International Journal of Life Sciences 10 (1) : 2016; 10 - 16 Founded 2007 An Independent, Open Access, Peer Reviewed, Non-Profit Journal International Journal of InternationalInternational JournalJournal ofof LifeLife SciencesSciences ISSN 2091-0525 website: http://nepjol.info/index.php/IJLS/index ifeSciences LCopyright © International Journal of Life Sciences

Research Article Five Species of Plant Parasitic (Order: Tylenchidae) Affecting Vegetable Crops of Nepal Arvind K. Keshari1, Ranjana Gupta2

1Department of Zoology, Patan M. Campus, Patan, Lalitpur, Nepal. 2Head, CentralDepartment of Zoology, Tribhuvan University, Kirtipur, Nepal.

Article Information ABSTRACT Key words: During a survey for plant parasitic nematodes affecting various vegetable crops grown in three hilly Plant parasitic districts surrounding Kathmandu Valley, Nepal, five species of order Tylenchidae are reported for the nematodes; first time from Nepal.The species are , Tylenchorhynchus mashhoodi, tylenchidae; Helicotylenchus incisus, Microposthonia paraxestis and Hemicriconemoides cocophilus.All the species ; are illustrated with line diagrams and described with their morphometric data along with localities and species; new records host plants.

INTRODUCTION isolated by modified by Cobb's (1918) sieving and decantation and modified Baermann's (1917) funnel techniques were killed and fixed in hot FA (4:1) for 24 A nematological literature available so far reveals that hours (Seinhorst 1962). After fixation, the nematodes very little taxonomic work is done in Nepal. The first were transferred to glycerine–alcohol (5 parts glycerine survey work on generic level was done by Amatya and and 30% alcohol) and kept in a desiccator for slow Shrestha (1968) reporting 34 genera of plant parasitic dehydration. Permanent mounts were made in nematodes from Nepal. Besides some fragmentary anhydrous glycerine by using wax ring technique. works, no significant progress is done. The present Measurements, morphological observations were made taxonomic work is an attempt to identify the nematodes using an Olympus microscope while drawing with upto the species level. camera lucida. The specimens on slides are deposited in Central Department of Zoology, Tribhuvan University, The present paper is based on the collection of soil Kirtipur, Kathmandu, Nepal. samples from ten vegetable crops of three hilly districts- Bhaktapur, Kavre and Dhadhing surrounding the Kathmandu Valley, Nepal. The analysis of samples yielded RESULTS AND DISCUSSION varieties of plant parasitic nematodes. Among them, five species of order Tylenchidae are reported for the first Descriptions and illustrations of Hoplolaimus indicus, time from Nepal. Brief descriptions, illustrations and Tylenchorhynchus mashhoodi, Helicotylenchus incisus, information on host, locality of these species are Microposthonia paraxestis and Hemicriconemoides provided here. cocophilus are provided herewith.

Hoplolaimus indicus Sher 1963 MATERIALS AND METHODS (Fig. 1; A–D)

Anematological survey was carried out during 2012- Measurements: 2013 from three major vegetable producing districts of Nepal–Bhaktapur, Kavre and Dhadhing at altitudes of Females: L = 1.04–1.26 mm; a = 31.4–36.9; b = 8.7–10.6; 1319 m, 1439 m and 1518 m, respectively. Nematodes b' = 7.0–8.5; c = 63.3–76.7; c' = 0.55–0.76; V = 52.8–57.3;

*Corresponding author Email address: [email protected]

Copyright reserved © International Journal of Life Sciences DOI:dx.doi.org/10.3126/ijls.v10i1.14515 International Journal of Life Sciences 10 (1) : 2016; 10 - 16 stylet = 33.2–37.3µm; conus = 16.7–18.2µm; oesophagus Males: L = 0.63–0.70 mm; a = 31.2–36.0; b = 5.0–5.5; c = = 145–158µm; excretory pore = 94–114µm; tail = 13.2–14.7; c' = 3.0–3.7; stylet = 18.0–19.4 µm; conus = 8.6 15.2–19.2µm. µm; oesophagus = 116–129 µm; excretory pore = 100.0–107.5 µm; spicules = 22–27 µm; gubernaculum = Males: L = 0.92–1.12 mm; a = 32.8–38.6; b = 8.7–10.7; b' = 16.5–18.3 µm; bursa = 64–80 µm; tail = 41.5–52.5 µm. 6.7–7.8; c = 33.5–43.5; c' = 1.3–1.5; stylet = 30.2–34.5µm; conus = 15.2–18.2µm; oesophagus = 126–168µm; Descriptions: excretory pore = 89.0–96.0µm; spicules = 33.0–40.4µm; gubernaculum = 18.0–21.2µm; bursa = 50–63 µm; tail = Body curved in loose spiral shape upon fixation. Cuticle 24.2–28.7µm. marked with distinct transverse striae, 1–2 µm apart. Lip region continuous with body contour, bearing 3–4 Description: annules. Cephalic framework lightly sclerotized. Stylet 18–19 µm long with rounded basal knobs which may be Females: Body 1.04–1.26 mm, slightly ventrally arcuate slightly flattened anteriorly. Dorsal oesophageal gland upon fixation. Cuticle transversely striated. Cephalic orifice 10− 13µm behind spear base. Excretory pore framework strongly sclerotized. Lip region slightly posterior to level of nerve ring. Hemizonid just hemispherical, set off by deep constriction, 7.5–9.0 µm anterior to excretory pore. Gonads amphidelphic, high. Labial disc slightly elevated, elongate–ovoid. Stylet outstretched, oocytes arranged in a single row. Vulva with strong, 33–37 µm long, basal knobs –shaped. Dorsal a transverse slit, vagina about ½ of body width. oesophageal gland opening 4.5 µm behind spear base. Spermatheca rounded, functional. Tail 45–56 µm long, Oesophagus 145–158 µm long. Nerve ring 86–102 µm cylindrical to subcylindrical in shape, slightly narrowing from anterior end. Basal oesophagus glands overlapping behind anus, rounded terminus. Phasmids in anterior intestine dorso–laterally,basal glandular part with six half of tail. Male tail elongate–conoid with acute or gland nuclei. Excretory pore near oesophago–intestinal subacute terminus. junction, 95–115µm from anterior end. Hemizonid two to ten annules posterior to excretory pore.Female Host and locality: Soil around the roots of tomato reproductive system amphidelphic, outstretched. (Lycopersicum esculentum) and cucumber (Cucumis Oocytes arranged in single row. Epiptygma double. Vulva sativus) from Bhaktapur, Nepal. a transverse slit. Tail short measuring 15–19µm, hemispherical. Scutellum circular, 4.5 µm in diameter. Remarks: Tylenchorhynchus mashhoodi is a commonly distributed species. Baqri and Jairajpuri (1970) studied Males: Similar to females but slightly smaller in length. intraspecific variations of the species in terms of length Spicules strong, 33–40µm long, arcuate, gubernaculum and shape of tails, number of tail annules, length of spear simple. Bursa striated, enveloping tail. and length and shape of spicules. The measurements and description of the present species agree well within the Host and locality: Soil around the roots of cauliflower range given by Siddiqi and Basir1959. (Brassica oleracea) and brinjal (Solanum melagena) from Kavre, Nepal. Helicotylenchus incisus Darker and Khan 1978 (Fig. 2; A-D) Remarks: H. indicus is most widely distributed species (Khan and Chawla 1975). Since Sher (1963) described Measurements: this species from India, many workers (Hussain and Rashid 1969) and Khan and Chawla 1975 have studied Females: L = 0.65–0.93 mm; a = 33.5–44.3; b= 6.3–7.6; b' variations in species. Anderson (1983) reported this = 5.6–6.4; c = 39.8–56.0; c' = 0.91–1.11; v = 59.3–65.0; species from Canada. The measurements and description stylet = 22.5–27.0µm; conus = 12.0µm; oesophagus = of the present species agree well with previous 114–156µm; excretory pore = 87–129µm; tail = descriptions. 15–18µm.

Tylenchorhynchus mashhoodi Siddiqi and Basir 1959 Descriptions: (Fig. 1; E–H) Female: Body C-shaped to spirally coiled. Cuticle finely Measurements: transversely striated, each striae about 1.5 µm apart at midbody. Lateral fields with four incisures. Lip region Females: L = 0.62–0.74 mm; a = 33.2–35.2; b = 5.2–5.8; c continuous, smooth, truncated. Lip annules absent. = 12.2–15.0; c' = 3.2–4.0; V = 47.3–56.5; stylet = 18.3–19.4 Cephalic framework strongly sclerotized. Stylet well µm; conus = 7.5–9.0 µm; oesophagus = 115–128 µm; developed, conus conoid, 48–53% of stylet length with excretory pore = 96.0–106.5µm; tail = 45–56 µm. basal knobs spherical. Dorsal oesophageal gland opening 7.5–9.0 µm behind spear base. Oesophagus with tubular

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36–57 µm long procorpus. Metacorpus spherical, 12–15 Remarks: Dimensions and morphological characters of µm in diameter. Isthmus narrow, 12–21 µm long. Basal thepresent specimens conform to those descriptions oesophagus glandular part overlapping intestine dorso- given by Dhanachand and Renubala 1991. It is the genus ventrally, more on ventral side. Nerve ring at 74–105 µm reported for the first from Nepal. from anterior end. Excretory pore at the base of isthmus. Hemizonid anteriorly adjacent to excretory pore. Tail Hemicriconemoides cocophilus (Loos 1949) 15–18 µm, sub-clavate to hemispherical. Phasmids Chitwood and Birchfield 1957 12–16 annules anterior to anus. Gonads amphidelphic, (Fig. 3; A– E) outstretched. Oocytes arranged in single row. Uterus bipartite, muscular part followed by glandular part. Measurements: Spermatheca absent. Intestine not overlapping rectum. Anus pore like. Females : L = 0.33–0.38 mm; a = 14–17; b = 4.0–4.5; c = 15–23; V = 92.0–93.9); stylet = 45–51µm; conus = Male: Not found. 37.5–43.5 µm; tail = 20.0– 27.5 µm; R = 122–138; Rst = 16–18; Roes = 25–32; Rex = 21–31; Rv = 8–10; Rvan = Host and locality: Soil around roots of tomato 1–2; Ran = 7–9; VL/VB = 1.0-1.4; St%L = 13.5–16.4. (Lycopersicum esculentum) from Dhadhing, Kathmandu. Description: Remarks: The measurement of present specimens fits well within the range given by Darekar and Khan 1978. Female: Body stout, ventrally arcuate upon fixation narrowing towards extremities. Outer cuticular sheath Macroposthonia paraxestis (Dhanachand and tightly enclosing body and well separated on tail. Lip Renubala 1991) Ahmad 1996 region slightly set off with two distinct lip annules, first (Fig.2; E–G) annule 10.5 µm wide, second 13.5 µm wide. Labial disc more or less round, oral disc circular, raised, with a fine Measurements: rim like collar. Cephalic framework moderately sclerotized, stylet well developed, metenchium 82–85% Females: L = 0.33–0.40mm; a = 9–11; b = 3.6–4.0; c = of stylet length, telenchium 7.5–9.0 µm long. Basal knobs 35–43; V = 94–96; Stylet = 62–67μm; Tail = 9.0–11.2μm; R massive, long, anchor shaped stylet knobs. Dorsal = 54–64; Rst = 9–12; Roes = 16–19; Rex = 16–20; Rv = 4–6; oesophageal gland opening 3µm behind base of stylet. Rvan = 2–3; Ran = 2–3; St%L = 15.4–19.8; St% Pharynx = Oesophagus 78–87µm long. Excretory pore 81–90 µm 61–68. and nerve ring 67.5–75.0 µm from anterior end.

Description: Reproductive system mono-prodelphic, ovary out- stretched. Vulva an open transverse slit, vulval flaps two Females: Body short, slightly arcuate ventrally upon annules long. Vagina anteriorly directed, 10.5–13.5 µm fixation, anteriorlyblunt and posteriorly tapering to a long. Anus small, pore like, located adjacent to vulva. Tail blunt terminus. Bodyannules finely crenate, annular conoid and tail tip finely rounded. anastomoses absent. Lip region with two distinct annuli, first annules 6-15μm wide with a prominent labial Male: Not found. disc.Cephalic framework weakly developed, extending through first bodyannule. Stylet knobs strongly Host and locality: Soil around the roots of cauliflower developed, forward directed. Pharynx typically (Brassica oleracea) from Bhaktapur, Nepal. criconematoid, consisting of a broad procorpus, amalgamated with a large median bulb, a very short and Remarks: It is the widely distributed species. Loos 1949 broad isthmus and reduced basal pharyngeal bulb. first described this species from Sri Lanka. Later Siddiqi Dorsal pharyngeal gland opening2μm behind base of 1961, Edward and Mishra 1963 and Decraemer and stylet. Excretory pore 25–28 annules and nerve ring Geraert 1992 redescribed this species. The measure- 71–75μm from anterior end of the body. Reproductive ments of this specimens closely agree with previous system mono-prodelphic, ovary outstretched. Vulva descriptions and measurements given by Loos 1949. transverse, open. Oocytes arranged irregularly. Spermatheca indistinct. Tail short, conoid towards tip.

Male: Not found. ACKNOWLEDGEMENT

Host and locality: Soil around the roots of brinjal We sincerely thank all the members of Nematology (Solanum melagena) from Dhadhing, Nepal. section, Dept. of Zoology, Aligarh Muslim University, Aligarh, India for the laboratory facilities and identification.

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