Reactions of Several Cultivars of Dactylis Glomerata to an Infection by Cocksfoot Mottle Virus
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日 植 病 報 49: 720-723 (1983) 短 報 Ann. Phytopath. Soc. Japan 49: 720-723 (1983) Phytopathological Notc Reactions of Several Cultivars of Dactylis glomerata to an Infection by Cocksfoot Mottle Virus Shigemitsu TORIYAMA*, Yoshiro MIKOSHIBA**, Syutaro KAWABATA*** and Yoji DOI* 鳥 山 重 光 *・御 子 柴 義 郎 **・川 端 習 太 郎 ***・土 居 養 二 *:コ ッ ク ス フ ッ トモ ッ トル ウ イ ル ス に 対 す る オ ー チ ャ ー ド グ ラ ス(Dactylis glomerata)品 種 の 感 受 性 Key Words: cocksfoot mottle virus, Dactylis glomerata, cultivars, susceptibility. Mottling diseases of cocksfoot (Dactylis glomerata), which are caused by cocksfoot mottle virus (CfMV), are prevalent in cocksfoot pastures of Japan1). Most diseased cocksfoot plants in Japan showed rather mild mosaic symptoms but some showed severe mottling symptoms as described by Serjeant2). The disease is spread rapidly in the field by cutting implements and grazing animals and its incidence appears to increase with age of the crop2,3,4). The virus disease may be a major factor of the pasture's decline3). Resistant and susceptible cultivars of cocksfoot have been reported in England2,3). This paper is a report of the incidence of the mottling disease in five cultivars of Dactylis glomerata in an experimental field, and the susceptibility of nine cultivars to CfMV by artificial inoculation. Five cultivars (Aonami, Kitamidori, Frode, Masshardy and Aberystwyth S26) of cocksfoot, which had been grown over 3-4 years in five different localities of Japan (Konsen, Hokkaido; Iwadeyama, Miyagi Prefecture; Fujinomiya, Shizuoka Pref.; Aso, Kumamoto Pref.; Nishinasuno, Tochigi Pref.) were transplanted on September 1975 to the experimental field of the National Grassland Research Institute, Nishinasuno- cho, Tochigi Pref. for selection of excellent agronomical characters. Each of the 1100 tussocks was divided into three and planted at random to three blocks. Our inspection of cocksfoot mottling diseases in these 3300 tussocks was carried out on October 1981, 8-9 years after sowing. An index of the severity of the disease was set up by assigning plants to four grades depending on visible symptoms: mottling or obvious mosaic ($, in figure 1), mild mosaic (+), mild mosaic but only partly infected (;) and no symptom (no sign). For serological tests, leaf pieces of diseased plants were * Faculty of Agriculture , The University of Tokyo, Bunkyo-ku, Tokyo 113, Japan 東 京 大 学 農 学 部 ** Present Address: Tohoku National Agricultural Experiment Station , Morioka, Iwate 020-01, Japan 東 北 農 業 試 験 場 *** National Grassland Research Institute , Nishinasuno-cho, Tochigi 329-27, Japan 草 地 試 験 場 1) Toriyama, S. (1982). Ann. Phytopath. Soc. Japan 48: 514-520. 2) 5erjeant, E.P. (1963). Plant Pathol. 13: 23-24. 3) Serjeant, E.P. (1963). Rep. Rothamsted Exp. Sta. for 1962. p. 112. 4) Upstone, M.E. (1969). Ann. appl. Biol. 64: 49-55. Ann. Phytopath. Soc. Japan 49 (5). December, 1983 721 Fig. 1. The incidence of mottling diseases in five cultivars of D. glomerata in the field, which is surrounded by two rows of cv. Aonami. A diagram of the distribution of each cultivar in the three blocks (I-III) is shown under the surveyed field map. $, a plant showed obvious mosaic or mottling with necrosis; +, a plant with mild mosaic; ;, a plant, mild mosaic but partly infected; no sign, a plant with no symptom. ground in ca. 1ml of 0.03M borate buffer, pH 6.5, and sap was mixed with ca. 0.5ml of chloroform and centrifuged in Eppendorf plastic tubes at low speed. The virus in the aqueous supernatant was examined in 1% agar gel diffusion test as described previously4,5). Antisera to CfMV and ryegrass mottle virus (RMotV)6) were provided by Toriyama1) and Toriyama et al.6) Fig. 1 and 2 show that the mottling diseases are most prevalent (70%) in cv. Aonami and least (20%) in cv. Aberystwyth S26 in three blocks. Disease incidence in cvs. Masshardy, Kitamidori and Frode was 45%, 40% and 30%, respectively 5) Catherall, P.L. and Wilkins, P.W. (1970).Ann. Phytopathol. 9: 245-248. 6) Toriyama, S., Mikoshiba, Y. and Doi, Y. (1983).Ann. Phytopath. Soc. Japan 49: 610-618. 722 日本 植物 病理 学 会 報 第49巻 第5号 昭 和58年12月 Fig. 2. Disease incidence (%) in five cultivars in the three blocks, displayed by different symptoms. • , lacked tussock; •¡, obvious mosaic or mottling; • , mild mosaic; •¡, mild mosaic but partly infected; • , no symptom. (Fig. 1 and 2). As shown in Fig. 2, symptoms were most obvious or severe in cv. Aonami and mildest in cv. Aberystwyth S26. The differences in disease incidence are statistically significant for the five cultivars surveyed (P=0.01) but not for the three blocks and localities. The lack of significance between incidence in plants (cultivars) from different localities also suggests that the mottling diseases are spread uniformly within the experimental field. Mottling symptoms observed in some plants of the field were suspicious of those of cocksfoot plants infected with RMotV6). However, all sap preparations of 52 plants with obvious mosaic or mottling symptoms, which were collected at random from all cultivars in the field, reacted to antiserum to CfMV but not to antiserum to RMotV. Table 1 shows the results of mechanical inoculation of CfMV to nine cultivars of cocksfoot. The percentage of infected plants of cv. Aonami was 30-50%, which is lower than the percentage of infected plants in the field (Fig. 2). The percentage of infected plants was c. 20% in cv. Aberystwyth S26, 20-30% in cvs. Akimidori, Kitamidori, Makibamidori, Okamidori and Aberystwyth S143, and 30-40% in cvs. Frode and Masshardy. Cv. Okamidori seemed to be resistant in a preliminary experiment1). In four inoculations (Table 1), however, the susceptibility of cv. Okamidori was lower than that of cv. Aonami, but much the same as that for cvs. Makibamidori and Kitamidori. The proportion of infected plants of five cultivars in Fig. 2, examined by mechanical inoculation tests, differed between experiments and in some experiments was lower than that of the disease incidence as shown in Fig. 2. In the field, plants are inoculated frequently, due to repeated cutting and mechanical contamination with exudates from the cut surface of adjacent CfMV-infected leaves. In England, cvs. Aberystwyth S26, S37 and S143 are susceptible to CfMV and cvs. Conrad and Ann. Phytopath. Soc. Japan 49 (5). December, 1983 723 Table 1. Inoculations of cocksfoot mottle virus to nine cultivars of Dactylis glomerataa) a) Infected leaves of cocksfoot were ground in 0.03M borate buffer pH 6.5 and mechanically inoculated to cocksfoot seedlings. Plants were inspected for symptoms for at least 3 months after inoculation. b) Date of inoculation. c) No. of infected plants/no. of inoculated plants: Percentage of infected plants was shown in parenthesis. d) A figure in parenthesis of n (): no. of plants which showed necrotic mottling symptoms. Cambria are highly resistant2,5). In this study cv. Aberystwyth S26 was less susceptible than the other eight cultivars tested but cv. Aonami particulary was highly susceptible. From the results reported here it seems unlikely that it will be possible to select resistant clones from the uninfected plants of any of the cultivars in the experimental field. Thus, a highly resistant variety, such as cvs. Conrad and Cambria, may be needed as a genetic source for breeding a variety resistant to CfMV in Japan. In mechanically infected plants, the symptoms also varied as observed in field. As shown in Table 1, typical necrotic mottling was prominent in cvs. Aonami, Makibamidori, Okamidori, Masshardy and Aberystwyth S143. In Exp. 1, however, necrotic moottling symptoms appeared only cv. Okamidori. Appearance of necrotic symptoms seem to vary with time of infection (seasonal temperature). In some plants with necrotic mottling symptoms, conspicuous, spindle-shaped, greyish necrotic lesions often appeared on otherwise green healthy looking leaves. Such lesions coalesced and quickly caused wilting and death of the leaves. This symptom is very similar to that of the diseased cocksf oot plants, found in the field at Morioka, Japan, as described previously1). (Received May 9, 1983).