As the international trade increases, invasion of new pests are increasing (Kiritani, 2001). Invasive species can adopt, spread quickly because they don’t have any natural enemies in the new environment (Howard, 2004). In order to prevent the invasion of these pests, it needs the thorough inspection, collection of the information on pests and rapid identification of detected exotic pests (Rugman-Jones, 2006).
Root-lesion nematode, the species belonging to the genus Pratylenchus Filipjev, 1936, is frequently detected in plant quarantine. There are more than 70 species around the world (Siddiqi, 2000; Palomares-Rius et al., 2010), 14 species were recorded in the Republic of Korea (Park, 1996; Park et al., 2002; Choi et al., 2006; Kim et al., 2006; Park et al., 2009). Only four species, P. brachyurs, P. crenatus, P. fallax, and P. zeae are listed as the regulated pest in announcement of Animal and Plant Quarantine Agency (No. 2013-137). These nematodes are migratory parasite that invade cortex of root, tubers, and other below-ground organs (Bridge and Starr, 2007). Their feeding activities results in destruction of host cell with formation of necrotic lesion (Baxter and Blake, 1968; Corbett, 1972; Acosta and Malek, 1981). It causes reduction of yields (Lownsberry, 1956; McKenry, 1989) or lowering of the market value of crops (Bernard and Laughlin, 1976). Their host range is wide, various crops are affected by root-lesion nematode worldwide (Vanstone et al., 1995; Kimpinski et al., 1989; Vanstone et al. 1998). In Korea, they cause the wilt disease in Chrysanthemum with soil pathogens (Lee et al., 2008), it affects export of Chrysanthemum. Also they were detected on several crops such as Perilla (Kim et al, 2000) and Tea (Park et al., 2002).
Identification of Pratylenchus species is usually based on shape and measurement of female (Corbett, 1969; Frederick and Tarjan, 1989; Handoo and Golden, 1989). Small number of diagnostic characters and the intra specific variability makes the species identification difficulty (Taylor and Jenkins, 1957; Roman and Hirschmann, 1969). And we have little information of exotic species. Therefore the goal of this study is to provide identification information about exotic nematodes. Finally it will contribute to improving the identification rate of intercepted exotic root-lesion nematodes.
Materials and Methods
Total 18 exotic examples are used to identification (Table 1). The nematode was collected from the roots of imported crops. The plant cut into small bits and soak in water over 12 hours. After then the nematodes were extracted by Cobb’s sieving and gravity method (Cobb, 1918; Sauthey, 1986) and modified Bermann funnel method. Extracted nematodes were into the water and killed by heating. They were dehydrated by rapid lactophenol method (Franklin and Goodeyi, 1949). Fixed nematodes were brought into glycerin and sealed by paraffin ring method (De Maesener and De Herde, 1963). Identification of species conducted under the Zeiss M1 light microscope. Photographs and measurements from the specimens were conducted using Axiovision 4.7 microscopy software. Dichotomous key suggested by Pablo Castilo and Nicola Vovlas were used (Castilo and Vovlas, 2007). Measurements are described De Man (1880) formula (Table 2).
Results
Pratylenchus crenatusLoof, 1960 (Fig. 1)
Measurements
Female (n=2): L=511-574 μm; a=22.8-24.8; b=7.3; b’=5.5; c=19.5-20.8; c’=1.9; V=81.5-81.7%; stylet=16.8 μm
Diagnosis
Labial region with three annuli. Body annulation prominent. Excretory pore 81.2 μm from anterior end. Spermatheca rounded, empty. Lateral field with four lines. Post-vulval uterine sac short (16.8-19.6 μm). Distance between vulva and anus 60.9- 88.9 μm. Tail broadly rounded, often spatulate, 24.5-29.4 μm long. Tail tip coarsely and distinctly annulated with 24-26 annuli.
Host
Dicentra spectabilis (bulb)
Remarks
The length of post-vulval uterine sac was differently described by several nematologists. Loof(1960) indicated that their post-vulval uterine sac is short and Sher & Allen(1953) reported that it is two body diameter long. This study is based on Loof’s concept.
Pratylenchus fallax Seinhorst, 1968 (Fig. 2)
Measurements
Female (n=4): L=469-560 μm; a=21.6-22.7; b=5.2-5.8; c=17.0-20.0; c’=2.1-2.4; V=79-81%; stylet=16.8-17.5 μm
Diagnosis
Labial region with three annuli. Stylet knobs anteriorly flattened or pointing forward. Lateral field with four lines. Excretory pore 79.8 μm from head end. Post-vulval uterine sac 11.2-20.3 μm long. Spermatheca round, filled with sperm. Distance between vulva and anus 77-80.5 μm. Tail conical, 26-32.2 μm long, annulated with 19-24 annuli. Tail tip rounded, slightly crenate.
Host
Dicentra spectabilis (bulb)
Remarks
The species is close to P. penetrans, but it can be differentiated by crenate tail terminus
Pratylenchus kumamotoensis Mizukubo, Sugimura & Uesugi, 2007 (Fig. 3)
Measurements
Female (n=3): L=581.8-612.3 μm; a=24.08-30.86; b’=4.7; c=20.0; c’=2.2; V=77.9-78.7; stylet=13.6-15.4 μm
Male (n=2): L =469.5-490.8 μm; a=24.45-27.65; spicules= 18.78 μm
Diagnosis
Female: Lip region with three annuli. Stylet knobs rounded. Lateral field with four lines. Pharyngeal gland lobes overlapping intestine ventrally or dorsally. Post-vulval uterine sac 42-49.6 μm long. Spermatheca oval, filled with sperm. Tail conoid, 30.66 μm long.
Host
Citrus unshiu (seedling)
Remarks
The species showed variation in overlapping with pharyngeal gland lobe and intestine. The nematodes of belonging to the genus Pratylenchus overlap intestine ventrally, but P. kumamotoensis sometimes overlaps intestine dorsally. Our specimens show that these characters, we identified them as P. kumamtoensis. It is recorded on Chrysanthemum morifolium only, but we detected from Citrus unshiu.
Pratylenchus panamaensis Siddiqi, Dabur & Bajaj, 1991 (Fig. 4)
Measurements
Female (n=4): L =470-580 μm; a=16.8-28; b=5.3-7.3; b’= 4.3-5.1; c=16.9-23.1; V=76-85%; stylet=16.5-18.2 μm
Diagnosis
Body straight or slightly curved ventrally when heat-relaxed. Labial region flat, low and offset from body with two annuli. Stylet knobs anteriorly flattened. Lateral fields with four equidistance lines. Opening of dorsal pharyngeal gland 2-3 μm from stylet base. Median bulb well developed. Pharyngeal gland lobe overlapping intestine for 1.5-2 body diameter long. Excretory pore 60-88 μm from anterior extremity. Spermatheca longitudinally oval filled with sperm. Post-vulval uterine sac 18-25 μm long. Tail sylindrical, with distinctly crenate terminus.
Host
Epimidium koreanum (bulb)
Remarks
The examined specimens were collected Epimidium koreanum in U.S.A.. According to Siddiqi, Dabur and Bajaj (1991), however, the species is distributed in Panama and parasites on Coffee sp..
Pratylenchus penetrans (Cobb, 1917) Filipjev & Schuurmans Stekhoven, 1941 (Fig. 5)
Measurements
Female (n=2): L=609-647.5 μm; a=23.5-25.6; b=5.8-7.5; b’=4.6-5.4; c=18.9-23.1; c’=2.1; V=78.3-80%; stylet=16.8- 17.5 μm
Male (n=1): body length=448 μm; a=25.6; c=17.7; stylet= 15.7 μm; spicules=15.4 μm; gubernaculums=4.2 μm
Diagnosis
Female: Labial region slightly offset from body, low, with three annuli. Excretory pore 92.4-93.8 μm from anterior end. Pharynx overlapping ventrally or laterally. Spermatheca rounded, filled with sperm. Post-vulval uterine sac short and undifferentiated. Tail generally rounded, 28-32.2 μm long. Tail tip smooth.
Host
Dicentra spectabilis (bulb)
Remarks
This species characterized by labial region with three annuli, short post-vulval uterine sac and round tail.
Discussion
The species identification of nematodes is based on morphological characters, but, due to wide range of variation in diagnostic characters (Andres et al., 2000; Yu et al., 2012), variable features must be integrated for identification. Thus, it caused serious time-consuming and requires expert knowledge (Kumari et al., 2004; Devran and Sogut, 2009). In quarantine inspectsion, especially, detected nematodes were mostly nonnative species, and it was difficult for full identification.
Pratylenchus species are parasites on roots of economic plants, and often detected during quarantine inspection (Handoo et al., 2001; Ranjan, 2005; Zhang et al., 2011). In this study, five species, P. crenatus, P. fallax, P. kumamotoensis, P. panamaensis, and P. penetrans were found from imported plants into Korea. In distribution, only one species, P. penetrans is inhabited in Korea (Castilo and Vovlas, 2007). And in species identification, it was known that P. kumamotoensis showed variation in diagnostic characters of genus level (Mizukubo et al., 2007). For resolving of these problems, morphological diagnoses of above five species were clearly described. Therefore, our results might lead to increase full identification rate and decrease time-consuming.