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Summary

Rapid diagnosis tools for detection of root-knot nematodes play an important role in the disease control and eradication programme. Recombinase polymerase amplification (RPA) assays were developed targeting the IGS rRNA gene of the pacara earpod tree root-knot nematode, Meloidogyne enterolobii. The RPA assays using TwistAmp® Basic and TwistAmp® exo kits allowed detection of M. enterolobii from gall tissues and crude nematode extracts of all stages of target species without a DNA extraction step. The results of real-time RPA assays using a real-time fluorescent detection of a series of crude nematode extracts showed reliable detection with sensitivity of 1/10 of a second-stage juvenile in a RPA reaction tube after 15-20 min. The RPA assay provides affordable, simple, fast and sensitive detection of M. enterolobii.

Open Access
In: Nematology
In: Advances in Entomopathogenic Nematode Taxonomy and Phylogeny
The sheath nematodes belonging to the superfamily Hemicycliophoroidea are unique amongst all plant parasitic nematodes known to man due to the presence of an extra cuticular covering or sheath over the inner cuticle and body of all juvenile and adult life stages. These plant-parasitic nematodes include species of agricultural and quarantine importance.
In Systematics of the Sheath Nematodes of the Superfamily Hemicycliophoroidea John Chitambar and Sergei Subbotin provide a detailed review of the taxonomy and diagnosis of the superfamily, its member genera and 153 related species based on their morphological and molecular analyses, as well as a further understanding of the relationships within the superfamily using molecular phylogenetics. In addition, Chitambar and Subbotin also give detailed information on the global distribution, biology, host-parasite relationships and ecology of sheath nematodes.

Summary

During surveys for stubby root nematodes in the natural vegetation of California, Oregon and Washington, USA, three known species, viz., Trichodorus californicus, T. intermedius and T. obscurus were recovered together with ten unidentified Trichodorus species. The three known and one new species were studied using an integrated approach. Trichodorus pseudoaequalis n. sp. is characterised by a medium-sized body about 800 μm long on average, male with two ventromedian cervical papillae anterior to secretory-excretory pore and three precloacal supplements all located anterior to the retracted spicules; spicules 39 μm long (average), slightly ventrally curved, more so in head region and blade with slight indentation mid-way and striation more pronounced in posterior half. Females possess a rather short (average 33% of corresponding body diam.) pear-shaped vagina with small vaginal sclerotised pieces (ca 1.5 μm long), rounded triangular in shape, obliquely orientated and close together; one pair of sublateral body pores anterior (about four vulval body diam.) to vulva and one pair of post-advulvar sublateral body pores. Trichodorus pseudoaequalis n. sp. differs from the most similar species, T. aequalis, in male characters like general spicule shape and ornamentation and in the position of the dorsal pharyngeal gland nucleus in both sexes (at same level of posterior ventrosublateral pair vs clearly separated). The phylogenetic relationships of the recovered species were reconstructed using the ITS2 rRNA and the D2-D3 expansion segments of 28S rRNA gene sequences.

In: Nematology

Summary

Molecular characterisation of two species of Meloinema: M. chitwoodi from Oregon, USA, and M. odesanens from South Korea, is given based on the partial 18S rRNA, the D2-D3 of 28S rRNA, ITS rRNA, and COI gene sequences. In the phylogenetic trees, Meloinema clustered with Meloidogyne, in a basal position and more closely with Meloidogyne indica and M. nataliei. The Shimodaira-Hasegawa (SH) maximum likelihood testing of an alternative topology with two gene fragments (D2-D3 of 28S rRNA and 18S rRNA genes) did not reject a sister relationship of Meloidogyne and Meloinema. Molecular results confirmed the view of Siddiqi (2000) that Meloidogyne and Meloinema evolved from a Pratylenchidae-type ancestor. The clade Meloinema + Meloidogyne + Nacobbus was rejected by the SH test of the D2-D3 of 28S rRNA gene sequence dataset. The molecular results suggested that the genus Nacobbus should be placed not in the Meloidogynidae, but in a separate subfamily, the Nacobbinae, under the family Pratylenchidae.

In: Nematology
Volume 8 in the series, appearing in two parts, A and B, deals with the systematics of cyst nematodes of the subfamily Heteroderinae. Cyst nematodes are important pathogens causing extensive damage and significant yield loss to many crops in both temperate and tropical regions. Part A presents summarised information on various aspects of morphology, biology, ecology, pathology, biogeography, control measures and diagnostics of these nematodes. Molecular diagnostic tables and comprehensive tabular and dichotomous keys for species identification, together with descriptions of protocols for extraction, light and electron microscopy studies, and molecular diagnostics are also included. The taxonomic section of part A provides diagnoses for six genera, viz., Globodera, Punctodera, Cactodera, Dolichodera, Betulodera and Paradolichodera, and includes descriptions and morphometrics of 30 valid species. The work is illustrated by 101 drawings and photos. The genus Heterodera is mainly covered in part B, complete descriptions and morphometrics of the 80 known valid species being provided and the species illustrated by 159 line drawings and photos. Identification of the species using morphological and molecular techniques is also covered. Each part includes an extensive bibliography.

Pin nematodes of the genus Paratylenchus are widely distributed across the world and associated with many plant species. Morphological identification of Paratylenchus species is a difficult task because it relies on many characters with a wide range of intraspecific variation. In this study we provide morphological and molecular characterisation of several pin nematodes: Paratylenchus aquaticus, P. dianthus, P. hamatus, P. nanus and P. straeleni, collected in different states of the USA and South Africa. Paratylenchus aquaticus is reported from South Africa and Hawaii and P. nanus is found from South Africa for the first time. Morphological descriptions, morphometrics, light and scanning electron microscopic photos and drawings are given for these species. Molecular characterisation of nematodes using the D2-D3 of 28S rRNA and ITS rRNA gene sequence revealed that samples morphologically identified as P. aquaticus, P. hamatus and P. nanus indeed represent species complexes containing several species. Sequences of the rRNA genes are also provided for several unidentified Paratylenchus. Phylogenetic relationships within the genus Paratylenchus are given as inferred from the analyses of the D2-D3 of 28S rRNA and ITS rRNA gene sequences. We present here the most complete phylogenetic analysis of the genus.

In: Nematology

Abstract

A method for rapid identification of juveniles and cysts of the soybean cyst nematode based on PCR with species specific primers is described. The PCR assay was tested on 53 populations originating from China, Russia, USA and Brazil. A single cyst or second stage juvenile of Heterodera glycines alone or in a mixture with other soil inhabiting nematodes was detectable.

In: Nematology

This contribution presents a study of Iberian and Californian populations of Aporcelaimellus simplex, including morphological, morphometric and molecular data. The species is characterised and distinguished by its lip region offset by an expansion, pars refringens vaginae absent, and conical tail with a small but distinct hyaline terminal portion. No significant difference exists between American and European populations. Molecular data and the derived evolutionary tree show a topology in which this species forms a well-supported group with members of Discolaiminae, far from other representatives of Aporcelaimellus. Putting special emphasis in the absence of pars refringens vaginae, A. simplex is transferred to the genus Aporcella. The taxonomy of this genus is revised in depth, with the proposal of an emended diagnosis, the provision of a list of 13 valid species (mostly new combinations from Aporcelaimellus) as well as a key to their identification and a compendium of their main morphometrics. Aporcella debruinae sp. n. is proposed for Aporcelaimellus papillatus apud de Bruin & Heyns, 1992.

In: Nematology