Mitsuro Kameya-Iwaki

Biological Mode of Action of the Fungicide, Flusulfamide, Against Plasmodiophora brassicae (clubroot)

Flusulfamide (2′, 4-dichloro-α,α,α-trifluoro-4′-nitro-m-toluenesulfonanilide) was investigated for its mode of action against Plasmodiophora brassicae Woronin. Seedlings of Chinese cabbage (Brassica rapa L. subsp. pekinensis) were grown for 14 and 21 days in soil infested with P. brassicae and then transplanted into soil containing flusulfamide (0.9 µg a.i. g−1 dry soil). Clubroot was not suppressed by this treatment, indicating that the fungicide is ineffective against P. brassicae established within cortical cells of the host root. Where seedlings were grown in soil infested with resting spores which had previously been treated with flusulfamide, root-hair infection and club formation were suppressed. This indicates that flusulfamide directly acts against resting spores. When placed in root exudates of Chinese cabbage, untreated resting spores germinated at a high frequency while flusulfamide-treated resting spores hardly germinated at all. Use of the Evans blue staining assay indicated that flusulfamide-treated resting spores remained viable. Flusulfamide was detected by high performance liquid chromatography on resting spores treated with flusulfamide for 30 min. This indicates that the chemical is adsorbed onto resting spores. These results suggest that flusulfamide suppresses clubroot disease by inhibiting germination of P. brassicae resting spores through adsorption onto their cell walls.

Comparison of the S RNA segments among Japanese isolates and Taiwanese isolates of Watermelon silver mottle virus

Summary. The complete nucleotide sequences of the S RNA of two Japanese isolates of Watermelon silver mottle virus were determined. One was isolated from naturally infected watermelon and causes malformation on upper leaves of Tetragonia expansa. The other was isolated from melon and causes characteristic yellow necrotic lesions on upper leaves of T. expansa. The total nucleotide sequences of the S RNA of WS-O and WS-Y were 3553 nt and 3558 nt long, respectively. Both the nucleotide sequences and the deduced amino acid sequences of WS-O and WS-Y were quite similar even though the symptoms on T. expansa are quite different. They were also significantly similar to those of the Taiwanese isolates, Topso-W and Tospo-To. These results suggested that the Japanese isolates and the Taiwanese isolates of WSMoV were classified as one group not only serologically but also genetically. Within the S RNA sequences, the most variable region was the intergenic region between the N gene and the NSs gene. This was due to a 20 nt insertion between the Japanese isolates and the Taiwanese isolates.

Detection of Thiophanate-methyl-resistant Strains in Diplocarpon mail, Causal Fungus of Apple Blotch

Isolates of Diplocarpon mali, causal fungus of apple blotch, collected from four prefectures in Japan in 1997–1998 were tested for sensitivity to thiophanate-methyl. Results from mycelial growth tests showed that MIC values of the fungicide were 0.19 μg/ml against all isolates from Akita, Nagano and Saga prefectures but 100 or 200 μg/ml against all isolates from the Tokusa area in Yamaguchi prefecture. Detached apple leaves sprayed with the fungicide developed severe symptoms when inoculated with the isolate from Tokusa, but developed no symptoms with the isolate from Nagano. These results are the first confirmation of thiophanate-methyl-resistant strains in D. mali.

Cycas necrotic stunt virus isolated from gladiolus plants in Japan

An undescribed spherical virus ca. 30 nm in diameter was isolated from gladiolus (Gladiolus spp.) plants in Japan. The virus had a moderate host range within eight families. Purified virus preparations contained two large RNA components and one coat protein with mobility similar to Cycas necrotic stunt virus (CNSV) from cycas (Cycas revolute). The virus was serologically closely related to CNSV. Its nucleotide sequence of the coat protein gene had 89% common identity with that of CNSV. These results indicated that the virus isolated from gladiolus is a new strain of CNSV.

Semipersistency of Myzus persicae Transmission of Cucumoviruses Systemically Infecting Leguminous Plants

Sequential transmission tests of Peanut stunt virus (PSV) and Cucumber mosaic virus (CMV) systemically infecting common bean, Phaseolus vulgaris, were conducted using Myzus persicae allowed to fast for 2 hr and then to acquisition feed on infected common bean plants or purified virus for 10 min. In the sequential transmission tests using either one or 10 aphids per assay plant, three isolates of PSV (J,S,Y5) and one of CMV (V) were transmitted from and to common bean up to a third or fourth inoculation access. Many aphids transmitted these viruses to two or three plants. Purified viruses of PSV-S and CMV-V were also transmitted up to a third or second inoculation access at low percentage. On tobacco, Nicotiana tabacum, aphids transmitted PSV-S and CMV-V only in the first inoculation access, although PSV-S was transmitted to only one plant in the fourth and fifth inoculation access. These viruses may be transmitted in two phases by aphids, depending on the plant species.

A Cucumber mosaic virus Isolated from Silene armeria

A Cucumber mosaic virus was newly isolated from Silene armeria and was characterized by biological, serological and molecular biological methods.

Effects of susceptibility of test plants on modes of cucumovirus transmission by Myzus persicae

In sequential transmission tests of Peanut stunt virus S (PSV-S) and Cucumber mosaic virus V (CMV-V) using Myzus persicae, these viruses behave as a semipersistent virus in Phaseolus vulgaris, but as a nonpersistent virus in Nicotiana tabacum, regardless of the species of plant used as the virus source. In addition, viruliferous aphids retained virus infectivity and transmitted it to P. vulgaris, even after they lost infectivity to N. tabacum. Apparently, the mode of transmission by the aphids differs depending on the plant species used for the assay. After mechanical inoculation with purified PSV-S or CMV-V, P. vulgaris appeared more susceptible to PSV-S than N. tabacum. However, the susceptibility to CMV-V appeared similar in both assay plants.

Characteristics of Epitopes on the Tomato Strain of Tobacco Mosaic Virus Detected by Monoclonal Antibodies

Eleven murine monoclonal antibodies (MABs) specific for L11A, a tomato strain of tobacco mosaic virus (TMV-L11A), were produced in culture supernatants and in ascitic fluids. Reactivities of all the 11MABs with tobamovirus strains were investigated by two indirect ELISA procedures, competitive ELISA, precipitin ring interface test and immuno-double-diffusion test. Five MABs among them reacted only with tomato strains (L11A, L and CH2) and the others reacted not only with tomato strains but also with other strains [TMV-OM, P, W and cucumber strain of cucumber green mottle mosaic virus (CGMMV-Cu)]. Based on these results, at least eight epitopes were distinguished on the surface of the particles of L11A. The epitopes, except for one detected by one of the MABs (6F11), were all neotopes, and 10 out of 11MABs were anti-neotope antibodies including a heterospecific antibody (8C8). A superior epitope which reacted mainly in the antiserum was identified within these epitopes. The high specificity of the MABs showed that they were useful for the differentiation of tobamovirus strains. The titers of three MABs in ascitic fluids were 1:2, 560 and they could be substituted for rabbit antisera. This study showed that the antigenic specificity of the strains was mainly dependent on the neotopes.