Plant resistance is currently the most effective and environmentally safe method to control plant parasitic nematodes (PPNs). Resistance genes generally act against sedentary PPNs by inducing a hypersensitive reaction that prevents the parasite installation and/or reproduction. However, the recent emergence of virulent biotypes able to overcome the plant resistance genes may constitute a severe limitation to this control strategy. In selection experiments conducted under controled environment, the genetic variation, specificity and inheritance of nematode virulence have been demonstrated. Moreover, the occurrence of gene-for-gene interactions has been shown in a few cases. Moleculars markers have been extensively used to investigate the genetic variability of PPNs, but so far, the genomic polymorphisms observed are largely independent of virulence. Such data suggest that, within a species, virulent isolates do not share a common origin, but are probably the result of independent mutational events. To understand the molecular mechanisms responsible for virulence in PPNs, several strategies have been developed, in relation with their mode of reproduction (parthenogenesis versus amphimixis). As an example, recent results obtained in our laboratory on the root-knot nematodes Meloidogyne spp. are presented. On a more general point of view, factors that may induce stable genome variability in PPNs, e.g. Transposition of mobile elements and chromosomal rearrangements (leading to polyploidy, aneuploidy, etc) will also be considered. Advances in knowledge in these areas should have important consequences for the management and durability of natural resistance genes, and for the engineering of new forms of resistance.

Genetic variability of nematodes: a threat to the durability of plant resistance genes?

Subbotin, S. A., Madani, M., Krall, E., Sturhan, D., and Moens, M. 2005. Molecular diagnostics, taxonomy, and phylogeny of the stem nematode Ditylenchus dipsaci species complex based on the sequences of the internal transcribed spacer-rDNA. Phytopathology 95:1308-1315. The stem nematode Ditylenchus dipsaci is of great economic importance worldwide as a parasite of agricultural crops and horticultural plants. The internal transcribed spacer (ITS) of rDNA from 23 populations of the D. dipsaci complex from various host plants were amplified and sequenced. Seven previously studied populations were also included in the study. The phylogenetic analysis of the full ITS and ITS2 sequence alignments using minimum evolution, maximum parsimony, and Bayesian inference under the complex model of DNA evolution revealed trees with two main clades: (i) D. dipsaci sensu stricto with diploid chromosome numbers and comprising most isolates from agricultural, ornamental, and several wild plants, and (ii) Ditylenchus spp. with polyploid chromosome numbers, reproductively isolated from diploid populations, and subdivided into six subclades (“giant race” from Vicia faba, Ditylenchus species parasitizing various Asteraceae, and a Ditylenchus sp. from Plantago maritima). Using the energy minimization approach and comparative sequence analysis, it has been found that the secondary structure of ditylenchid ITS2 is organized in three main domains. The importance of knowledge on the RNA structure for phylogenetic analysis is discussed. Conventional polymerase chain reaction (PCR) and real-time PCR with SYBR green dye I with a species specific primer have been developed for detection and quantification of D. dipsaci sensu stricto. Validation tests revealed a rather high correlation between real numbers of fourth-stage juveniles of the stem nematodes in a sample and expected numbers detected by real-time PCR. Problems of accuracy of quantification are discussed.

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Molecular Diagnostics, Taxonomy, and Phylogeny of the Stem Nematode Ditylenchus dipsaci Species Complex Based on the Sequences of the Internal Transcribed Spacer-rDNA.

The potential phytosanitary importance of all named plant-parasitic nematode species was determined by evaluating available information on species characteristics, association with economically-important crop hosts, and ability to act as vectors of viruses or form disease complexes with other pathogens. Most named species of plant-parasitic nematodes (PPN) are poorly known, recorded from a single location only, not associated with economically-important crops, and not known to be associated with other plant disease organisms. However, 250 species from 43 genera fulfilled one or more of the criteria to be considered to present a phytosanitary risk. The genera and number of species (in parentheses) considered as posing phytosanitary risk included: Achlysiella (1), Anguina (8), Aphasmatylenchus (1), Aphelenchoides (12), Aphelenchus (1), Belonolaimus (2), Bitylenchus (3), Bursaphelenchus (4), Cactodera (3), Ditylenchus (8), Dolichodorus (1), Globodera (3), Helicotylenchus (7), Hemicriconemoides (3), Hemicycliophora (3), Heterodera (25), Hirschmanniella (5), Hoplolaimus (5), Ibipora (3), Longidorus (10), Macroposthonia (2), Meloidogyne (38), Merlinius (3), Nacobbus (1), Neodolichodorus (2), Paralongidorus (2), Paratrichodorus (11), Paratylenchus (3), Pratylenchus (24), Punctodera (3), Quinisulcius (3), Radopholus (5), Rotylenchulus (3), Rotylenchus (1), Scutellonema (5), Sphaeronema (1), Subanguina (3), Trichodorus (5), Tylenchorhynchus (8), Tylenchulus (2), Vittatidera (1), Xiphinema (15) and Zygotylenchus (1). For each of the 250 species main hosts and yield loss estimates are provided with an extensive bibliography. Of the 250 species, only 126 species from 33 genera are currently listed as regulated pests in one or more countries worldwide. Almost all of these 250 species were also associated with economically important crops and some also acted as vectors for viruses. © 2013 The Authors. Journal compilation

Plant-parasitic nematodes of potential phytosanitary importance, their main hosts and reported yield losses

The soil as a reservoir for natural enemies of pest insects and mites with emphasis on fungi and nematodes

Morphologial, biochemical, molecular and karyological analyses of different populations and races of the stem and bulb nematode Ditylenchus dipsaci have suggested that it represents a species complex, of which only D. dipsaci sensu stricto and its morphologically larger variant, known as the giant race of the stem and bulb nematode, are plant parasites of economic importance. The present study singles out the giant race from this complex, herein described as a new species named Ditylenchus gigas n. sp., on the basis of morphological and molecular data obtained from several populations collected from broad beans in southern Italy, southern Spain and Lebanon. The new species epithet, which refers to the large body size of the nematode with respect to the normal races, must be considered to be conspecific with the D. dipsaci ‘giant race’ from Fabaceae in recent literature. Morphologically, the new species is characterized by a body size 1AE5‐2 times longer than the ‘normal race’, stylet delicate (11AE5‐13AE0 lm long) with knobs distinctly sloping backwards, and long post-vulval uterine sac (81‐150 lm long). Results of molecular analysis of rDNA sequences including the ITS1-5.8S-ITS2 region, the D2‐D3 fragment of the 28S gene, the small 18S subunit, the partial mitochondrial gene for cytochrome c oxidase I (mtCOI), and hsp90 gene sequences, support the new taxonomic species status for the former D. dipsaci giant race from Vicia faba, and clearly distinguish D. gigas n. sp. from D. dipsaci sensu stricto.

Ditylenchus gigas n. sp. parasitizing broad bean: a new stem nematode singled out from the Ditylenchus dipsaci species complex using a polyphasic approach with molecular phylogeny

The wide geographic distribution of species of this genus may be interpreted as evidence of an ancient origin of the genus. Stem nematode, stem and bulb nematode 1 A total of 13 nominal species have been synonymized. Most species were generally characterized and distinguished from other nominal stem nematode species by certain host preferences. Stem nematodes in, for example, flower bulbs and seeds of lucerne, field beans, onions, and other Allium species are on the list of quarantine organisms of many countries all over the world. Stem nematodes were found in leaf galls of the false dandelion H.radicata and species of the genus in several European countries and in the western USA, where they are dispersed by seeds of the weed. Leaves are often distorted, discolored and smaller, with reduced and twisted leaf stems or they may obtain a shape morphologically quite different from the normal; in many plant species galls or necrotic spots may develop.

Michał W. Brzeski

Papers

Stem and bulb nematodes, Ditylenchus spp.