Nematode Colonization Research Articles

Pinus thunbergii Parl. Somatic Plants' Resistance to Bursaphelenchus xylophilus Depends on Pathogen-Induced Differential Transcriptomic Responses.

Pinus thunbergii Parl. is an economically and medicinally important plant, as well as a world-renowned horticultural species of the Pinus genus. Pine wilt disease is a dangerous condition that affects P. thunbergii. However, understanding of the genetics underlying resistance to this disease is poor. Our findings reveal that P. thunbergii's resistance mechanism is based on differential transcriptome responses generated by the early presence of the pathogen Bursaphelenchus xylophilus, also known as the pine wood nematode. A transcriptome analysis (RNA-seq) was performed to examine gene expression in shoot tissues from resistant and susceptible P. thunbergii trees. RNA samples were collected from the shoots of inoculated pines throughout the infection phases by the virulent Bursaphelenchus xylophilus AMA3 strain. The photosynthesis and plant-pathogen interaction pathways were significantly enriched in the first and third days after infection. Flavonoid biosynthesis was induced in response to late infestation (7 and 14 days post-infestation). Calmodulin, RBOH, HLC protein, RPS, PR1, and genes implicated in phytohormone crosstalk (e.g., SGT1, MYC2, PP2C, and ERF1) showed significant alterations between resistant and susceptible trees. Furthermore, salicylic acid was found to aid pine wood nematodes tolerate adverse conditions and boost reproduction, which may be significant for pine wood nematode colonization within pines. These findings provide new insights into how host defenses overcame pine wood nematode infection in the early stage, which could potentially contribute to the development of novel strategies for the control of pine wilt disease.

Local climate modulates the development of soil nematode communities after glacier retreat

AbstractThe worldwide retreat of glaciers is causing a faster than ever increase in ice‐free areas that are leading to the emergence of new ecosystems. Understanding the dynamics of these environments is critical to predicting the consequences of climate change on mountains and at high latitudes. Climatic differences between regions of the world could modulate the emergence of biodiversity and functionality after glacier retreat, yet global tests of this hypothesis are lacking. Nematodes are the most abundant soil animals, with keystone roles in ecosystem functioning, but the lack of global‐scale studies limits our understanding of how the taxonomic and functional diversity of nematodes changes during the colonization of proglacial landscapes. We used environmental DNA metabarcoding to characterize nematode communities of 48 glacier forelands from five continents. We assessed how different facets of biodiversity change with the age of deglaciated terrains and tested the hypothesis that colonization patterns are different across forelands with different climatic conditions. Nematodes colonized ice‐free areas almost immediately. Both taxonomic and functional richness quickly increased over time, but the increase in nematode diversity was modulated by climate, so that colonization started earlier in forelands with mild summer temperatures. Colder forelands initially hosted poor communities, but the colonization rate then accelerated, eventually leveling biodiversity differences between climatic regimes in the long term. Immediately after glacier retreat, communities were dominated by colonizer taxa with short generation time and r‐ecological strategy but community composition shifted through time, with increased frequency of more persister taxa with K‐ecological strategy. These changes mostly occurred through the addition of new traits instead of their replacement during succession. The effects of local climate on nematode colonization led to heterogeneous but predictable patterns around the world that likely affect soil communities and overall ecosystem development.

Whale falls as chemosynthetic refugia: a perspective from free-living deep-sea nematodes

Whale carcasses create habitats in the deep sea which are colonized by organisms related to other chemosynthetic environments suggesting that whale falls may act as intermediate refugia for the dispersal and evolution of deep-sea organisms. Such evidence comes mainly from macrofaunal organisms whereas for the smaller meiofauna, data on whalebone assemblages is lacking. In this study, we investigated nematode colonization of whalebones experimentally deployed at 1500 and 3300 m depth and bones from a natural whale carcass found at 4204 m, in the Southwest Atlantic Ocean, off Brazil. By comparing whalebone assemblages to other deep-sea environments, we tested the hypothesis that whale fall nematode assemblages are more similar to those from other cognate chemosynthetic-based habitats, rather than those from sediments surrounding the carcass or other deep-sea habitats. Our results showed that whalebone nematode assemblages resemble those of hydrothermal vents and cold seeps and suggest that organic falls may act as intermediate refugia for meiofauna from chemosynthetic environments. It also showed that oceanographic conditions and the age of the carcasses on the ocean floor could influence nematode assemblage composition and richness. Such findings highlight the importance of organic falls for understanding the connectivity and phylogeny of benthic organisms, including representatives of the meiofauna.

Xenorhabdus nematophila bacteria shift from mutualistic to virulent Lrp-dependent phenotypes within the receptacles of Steinernema carpocapsae insect-infective stage nematodes.

Xenorhabdus nematophila bacteria are mutualists of Steinernema carpocapsae nematodes and pathogens of insects. Xenorhabdus nematophila exhibits phenotypic variation between insect virulence (V) and the mutualistic (M) support of nematode reproduction and colonization initiation in the infective juvenile (IJ) stage nematode that carries X. nematophila between insect hosts. The V and M phenotypes occur reciprocally depending on levels of the transcription factor Lrp: high-Lrp expressors are M+V- while low-Lrp expressors are V+M-. We report here that variable (wild type) or fixed high-Lrp expressors also are optimized, relative to low- or no-Lrp expressors, for colonization of additional nematode stages: juvenile, adult and pre-transmission infective juvenile (IJ). In contrast, we found that after the bacterial population had undergone outgrowth in mature IJs, the advantage for colonization shifted to low-Lrp expressors: fixed low-Lrp expressors (M-V+) and wild type (M+V+) exhibited higher average bacterial CFU per IJ than did high-Lrp (M+V-) or no-Lrp (M-V-) strains. Further, the bacterial population becomes increasingly low-Lrp expressing, based on expression of an Lrp-dependent fluorescent reporter, as IJs age. These data support a model that virulent X. nematophila have a selective advantage and accumulate in aging IJs in advance of exposure to insect hosts in which this phenotype is necessary.

Decreased Root-Knot Nematode Gall Formation in Roots of the Morning Glory Ipomoea tricolor Symbiotic with Ergot Alkaloid-Producing Fungal Periglandula Sp.

Many species of morning glories (Convolvulaceae) form symbioses with seed-transmitted Periglandula fungal endosymbionts, which produce ergot alkaloids and may contribute to defensive mutualism. Allocation of seed-borne ergot alkaloids to various tissues of several Ipomoea species has been demonstrated, including roots of I. tricolor. The goal of this study was to determine if infection of I. tricolor by the Periglandula sp. endosymbiont affects Southern root-knot nematode (Meloidogyne incognita) gall formation and host plant biomass. We hypothesized that I. tricolor plants infected by Periglandula (E+) would develop fewer nematode-induced galls compared to non-symbiotic plants (E-). E+ or E- status of plant lines was confirmed by testing methanol extracts from individual seeds for endosymbiont-produced ergot alkaloids. To test the effects of Periglandula on nematode colonization, E+ and E- I. tricolor seedlings were grown in soil infested with high densities of M. incognita nematodes (N+) or no nematodes (N-) for four weeks in the greenhouse before harvesting. After harvest, nematode colonization of roots was visualized microscopically, and total gall number and plant biomass were quantified. Four ergot alkaloids were detected in roots of E+ plants, but no alkaloids were found in E- plants. Gall formation was reduced by 50% in E+ plants compared to E- plants, independent of root biomass. Both N+ plants and E+ plants had significantly reduced biomass compared to N- and E- plants, respectively. These results demonstrate Periglandula's defensive role against biotic enemies, albeit with a potential trade-off with host plant growth.

The Mortality of Nematodes in Drinking Water in the Presence of Ozone, Chlorine Dioxide, and Chlorine

ABSTRACTNematodes are among the organisms most commonly found in treated water. Although they are believed to be harmless, colonization of nematodes by pathogenic bacteria may transform them into harmful contaminants of the drinking water. Thus, efficient disinfectants must be applied to inactivate and remove nematodes. In the present study, nematode inactivation using different concentrations of ozone, chlorine dioxide, and chlorine for different exposure times are described. Three different disinfection models were tested. Based on experimental data, the best statistically significant relationship between mortality of nematodes in probits (Y), concentrations of disinfectants mg/L (C), and the exposure times in minutes (t) was established as Y = a*log(C) + b*log(t) + c. The estimated values of regression coefficients a, b, and c were 6.89, 0.76, and 1.13 for ozone, 0.66, 1.23, and 5.28 for chlorine dioxide and 1.59, 0.92, and 3.06 for chlorine, respectively. Comparison of the predicted LC(90) molar concentrations of the investigated disinfectants, chlorine dioxide, ClO2, was found to be the most effective disinfectant, followed by ozone, O3, while chlorine, Cl2, was the least efficient disinfectant for inactivation of nematodes.

Rapid Changes in Soil Nematodes in the First Years after Technosol Construction for the Remediation of an Industrial Wasteland

Technosol construction is an emergent technology that uses an assemblage of technogenic materials for the ecological reclamation of derelict land and waste recycling. Knowledge about the colonisation of Technosols by soil biota is limited, despite the latter’s central role in ecosystem functioning. In this four-year field (2008 to 2011) study, we characterized the development over time of the diversity and the abundance of soil nematodes in two types of Technosols in North-Eastern France. We also studied the nematode community structure, abundance of taxa and functional groups in both Technosol profiles in the third year of the study. Samples were collected from the top soil layer (0–20 cm) each year in the spring (April), on a one ha. field experiment that had spatially divided in 24 sampling areas. For soil profiles, three samples were collected in three horizons within six pits (three pits per Technosol). Nematodes were extracted from soil and identified at the family or genus level and then classified into functional feeding guilds. In the first year, the community was dominated by opportunistic bacterial feeders. The taxonomic and functional nematode diversity increased with time, with a dominance of non-opportunistic bacterial feeders after four years, but also the significant presence of fungal feeders, omnivorous and carnivorous, as well as plant parasites and insect parasites. No significant difference was observed between the two Technosols. Each layer showed distinct communities, with nematode diversity and abundance decreasing with depth. Abundance and diversity, coupled with the analysis of several indexes, commonly used for nematodes, including Maturity index (MI), Enrichment index (EI), Structure index (SI) and Nematode channel ratio (NCR), lead to the conclusion that the high organic matter content, particularly in the upper horizon of both Technosols, guaranteed nematode colonization and progressive diversification, and is likely to be the key for successful biodiversity reclamation.

The C-terminal coiled-coil domain of Corynebacterium diphtheriae DIP0733 is crucial for interaction with epithelial cells and pathogenicity in invertebrate animal model systems

BackgroundCorynebacterium diphtheriae is the etiologic agent of diphtheria and different systemic infections. The bacterium has been classically described as an extracellular pathogen. However, a number of studies revealed its ability to invade epithelial cells, indicating a more complex pathogen-host interaction. The molecular mechanisms controlling and facilitating internalization of C. diphtheriae still remains unclear. Recently, the DIP0733 transmembrane protein was found to play an important role in the interaction with matrix proteins and cell surfaces, nematode colonization, cellular internalization and induction of cell death.ResultsIn this study, we identified a number of short linear motifs and structural elements of DIP0733 with putative importance in virulence, using bioinformatic approaches. A C-terminal coiled-coil region of the protein was considered particularly important, since it was found only in DIP0733 homologs in pathogenic Corynebacterium species but not in non-pathogenic corynebacteria. Infections of epithelial cells and transepithelial resistance assays revealed that bacteria expressing the truncated form of C. diphtheriae DIP0733 and C. glutamicum DIP0733 homolog are less virulent, while the fusion of the coiled-coil sequence to the DIP0733 homolog from C. glutamicum resulted in increased pathogenicity. These results were supported by nematode killing assays and experiments using wax moth larvae as invertebrate model systems.ConclusionsOur data indicate that the coil-coiled domain of DIP0733 is crucial for interaction with epithelial cells and pathogenicity in invertebrate animal model systems.

Strains of Photorhabdus spp. associated with polish Heterorhabditis isolates: their molecular and phenotypic characterization and symbiont exchange.

The relationships between six bacterial symbionts of the entomopathogenic nematodes Heterorhabditis bacteriophora and Heterorhabditis megidis from Poland to species and subspecies of the genus Photorhabdus were evaluated. This study was based on phylogenetic analysis of sequence data of five genes: 16S rRNA, gyrB, recA, gltX, and dnaN. The bacteria were also characterized phenotypically by biochemical and physiological tests. Our results have revealed that the Photorhabdus strains isolated from H. megidis belong to P. temperata, subsp. temperata and subsp. cinerea. Isolates from H. bacteriophora represent P. luminescens subs. kayaii and P. temperata subs. cinerea. This study for the first time provides evidence for H. bacteriophora and P. temperata subsp. cinerea symbiotic association. In addition, we tested whether the microsymbionts of the Polish H. bacteriophora and H. megidis isolates support the development of non-native nematode host population and colonization of their infective juveniles. It has been shown that the studied Photorhabdus strains can readily swap their nematode host, both at intra- and interspecies level. It supports the hypothesis of different symbiotic associations in the Heterorhabditis-Photorhabdus lineage.

Pathogen-nematode interaction: Nitrogen supply of Listeria monocytogenes during growth in Caenorhabditis elegans.

Listeria monocytogenes is a Gram-positive facultatively intracellular human pathogen. Due to its saprophytic lifestyle, L. monocytogenes is assumed to infect and proliferate within soil organisms such as Caenorhabditis elegans. However, little is known about the nutrient usages and metabolite fluxes in this bacterium-nematode interaction. Here, we established a nematode colonization model for L. monocytogenes and a method for the efficient separation of the pathogen from the nematodal gut. Following (15)N labelling of C. elegans and gas chromatography-mass spectrometry-based (15)N isotopologue analysis, we detected a high basal metabolic rate of the nematode, and observed a significant metabolic flux from nitrogenous compounds of the nematode to listerial proteins during proliferation of the pathogen in the worm's intestine. For comparison, we also measured the N fluxes from the gut content into listerial proteins using completely (15)N-labelled Escherichia coli OP50 as food for C. elegans. In both settings, L. monocytogenes prefers the direct incorporation of histidine, arginine and lysine over their de novo biosynthesis. Our data suggest that colonization of nematodes is a strategy of L. monocytogenes to increase its access to N-rich nutrients.

Method for fluorescent marker swapping and its application in Steinernema nematode colonization studies

An allelic exchange vector was constructed to replace gfp by mCherry in bacteria previously tagged with mini-Tn5 derivatives. The method was successfully applied to a gfp-labeled Yersinia pseudotuberculosis strain and the re-engineered bacterium was used to study the colonization of Steinernema nematodes hosting their Xenorhabdus symbiont using dual-color confocal microscopy.

The importance of microphytic composition on coarse woody debris for nematode colonization: a case study in a fluvial floodplain environment

Coarse woody debris (CWD) are an important structural feature of many aquatic ecosystems. This study focused on the relationships between nematode colonization and microphytic community structure on CWD. In situ investigations were performed using white willow branches introduced into the floodplain lake, a part of the large natural floodplain in the middle section of the Danube River (Europe) impacted by strong sandy and silt accumulations. Our results showed that CWD support rapid colonization and high species richness of microphytes, regardless of seasonality and flood-induced environmental changes. Successional stages during the accrual of microphytic communities were expressed. Nematode functional feeding group compositions closely followed microphytic composition. The epistrate feeder Chromadorina bioculata over-dominated in diatom-dominated communities with adnate/prostrate and stalk-forming species in spring and autumn. The complex structure of cyanobacterial mats associated with green algae and diatom assemblages in summer attracted suction feeders, represented by the Dorylaimidae family. The obtained results indicated that nematodes are involved in strong trophic coupling with a microbial habitat on CWD. The retention of CWD in aquatic environments or addition of CWD to systems with little or no wood may have important implications for the primary and secondary production in these ecosystems.

NilD CRISPR RNA contributes to Xenorhabdus nematophila colonization of symbiotic host nematodes.

The bacterium Xenorhabdus nematophila is a mutualist of entomopathogenic Steinernema carpocapsae nematodes and facilitates infection of insect hosts. X. nematophila colonizes the intestine of S. carpocapsae which carries it between insects. In the X. nematophila colonization-defective mutant nilD6::Tn5, the transposon is inserted in a region lacking obvious coding potential. We demonstrate that the transposon disrupts expression of a single CRISPR RNA, NilD RNA. A variant NilD RNA also is expressed by X. nematophila strains from S. anatoliense and S. websteri nematodes. Only nilD from the S. carpocapsae strain of X. nematophila rescued the colonization defect of the nilD6::Tn5 mutant, and this mutant was defective in colonizing all three nematode host species. NilD expression depends on the presence of the associated Cas6e but not Cas3, components of the Type I-E CRISPR-associated machinery. While cas6e deletion in the complemented strain abolished nematode colonization, its disruption in the wild-type parent did not. Likewise, nilD deletion in the parental strain did not impact colonization of the nematode, revealing that the requirement for NilD is evident only in certain genetic backgrounds. Our data demonstrate that NilD RNA is conditionally necessary for mutualistic host colonization and suggest that it functions to regulate endogenous gene expression.

Survival and colonization of nematodes in a composting process

AbstractNematodes are omnipresent in composts and are active in virtually all stages of the composting process. Major shifts in species composition, life strategies, and feeding behavior occur during the composting process. Due to the heat peak, nematodes can be virtually absent, but several taxa appear immediately when the temperatures drop. These comprise both taxa present before the heat peak and new taxa. However, it is not known how nematodes populate the compost. In this study, we aimed to assess the survival and colonization capacity of nematodes in compost. Our results showed that composting processes inaccessible to insects or not in contact with soil did not significantly influence nematode succession during composting. However, differences between treatments were found for some specific taxa (i.e., for Acrostichus sp., Neodiplogasteridae sp., Nygolaimoides sp., and Rhabditidae sp. 1), illustrating the importance of insects for the dispersal of nematodes to compost. Experiments in the lab with the blue bottle fly as a possible carrier demonstrated actual transport of nematodes isolated from compost by the fly (i.e., Halicephalobus cfr. gingivalis, Diploscapter coronatus, Diplogasteritus sp., Acrostichus sp., and Mesorhabditis sp.). Juveniles and dauer stages of Aphelenchoides sp., Panagrolaimus sp., and rhabditids survived an experimentally induced temperature peak, while members of Tylenchidae did not. In conclusion, our results indicate that the rapidly changing nematode community in compost is the result of both differential survival and colonization capacities.

Impact of nematode-induced galls on Miconia prasina (Sw.) DC (Melastomataceae) traits in the Atlantic forest of northeastern Brazil

We tested the hypothesis that parasitic Ditylenchus sp., leaf gall inductor, decrease plant performance by reducing the number of leaves, shoots, and plant height, increasing herbivory damage and nematode colonization in aggregate plants. We compared the effects on 30 paired healthy and attacked individuals of Miconia prasina. Nematode infection did not affect the number of leaves and shoots and plant height. The damage by free-feeding herbivores was greater in healthy plants, while the number of insect galls was higher in attacked plants. The number of leaf miners did not vary. Healthy plants were more grouped than the attacked plants, and the mean plant distance and the index of nematode attack were positively related.

Mutational Analyses Reveal Overall Topology and Functional Regions of NilB, a Bacterial Outer Membrane Protein Required for Host Association in a Model of Animal-Microbe Mutualism

The gammaproteobacterium Xenorhabdus nematophila is a mutualistic symbiont that colonizes the intestine of the nematode Steinernema carpocapsae. nilB (nematode intestine localization) is essential for X. nematophila colonization of nematodes and is predicted to encode an integral outer membrane beta-barrel protein, but evidence supporting this prediction has not been reported. The function of NilB is not known, but when expressed with two other factors encoded by nilA and nilC, it confers upon noncognate Xenorhabdus spp. the ability to colonize S. carpocapsae nematodes. We present evidence that NilB is a surface-exposed outer membrane protein whose expression is repressed by NilR and growth in nutrient-rich medium. Bioinformatic analyses reveal that NilB is the only characterized member of a family of proteins distinguished by N-terminal region tetratricopeptide repeats (TPR) and a conserved C-terminal domain of unknown function (DUF560). Members of this family occur in diverse bacteria and are prevalent in the genomes of mucosal pathogens. Insertion and deletion mutational analyses support a beta-barrel structure model with an N-terminal globular domain, 14 transmembrane strands, and seven extracellular surface loops and reveal critical roles for the globular domain and surface loop 6 in nematode colonization. Epifluorescence microscopy of these mutants demonstrates that NilB is necessary at early stages of colonization. These findings are an important step in understanding the function of NilB and, by extension, its homologs in mucosal pathogens.

Evidence of differences between the communities of arbuscular mycorrhizal fungi colonizing galls and roots of Prunus persica infected by the root-knot nematode Meloidogyne incognita.

Arbuscular mycorrhizal fungi (AMF) play important roles as plant protection agents, reducing or suppressing nematode colonization. However, it has never been investigated whether the galls produced in roots by nematode infection are colonized by AMF. This study tested whether galls produced by Meloidogyne incognita infection in Prunus persica roots are colonized by AMF. We also determined the changes in AMF composition and biodiversity mediated by infection with this root-knot nematode. DNA from galls and roots of plants infected by M. incognita and from roots of noninfected plants was extracted, amplified, cloned, and sequenced using AMF-specific primers. Phylogenetic analysis using the small-subunit (SSU) ribosomal DNA (rDNA) data set revealed 22 different AMF sequence types (17 Glomus sequence types, 3 Paraglomus sequence types, 1 Scutellospora sequence type, and 1 Acaulospora sequence type). The highest AMF diversity was found in uninfected roots, followed by infected roots and galls. This study indicates that the galls produced in P. persica roots due to infection with M. incognita were colonized extensively by a community of AMF, belonging to the families Paraglomeraceae and Glomeraceae, that was different from the community detected in roots. Although the function of the AMF in the galls is still unknown, we hypothesize that they act as protection agents against opportunistic pathogens.

Seed treatment affects Meloidogyne incognita penetration, colonization and reproduction on cotton

The effect of abamectin, applied as seed treatment, on Meloidogyne incognita penetration, colonization, and reproduction on cotton was evaluated in two greenhouse experiments. Second stage juveniles (J2) penetration was evaluated 3, 9 and 15 days after germination (dag), whereas the nematode colonization was evaluated 27dag, by counting the total number of galls and egg masses per root system. Eggs and J2 were extracted from the roots to assess the nematode reproduction at 50 and 100dag. The seed treatment caused a decrease in J2 penetration in the roots, resulting in lower colonization and reproduction of M. incognita. These findings are discussed considering the environmental aspects of abamectin in the soil and the current practices for the crop management.

Examination of Xenorhabdus nematophila Lipases in Pathogenic and Mutualistic Host Interactions Reveals a Role for xlpA in Nematode Progeny Production

Xenorhabdus nematophila is a gammaproteobacterium and broad-host-range insect pathogen. It is also a symbiont of Steinernema carpocapsae, the nematode vector that transports the bacterium between insect hosts. X. nematophila produces several secreted enzymes, including hemolysins, lipases, and proteases, which are thought to contribute to virulence or nutrient acquisition for the bacterium and its nematode host in vivo. X. nematophila has two lipase activities with distinct in vitro specificities for Tween and lecithin. The gene encoding the Tween-specific lipase, xlpA, has been identified and is not required for X. nematophila virulence in one insect host, the tobacco hornworm Manduca sexta. However, the gene encoding the lecithin-specific lipase activity is not currently known. Here, we identify X. nematophila estA, a gene encoding a putative lecithinase, and show that an estA mutant lacks in vitro lipase activity against lecithin but has wild-type virulence in Manduca sexta. X. nematophila secondary-form phenotypic variants have higher in vitro lecithinase activity and estA transcript levels than do primary-form variants, and estA transcription is negatively regulated by NilR, a repressor of nematode colonization factors. We establish a role for xlpA, but not estA, in supporting production of nematode progeny during growth in Galleria mellonella insects. Future research is aimed at characterizing the biological roles of estA and xlpA in other insect hosts.

1-Aminocyclopropane-1-carboxylic acid (ACC) concentration and ACC synthase expression in soybean roots, root tips, and soybean cyst nematode (Heterodera glycines)-infected roots

Colonization of plant roots by root knot and cyst nematodes requires a functional ethylene response pathway. However, ethylene plays many roles in root development and whether its role in nematode colonization is direct or indirect, for example lateral root initiation or root hair growth, is not known. The temporal requirement for ethylene and localized synthesis of ethylene during the life span of soybean cyst nematode (SCN) on soybean roots was further investigated. Although a significant increase in ethylene evolution was not detected from SCN-colonized roots, the concentration of the immediate precursor to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC), was higher in SCN-colonized root pieces and root tips than in other parts of the root. Moreover, expression analysis of 17 ACC synthase (ACS) genes indicated that a select set of ACS genes is expressed in SCN-colonized root pieces that is clearly different from the set of genes expressed in non-colonized roots or root tips. Semi-quantitative real-time PCR indicated that ACS transcript accumulation correlates with the high concentration of ACC in root tips. In addition, an ACS-like sequence was found in the public SCN nucleotide database. Acquisition of a full-length sequence for this mRNA (accession GQ389647) and alignment with transcripts for other well-characterized ACS proteins indicated that the nematode sequence is missing a key element required for ACS activity and therefore probably is not a functional ACS. Moreover, no significant amount of ACC was found in any growth stage of SCN that was tested.