Molecular phylogeny of the Tylenchina and evolution of the female gonoduct (Nematoda: Rhabditida)

Tylenchina are a morphologically and functionally diverse group of nematode species that range from free-living bacteriovores, over transitory grazing root-hair feeders to highly specialized plant-parasites with complex host associations. We performed phylogenetic analyses of small subunit rDNA sequences from 97 species including an analysis that account for the RNA secondary structure in the models of evolution. The present study confirms the sister relationship of the bacteriovore Cephalobidae with the predominantly plant-parasitic Tylenchomorpha. All analyses appoint the fungal-feeding Aphelenchidae and Aphelenchoididae as being polyphyletic but the morphology based hypothesis of their monophyly could not be significantly rejected. Within the Tylenchomorpha, the families that exclusively parasitize higher plants are joined in a single clade. However, only the monophyletic position of the (super)families Hoplolaimidae and Criconematoidea were supported; Anguinidae, Tylenchidae, Belonolaimidae and Pratylenchidae appeared to be paraphyletic or polyphyletic. Parsimony and likelihood ancestral state reconstruction revealed that burrowing endoparasitism and sedentary endoparasitism each evolved, respectively, at least six and at least three times independently, mostly from migratory ectoparasitic ancestors. Only root-knot nematodes have evolved from burrowing endoparasitic nematodes. Traditional classifications are partially misled by this convergent evolution of feeding type and associated morphology. Contrastingly, mapping attributes of the gonoduct cellular architecture, including newly obtained data of 18 species belonging to the Aphelenchoidea, Criconematoidea, Anguinidae and Panagrolaimidae, revealed a broad congruence of the gonoduct characters and the molecular phylogenetic hypothesis. Yet, the presence of an offset spermatheca and proliferation of uterus cells has evolved multiple times, the latter associated with derived endoparasitic feeding specialization and resulting reproduction mode. Ancestral state reconstruction further revealed that the gonoduct of the morphologically and ecologically dissimilar tylenchid and cephalobid nematodes evolved from a common ancestor.     

Nematodes of the Order Rhabditida from Andalucía Oriental,Spain. The Genera Nothacrobeles Allen &Noffsinger, 1971 and Zeldia Thorne, 1937

A new species of the genus Nothacrobeles is described from natural areas (a salt lake) in the Southeast Iberian Peninsula. Nothacrobeles lanceolatus sp. n. is characterized by its body length, two rows of cuticular punctations per annulus, labial probolae bifurcate with divergent prongs, pharyngeal corpus 2.4 to 3.5 times isthmus length, spermatheca length, postuterine sac 0.5 to 1.1 times the corresponding body diameter ratio, female tail conical and bearing a spindle-shaped or conical mucro with acute terminus, phasmid at 8 to 17 µm posterior to the anus, male tail conical with acute mucro, spicules length, and gubernaculum length. In addition, Nothacrobeles cf. lunensis and Zeldia punctata are studied. Cervidellus capricornis is transferred to genus Nothacrobeles. A key to species of Nothacrobeles is also provided.     

Morphological Comparison of Meloidogyne Female Head Structures,Perineal Patterns,and Stylets

The external morphology of female heads of three populations of each of two cytological races of Meloidogyne hapla (race A-meiotic, race B-mitotic) and single populations of M. arenaria, M. incognita, and M. javanica was compared by light (LM) and scanning electron microscopy (SEM). Perineal patterns of all nine populations were observed with a LM and then examined with a SEM. In addition, female stylets of each population were excised, viewed with a SEM, and compared with observations made with a LM. Head morphology of the females, including shape of medial and lateral lips, expression of sensilla, and head annulation, was distinct for each species, each race of M. hapla, and each population of M. hapla race A. The morphology of a given perineal pattern appeared similar with the SEM and the LM. The SEM emphasized surface details, whereas the LM revealed subcuticular structure as well. Stylet morphology was unique for each species but similar in all populations of M. hapla. There were differences between species in the shape of the cone, shaft, and knobs and in the distance of the dorsal esophageal gland orifice from the stylet knob base. Several of the morphological characters first detected in the SEM were seen subsequently with the LM and are helpful in species identification.     

Evolution of specialized spermatheca morphology in ant queens: Insight from comparative developmental biology between ants and polistine wasps

In many ant species, the queens can keep spermatozoa alive in their spermatheca for several years, which goes along with unique morphological characteristics of the queen's spermatheca. The relative spermatheca size in ant queens is prominently larger than that in social wasps. Furthermore, the epithelium lining the spermatheca reservoir of ants consists of columnar cells in the hilar region and squamous cells in the distal region, whereas it is formed by columnar cells only in social wasps. To study the evolution of the unique spermatheca morphology in ant queens, we compared the various processes during spermatheca development between two ponerine ant species of the genus Pachycondyla (= Brachyponera) and three polistine wasp species of the genus Polistes. From histological observations, we can define four developmental events in the ant queens: (1) invagination of the spermatheca primordium, (2) the reservoir wall thickness becomes unequal, (3) the reservoir diameter doubles as the lining epithelial cells become flattened except for the hilar region, and (4) the increase in thickness of the reservoir epithelium is limited to the hilar region which doubles in thickness. In polistine wasps, the second and the third developmental events are absent and the entire epithelium of the spermatheca wall becomes thick in the final step. We therefore conclude that for ant queens the second and third steps are crucial for the enlargement of the spermatheca size, and that the second to the fourth steps are crucial for the specialization of the reservoir wall structure.     

A new species of Alopoglossus lizard (Squamata,Gymnophthalmidae) from the tropical Andes,with a molecular phylogeny of the genus

We describe a new species of Alopoglossus from the Pacific slopes of the Andes in northern Ecuador based on morphological and molecular evidence. The new species differs most significantly from all other congeners in having a double longitudinal row of widened gular scales, lanceolate dorsal scales in transverse rows, 29–32 dorsal scales in a transverse row at midbody, and 4 longitudinal rows of ventrals at midbody. It is most similar in morphology to A. festae, the only species of Alopoglossus currently recognized in western Ecuador. We analyze the phylogenetic relationships among species of Alopoglossus based on the mitochondrial gene ND4. Cis-Andean [east of the Andes] and Trans-Andean [west of the Andes] species are nested in two separate clades, suggesting that the uplift of these mountains had an important effect in the diversification of Alopoglossus. In addition, we present an updated key to the species of Alopoglossus.     

Larinus berti sp. n. (Coleoptera,Curculionidae, Lixinae) from North Africa

A new species, Larinus berti sp. n. is described from Morocco and assigned to subgenus Cryphopus Petri, 1907 (Curculionidae: Lixinae; Lixini). Diagnostic characters of the new species are large size, elongate-ovate body, bisulcate sub-quadrangular rostrum, triangularly raised dorsum of rostrum, flat subgena and submentum, Y-shaped apodeme of sternite VIII of female and thin nodulus of spermatheca.     

In Vitro Parasitism of Rotylenchus robustus by Isolates of Hirsutella rhossiliensis

We tested the hypothesis that isolates of Hirsutella rhossiliensis from host nematodes in the family Hoplolaimidae (Rotylenchus robustus and Hoplolaimus galeatus) would be more virulent to R. robustus than would isolates from host nematodes not in the Hoplolaimidae (Heterodera schachtii and Criconemella xenoplax). Nematodes were touched to 10-20 spores of different isolates and incubated at 20 C in 4.5 mM KC1; the percentage of nematodes colonized (filled with hyphae) was determined after 2, 5, 10, 20, and 30 days. The hypothesis was rejected because isolates from H. schachtii and C. xenoplax were equivalent or better at parasitizing R. robustus than were isolates from R. robustus and H. galeatus. In addition, the R. robustus and H. galeatus isolates were as pathogenic to C. curvata as they were to R. robustus, but produced fewer spores per colonized nematode (H. schachtii) than did the other isolates.     

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.     

Seasonal Fluctuations in the Spatial Distribution of Nematode Populations in a California Vineyard

Distribution of Xiphinema americanum and four Meloidogyne spp, was studied in a vineyard over a 13-mo. period. The X. arnericanum population was concd in the upper 60-cm of undisturbed soil in the vine row, whereas the Meloidogyne species were distributed both in and between rows and to greater depths, similar to the distribution of the root system. Samples for assessment of X. americanum densities had least variation when taken in the vine row from the upper 60-cm of soil. Sampling error is reduced in Meloidogyne populations by sampling within 40 cm of the vine both within and/or between rows.     

Histological Comparisons of Fergusobia/Fergusonina-Induced Galls on Different Myrtaceous Hosts

The putative mutualism between different host-specific Fergusobia nematodes and Fergusonina flies is manifested in a variety of gall types involving shoot or inflorescence buds, individual flower buds, stems, or young leaves in the plant family Myrtaceae. Different types of galls in the early-to-middle stages of development, with host-specific species of Fergusobia/Fergusonina, were collected from Australian members of the subfamily Leptospermoideae (six species of Eucalyptus, two species of Corymbia, and seven species of broad-leaved Melaleuca). Galls were sectioned and histologically examined to assess morphological changes induced by nematode/fly mutualism. The different gall forms were characterized into four broad categories: (i) individual flower bud, (ii) terminal and axial bud, (iii) ''basal rosette'' stem, and (iv) flat leaf. Gall morphology in all four types appeared to result from species-specific selection of the oviposition site and timing and number of eggs deposited in a particular plant host. In all cases, early parasitism by Fergusobia/Fergusonina involved several layers of uninucleate, hypertrophied cells lining the lumen of each locule (gall chamber where each fly larva and accompanying nematodes develop). Hypertrophied cells in galls were larger than normal epidermal cells, and each had an enlarged nucleus, nucleolus, and granular cytoplasm that resembled shoot bud gall cells induced by nematodes in the Anguinidae.     

Temporal patterns in effective quantum yield of individual zooxanthellae expelled during bleaching

Bleaching is a worldwide phenomenon affecting coral reefs. During elevated temperature and light conditions (bleaching), expelled zooxanthellae show distinct patterns in photosynthetic health. An innovative new device was used to collect individual expelled zooxanthellae, when a coral was exposed to bleaching conditions. This has provided new insight into the photosynthetic condition and abundance of expelled zooxanthellae. It has been assumed that expelled zooxanthellae were dead or moribund; however, we have found individual cells can have healthy effective quantum yields (?_PSII) >0.65 after 8 h of bleaching conditions (500 μmol photons m⁻² s⁻¹, 33 °C). The population of expelled zooxanthellae from Cyphastrea serailia and Pocillopora damicornis showed distinct patterns in the frequency distribution of ?_PSII over time and between locations (sun versus shade) within a colony. During the first 4 h of exposure to bleaching conditions, only 5% of expelled individual cells from P. damicornis were photosynthetically inactive (?_PSII<0.05), whereas for C. serailia, this was 30%. The overall photosynthetic health of expelled zooxanthellae from C. serailia was better than P. damicornis (0.53±0.13 and 0.38±0.13 after 8 h, respectively). This was generally reflected by the in hospite measurement of the coral, yet, the in hospite cells always had a higher ?_PSII than expelled cells, suggesting that host tissue provided added photoprotection for the zooxanthellae.     

Postembryonic Morphology in Epsilonematidae, with a Discussion on the Variability of Caudal Gland Outlets

A new species of Akanthepsilonema and the first-stage juvenile of Glochinema trispinatum are described. Furthermore, additional morphological information is provided for Triepsilonema tripapillata. Animals originate from a cold-water coral degradation zone in the Porcupine Seabight area (North-East Atlantic Ocean). Akanthepsilonema sinecornibus sp. n. differs from A. heUeouetae in number of body annules, sexual dimorphism in amphid size, absence of copulatory thorns in males, absence of large spines and horns, shape of the copulatory apparatus, and position of ambulatory setae relative to vulva in females. The genus diagnosis for Akanthepsilonema is adjusted to incorporate the new species. Akanthepsilonema mainly differs from every other genus in the family by the combination of six rows of ambulatory setae situated around the vulva in females and eight subcephalic setae not displaced toward the anterior part of the head capsule. Small differences between the Papua New Guinea and the Porcupine Seabight populations of T. tripapillata indicate minimal intraspecific variability. Second-stage juveniles from Papua New Guinea have two rows of three ambulatory setae, whereas Porcupine Seabight specimens have two rows of four ambulatory setae. First- and fourth-stage juveniles of T. tripapillata are described for the first time. Literature data and personal observations showed that the molting of first-stage juveniles into second-stage juveniles and of third-stage juveniles into fourth-stage juveniles involves a decrease in the number of body rings, resulting in a loss of flexibility which is possibly compensated for by the development (I-II) or the doubling of the number of rows (III-IV) of ambulatory setae. This decrease is also linked with the formation of the head capsule and the smooth tail tip, although intergeneric variability is evident. The molting of second-stage juveniles into third-stage juveniles and of fourth-stage juveniles into adults is also subject to intergeneric variability. The variability in the number and orientation of caudal gland outlets among different nematode taxa is discussed. The presence of separate outlets for the caudal glands seems to be widespread within the family Epsilonematidae and has also been observed in various other, unrelated taxa of free-living aquatic nematodes, although their arrangement in Epsilonematidae is opposite. This aberrant arrangement is probably related to the aberrant locomotory pattern in this family.     

Hirschmanniella pomponiensis n. sp. (Nemata: Pratylenchidae), Parasitic on Bulrush,Scirpus robustus Pursh

A new species of Hirschmanniella was found in bulrush roots; LM and SEM morphological studies revealed that it is distinct from other species in the genus. Therefore, it is designated Hirschmanniella pomponiensis n. sp. Six lips are fused to form a hexagonal labial plate, six inner sensilla encircle the stoma opening, and four cephalic sensilla open in the corners of subdorsal and subventral lips. Cephalic lip region consists of six or seven annuli. The female has incomplete areolation in the lateral field, the intestine overlaps the rectum, the tail tip is pointed and without annulation.     

Description of Pratylenchus gutierrezi n. sp. (Nematoda: Pratylenchidae) from Coffee in Costa Rica

A lesion nematode, Pratylenchus gutierrezi n. sp., collected from the roots of coffee in the Central Plateau of Costa Rica, is described and illustrated. Its relationships to Pratylenchus flakkensis, P. similis, and P. gibbicaudatus, the only other species of the genus having two head annules, males, or spermatheca with sperm, and an annulated tail terminus, is discussed. Other distinctive characters are its posterior vulva (mean of 80%); its prominently rounded stylet knobs, low head, and subcylindrical tail. SEM observations provide additional details of females and males, especially face views, which show for the first time sexual dimorphism.     

Redescription of Three Ektaphelenchus Species (Nematoda: Aphelenchina)

Three Ektaphelenchus species with unusual spicules are redescribed. Ektaphelenchus obtusus and E. riograndensis are redescribed from newly discovered material in the collection of the USDA Forest Service RMRS. Lectotype slides are designated for both species. Ektaphelenchus obtusus is characterized by a set-off head, lips of unequal size, double rows of oocytes and spermatocytes, and a hooked spicule terminus. Ektaphelenchus riograndensis is distinguished by a rounded head with six equal lips, and spicules with a large, rounded apex, and recurved terminus. A bursa and gubernaculum are absent in both species. Ektaphelenchus scolyti is redescribed from type material deposited in the collection of Rothamsted Experimental Station. It is characterized by a well set-off head, double rows of oocytes and spermatocytes, and spicules with a hooked terminus. A bursa and gubernaculum are absent.     

Leptosomatides brevicaudatus n. sp. and a Redescription of Leptosomatides marinae Platonova, 1967 (Enoplida: Leptosomatidae)

The free-living marine nematodes Leptosomatides brevicaudatus n. sp. and L. marinae were described and redescribed, respectively, from material collected in the northwest Pacific. Leptosomatides brevicaudatus n. sp. from Simushir Island differs from L. marinae in the ratio c8 (body length divided by tail length measured on the chord) and the length of the spicules. Leptosomatides marinae is redescribed from light microscopy (LM) observations of the type specimens and LM and scanning electron microscopy (SEM) observations of specimens from Hokkaido, Japan. It appears to be impossible to distinguish among some species of Leptosomatides because they are either insufficiently described or known only from females. Secondary sexual characters of males are essential for purposes of identification.     

Batwanema gen. n. and Chokwenema gen. n. (Oxyurida,?Hystrignathidae), new nematode genera?as parasites of Passalidae (Coleoptera) from the Democratic Republic of Congo

Two new genera and species parasitizing passalid beetles from the Democratic Republic of Congo are described. Batwanema congo gen. n. et sp. n. is characterized by having females with the cervical cuticle armed with scale-like projections, arranged initially in rows of eight elements that gradually divide and form pointed spines toward the end of the spiny region, two cephalic annuli, clavate procorpus and genital tract monodelphic-prodelphic. Two Malagasian species of Artigasia Christie, 1934 were placed in this genus as B. latum (Van Waerebeke, 1973) comb. n. and B. annulatum (Van Waerebeke, 1973) comb. n. Chokwenema lepidophorum gen. n. et sp. n. is characterized by having females with the cervical cuticle armed with scale-like projections, arranged initially in rows of eight elements (similar to Batwanema) that divide gradually, forming spines; a single cephalic annule cone-like, truncated, moderately inflated; procorpus sub-cylindrical and genital tract didelphic-amphidelphic.     

Arabinogalactan protein-rich cell walls,paramural deposits and ergastic globules define the hyaline bodies of rhinanthoid Orobanchaceae haustoria

Background and Aims

Parasitic plants obtain nutrients from their hosts through organs called haustoria. The hyaline body is a specialized parenchymatous tissue occupying the central parts of haustoria in many Orobanchaceae species. The structure and functions of hyaline bodies are poorly understood despite their apparent necessity for the proper functioning of haustoria. Reported here is a cell wall-focused immunohistochemical study of the hyaline bodies of three species from the ecologically important clade of rhinanthoid Orobanchaceae.

Methods

Haustoria collected from laboratory-grown and field-collected plants of Rhinanthus minor, Odontites vernus and Melampyrum pratense attached to various hosts were immunolabelled for cell wall matrix glycans and glycoproteins using specific monoclonal antibodies (mAbs).

Key Results

Hyaline body cell wall architecture differed from that of the surrounding parenchyma in all species investigated. Enrichment in arabinogalactan protein (AGP) epitopes labelled with mAbs LM2, JIM8, JIM13, JIM14 and CCRC-M7 was prominent and coincided with reduced labelling of de-esterified homogalacturonan with mAbs JIM5, LM18 and LM19. Furthermore, paramural bodies, intercellular deposits and globular ergastic bodies composed of pectins, xyloglucans, extensins and AGPs were common. In Rhinanthus they were particularly abundant in pairings with legume hosts. Hyaline body cells were not in direct contact with haustorial xylem, which was surrounded by a single layer of paratracheal parenchyma with thickened cell walls abutting the xylem.

Conclusions

The distinctive anatomy and cell wall architecture indicate hyaline body specialization. Altered proportions of AGPs and pectins may affect the mechanical properties of hyaline body cell walls. This and the association with a transfer-like type of paratracheal parenchyma suggest a role in nutrient translocation. Organelle-rich protoplasts and the presence of exceptionally profuse intra- and intercellular wall materials when attached to a nitrogen-fixing host suggest subsequent processing and transient storage of nutrients. AGPs might therefore be implicated in nutrient transfer and metabolism in haustoria.