Meloidogyne haplanaria
Eisenback et al., 2003
Photo Gallery- Specimens in culture
Description
Holotype (female in glycerin): Body length including neck = 921.0 um; body width 576.0 um; neck length = 351.9 um; stylet length = 14.5 um; stylet knob height = 2.2 um; stylet knob width = 4.9 um; dorsal esophageal gland orifice to stylet base = 5.5 um; excretory pore to head end = 69.9 um. Females as in general description; perineal region not visible.
Female (n = 30, in 2% glutaraldehyde in 0.1 M cacodylic acid buffer pH 7.2 and perineal patterns in glycerin); measurements are listed in Table 1.
    Body translucent white, variable in size, pear-shaped with short neck, posteriorly rounded, without tail protuberance.  In SEM, stoma slit-like, located in ovoid prestoma, surrounded by pit-like openings of six inner labial sensilla.  Labial disc fused with medial lips; dumbell-shaped in face view.  Medial lips crescent-shaped.  Lateral lips large, triangular, separated from medial lips and head region.  Head region not set off from regular body annules.  In LM, cephalic framework distinct, hexaradiate; lateral sectors enlarged.  Vestibule and extensions prominent.  Cephalids and hemizonid not observed.  Distance of excretory pore to head end variable in distance (30.9-118.2 um); located in most specimens midway between anterior end and metacorpus; teminal excretory duct very long.  Stylet long and robust; cone of same size as shaft, tip straight or slightly curved dorsally, widening gradually poteriorly; junction of cone and shaft eneven.  Shaft cylindrical and same width throughout, or widening slightly near junction with knobs; knobs broad laterally, set off from shaft, distinctly separated from each other; knobs very slightly indented anteriorly.  Distance between stylet base and dorsal esophageal gland orifice (DGO) moderately long (4.7-6.3 um); gland orifice branched into three channels; dorsal gland ampulla large; subventral gland orifices branched, located posteriorly to enlarged triradiate lumen lining of metacorpus.  Esophageal lumen lining with small rounded vesicles anterior to triradiate lumen lining.  Eosophageal glands large, trilobed; dorsal lobe largest, unicleate; two subventral nucleated lobes variable in size, shape, and position; located posterior to dorsal gland lobe.  Esophago-intestinal cells two, small, rounded, nucleated, located between metacorpus and intestine.  Two ovaries and six rectal glands as characteristic of genus.
    Perineal patterns extremely rounded to oval-shaped.  Dorsal arch high and rounded except for striae near vulva, which are low with rounded shoulders.  Lateral field with distinctly forked striae.  Ventral striae vary from wavy to coarse.  Tail tip area well defined, free of striae; often with a few to seveal subcuticular punctations.  Subcuticular punctations located randomly within the pattern area.  Perivulval region not striated, rarely striae near lateral edges of vulva.  Vulva located in depression, surrounded by wide cuticular ridge.  Phasmidial ducts distinct, phasmid surface structure often obscured by striae in SEM.  Anus distinct, surrounded by a thick cuticular layer.

Allotype (male in glycerin):  Body length = 1,873 um; body width = 38.2 um; stylet length = 18.9 um; stylet knob height = 3.3 um; stylet knob width =  5.2 um; dorsal esophageal gland orifice to stylet base = 5.8 um; excretory pore to head end = 163.5 um; tail length = 14.2 um; gubernaculum length = 7.6 um; testis length = 788.0 um; a = 41.4 um; c = 102.9 um; and T = 44%.  Male as in general description.
Males: (n = 30, in fresh tap water stored in refrigerator for at least 48 hours and killed by gentle heat) measurements are listed in Table 2.
Body translucent white, vermiform; body tapering anteriorly, bluntly rounded posteriorly; tail twisting through 90O in heat-killed specimens.  Head cap high in lateral view, extending posteriorly onto distinctly set-off head region.  Head region high in lateral view, tapering posteriorly, distinctly set off from body.  Hexaradidate cephalic framework well sclerotized; vestibule and extension distinct.  Prestoma large, hexagonal.  Stoma slit-like, located in large, hexagonal prestomatal cavity, surrounded by pore-like openings of six inner labial sensilla.  In SEM, labial disc rounded, very large; often separated from medial lips by a shallow groove.  Medial lips very wide, outer margins crescent-shaped, sloping posteriorly.  Labial disc and medial lips may or may not be fused to form elongate and wide lip structure extending posteriorly onto head region.  Four cephalic sensilla marked on medial lips by shallow, elongate, ovoid, depressions.  Amphidial apertures large, elongated, slit-like between labial disc and lateral sectors of head region.  Lateral lips absent.  Head region smooth, annulation absent. Body annules large, distinct.  Lateral field with four incisures, two beginning near level of stylet knobs and two near level of metacorpus; lateral field areolated, encircling tail.  Stylet robust, large, cone straight, pointed, gradually increasing in diameter posteriorly; opening located several micrometers from stylet tip; cone of same size as shaft.  Shaft cylindrical, posterior end wider than anterior end.  Knobs large, wide, rounded, set off from shaft.  Dorsal esophageal gland orifice to stylet base variable in distance (3.7-6.4 um), dorsal gland duct branched into three channels, gland ampulla indistinct.  Procorpus indistinctly outlined, indistinctly outlined metacorpus elongated, oval-shaped with valve enlarged, triradiate cuticular lumen lining; subventral esophageal gland orifices branched, located posteriorly to metacorpus.  Esophago-intestinal junction indistinct.  Gland lobe varible in length, with indistinct nuclei rarely visible.  Excretory pore distinct, variable in position (150.0-180.9 um), terminal duct long.  Hemizonid located anterior to excretory pore.  Intestinal caecum short, extends anteriorly on dorsal side to base of metacorpus.  Usually one testis, rarely two testes, outstretched, or reflexed anteriorly. Spicules long, slender, slightly arcuate with single tip, short head, wide vellum, and indistinct shaft.  Gubernaculum distinct, crescent shaped.  Tail short and rounded.  Phasmids slit-like opening near level of cloaca.

Second-stage juveniles: (n = 30 J2 in fresh tap water killed by gentle heat) measurements are listed in Table 3.
Body translucent white, long, slender, tapering anteriorly but more so posteriorly.  Body annules distinct, increase in size and become irregular in posterior tail region.  Lateral field starts approximately at middle of procorpus and extends to near phasmids, with four incisures, areolated in some specimens.  Stoma slit-like, located in oval-shaped prestomatal depression, surrounded by pore-like openings of six inner labial sensilla.  Head cap high, narrower than head region.  Labial disc elongated, round-shaped, completely fused with medial lips.  Medial lips with outer margins crescent-shaped, smooth.  Medial lips and labial disc dumbell-shaped.  Lateral lips distinct, lower than medial lips, margins crescent-shaped.  Head region smooth without annulation.  Amphidial apertures elongate, located between labial disc and lateral lips.  Head region high, distinctly set off from body.  Hexaradiate framework weakly sclerotized in LM, vestibule and vestibule extension distinct.  Stylet moderately long (9.1-12.3 um); but  delicate, stylet cone sharply pointed, increases in width gradually posteriorly; shaft cylindrical, may widen slightly posteriorly, knobs rounded.  Distance of dorsal esophageal gland orifice to stylet base moderately long (1.8-3.6 um); orifice branched into three channels; ampulla poorly defined.  Procorpus faintly outlined, metacorpus ovoid with distinct valve; subventral esophageal gland orifices posterior to valve; ampulla distinct.  Esophagus-intestinal junction indistinct, at the level of nerve ring.  Esophageal gland lobe variable in length with three small nuclei of same size.  Excretory pore distinct, variable in postion (74.5-105.5 um), terminal duct very long.  Hemizonid distinct, located anteriorly to excretory pore.  Tail slender, ending in slightly round tip; tail annules larger and irregular posteriorly.  Hyaline tail end long, variable in size (10.9-16.4 um).  Rectal dilatation large.  Phasmids small, indistinct, located at one edge of lateral field, below level of anus.

Eggs (n = 30 in fresh tap water): Length = 93-110 um; (mean 98.8 um + 4.45 standard, error of mean at 95% confidence interval); width = 51-57 um (mean 53.4 um  + 2.99 standard); length/width ratio = 1.7-2.0 um (mean 1.9 + 0.08 standard).  Morphology similar to other Meloidogyne species.

Diagnosis: Meloidogyne haplanaria can be distinguished from the four most common species of root-knot nematode (M. arenaria, M. hapla, M.incognita and M. javanica) (Eisenback et al., 1981) by the unique form of perineal pattern, shape, and morphology of head and stylet, shape and constitution of metacorpus, and position of excretory pore in females; and shape and morphology of head and stylet in males. The perineal pattern is similar to M. arenaria in overall shape but similar to M. hapla because of the punctations that are present in the tail tip area.
Meloidogyne haplanaria can be easily diagnosed and separated from the four most common root-knot nematodes by the inability to infect cotton, tobacco, pepper, and watermelon.  Pepper, wateremelon, and cotton are good hosts for M. incognita; watermelon is also a good host for M. javanica; pepper is a good host for M. arenaria and M. hapla (Hartman and Sasser, 1985) but not M. haplanaria . Peanut is good host for M. arenaria and M. hapla as well as M. haplanaria.
(Description- Eisenback et al., 2003)

DNA Sequences Obtained
Specimen: Collected:
Texas A&M 7 in culture- Texas A&M
Texas A&M 8 in culture- Texas A&M