J Gen Plant Pathol (2004) 70:192–193 DOI 10.1007/s10327-004-0104-0
© The Phytopathological Society of Japan and Springer-Verlag Tokyo 2004
DISEASE NOTE Fumihiro Terami · Fumiyoshi Fukumoto Kaoru Hanada
Cucumber mosaic virus isolated from Amazon lily (Eucharis grandiflora)
Received: August 25, 2003 / Accepted: November 14, 2003
Abstract Cucumber mosaic virus (CMV) was isolated from a mosaic diseased plant of Eucharis grandiflora. The virus caused mosaic symptoms on leaves and slight distortion of flower petals in E. grandiflora by either mechanical or aphid inoculation. The virus was identified as a strain of CMV subgroup I from its biological and serological characteristics. Key words Eucharis grandiflora · Mosaic disease · Cucumber mosaic virus
Mosaic diseases of Eucharis grandiflora are caused by several viruses, including Hippeastrum mosaic virus, Nerine latent virus, and Amazon lily mosaic virus (ALiMV) (Jayasinghe and Dijkstra 1979; Terami et al. 1995). Cucumber mosaic virus (CMV) was detected by serological methods from a mosaic-diseased E. grandiflora plant together with the other three filamentous viruses mentioned (Terami et al. 1995). Although CMV has not been isolated previously in E. grandiflora, the virus was successfully isolated from the E. grandiflora plant by serial local-lesion transfer to Gomphrena globosa, Tetragonia tetragonoides (twice), and Chenopodium quinoa (twice). The virus was then propagated in T. tetragonoides. The isolated virus was extracted and purified from inoculated leaves of T. tetragonoides as described by Tochihara (1970). Virus-free plants of E. grandiflora obtained from mericlonal plants were mechanically inoculated with the purified virus. Chlorotic spots appeared in the first developing leaves 2 to 3 months after inoculation, and the subse-
F. Terami (*) · F. Fukumoto National Agricultural Research Center for Hokkaido Region, 1 Hitsujigaoka, Toyohira-ku, Sapporo 062-8555, Japan Tel. ⫹81-11-857-9141; Fax ⫹81-11-859-2178 e-mail:
[email protected] K. Hanada National Institute of Agrobiological Sciences, Ibaraki, Japan
quent leaves had mosaic or yellow mosaic symptoms (Fig. 1A). The petals of flowers of inoculated plants were only slightly distorted (Fig. 1B). These symptoms were much less severe than those of the plant from which the virus was originally isolated. The host range of the virus was examined by inoculating 37 plant species from 11 families with the crude sap of infected leaves of T. tetragonoides. The following 14 species were systemically infected and developed mosaic symptoms: Cucumis melo var. reticulates (Earl’s Favorite), C. melo var. makuwa (New melon), C. sativus (Sagami Hanjiro), Nicotiana benthamiana, N. glutinosa, N. clevelandii, N. rustica, N. tabacum (Xanthi-nc, Bright Yellow), Lycopersicon esculentum (Fukuju No. 2), Petunia ⫻ hybrida (Cascade), Spinacia oleracea, Vicia faba, Vigna unguiculata (Kurodane Sanjaku, Blackeye), Zea mays (Honey Bantam), and Zinnia elegans. Necrotic or chlorotic lesions were also produced on inoculated leaves of N. tabacum, S. oleracea, V. faba, V. unguiculata, and Z. mays. Cucurbita maxima (Tokyo) had chlorotic spots on systemically infected leaves. Gomphrena globosa had systemic necrosis followed by the appearance of necrotic spots in inoculated leaves. Chrysanthemum coronarium was infected systemically without symptoms. The virus caused necrotic or chlorotic local lesions without systemic infection in Abelmoschus esculentus, Beta vulgaris, C. amaranticolor, C. quinoa, Citrullus lanatus (Ibuki), C. Pepo (Kinshi), C. moschata (Shirokikuza), C. melo var. conomon (Dai Tokyo), Luffa aegyptiaca, Sesamum indicum, and T. tetragonoides. Latent infection in inoculated leaves was observed in Glycine max (Tsurunoko) and Solanum melongena (Senryou) but not systemic infection. The following seven species were not infected with the virus: Brassica campestris (Kanemachikokabu), Datura stramonium, Phaseolus angularis (Dainagon), P. vulgaris (Honkintoki, Top Crop, Yamashirokurosando), Pisum sativum (Fukkoku), Trifolium repens, and Raphanus sativus (Kameido). The virus was transmitted nonpersistently by Myzus persicae to E. grandiflora and N. glutinosa from an infected plant of the corresponding species. Symptom appearance in
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A
B
Fig. 1. Symptoms of Eucharis grandiflora inoculated with the isolated cucumber mosaic virus (CMV). A Mosaic symptoms of a systemically infected leaf. B Slightly distorted flower of the CMV-inoculated plant (top) and a flower of a virus-free plant (middle). A markedly malformed flower (bottom) appeared on a plant with mosaic from which CMV was isolated. The diseased plant was also infected with Amazon lily mosaic virus, Hippeastrum mosaic virus, and Nerine latent virus
Fig. 2. Agar gel diffusion test of the CMV from isolate Eucharis grandiflora. Y, purified preparation of CMV-Y; E, purified preparation of the isolated strain of CMV; P, Purified preparation of CMV-P; Yas, antiserum to CMV-Y; Pas, antiserum to CMV-P
data indicate that the virus is a strain of CMV subgroup I and is related to the legume strain (Tomaru and Udagawa 1967).
References E. grandiflora by aphid inoculation was also slow, as in the case of mechanical inoculation; and the inoculated plants developed mosaic symptoms 2 months after inoculation. In N. glutinosa the first symptoms appeared within a week. Spherical virus particles (about 30 nm diameter) with typical characteristics of cucumoviruses were observed in the leaf sap of E. grandiflora with mosaic symptoms and purified preparations of the virus. The coat protein of the virus was of a size similar to that of CMV Y strain (subgroup I) (Tomaru and Hidaka 1960; Wahyuni et al. 1992) seen by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The virus reacted strongly to both Y strain and P strain (subgroup II) (Tochihara and Tamura 1976; Wahyuni et al. 1992) antisera in an agar gel diffusion test. Precipitation lines of the virus and Y strain were fused in the reaction to Y strain antiserum, and a spur formed between the virus and P strain in the case of P strain antiserum (Fig. 2). These
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