Morphological,Molecular, and Differential-Host Characterization of Meloidogyne floridensis n. sp. (Nematoda: Meloidogynidae), a Root-Knot Nematode Parasitizing Peach in Florida

A root-knot nematode, Meloidogyne floridensis n. sp., is described and illustrated from peach originally collected from Gainesville, Florida. This new species resembles M. incognita, M. christiei, M. graminicola, and M. hispanica, but with LM and SEM observations it differs from these species either by the body length, shape of head, tail and tail terminus of second-stage juveniles, body length and shape of spicules in males, and its distinctive female perineal pattern. This pattern has a high to narrowly rounded arch with coarsely broken and network-like striae in and around anal area, faint lateral lines interrupting transverse striae, a sunken vulva and anus, and large distinct phasmids. Molecular data from ribosomal IGS illustrate that M. floridensis n. sp. is different from the mitotic species M. arenaria, M. incognita, and M. javanica. Data from RAPDs confirm it and suggest that this new species lies in an intermediate phylogenetic position between the previous species and the meiotic species M. hapla, M. fallax, and M. chitwoodi. Differential host tests based on annual crops and on Prunus accessions are reported.     

Meloidogyne platani n. sp. (Meloidogynidae), a Root-knot Nematode Parasitizing American Sycamore

Meloidogyne platani n. sp. is described and illustrated from specimens obtained from roots of American sycamore, Platanus occidentalis, in Virginia. This new species shows certain similarities with M. arenaria but differs from it by a number of distinctive characters. The perineal pattern of females is rounded with fine, wavy to zig-zag striae and raised, convoluted striae in the inner lateral line regions. The stylet of females is 16.5 μm long with large, rounded stylet knobs set off from the shaft. Males have a low head cap and smooth head region. The styler length is 22.0 μm, and the stylet knobs are rounded and set off from the shaft. Mean second-stage juvenile length is 443.0 μm, and stylet length is 12.2 μm. The head region of juveniles is not annulated, and the tail has a definite terminus. This nematode causes severe galling and reproduces well on sycamore. Other good hosts include white ash and tobacco cv. NC 95. M. platani n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of approximately 45 (2n).     

Meloidogyne microcephala n. sp. (Meloidogynidae), a Root-knot Nematode from Thailand

Meloidogyne microcephala n. sp. is described and illustrated from specimens obtained from tobacco (Nicotiana tabacum L.) in Thailand. The female perineal pattern usually has a low dorsal arch, coarse striae, and a series of small cuticular flaps around the tail terminus. The stylet of the female is 14.4 μm long, with large, square to rectangular stylet knobs, The distinctive male head region is narrow, small, and truncate with a low, flattened head cap. The stylet length is 20.6 μm, and the knobs are small, angular, and set off from the shaft. Mean length of second-stage juveniles is 457.5 μm, and stylet length is 9.3 μm. The tail tip in the juveniles is set off from the rest of the tail as a small finger-like projection. M. microcephala reproduces by mitotic parthenogenesis, and has a chromosome number of 2n = 36.     

Meloidogyne trifoliophila n. sp. (Nemata: Meloidogynidae), a Parasite of Clover from Tennessee

Meloidogyne trifoliophila n. sp. is described from white clover collected at Ames Plantation, Fayette County, Tennessee. The perineal pattern is rounded, with long, smooth striae and rounded arch, and without distinct lateral lines or perivulval striae. The female stylet is 12.6-15.5 μm long, the excretory pore is level with or up to one stylet length posterior to the stylet knobs, and the vulva is subterminal. The posterior terminus is weakly protuberant. The male lateral field is composed of approximately eight repeatedly broken or forked incisures. The male stylet is 17.0-18.9 μm long, the stylet knobs are rounded and sloping, gradually merging with the shaft, and the head region consists of one large annule. Second-stage juveniles are 357-400 μm long, with a stylet length of 11.9-13.6 μm and one head annule. The tail tapers to a slender tip. This new species is similar to M. graminicola and M. triticoryzae but differs from them in perineal pattern and lateral field morphology, and numerous morphometric characters.     

Meloidogyne morocciensis n. sp. (Meloidogyninae), a Root-knot Nematode from Morocco

Meloidogyne morocciensis n. sp. is described from specimens parasitic on peach rootstock from Morocco. This species exhibits a combination of morphological characters similar to M. arenaria, M. incognita, and M. javanica. The perineal pattern of females is oval to squarish with a moderately high to high dorsal arch, and widely spaced, smooth striae; lateral lines are absent. The stylet, 16.5 μm long, has transversely ovoid, set-off knobs. Males have a set-off, annulated head region. The large, rounded labial disc is distinctly demarcated from the crescent-shaped medial lips; lateral lips are absent. The robust stylet, 24.6 μm long, has large, rounded knobs that taper slightly posteriorly. Mean second-stage juvenile (J2) length is 401 μm. The set-offhead region has incomplete annulations; the lip structures are dumbbell shaped. The stylet, 12.3 μm long, has rounded knobs that slope posteriorly. The J2 tail, 52.6 μm long, has irregularly sized annules in the posterior region and ends in a bluntly rounded tip. Tomato, tobacco, pepper, and watermelon are good hosts; cotton and peanut are not hosts. Meloidogyne morocciensis n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 47-49. Its esterase phenotype is identical with the three-banded phenotype (A3) of M. arenaria.     

Description of Globodera ellingtonae n. sp. (Nematoda: Heteroderidae) from Oregon

A new species of cyst nematode, Globodera ellingtonae, is described from soil collected from a field in Oregon. Second-stage juveniles (J2) of the species are characterized by body length of 365-515 μm, stylet length of 19-22.5 μm, basal knobs rounded posteriorly and pointed anteriorly, tail 39-55 μm, hyaline tail terminus 20-32.5 μm, and tail tapering uniformly but abruptly narrowing and constricted near the posterior third of the hyaline portion, ending with a peg-like, finely rounded to pointed terminus. Cysts are spherical to sub-spherical, dark to light brown and circumfenestrate and cyst wall pattern is ridge-like with heavy punctations. Males have a stylet length of 21-25 μm and spicule length of 30-37 μm with a pointed thorn-like tip. Females have a stylet length of 20-22.5 μm, one head annule and labial disc, heavy punctations on the cuticle, and short vulval slit 7.5-8 μm long. Morphologically this new, round-cyst species differs from the related species G. pallida, G. rostochiensis, G. tabacum complex and G. mexicana by its distinctive J2 tail, and by one or another of the following: shorter mean stylet length in J2, females and males; number of refractive bodies in the hyaline tail terminus of J2; cyst morphology including Granek’s ratio; number of cuticular ridges between the anus and vulva; and in the shape and length of spicules in males. Its relationship to these closely related species are discussed. Based upon analysis of ribosomal internal transcribed spacer (ITS) sequences, G. ellingtonae n. sp. is distinct from G. pallida, G. rostochiensis, G. tabacum and G. mexicana. Bayesian and Maximum Parsimony analysis of cloned ITS rRNA gene sequences indicated three clades, with intraspecific variability as high as 2.8%. In silico analysis revealed ITS restriction fragment length polymorphisms for enzymes Bsh 1236I, Hinf I, and Rsa I that overlap patterns for other Globodera species.     

Meloidogyne lusitanica n. sp. (Nematoda: Meloidogynidae), a Root-knot Nematode Parasitizing Olive Tree (Olea europaea L.)

A root-knot nematode from Portugal, Meloidogyne lusitanica n. sp., is described and illustrated from specimens obtained from olive trees (Olea europaea L.). Females of the new species have a characteristic perineal pattern with medium to high trapezoidal dorsal arch with distinct punctuations in the tail terminus area. The excretory pore is located posterior to the stylet, about 1.5-2.5 stylet lengths from the anterior end. The stylet is 17.1 μm long with pear-shaped knobs. Males have a rounded, posteriorly sloping head cap and head region not annulated. The robust stylet, 24.5 μ long, has large, elongate knobs. Mean length of the second-stage juveniles is 449.5 μm, stylet length 14.2 μm, and tail length 44.1 μm. Scanning electron microscope observations provide further details of perineal patterns and head and stylet morphology of females, males, and second-stage juveniles. Meloidogyne lusitanica n. sp. did not reproduce on any of the differential hosts used to separate the four most common Meloidogyne species. The common name "olive root-knot nematode" is proposed for M. lusitanica n. sp.     

Morphological Comparison of Head Shape and Stylet Morphology of Second-stage Juveniles of Meloidogyne Species

Head shape and stylet morphology of second-stage juveniles of one population each of M. incognita, M. javanica, M. arenaria, and M. hapla were compared by light microscopy. Excised stylets of each species were also compared by scanning electron microscopy (SEM). Differences in head morphology were observed only between M. hapla and the other three species. In SEM, differences in stylet size, shape, and relative distance of the dorsal esophageal gland orifice to the base of the stylet were evident. Differences in stylet morphology between M. incognita and M. javanica could not he detected by light microscopy, but M. arenaria and M. hapla could be distinguished from each other and from the other two species. Head shape and styler morphology of second-stage juveniles are considered useful taxonomic characters.     

Description and SEM Observations of Meloidogyne chitwoodi n. sp. (Meloidogynidae), a Root-knot Nematode on Potato in the Pacific Northwest

Meloidogyne chitwoodi n. sp. is described and illustrated from potato (Solanum tuberosum) originally collected from Quincy, Washington, USA. This new species resembles M. hapla, but its perineal pattern is basically round to oval with distinctive and broken, curled, or twisted striae around and above the anal area. The vulva is in a sunken area devoid of striae. Vesicles or vesicle-like structures are present in the median bulb of females. The larva tail, being short and blunt with a hyaline tail terminal having little or no taper to its rounded terminus, is distinctively different from M. hapla. SEM observations revealed the nature of the perineal pattern and details of the head of larvae and males, and showed the spicules to have dentate tips ventrally. Hosts for M. chitwoodi n. sp. include potato, tomato, corn, and wheat but not strawberry, pepper, or peanut. The latter three crops are excellent hosts for M. hapla. The known distribntion of this new root-knot species presently involves certain areas of Idaho, Washington, and Oregon. The common name "Columbia root-knot nematode" is proposed for M. chitwoodi n. sp.     

Meloidogyne mayaguensis n. sp. (Meloidogynidae), a Root-knot Nematode from Puerto Rico

Meloidogyne mayaguensis n. sp. is described and illustrated from specimens obtained from galled roots of eggplant, Solanum melongena L., from Puerto Rico. The perineal pattern of females is round to ovoid with fine, widely spaced striae. It has occasional breaks of striation laterally and a circular tail tip area lacking striae. The stylet, 15.8 μm long, has reniform knobs that merge gradually with the stylet shaft. Males have a high, rectangular, smooth head region, not set off from the body contour. The labial disc is continuous with the medial lips which do not slope posteriorly. The styler, 22.9 μm long, has large rounded backward sloping knobs; the shaft is of uneven diameter. Mean body length of second-stage juveniles is 453.6 μm. The truncate head region is not annulated, and the rounded, slightly raised labial disc and the crescentic medial lips form dumbbell-shaped lip structures. The stylet, 11.6 μm long, has rounded, posteriorly sloping knobs. The slender tail, 54.4 μm long, gradually tapers to a bluntly pointed tip. Tomato, tobacco, pepper, and watermelon are good hosts; cotton and peanut are not hosts. M. mayaguensis n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 2n = 44-45. The enzyme patterns are unique among Meloidogyne species.     

Description and SEM Observations of Meloidogyne sasseri n. sp. (Nematoda: Meloidogynidae), Parasitizing Beachgrasses

Meloidogyne sasseri n. sp. is described and illustrated from American beachgrass (Ammophila breviliffulata) originally collected from Henlopen State Park and Fenwick Island near the Maryland state line in Delaware, United States (6). Its relationship to M. graminis, M. spartinae, and M. californiensis is discussed. Primary distinctive characters of the female perineal pattern were a high to rounded arch with shoulders, widely spaced lateral lines interrupting transverse striations, a sunken vulva and anus, and coarse broken striae around the anal area. Second-stage juvenile body length was 554 μm (470-550), stylet length 14 μm (13-14.5), tail length 93 μm (83-115), tapering to a finely rounded terminus. Male stylet length 20 μm (19-21.5), spicule length 33 μm (30-36). Scanning electron microscope observations provided additional details of perineal patterns and face views of the female, male, and J2 head. Wheat, rice, oat, Ammophila sp., Panicum sp., bermudagrass, zoysiagrass and St. Augustinegrass were tested as hosts. Distribution of the species was the coasts of Delaware and Maryland. The common name "beachgrass root-knot" is proposed for M. sasseri n. sp.     

Scanning Electron Microscopy of Perineal Patterns of Three species of Meloidogyne

The perineal pattern of Meloidogyne graminicola is oval, characterized by the anastomosing of cuticular striae converging, and elevating at the tail terminus. A deep lateral incisure forming a low arch traverses the rounded perineal pattern of Meloidogyne hapla. Punctations at the tail region of M. hapla are not observed with the scanning electron microscope. Minute crosslines invaginate with the vulvar lips of Meloidogyne incognita. Punctate indentations around the anal opening and bordering the lateral sides of the vulvar slit of M. incognita are revealed.     

Meloidogyne paranaensis n. sp. (Nemata: Meloidogynidae), a Root-Knot Nematode Parasitizing Coffee in Brazil

A root-knot nematode parasitizing coffee in Paran  State, Brazil, is described as Meloidogyne paranaensis n. sp. The suggested common name is Paraná coffee root-knot nematode. The perineal pattern is similar to that of M. incognita; the labial disc and medial lips of the female are fused and asymmetric and rectangular; the lateral lips are small, triangular, and fused laterally with the head region. The female stylet is 15.0-17.5 μm long, with broad, distinctly set-off knobs; the distance from the dorsal esophageal gland orifice (DGO) to the stylet base is 4.2-5.5 μm. Males have a high, round head cap continuous with the body contour. The labial disc is fused with the medial lips to form an elongate lip structure. The head region is frequently marked by an incomplete annulation. The stylet is robust, 20-27 μm long, usually with round to transversely elongate knobs, sometimes with one or two projections protruding from the shaft. The stylet length of second-stage juveniles is 13-14 μm, the distance of the DGO to the stylet base is 4.0-4.5 μm, and the tail length is 48-51 μm. Biochemically, the esterase (F₁) and malate dehydrogenase (N₁) phenotypes are the most useful characters to differentiate M. paranaensis from other species. However, the esterase phenotype appears similar to that of M. konaensis. Reproduction is by mitotic parthenogenesis, 3n = 50-52. In differential host tests, tobacco, watermelon, and tomato were good hosts, whereas cotton, pepper, and peanut were nonhosts.     

Meloidogyne hispanica n. sp. (Nematoda: Meloidogynidae), the 'Seville Root-Knot Nematode'

Meloidogyne hispanica n. sp. is described and illustrated from specimens obtained from peach rootstock, Prunus persica silvestris Batsch, from the Seville district of Spain. The perineal pattern of the female is oval shaped to rectangular with low dorsal arch and often widely spaced lateral lines with fringe-like striae. The stylet, 14.1 μm long, has broad, distinctly set off knobs. Males have a high, rounded head cap that slopes posteriorly. Labial disc and medial lips are fused to form elongate lip structures. The robust styler, 23.5 μm long, has large, rounded knobs that are slightly set off from the shaft. Mean second-stage juveniles length is 392.6 μm. The truncate head region is generally not annulated. The distinctly rounded and raised labial disc and the crescent-shaped medial lips form dumbbell-shaped lip structures. The stylet, 11.1 μm long, has rounded, posteriorly sloping knobs. The slender tail, 46.4 μm long, has large irregular-sized annules in the posterior region and ends in a bluntly rounded tip. Tomato was a good host; tobacco, pepper, and watermelon were poor hosts; cotton and peanut were nonhosts. Meloidogyne hispanica n. sp. reproduces by mitotic parthenogenesis and has a somatic chromosome number of 2n = 33-36. The esterase pattern is unique among Meloidogyne species.     

Morphological and Morphometrical Characterization of Meloidogyne incognita from Different Host Plants in Four Districts of Punjab,India

The population of M. incognita, the root knot nematode (RKN) was found infesting five different host plants (okra, banana, sunflower, bottle gourd, and brinjal) out of 24 examined from four districts of Punjab, India (Gurdaspur, Ludhiana, Patiala, and Hoshiarpur). Morphological and morphometrical characterization indicated that in the case of mature female, the characters of body length and width, neck length, ratio ‘a’, anus to tail terminus (ATT), interphasmid distance (IPD), and perineal pattern were recorded as stable characters. These taxonomic characters can be reliable for identification. All characters of second-stage juvenile (J2) such as body length, stylet length, head to median bulb length (H-MB), distance from median bulb to excretory pore (MB-EP), tail length, anal body width (ABW), and ratios C and C’ were highly variable. Analysis of interpopulation morphometric characters of mature female of M. incognita, namely, body length, width, and ratio ‘a’ were moderately variable characters (CV 0.26% to 20%) and stylet length, neck length, length of median bulb (LMB), and width of median bulb (WMB) were highly variable (CV 1.0% to 36.1%). In the perineal pattern, the two characters ATT and IPD were moderately variable (CV 8.8% to 17.6%) and two characters, anus to vulval slit (AVS) and length of vulval slit (LVS), were highly variable (CV 2.1% to 40.5%). In J2, body length, stylet length, H-MB, MB-EP, ABW, tail length, ratios C, and C’ were highly variable characters (CV > 12%).     

Morphology of Second-stage Juveniles and Males of Globodera tabacum tabacum,G. t. virginiae,and G. t. solanacearum (Nemata: Heteroderinae)

Morphological comparisons with light microscopy and scanning electron microscopy were made among second-stage juveniles (J2) and males of several isolates of the three subspecies of the tobacco cyst nematode complex, Globodera tabacum sspp. tabacum, virginiae, and solanacearum. Observations focused on the anterior region, (including head shape, lip pattern, and stylet morphology) and the tail region (including tail shape in J2 and spicules in males). The three subspecies could not be separated on the basis of any of these characters.     

Merlinius paniculoides n. sp. from Italy (Tylenchina: Belonolaimidae)

Merlinius paniculoides n. sp. is described from a bisexual Italian population found in northern Italy. This species has a coarsely annulated cuticle with approximately 30 longitudinal striae in addition to the six lateral field incisures; consequently the cuticle appears tessellate. The female tail is subcylindrical with a bluntly, rounded, non-annulated terminus. Merlinius paniculoides shares the tessellate cuticular ornamentation and the offset labial region with M. lenorus (Brown, 1956) Siddiqi, 1970 and M. tartuensis (Krall, 1959) Siddiqi, 1970. It differs from M. lenorus by the higher c and lower c'' ratios (c = 16-19 vs. 12-15 and c'' = 2.2-2,8 vs. 3.7) and greater number of longitudinal striae (28-32 vs. 24). It differs from M. tartuensis by a fewer number of annuti from stylet knob base to lip constriction (8-9 vs. 21), shorter stylet (19-21 vs. 24 μm), presence of males, and higher c'' value (c'' = 2.2-2.8 vs. 1.9).     

Meloidogyne californiensis n. sp. (Nemata: Meloidogyninae), Parasitic On Bulrush,Scirpus robustus Pursh

Meloidogyne californiensis n. sp. is described and illustrated from bulrush Scirpus robustus in California. LM and SEM studies revealed that this species differs from other known species in the genus Meloidogyne especially by the prominent posterior cuticular protuberances in the female, the distinct shape of the perineal pattern which is marked by one prominent stria in the perineum, indistinct lateral lines, many broken discontinuous striae on both sides of the arch, and the excretory pore being located posterior to stylet base. Second-stage juveniles 448-628 μm long, stylet length 11-13 μm, styler delicate, with small knobs sloping posteriorly, cephalic region with 2 or 3 annuli, and inflated rectum. Males vary greatly in size (712-1,952 μm), stylet length 18-28 μm (mean 22 μm), cephalic region slightly set off the body with two or three annuli, spear heavy with massive rounded knobs, lateral field marked by four areolated incisures as seen by SEM.     

Identification of Meloidogyne Species on the Basis of Head Shape and,Stylet Morphology of the Male

Head shape and stylet morphology of males of 90 populations of M. arenaria, M. hapla, M. incognita, and M. javanica from geographic regions of the world were compared by light microscopy (LM). In addition, stylets of one population each of M. arenaria, M. incognita, and M. javanica and three different chromosomal forms of M. hapla race A and two of race B were excised and examined with a scanning electron microscope (SEM). Differences among species occurred in both head and stylet morphology. Head morphology differed in size and shape of the head cap, annulation of the head region, and width of the head region relative to the first body annule. Differences in stylets occurred in size and shape of the cone, shaft, and knobs. All populations of M. hapla, except one, had similar head morphology, but stylet morphology was different between cytological races A and B. Populations of M. javanica varied with respect to the presence of head annulations. Head shape and stylet morphology of males are recommended as additional characters useful in the identification of root-knot nematodes.     

Description of the Blueberry Root-knot Nematode,Meloidogyne carolinensis n. sp.

Meloidogyne carolinensis n. sp. is described from cultivated highbush blueberry (cultivars derived from hybrids of Vaccinium corymbosum L. and V. lamarckii Camp) in North Carolina. The perineal pattern of the female has a large cuticular ridge that surrounds the perivulval area, and the excretory pore is near the level of the base of the stylet. The stylet is 15.9 μm long and the knobs gradually merge with the shaft. The head shape and stylet morphology of the male are quite variable. The typical head and four variants, as well as the typical stylet and two variants, are described. The labial disc, medial lips, and lateral lips of second-stage juveniles are fused and in the same contour. The head region is not annulated. Mean juvenile length is 463.7 μm, stylet length is 11.9 μm, and tail length is 42.5 μm.