitive and specific conjugates, four MABs were used for compounding cocktail conjugate. When the MABs which did not compete in the reaction with antigens were compounded, the highest sensitivity was obtained. When the cocktail γ-globulin and cocktail conjugates were used for detection of TMV-W from infected plants in the DAS-ELISA, the sensitivity was almost the same as that used polyclonal antibodies. Furthermore, the cocktail MABs distinguished TMV-W from crucifer strain of TMV, the differentiation of which was difficult by using polyclonal antibodies.
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  • Motoichiro KODAMA, Toshimasa SUZUKI, Hiroshi OTANI, Keisuke KOHMOTO, S ...
    1990 Volume 56 Issue 5 Pages 628-636
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Alternaria alternata tobacco pathotype, the causal organism of brown spot disease of tobacco, produced a host-selective toxin (named AT-toxin) in culture filtrates and spore-germination fluids. The toxin was purified from culture filtrates of this pathogen by a serial use of ion exchange, silicic acid and gel filtration columns, and thin layer chromatography. Purified toxin inhibited the seedling root growth of both susceptible and moderately resistant cultivars of tobacco at 0.2μg/ml, whereas the toxin induced necrosis on each leaf at 1 and 20μg/ml, respectively. The toxin selectively affected the viability of protoplasts derived from tobacco leaves, and caused an increase in the oxygen uptake by leaf tissues. Among several plants tested, only species belonging to the genus Nicotiana were sensitive to the toxin. All isolates of A. alternata that produced the toxin in spore-germination fluids caused brown spot disease of tobacco. Successful infection by nonpathogenic Alternaria on tobacco leaves was obtained when spores were inoculated with the toxin. These results indicate that AT-toxin is a host recognition factor in the genus Nicotiana-A. alternata pathosystem.
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  • Motoichiro KODAMA, Pyoyun PARK, Takashi TSUJIMOTO, Hiroshi OTANI, Keis ...
    1990 Volume 56 Issue 5 Pages 637-644
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    The primary target organelle of AT-toxin which is a host-selective toxin from the tobacco pathotype of Alternaria alternata causing brown spot disease of tobacco, was determined by electron microscopy and morphometrical analysis for the ultrastructure. The first change in the ultrastructure of toxin-treated susceptible leaf cells was swelling and matrical lucency of mitochondria, that were evident by 24hr after the onset of toxin exposure. The mitochondria showed a remarkable decrease in volume of cristae and the matrical cavitation by 48hr and 72hr after the onset of treatment. Besides, dense granules disappeared, and the bulge formation and attenuation of mitochondrial membranes were found. The mitochondrial alterations became more prominent with the time after treatment. But the other organelles appeared to be normal. These results indicate that mitochondria are the primary target organelle of AT-toxin.
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  • Hiroshi KAMIUNTEN
    1990 Volume 56 Issue 5 Pages 645-650
    Published: December 25, 1990
    Released: February 19, 2009
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    Pseudomonas syringae pv. eriobotryae is a causal agent of stem canker of loquat. All of the strains which produced dark brown pigment harboured three plasmids of approximately 32, 39, and 85 megadaltons (Mdal). Twenty-eight derivative strains cured of the 85 Mdal plasmid were obtained from NAE57 and NAE34 by five or six times subcultures as the elevated temperature (32C). All of the cured strains induced symptoms with no callus-like tissue on the inoculated stems of loquat, whereas 28 derivative strains that still harboured 85 Mdal plasmid induced symptoms with the development of the similar callus-like tissue as those of parent strains. Since the loss of the 85 Mdal plasmid was always accompanied by change in symptoms, it is strongly suggested that the 85 Mdal plasmid in P. syringae pv. eriobotryae plays an important role in the development of symptoms.
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  • Tetsuji YAMADA, Hiroshi TSUKAMOTO, Tomonori SHIRAISHI, Shinji KAWAMATA ...
    1990 Volume 56 Issue 5 Pages 651-657
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Taphrina wiesneri and T. deformans, causal agents of plant hyperplastic diseases, produce indoleacetic acid (IAA) via indolepyruvic acid and indoleacetaldehyde from L-tryptophan as intermediates. They also convert indoleacetonitrile into IAA. We have isolated Iaa- mutants of these Taphrina spp. by the treatment with clofibric acid. These mutants are Pur- (deficient in purine biosynthesis) and exhibit unique morphology including the smaller conidium size. They can no longer accumulate IAA in culture filtrate but accumulate indoleethanol with the addition of tryptophan. They show significantly different chromosome length polymorphism from the parental ones. However, the length of the three chromosomal DNA in Iaa- mutants of T. wiesneri and T. deformans is identical.
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  • Seisaku UMEMOTO
    1990 Volume 56 Issue 5 Pages 658-664
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    The significance of two kinds of Japanese pear scab infection sources, ascospores and conidia, in primary infection was investigated. Throughout three years experiments, the discharge of ascospores from pseudothecia produced on fallen leaves began from late March to early April, and finished in the end of May. Ascospores were first trapped at 10 to 22 days before the full blooming on cv. Kosui. Dispersion of conidia produced on lesions of scales and basal parts of flower clusters generally started in mid April and the number of dispersed conidia into the rain water tended to increase with the lapse of days. The germination ratio of these conidia was high. Scab occurrence on cv. Chojuro was light when trees were exposed to infection with conidia in the beginning of blooming but severe when infection occurred after the end of blooming. In the comparative experiments using two kinds of infection sources, the scab occurrence on leaves infected with ascospores was earlier and severer than those infected with conidia. From these results, it was concluded that ascospores were highly involved in the primary infection with Japanese pear scab.
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  • Toshihiro IWAKUMA, Yuhji ATAKA, Nobuaki MATSUYAMA, Satoshi WAKIMOTO
    1990 Volume 56 Issue 5 Pages 665-670
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Infection of the rice blast fungus Pyricularia oryzae elicited the diterpenoid phytoalexin (PA) production in rice leaves. Striking difference in the time course of PA production was observed between incompatible and compatible race-cultivar relations. The PA production was firstly observed at 36-48hr, in general, after inoculation in incompatible case and no production was observed at this time in compatible case. The production was sometimes detected at 12hr in incompatible case. Such difference, however, disappeared in the case of the treatment with heat-killed spores of the fungus and PA production was observed at 48hr in incompatible and compatible cases. No elicitation of PA production was documented at the treatment with protoplasts originated from the hyphal cells of Pyricularia oryzae. Hyphal cell-wall components, especially water insoluble fraction, obtained by the fractionation of the fungal cells of compatible and incompatible races elicited excellently PA production. These results clearly verified the existence of the elicitor in hyphal cell wall.
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  • Kinji TANAKA, Fukuji NONAKA
    1990 Volume 56 Issue 5 Pages 671-673
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Onions grown in Ariake state-operated reclamation in Saga Prefecture were found to be more susceptible to the black mold caused by Aspergillus niger van Tieghem than those grown in Shiroishi-machi and Genkai-machi in which onions have been continuously cultivated for more than twenty years. The content of inorganic substances in the cuticular layer of onion leaf was investigated by a transmission electron microscope equipped with an energy dispersive X-ray analyser. Among of inorganic substances, the calcium contents in cuticular layer were constantly lower in the onions grown in the polder land than those grown in traditionally cultivated areas. These results suggest that the calcium in onion tissues may play an important role in the mechanism of resistance to the disease.
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  • Mabito IWASAKI, Tadaoki INABA
    1990 Volume 56 Issue 5 Pages 674-676
    Published: December 25, 1990
    Released: February 19, 2009
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  • Norio KONDO, Kippei MURATA, Hisanori SHIMADA, Shigehisa SHIRAI, Fujio ...
    1990 Volume 56 Issue 5 Pages 677-679
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
  • Jun SHIMOYAMA, Kaoru HANADA, Mitsuro KAMEYA-IWAKI, Hiroshi TOCHIHARA, ...
    1990 Volume 56 Issue 5 Pages 680-683
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Konjak (Amorphophallus konjac) plants showing necrosis in leaflets and stunting of petiole have been found in Gunma Prefecture since around 1960. The causal virus was transmissible by aphids and by mechanical inoculation, but not through corms and cormlets of konjak. The virus was identified to be cucumber mosaic virus (CMV) by immunodiffusion tests. Five CMV isolates obtained from different konjak fields were compared serologically, and three of them were indistinguishable from CMV-Y whereas the other two were identical to CMV-P. All these isolates caused similar symptoms on konjak plants to those observed in the fields. We propose the name of the disease, konjak necrotic stunt disease.
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  • Taeko TAKEUCHI
    1990 Volume 56 Issue 5 Pages 684-686
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    All of the ten isolates of Pseudoperonospora cubensis sampled in greenhouses at Asahi area, Chiba Prefecture, where the effectiveness of the application of phenylamide fungicides for the control of downy mildew of cucumber appeared to have declined since the spring of 1989, were found to be resistant to the fungicides at 100ppm concentration, in assay using the cucumber leaf disc method. Both the preventive and curative effect of the mixtures of phenylamide and mancozeb or copper compounds on downy mildew caused by the resistant isolates were found to decrease in experiments using potted cucumber plants.
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  • Hideaki NEGISHI, Tetsuji YAMADA, Tomonori SHIRAISHI, Hachiro OKU, Hiro ...
    1990 Volume 56 Issue 5 Pages 687-690
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
  • Masamichi NISHIGUCHI, Fusao MOTOYOSHI, Nobuyuki OSHIMA
    1990 Volume 56 Issue 5 Pages 691-694
    Published: December 25, 1990
    Released: February 19, 2009
    JOURNALS FREE ACCESS
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