Japanese Journal of Phytopathology
Online ISSN : 1882-0484
Print ISSN : 0031-9473
ISSN-L : 0031-9473
Volume 58 , Issue 2
Showing 1-22 articles out of 22 articles from the selected issue
  • Hidetoshi UEKUSA, Makoto ISHII, Tohru TERAOKA, Daijirou HOSOKAWA, Mino ...
    1992 Volume 58 Issue 2 Pages 181-187
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    When cucumber cotyledons that had been inoculated with tobacco mosaic virus (TMV) were challenge-inoculated with cucumber green mottle mosaic virus (CGMMV), the multiplication of TMV and the size and number of starch-lesions in the cotyledons remained the same as in single inoculation with TMV. Inoculation with CGMMV before TMV-infection, however, decreased the amount of TMV and the number of starch-lesions. The same results were obtained when cucumber mosaic virus (CMV) was inoculated before or after TMV. When cucumber cotyledons were inoculated with zucchini yellow mosaic virus (ZYMV) 4 days before TMV inoculation or jointly inoculated with ZYMV and TMV, TMV concentration increased roughly 7 and 4 times, respectively. In this case, TMV was widely distributed in horizontal and vertical directions in the cotyledon tissues, but eventually localized. In cucumber cotyledons co-infected with ZYMV and CGMMV, CGMMV concentration was not affected by ZYMV infection. Factors involved in the localization of TMV are discussed based on these observations.
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  • Kenichi YAMAGUCHI, Takao SANO, Masanobu ARITA, Masayoshi TAKAHASHI
    1992 Volume 58 Issue 2 Pages 188-194
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Fusarium oxysporum MT0062 isolated from the root of tomato induced suppressiveness to Fusarium wilt of tomato. F. oxysporum MT0062 was not pathogenic to several plants known as the host of F. oxysporum. The bud cell of F. oxysporum MT0062 was easily produced on the novel semi-synthetic medium based on black strap molasses in a jarfermentor. Protective effect of F. oxysporum MT0062 formulated with a zeolite material rose to the maximum when it was pre-treated on the surface of the nursery soil 7-14 days before transplanting the tomato seedlings to the soil infested with F. oxysporum f. sp. lycopersici race J1. The magnitude of protective effect was corresponding to the frequency of isolation of F. oxysporum MT0062 from the root of tomato. In the case of tomato seedlings double-treated with the MT0062-formula 10 days before and at the time of transplanting, protective effect against Fusarium wilt of tomato and Verticillium wilt of eggplant was demonstrated for more than 60 days after transplanting into the infested soil with F. oxysporum f. sp. lycopersici under greenhouse condition. Furthermore, F. oxysporum MT0062 showed protective effect on Verticillium wilt of eggplant. These results suggested that non-pathogenic F. oxysporum MT0062 was effective in reducing some of the wilt incidences in solanaceous crops such as Fusarium wilt of tomato and Verticillium wilt of eggplant.
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  • Hiroyuki IEKI, Hiroyuki SAWADA
    1992 Volume 58 Issue 2 Pages 195-199
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Inoculation method for evaluating the degree of resistance to crown gall disease in grapevine, criteria for the evaluation of the degree of resistance, and classification of the degree of resistance in the major 60 cultivars of grapevine were examined. Second and third internodes of current shoots developed from nursery plants were inoculated at several sites by the needle prick inoculation method with a suspension (ca. 2×108cfu/ml) of A. tumefaciens biovar 3, and were subsequently kept at 23°C for about 30 days. The diameter (2r) of the crown galls formed on the shoot was measured and the r2π value was calculated. Resistant “Delaware”, moderately resistant “Rose Ciotat”, and susceptible “Kyoho” cultivars should be inoculated at the same time when the degree of resistance in grapevine was evaluated. Then criteria for the evaluation of the degree of resistance were made as follows. When the index number against the r2π value of the crown galls formed on susceptible “Kyoho” cultivar was set at 100, the cultivars with an index number ranging from 0 to 20 were scored as resistant, those with index numbers from 21 to 60 were scored as moderately resistant, and those with an index number exceeding 60 were scored as susceptible. Based on the degree of resistance, the major 60 cultivars of grapevine inoculated with A. tumefaciens biovar 3 were classified into resistant (11 cultivars), moderately resistant (30 cultivars) and susceptible (19 cultivars) groups.
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  • Kazunari NOMURA, Shinji KAWASAKI
    1992 Volume 58 Issue 2 Pages 200-207
    Published: April 25, 1992
    Released: February 19, 2009
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    A concentrated culture filtrate of the rice blast fungus, Pyricularia oryzae, showed strong activity to disrupt protoplasts of rice leaves. The proportion of disrupted protoplasts decreased linearly with the logarithm of the dilution. The protoplast-disrupting factor was labile above 60°C, and was precipitated with 10% TCA (Trichloroacetic acid) or 50% saturated ammonium sulfate solution, suggesting its proteinaceous character. The factor was also precipitated with 80% acetone or ethanol. It was adsorbed to a DEAE-Toyopearl column, and then eluted with NaCl at about 0.1M. Via gel filtration on Sephacryl S-300, the factor was eluted as a major protein peak of approximately 30kDa. The fungal culture filtrate also disrupted protoplasts of wheat and rye but not those from barley or the three dicotyledonous plants, suggesting some host species specificity. Protoplast-disrupting activity of the culture filtrate appeared to be slightly higher in the incompatible combinations of rice cultivar and race of blast fungus than in the compatible combinations. The role of this factor in hypersensitive cell death at the early stage of infection is discussed.
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  • Hiroshi OKAZAKI, Michihiko SAITO, Osamu TSURUTA
    1992 Volume 58 Issue 2 Pages 208-213
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    The population levels of Aspergillus flavus and A. parasiticus in field soil were examined in two geographically different areas of the Kyushu district. The levels of A. flavus populations in the Kumamoto and Miyakonojo areas were 30±9.7 and 11±2.3 propagules/g soil, respectively. Although the population levels were low, A. flavus seemed to be distributed widely in the field soils of both areas. No A. parasiticus was detected in this study. A. flavus populations were high in some soils. However, the influence of the crops planted on the population levels of A. flavus was not clear in corn, peanut and sweet potato crops. Five out of 28 A. flavus isolates from both areas produced aflatoxins.
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  • Yutaka ARIMOTO, Yasuo HOMMA
    1992 Volume 58 Issue 2 Pages 214-219
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    The relationship between development of Sphaerotheca fuliginea and chlorophyll in cucumber plants (Cucumis sativus L.) was studied. As chlorophyll mutant, yellow mutant cucumber and variegated cucumber with white mottles, which occurred by a natural mutation were used. The yellow mutant cucumber lacks chlorophyll, and white mottle parts of variegated cucumber leaves also lack chlorophyll. On normal cucumber leaves, the hyphae elongated to 1, 209μm in length 48hr after inoculation, and 63 haustoria were formed from each conidium within 96hr after inoculation. Conidiophores were formed 4 days after inoculation. On the white mottle parts of variegated cucumber leaves, the hyphae reached 173μm 48hr after inoculation, and 5 haustoria were produced from each conidium within 96hr. The conidiophores were observed 14 days after inoculation. On cotyledons of yellow mutant cucumbers, the hyphae elongated to only 74μm in length, and no second haustorium was formed in 72hr after inoculation. On yellow mutant cucumber whose roots had been soaked in a 2% sucrose solution, however the hyphae elongated 1, 100μm in length in the same period, and 48 haustoria were formed 96hr after the inoculation. The conidiophores were formed 4 days after inoculation. Chlorophyll is not necessarily requisite for the fungal growth, since S. fuliginea developed as readily on yellow mutant cotyledons as on green leaves, if sucrose was supplied to the plants.
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  • Hirotaka DATE, Katsumi OZAKI, Takashi SHIRAKAWA, Hideo NASU, Motomu HA ...
    1992 Volume 58 Issue 2 Pages 220-227
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    An unknown bacterial wilt of the cucumber plants grafted on pumpkin rootstocks was occurred under vinylhouse conditions in Okayama, Japan, in 1989. In the wilted plants, yellowing was observed at stems and vascular bundle of rootstocks but not those of cucumber scion. The bacteria isolated from infected stems induced the wilt of cucumber plant grafted on pumpkin rootstock which was similar to the symptom caused by the natural infection. They showed distinct pathogenicity to pumpkin and tomato, but not to cucumber plant with no grafting. The bacteriological characteristics of the bacterium coincided with those of the standard isolates of Pseudomonas solanacearum. On the basis of pathogenicity test and bacteriological characteristics, the bacterium was identified as Pseudomonas solanacearum (Smith 1896) Smith 1914. Bacterial wilt of pumpkin was proposed for the name of the disease.
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  • Siti Muslimah WIDYASTUTI, Fukuji NONAKA, Keisuke WATANABE, Nobumichi S ...
    1992 Volume 58 Issue 2 Pages 228-233
    Published: April 25, 1992
    Released: February 19, 2009
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    Two kinds of biphenyl phytoalexins accumulated in leaves of Photinia glabra Maxim (Rosaceae) in response to inoculation with spores of Entomosporium mespili, a pathogen of P. glabra, or to treatment with 10-4M HgCl2. The phytoalexins were identified as 2'-methoxyaucuparin and 4'-methoxyaucuparin. The contents of 2'-methoxyaucuparin and 4'-methoxyaucuparin were 3.578 and 1.233μg/g of leaf (fresh weight), respectively, 4 days after the inoculation with spores of E. mespili, whereas their contents in the uninoculated leaf were minute. The two phytoalexins exhibited toxicity to Pestalotia photiniae, a pathogenic fungus to P. glabra, and to Botrytis cinerea and Cochliobolus miyabeanus, the non-pathogens. The toxicity of 4'-methoxyaucuparin to P. photiniae and Botrytis cinerea was higher than that of 2'-methoxyaucuparin.
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  • Pyoyun PARK, Tohgo OHNO, Syoyo NISHIMURA, Hiroshi OTANI, Keisuke KOHMO ...
    1992 Volume 58 Issue 2 Pages 234-243
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    This paper presents the first comprehensive morphometry analysis of Japanese pear leaves treated with AK-toxin to examine the response of the cells after the treatment. The mean area-ratio of nuclei, chloroplasts, vacuoles, mitochondria, and microbodies to the cells was not significant between the toxin-treated and water-treated susceptible cells. On the contrary, in the toxin-treated susceptible cells, the area of nucleoli, RER, Golgi regions, and mitochondrial cristae increased. Furthermore, the number of pinocytotic Golgi vesicles increased within 6hr after the toxin treatment. However, in the susceptible leaves contain-ing necrotic cells, most organelles extremely decreased in their area or number 10hr after the toxin treatment. The resistant cells showed no significant changes in those organelles by the treatment with the toxin. The results suggested that the toxin caused temporary activation of both synthetic cellular metabolism and Golgi secretion in the susceptible cells within 6hr after the toxin treatment and, subsequently, the activation of those organelles gradually decreased and, finally, the cells were dead.
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  • Hiroyuki SAWADA, Hiroyuki IEKI, Shozo KOBAYASHI, Iwao OIYAMA
    1992 Volume 58 Issue 2 Pages 244-252
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Nineteen strains of Agrobacterium tumefaciens and 2 strains of A. rubi were tested for host range (stems of grapevine, peach, pear, tomato, sunflower, Kalanchoe daigremontiana and Nicotiana tabacum, and leaves of K. daigremontiana), tumor morphology on K. daigremontiana and grapevine, opine induction in tomato tumors, and agrocin 84 sensitivity. By principal component analysis using the mean diameter of 60 tumors formed by each strain on each of the test plants as a variable, the total of 21 strains were separated into the following seven groups named Groups 1-7. Group 1 (octopine and cucumopineinducing strains), Group 2 (vitopine-inducing strains) and Group 3 (opine class was not determined) consisted of only biovar 3 strains, showed strong pathogenicity to grapevine, but differed in pathogenicity to other plants. Group 4 (octopine, agropine and mannopine-inducing strains) consisted of only biovar 1, whereas Group 5 (nopaline-inducing strains) comprised biovars 1 and 2, A. rubi and unclassified strains isolated from cherry, indicating its heterogeneity in chromosomal markers. Groups 4 and 5 showed weak pathogenicity to grapevine, but strong pathogenicity to the other plants. Both of Group 6 (opine class was not determined) and Group 7 (nopaline-inducing strains) contained unclassified strains (NCPPB 1650 and two kiwifruit isolates, respectively), and showed weak pathogenicity to all the inoculated plants. Strains belonging to each group gave uniform reactions within the group in tumor morphology, opine induction and agrocin 84 sensitivity, except for Group 5 which was divided into 2 subgroups (A and B) based on agrocin 84 sensitivity. Agrocin 84-sensitive Group 5-A consisted of biovars 1, 2 and unclassified cherry isolates, and resistant Group 5-B comprised only A. rubi. All the strains except for Group 5-A were agrocin-84 resistant. Six Japanese strains of biovar 3 belonging to Group 2 induced vitopine, indicating that the vitopine strains have a wide distribution in Japan.
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  • Hideo NASU, Motomu HATAMOTO
    1992 Volume 58 Issue 2 Pages 253-258
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Botryosphaeria berengeriana f. sp. persicae was confirmed to cause circular to ovalshaped sunken lesions around lenticels of peach trees, in addition to blisters on shoots, and gummosis. The blisters developed on 1 or 2 yr old shoots, and sunken lesions were formed on shoots older than 3 yr. Among 1 yr old shoots, most of blisters were formed on water sprout shoots, followed by intermediated type of shoots, whereas few blisters were formed on weak shoots which will bear fruits in next year. On water sprout shoot, many blisters were formed at basal part. The gummosis was found in rainy season on shoots older than 2 yr. Inoculation with conidia of the pathogen to peach trees in May, June or July caused more serious infection than those in April, September, or October. The symptoms produced by inoculation on 1, 2 and 3 yr old shoots were identical with those on naturally infected trees. Branches inoculated in April, May, June or July developed symptoms from late August to early September, those inoculated in August or September developed symptoms from October to December, and those inoculated in October developed in February of the next year. Increase in blisters occurred twice, i.e., in autumn and from spring to early summer of the next year. As a result of the experiment using infected branches as the inoculum, positive correlation was found between number of blisters formed and the amount of rain fall, and also the number of conidia trapped.
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  • Kiyoshi ISHIGURO, Seiichi TAKECHI, Akira HASHIMOTO
    1992 Volume 58 Issue 2 Pages 259-266
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    To construct a mechanistic model of the behavior of tricyclazole on rice leaves and its efficacy for the prevention of leaf blast epidemics under field conditions, quantitative data of temporal changes of the amount of fungicide residues on rice leave at various positions and a concentration-response curve of protectant effect against blast fungus versus fungicide concentration were determined. Disease progression in the filed with or without application of the fungicide was assessed to evaluate feasibility of the data. Tricyclazole was applied in two different timing to a rice field. Temporal changes of tricyclazole residues on leaves obey a decay curve model. On the leaves that had fully developed at the time of application, estimates of initial deposits of the fungicide ranged from 4.39 to 19.69 μg/g fresh leaves, and estimates of half-life value ranged from 2.46 to 5.33 days, respectively. In leaves which had not yet emerged or partly emerged at foliar application, 0.16 or 1.08 μg/g of the fungicide was detected when leaves were fully developed, respectively. Foliar application of tricyclazole almost completely protected the completely developed leaves from new infection about for 2 weeks. Also, the leaves newly emerged after fungicide application were somewhat protected. In the greenhouse experiment, tricyclazole showed a completely protectant effect at a concentration above 0.34μg/g, while the effect was not significant at a concentration below 0.20μg/g. The median effective concentration (EC50) of the fungicide was estimated to be 0.24μg/g. Generally, the values estimated in this experiment agreed with the disease progression observed in the field.
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  • Masayasu KATO, Norio SATO, Ahmed A. MOSA, Kiroku KOBAYASHI, Akira OGOS ...
    1992 Volume 58 Issue 2 Pages 267-275
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    More than three hundred Phytophthora infestans isolates collected from potato crops in Japan from 1987 to 1989 were investigated for their cultural features and divided into two groups, designated as S and M types. On potato tuber slices the S type isolates formed thin and appressed colonies producing abundant sporangia, while the M type isolates formed cottony colonies producing few sporangia. On V-8 juice agar (V-8A) the S type isolates grew well, while fresh isolates of the M type could not grow well. Cultural features on tuber slices and V-8A often changed after subculturing and storage. On rye A agar both types of isolates always grew well. On oatmeal agar the S type isolates could not grow, while the M type isolates grew well, and the features were stable. The cultural types S and M coincided with the mating types Al and A2, respectively.
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  • Katsunori TAMURA, Yuichi TAKIKAWA, Shinji TSUYUMU, Masao GOTO, Minoru ...
    1992 Volume 58 Issue 2 Pages 276-281
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Xanthomonas campestris pv. phormiicola, the pathogen of bacterial streak of New Zealand flax, induces hypertrophy on potato tuber tissues at the inoculated site. The hypertrophic active substance isolated from the bacterial culture filtrate produced visible chlorotic lesions on the leaves of New Zealand flax, ryegrass and tobacco plant. Rf values on TLC and 1H NMR spectrum of the purified toxin coincided well with those of coronatine. Thus, the toxin produced by X. campestris pv. phormiicola was identified as coronatine. This bacterium is the first example of coronatine producing xanthomonads. Coronatine was also detected in the leaves of host plant infected by this bacterium. As no plasmid DNA was detected, the possible involvement of plasmid DNA in coronatine production was suggested to be unlikely in this bacterium.
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  • Akira MORITA
    1992 Volume 58 Issue 2 Pages 282-285
    Published: April 25, 1992
    Released: February 19, 2009
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    Thirty-two seedlings of one year old healthy loquat plants were planted in the field of the Nagasaki Fruit-Tree Experiment Station and inoculated with canker pathogen, Pseudomonas syringae pv. eriobotryae, at a part of their growing terminal buds of each test seedling in February, 1980. The effects of the applications of Bordeaux mixture during the former half, the latter half and the whole of nine years of the test period from March, 1980 until November 1989 on the disease incidence were compared. Original inoculum used in the test was a type resistant to streptomycin and belonging to the A group which does not form holo lesions on the leaves and does not produce brown pigments on Wakimoto's agar medium. At the end of the test period, the lesions were collected and identified the type of the pathogens isolated, to determine their origin. Most of the isolates were the type used for the original inoculation. The best control effect was shown in the plot treated with Bordeaux mixture during the whole test period followed by that treated during the former half, that treated during the latter half of the test period and the untreated plot. The results show the importance of the application at early growth stages of the loquat plants.
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  • Chung L. CHEN, Atsushi MIYASAKA, Shun-ichiro MIYASHITA, Yoshio EHARA, ...
    1992 Volume 58 Issue 2 Pages 286-291
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
    Plasmid-like DNAs (plDNAs) were found associated with mitochondrial DNA fraction following CsCl-bisbenzimide density-gradient ultracentrifugation for 15 isolates belonging to the anastomosis group 4 (AG-4) of Rhizoctonia solani. These 15 isolates were divided into two classes based on the size of plDNAs observed. Four isolates contained 2.4kb plDNA and 2 of them carried an additional plDNA of 4.8kb. Eleven isolates contained 2.7kb plDNAs and 4 of them carried an additional plDNA of 5.4kb. All these plDNAs of 2.4kb and 2.7kb hybridized strongly with the probes of 2.7kb plDNAs, pRS64-1, -2 and -3. Restriction mapping of 2.4kb plDNAs revealed the presence of two types of maps. Two isolates carried both types of 2.4kb plDNAs. Restriction mapping of 2.7kb plDNAs revealed the presence of three types of maps. One isolate contained two types of 2.7kb plDNAs, and other isolate contained three types corresponding with those of pRS64-1, -2, and -3, respectively. The isolates in AG-4 of R. solani were classified into two groups based on the presence of 2.4 and 2.7kb plDNAs.
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  • Tomoki MOTOMURA, Kinji TANAKA, Nobumichi SAKO, Fukuji NONAKA
    1992 Volume 58 Issue 2 Pages 292-297
    Published: April 25, 1992
    Released: February 19, 2009
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    The effect of light and culture media associated with buoyancy of sclerotia of Corticium rolfsii, causal agent of southern blight disease of water chestnut (Trapa bispinosa Roxb.) was examined. Sclerotia formed in field became buoyant in water, but sclerotia on culture media sunk in water. Microscopic study revealed that sclerotia formed in field had more intercelluler spaces in sclerotial tissues and thinner cell walls than sclerotia on culture media. Many sclerotia were formed under light conditions, although buoyancy of sclerotia depended on both the light conditions and the nutrient constituents of the medium. When diluted potato sucrose agar (PSA) or water chestnut leaves agar media were used for formation of sclerotia under light conditions, buoyant ability and internal structures were similar to that of sclerotia formed in field. On the other hand, fewer sclerotia were formed on each kind of media under dark conditions, but the sclerotia did not float in water. Light conditions and nutrient constituents play important roles for the formation of buoyant sclerotia of C. rolfsii.
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  • Seiichi OKUDA, Mutsuki HASEGAWA, Tomohide NATSUAKI, Kazuhiko KAJI, Kei ...
    1992 Volume 58 Issue 2 Pages 298-304
    Published: April 25, 1992
    Released: February 19, 2009
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    A virus, designated NFU, isolated from bean plants (Phaseolus vulgaris cv. Kentucky Wonder) with severe vine necrosis (Turugare-byo) in Fukushima Prefecture was compared with a seed-borne isolate of bean yellow mosaic virus (BYMV-SB) from broad bean. When mechanically inoculated onto test plants, NFU caused severe necrotic symptoms on bean, broad bean and pea, whereas BYMV-SB caused mosaic symptoms. Isolate NFU was transmitted in a nonpersistent manner by Myzus persicae but not through seed. Virus particles and inclusion bodies showed typical potyvirus morphology when examined by electron microscopy. Analysis of a purified preparation of NFU by SDS-PAGE revealed two protein bands with relative molecular weights of 34k and 37k. In immunoelectron microscopy with some antisera against BYMV and clover yellow vein virus (CYVV), NFU was serologically more closely related to CYVV than BYMV. In enzyme linked immunosorbent assay (ELISA), agar gel diffusion test, immunoelectron microscopy, and Western blotting, NFU and BYMV-SB were serologically related but antigenically different. Based on these results NFU was identified as a strain of CYVV. To find the weed-host plants of CYVV causing necrosis on bean, wild plants around bean fields were collected and assayed by ELISA and sap inoculation onto bean; eight wild plants (Plantago asiatica, Veronica persica, Glechoma hederacea, Lamium amplexicaule, Perilla frutescens, Trifolium repens, T. pratense, Iris sp.) were infected with severe CYVV and thought to be important primary sources of inoculum for bean vine necrosis.
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  • Norihiro SHIMOMURA, Pyoyun PARK, Hiroshi OTANI, Motoichiro KODAMA, Kei ...
    1992 Volume 58 Issue 2 Pages 305-309
    Published: April 25, 1992
    Released: February 19, 2009
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  • Eri MATSUMIYA, Ken SUZUKI, Yoshikazu UENO, Junya MIZUTANI
    1992 Volume 58 Issue 2 Pages 310-314
    Published: April 25, 1992
    Released: February 19, 2009
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  • Masatsugu OBUCHI, Tohru TERAOKA, Daijiro HOSOKAWA, Minoru WATANABE
    1992 Volume 58 Issue 2 Pages 315-318
    Published: April 25, 1992
    Released: February 19, 2009
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  • Shinya TSUDA, Keiko T. NATSUAKI, Keiichi TOMARU
    1992 Volume 58 Issue 2 Pages 319-324
    Published: April 25, 1992
    Released: February 19, 2009
    JOURNALS FREE ACCESS
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