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Survey of Entomopathogenic Nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Japan

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Survey of Entomopathogenic Nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Japan View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Horizon / Pleins textes Fumiam. appl. NemalOl., 1998, 21 (2), 185-198 Survey of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Japan Mutsuhiro YOSHIDA*, Alexander P. REID**, Bernard R. BRISCOE** and William M. HOMINICK** * NaLionallnsLiLuLe ofAgro-Environmental Sciences, Kannondai 3-1-1, Tsukuba, lbaraki 305,japan and .'f.* CABllnt.emaLionallnstitute ofParasiLology, 395A HaLfield Road, St. Albans, Heris. AL4 oXV, VK. Accepted for publication 8July 1997. Surnmary -A survey was conducted for entomopathogenic nematodes in the five climatic regions of Japan: subtropical islands, warm temperate coastal region along the Pacific, central mountainous region, temperate region, and cool temperate region. Over 1400 soil samples from 266 sites were tested for the presence of nematodes using the Galleria baiting technique. One hundred and twenry steinernematids and 29 heterorhabditids were detected in 62 sites and 24 sites, respectively. Eight steinernematid species were recognized: seven undescribed species and Sleinernema kushidai. Each species had a characteristic distribution: three wide-spread common, one coastal, one subtropical, and three mountainous species. In contrast, the hetero­ rhabditids were identified as Heœrorhabditis indica and H. megidis. The former was widely found in the coastal region from the subtropical to the warm temperate zone while the latter was found mainly in the eastern part of the warm temperate coastal region. © Elsevier - ORSTOM Résumé - Enquête sur les nématodes entomopathogènes (Rhabditida: Steinernematidae et Heterorhabditidae) du Japon - Une enquête a été menée concernant les nématodes entomopathogénes dans les cinq régions climatiques du Japon: les îles subtropicales, la région tempérée chaude des côtes du Pacifique, la région montagneuse australe, la région tempérée et la région tempérée froide. Plus de 1400 échantillons de sol provenant de 266 sites ont été testés pour la présence de nématodes à l'aide de Gallena-piéges. Cent vingt-et-un Steinernematides et 29 Heterorhabditides Ont été détectés dans, respectivement, 62 et 24 sites. Huit espéces de Steinernematides ont été reconnues: sept espéces non décrites et Sœinernema kushidai. Chacune des espéces montre une répartition particuliére : trois sont très répandues, une est côtière, une subtropicale et les trois autres montagnardes. En contraste, les Heterorhabditidae ont ètè identifiès comme Helerorhabidùis indica et H. megidis. La première espèce est largement rèpandue dans la région côtière depuis la zone subtropicale jusqu'à la zone tempérée chaude, tandis que la seconde est surtout rencontrée dans la partie est de la région côtière tempérée. © Elsevier - ORSTOM Keywords: distribution, entomopathogenic nematodes, Helerorhabdilis, Japan, species diversiry, Sleinemema, survey. Nematodes of the families Steinernematidae and (Mamiya & Ogura, 1989) and Kyushu district Heterorhabditidae have characteristics that make (Kimura & Ishibashi, 1991) revealed the presence of them extremely useful as biological control agents. other steinernematids. Numerous surveys have been conducted world-wide, The present study describes a survey conducted resulting in the isolation of many species and strains; between 1991 and 1994 in five climatic regions, from Australia (Akhurst & Bedding, 1986), North America subtropical to cool temperate regions, mostly in major (Akhurst & Brooks, 1984; Poinar et al., 1987; Mraëek forests and ranging in elevation from near sea level to & Webster, 1993; Rueda et a1., 1993; Nguyen & about 1600 m. This work was aimed at collecting Smart, 1994), Puerto Rico (Roman & Beavers, 1982), indigenous entomopathogenic nematodes adapted to South America (Doucet, 1986; Doucet & Doucet, the various climates for possible use in biological con­ 1990, Nguyen & Smart, 1990; Stock, 1995), Hawaii trol regimens. (Hara et al., 1991), Europe (Mraëek, 1980; Deseo et al., 1984; Husberg et al., 1988; Hominick & Bris­ coe, 1990; Griffin et al., 1991), Africa (Shamseldean Materials and methods & Abd-Elgawad, 1994), India (Poinar et al., 1992), The present survey was conducted from the south Sri .Lanka (Amarasinghe et al., 1994), Asia (Zhang part of Japan in a north-east direction along the et al., 1992; Choo et al., 1995). In Japan, Steinernema Pacifie coast to collect nematodes adapted to dry, hot, kushidai Mamiya, 1988 was first isolated in Shizuoka and warm environments, then from the south to the from a tree nursery where an outbreak and an infesta­ north of the central part (Izu Peninsula - Nagano tion of white grubs had occurred (Kushida et al., Pref.) to collect nematodes adapted to cool or cold 1987). Later surveys conducted in eastern Japan mountainous environments. Moreover, sorne other Fundam. appl. NemalOl. 1164-5571/98/02/© Elsevier - ORSTOM 185 M. Yoshida et al. regions sampled between 1991 and 1994 were added also by PCR-RFLP analysis of the internai tran­ to the present srudy. scribed spacer region from the ribosomal DNA repeat A total of 1416 soil samples was collected from 276 unit. Specimens for morphological study were pre­ sites, which represented the five climatic regions pared according to Minagawa and Mizukubo (1994). (Fig. 1) in the present study as follows: z) subtropical The extraction of genomic DNA from a single nema­ islands, four islands in Okinawa Pref. (Nansei-shoto tode was conducted according to Joyce el al. (1994c) Isis.), 309 samples from 81 sires; il) warm temperate and Reid el al. (1997). The ITS region was amplified coastal region along the Pacifie coast, the warm region by PCR with the 18S and 26S primer set (Vrain el al., affected by the warm current "Kuroshio", from Cape 1992). Subsequent RFLP analysis was performed as Sata-misaki (south end of Kyushu, Kagoshima Pref.) per Reid el al. (1997). Each isolate was assigned to an to Cape Inubo-zaki (Chiba Pref.), 732 samples from RFLP type based on the restriction fragment patterns 116 sites; iiz) central mountainous region, the cool yielded by Alu 1 and Hinf 1 (Sleinernema spp.) or Alu 1 highland at more than 500m elevation from lzu and Rsa 1 (Helerorhabditis spp.). Seventeen restriction Peninsula (Shizuoka Pref.) to Niigata Pref., 165 sam­ enzymes (Alu l, BstO l, Dde l, EcoR l, Hae III, Hha l, pies from 31 sites; iv) temperare region, Oki Isis. (Shi­ Hind III, Hinfl, Hpa II, Kpn l, Pst l, Pvu Il, Rsa l, Sal mane Pref.) and sorne regions in the eastern and l, Sau3A 1, Sau96 l, Xba 1) were used for the detailed northern part of Japan (Chiba, Ibaraki and Miyagi RFLP analysis of each representative of the RFLP Prefs), 129 samples from 26 sites; v) cool temperate types (Reid et a!., 1997). The detailed RFLP profiles region (Hokkaido Pref.), 44 samples from twelve were compared with the RFLP database of the nema­ sites. The climatic division was derermined based on todes at CABI International Institute of Parasitology, vegetation and elevation, referring to Maekawa (1977) UK. and Nakamura el al. (1996). Samples were collected from November to May before the luxuriant growth Results of weeds and bushes, with the exception of the Positive soil samples with entomopathogenic ne­ Okinawa Is. where samples were taken in June after matodes were obtained from ail sampling regions the rainy season. The number of samples taken from (Fig. 1), and represented 10.0% of the total number each sire was usually five (ranging from three to thirty ofsoil samples (142/1416) or 28.2% of the geographi­ samples/site) with approximately 600-700 g soil col­ cal sires (75/266). Steinernematids were recovered in lected for each sample to a depth of 5-20 cm. 8.5% of the soil samples (12011416), from 23.3% of 3 Approximately 500 cm of soil of each sample was the sites (62/266), and heterorhabditids in 2.0% of the placed in a polythene cup (110 mm diam., 50 mm soil samples (2911416), from 9.0% of the sites (24/ depth) covered with a lid and three ta five final instar 266). The incidence and species diversity of the larvae of Galleria mellonella were added (Bedding & nematodes in the four main sampling regions were Akhurst 1975). Each cup was incubated at room tem­ obviously different. A much higher prevalence was perature (20-25°C) for 2 weeks. Each container was noted in the central mountainous region (64.5% of checked for larval mortality every 2-3 days from 5 or 6 the sites [20/31]) than in the subtropical islands (8.6% days after setting the Galleria trap up to 14 days. Dead [7/81]), the temperate region (23.1 % [6/26]) and the larvae were placed individually on a polyurethane warm remperate coastal region (33.6% [391116]). sponge (20 mm length, 15 mm width and less than 10 Eighty-two steinernematid isolates from 47 positive mm thickness) in each cell of a twelve-well plate with sires were classified into eight species: seven undes­ water. The plates were kept at room temperature (20­ cribed species and S. kushidai, by means of morpho­ 25°C) and periodically examined for presence of nem­ logy and/or RFLP analysis. The undescribed Slei­ atodes. Cadaver from which infective juveniles began nernema species are herein referred as MY l, MY2, to emerge were individually transferred to a plastic MY3, MY4, MY5, MY6, and MY7, based on RFLP cup (60 mm diam., 35 mm depth) with the poly­ profiles (Fig. 2). The incidence and species diversity urethane sponge and distilled water. In the present of steinernematids increased as follows: subtropical study, an "isolate" represented the nematodes islands (1.2% of the sites [1/81], Steinernema sp. obtained from a single dead Galleria larva and not MY4) < temperate region (23.1 % [6/26], Steiner­ from one soil sample. Infective juveniles isolated in nema spp. MYl, MY3, and unidentified isolates) < this manner were rinsed in distilled water several warm temperate coastal region (27.6% [321116], times in the cup and stored at 5-7°C (Sleinernema spp. Sleinernema spp. MYl, MY2, MY3, S. kushidai, and except S.
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