Search Results (15)

Rhizosphere microbes, particularly bacteria, are essential for controlling plant-parasitic nematodes (PPNs) through various mechanisms. However, the plant’s age and the genetic composition of nematode populations can significantly influence the inhibitory effectiveness of these microbes against the beet cyst nematode Heterodera schachtii. In this study, rhizosphere microbes were isolated from 39-day-old and 69-day-old resistant oilseed radish plants to evaluate their impact on the penetration of the second-stage juveniles (J2s) originating from four genetically distinct H. schachtii populations. The suppression of J2s penetration by the attached microbes varied across the nematode populations, which displayed differing levels of aggressiveness toward the resistant oilseed radish. Furthermore, differences in the alpha and beta diversity of rhizosphere bacteria were observed between the 39-day-old and 69-day-old plants, leading to variations in the bacterial attachment among the four nematode populations. In summary, the effectiveness of resistant catch crops against H. schachtii is influenced by the pathogenicity of the nematode populations and their interactions with the rhizosphere microbial community shaped by the plant’s age. Full article

Cyst nematodes remain a major threat to global agricultural production, causing huge losses. To understand the parasitism of the cyst nematodes Heterodera trifolii (HT) and Heterodera schachtii (HS), we constructed whole-genome assemblies using short- and long-read sequencing technologies. The nematode genomes were 379 Mb and 183 Mb in size, with the integrated gene models predicting 40,186 and 18,227 genes in HT and HS, respectively. We found more than half of the genes predicted in HT (64.7%) and HS (53.2%) were collinear to their nearest neighbor H. glycines (HG). Large-scale duplication patterns in HT and segmental duplications of more than half of the orthologous genes indicate that the genome of HT is polyploid in nature. Functional analysis of the genes indicated that 65.6% of the HG genes existed within the HT genome. Most abundant genes in HT and HS were involved in gene regulation, DNA integration, and chemotaxis. Differentially expressed genes showed upregulation of cuticle structural constituent genes during egg and female stages and cytoskeletal motor activity-related genes in juvenile stage 2 (J2). Horizontal gene transfer analyses identified four new vitamin biosynthesis genes, pdxK, pdxH, pdxS, and fabG, of bacterial origin, to be first reported in HT and HS. Mitogenomes of HT, HS, and HG showed similar structure, composition, and codon usage. However, rates of substitution of bases in the gene nad4l were significantly different between HT and HS. The described genomes, transcriptomes, and mitogenomes of plant-parasitic nematodes HT and HS are potential bio-resources used to identify several strategies of control of the nematode. Full article

Plant pathogenic nematodes play an important role in crop production and are difficult to control. One of them is Heterodera schachtii—a cyst nematode, pathogenic to sugar beet. Nature suggests a natural way to resolve this problem by using nematode-trapping fungi, one of which is Pleurotus ostreatus. It is one of the most widespread mushrooms in the world. It is a valuable and widely cultivated edible mushroom with nematocidal properties. The mycelium of this mushroom produces toxin droplets that paralyze nematodes, after which the nematodes become infected with the hyphae filament, resulting in their death. This feature can be put to practical use as a natural nematocidal agent. In this paper, we have described studies on the variability of the nematocidal properties in the progeny of three wild strains of P. ostreatus (Po1, Po2, Po4), obtained by crossing dikaryons with monokaryons (Buller phenomenon). The toxicity of mycelium was tested against the model organism Caenorhabditis elegans and against sugar beet pest—H. schachtii. The results of the study allowed the selection of the progeny heterokaryotic mycelia with the best parameters useful for in vitro control of both nematodes. They differed in the activity against C. elegans and H. schachtii, and in the toxic knobs production. The results indicated that the progeny of the Po1 strain presented a good ability to produce hyphal knobs in the presence of C. elegans, and progenies of the Po4 strain presented good quality of growth in preferred temperatures (10–25 °C). Three strains Po1 5dix27, Po2 20dix21, and Po4 2dix1, as well as a maternal strain Po4 controlled H. schachtii by entwining cysts better than other strains. These strains were moderately effective against C. elegans. Strains Po2 15dix17, Po4 1dix18, and Po4 1dix30 may be considered good controlling isolates against both tested organisms. The results of the research also clearly suggest that the killing properties of P. ostreatus mycelia should not be assessed only by their ability to produce toxic hyphal knobs. Their nematocidal properties also depend on other mechanisms developed by mycelia, which is observed as the lethality of nematodes. The results of this research will lead to a natural way to protect plants against nematodes. The research also proved the nematocidal properties of the wild strains to reduce the H. schachtii population in the soil. Full article

Heterodera schachtii (Schmidt, 1871) (Nematoda: Heteroderidae) is one of the most widespread plant-parasitic nematodes (PPNs) associated with cabbages, which cause severe yield losses in cruciferous vegetables. This study aimed to improve the current understanding of the prevalence and detection of H. schachtii in the cabbage-growing areas of Niğde Province, Türkiye. Field surveys were conducted between November and December 2021, and 100 soil samples were collected immediately after cabbage harvesting. Heterodera schachtii populations were identified by morphological and morphometric methods along with Internal Transcribed Spacer of the ribosomal region (ITS-rDNA) and Cytochrome Oxidase Subunit (COI-mtDNA) sequencing. The mean body length of H. schachtii was 463 ± 7 μm, while stylet and hyaline length ranged between 20.7–27.8 μm and 20.1–32.1 μm, respectively. Nearly half of the surveyed areas were infested with H. schachtii with a 41% incidence rate. However, the Merkez District had the highest proportion of infested fields with an over 51% incidence rate. The population density was determined in 41 samples with a mean of 79.5 cysts per 250 g of soil. These results will help to determine the control and management strategies of H. schachtii. Full article

Cyst nematodes are plant parasitic nematodes infecting crops, causing extensive crop damage and annual losses, and affecting food production. The precise species identification is significant to initiate their control. The repeatable, less expensive, and less laborious distinguishing cyst nematode species using image processing and artificial intelligence can be advantageous. The objective of this study was to distinguish cyst nematodes belonging to the species Globodera pallida, Globodera rostochiensis, and Heterodera schachtii based on image parameters using artificial neural networks (ANN). The application of parameters selected from a set of 2172 textures of images in color channels L, a, b, X, Y, Z, R, G, B, V, U, and S to build classification models using a narrow neural network, medium neural network, wide neural network, trilayered neural network, WiSARD, multilayer perceptron, and RBF network is a great novelty of the present study. Algorithms allowed for distinguishing cyst nematode species with an average accuracy reaching 89.67% for a model developed using WiSARD. The highest correctness was obtained for H. schachtii and this species was distinguished from each other with the highest accuracy of 95–98% depending on the classifier. Whereas the highest number of misclassified cases occurred between G. pallida, G. rostochiensis belonging to the same genus Globodera. The developed procedure involving image parameters and artificial neural networks can be useful for non-destructive and objective distinguishing cyst nematode species. Full article

Genes of host plants and parasitic nematodes govern the plant–nematode interaction. The biological receptors and parasitism effectors are variable among plant species and nematode populations, respectively. In the present study, hatch testing and bioassays on cabbage, oilseed radish, and mustard were conducted to compare the biological characteristics among six populations of the beet cyst nematode Heterodera schachtii. Genetic patterns of the vap1 gene for the studied populations were distinct as shown by denaturing the gradient gel electrophoresis of PCR-amplified gene fragments. Concurrently, significant differences in the hatching rates, number of penetrated J2 in roots, and eggs/cyst ratios among the six nematode populations for the three cruciferous species were observed. In conclusion, analyzing the population genetic structure of H. schachtii plays a pivotal role in illustrating the variability in the plant–nematode interaction among its populations and plant species, which in its role leads to developing nematode management depending on plant resistance. Full article

Arabidopsis contains 317 genes for defensin-like (DEFL) peptides. DEFLs have been grouped into different families based mainly on cysteine motifs. The DEFL0770 group contains seven genes, of which four are strongly expressed in roots. We found that the expression of these genes is downregulated in syncytia induced by the beet cyst nematode Heterodera schachtii as revealed by RNAseq analysis. We have studied one gene of this group, At3g59930, in detail. A promoter::GUS line revealed that the gene is only expressed in roots but not in other plant organs. Infection of the GUS line with larvae of H. schachtii showed a strong downregulation of GUS expression in infection sites as early as 1 dpi, confirming the RNAseq data. The At3g59930 peptide had only weak antimicrobial activity against Botrytis cinerea. Overexpression lines had no enhanced resistance against this fungus but were more resistant to H. schachtii infection. Our data indicate that At3g59930 is involved in resistance to nematodes which is probably not due to direct nematicidal activity. Full article

Sugar beet cyst nematode (SBCN, Heterodera schachtii) is an important nematode that causes significant yield losses of 25–50% or more in most areas of sugar beet production worldwide. Rapid and accurate identification of this species is essential to support decisions on pest management. However, the difference between H. schachtii and other Heterodera spp. based on morphology is a challenging task. In the present study, a SCAR-PCR assay was developed to identify and differentiate H. schachtii in infected root and soil samples. H. schachtii-species-specific SCAR-PCR primers OPA06-HsF and OPA06-HsR were designed from the randomly amplified polymorphic DNA (RAPD) marker amplified with random primer OPA06. The developed primers specifically amplify a 922-bp fragment from the target populations but did not amplify DNA from non-target cyst nematodes including Heterodera, Globodera, Cactodera, and other related species tested in this study. The sensitivity detection indicated that 5 × 10⁻⁴ of a single cyst, 1/320 of a single second-stage juvenile (J2), or 10 pg of genomic DNA could be detected. The assay accurately identifies the different stages of H. schachtii in sugar beet and oilseed rape roots as well as a single J2 in 10 g of soil. Finally, the SCAR-PCR assay detected H. schachtii in seven samples out of the fifteen field samples. The assay will not only be useful for differentiating H. schachtii from mixed populations of Heterodera spp. but also for effective detection of the species directly from infested samples. The assay also requires no expertise in the taxonomy and morphology of the species but serves to improve the diagnosis of H. schachtii in infested fields. Full article

Heterodera schachtii is a well-known cyst nematode that causes serious economic losses in sugar beet production every year. Rapid and visual detection of H. schachtii is essential for more effective prevention and control. In this study, a species-specific recombinase polymerase amplification (RPA) primer was designed from a specific H. schachtii sequence-characterized amplified region (SCAR) marker. A band was obtained in reactions with DNA from H. schachtii, but absent from nontarget cyst nematodes. The RPA results could be observed by the naked eye, using a lateral flow dipstick (LFD). Moreover, we combined CRISPR technology with RPA to identify positive samples by fluorescence detection. Sensitivity analysis indicated that 10⁻⁴ single cysts and single females, 4⁻³ single second-stage juveniles, and a 0.001 ng genomic DNA template could be detected. The sensitivity of the RPA method for H. schachtii detection is not only higher than that of PCR and qPCR, but can also provide results in <1 h. Consequently, the RPA assay is a practical and useful diagnostic tool for early diagnosis of plant tissues infested by H. schachtii. Sugar beet nematodes were successfully detected in seven of 15 field sugar beet root samples using the RPA assay. These results were consistent with those achieved by conventional PCR, indicating 100% accuracy of the RPA assay in field samples. The RPA assay developed in the present study has the potential for use in the direct detection of H. schachtii infestation in the field. Full article

The cyst nematodes Heterodera schachtii and Heterodera trifolii, whose major hosts are sugar beet and clover, respectively, damage a broad range of plants, resulting in significant economic losses. Nematodes synthesize metabolites for organismal development and social communication. We performed metabolic profiling of H. schachtii and H. trifolii in the egg, juvenile 2 (J2), and female stages. In all, 392 peaks were analyzed by capillary electrophoresis time-of-flight mass spectrometry, which revealed a lot of similarities among metabolomes. Aromatic amino acid metabolism, carbohydrate metabolism, choline metabolism, methionine salvage pathway, glutamate metabolism, urea cycle, glycolysis, gluconeogenesis, coenzyme metabolism, purine metabolism, pyrimidine metabolism, and tricarboxylic acid (TCA) cycle for energy conversion (β-oxidation and branched-chain amino acid metabolism) energy storage were involved in all stages studied. The egg and female stages synthesized higher levels of metabolites compared to the J2 stage. The key metabolites detected were glycerol, guanosine, hydroxyproline, citric acid, phosphorylcholine, and the essential amino acids Phe, Leu, Ser, and Val. Metabolites, such as hydroxyproline, acetylcholine, serotonin, glutathione, and glutathione disulfide, which are associated with growth and reproduction, mobility, and neurotransmission, predominated in the J2 stage. Other metabolites, such as SAM, 3PSer, 3-ureidopropionic acid, CTP, UDP, UTP, 3-hydroxy-3-methylglutaric acid, 2-amino-2-(hydroxymethyl-1,3-propanediol, 2-hydroxy-4-methylvaleric acid, Gly Asp, glucuronic acid-3 + galacturonic acid-3 Ser-Glu, citrulline, and γ-Glu-Asn, were highly detected in the egg stage. Meanwhile, nicotinamide, 3-PG, F6P, Cys, ADP-Ribose, Ru5P, S7P, IMP, DAP, diethanolamine, p-Hydroxybenzoic acid, and γ-Glu-Arg_divalent were unique to the J2 stage. Formiminoglutamic acid, nicotinaminde riboside + XC0089, putrescine, thiamine 2,3-dihydroxybenzoic acid, 3-methyladenine, caffeic acid, ferulic acid, m-hydrobenzoic acid, o- and p-coumaric acid, and shikimic acid were specific to the female stage. Overall, highly similar identities and quantities of metabolites between the corresponding stages of the two species of nematode were observed. Our results will be a valuable resource for further studies of physiological changes related to the development of nematodes and nematode–plant interactions. Full article

Transcription factors are proteins that directly bind to regulatory sequences of genes to modulate and adjust plants’ responses to different stimuli including biotic and abiotic stresses. Sedentary plant parasitic nematodes, such as beet cyst nematode, Heterodera schachtii, have developed molecular tools to reprogram plant cell metabolism via the sophisticated manipulation of genes expression, to allow root invasion and the induction of a sequence of structural and physiological changes in plant tissues, leading to the formation of permanent feeding sites composed of modified plant cells (commonly called a syncytium). Here, we report on the AtMYB59 gene encoding putative MYB transcription factor that is downregulated in syncytia, as confirmed by RT-PCR and a promoter pMyb59::GUS activity assays. The constitutive overexpression of AtMYB59 led to the reduction in A. thaliana susceptibility, as indicated by decreased numbers of developed females, and to the disturbed development of nematode-induced syncytia. In contrast, mutant lines with a silenced expression of AtMYB59 were more susceptible to this parasite. The involvement of ABA in the modulation of AtMYB59 gene transcription appears feasible by several ABA-responsive cis regulatory elements, which were identified in silico in the gene promoter sequence, and experimental assays showed the induction of AtMYB59 transcription after ABA treatment. Based on these results, we suggest that AtMYB59 plays an important role in the successful parasitism of H. schachtii on A. thaliana roots. Full article

Plant parasitic nematodes, including the beet cyst nematode Heterodera schachtii, constitute a devastating problem for crops worldwide. The limited availability of sustainable management options illustrates the need for new eco-friendly control means. Plant metabolites represent an invaluable source of active compounds for the discovery of such novel antagonistic agents. Here, we evaluated the impact of eight plant terpenoids on the H. schachtii parasitism of Arabidopsis thaliana. None of the metabolites affected the plant development (5 or 10 ppm). Nootkatone decreased the number of adult nematodes on A. thaliana to 50%, with the female nematodes being smaller compared to the control. In contrast, three other terpenoids increased the parasitism and/or female size. We discovered that nootkatone considerably decreased the number of nematodes that penetrated A. thaliana roots, but neither affected the nematode viability or attraction to plant roots, nor triggered the production of plant reactive oxygen species or changed the plant’s sesquiterpene profile. However, we demonstrated that nootkatone led to a significant upregulation of defense-related genes involved in salicylic and jasmonic acid pathways. Our results indicate that nootkatone is a promising candidate to be developed into a novel plant protection agent acting as a stimulator of plant immunity against parasitic nematodes. Full article

Reactive oxygen species are a byproduct of aerobic metabolic processes but are also produced by plants in defense against pathogens. In addition, they can function as signaling molecules that control various aspects of plant life, ranging from developmental processes to responses to abiotic and biotic stimuli. In plants, reactive oxygen species can be produced by respiratory burst oxidase homologues. Arabidopsis contains 10 genes for respiratory burst oxidase homologues that are involved in different aspects of plant life. Plant pathogenic cyst nematodes such as Heterodera schachtii induce a syncytium in the roots of host plants that becomes a feeding site which supplies nutrients throughout the life of the nematode. In line with this function, the transcriptome of the syncytium shows drastic changes. One of the genes that is most strongly downregulated in syncytia codes for respiratory burst oxidase homologue B. This gene is root-specific and we confirm here the downregulation in nematode feeding sites with a promoter::GUS (β-glucuronidase) line. Overexpression of this gene resulted in enhanced resistance against nematodes but also against leaf-infecting pathogens. Thus, respiratory burst oxidase homologue B has a role in resistance. The function of this gene is in contrast to respiratory burst oxidase homologues D and F, which have been found to be needed for full susceptibility of Arabidopsis to H. schachtii. However, our bioinformatic analysis did not find differences between these proteins that could account for the opposed function in the interaction with nematodes. Full article

Arabidopsis contains 34 genes for glycosylphosphatidylinositol (GPI)-anchored LTPg proteins. A motif analysis has placed these into four groups. With one exception, all are produced with a signal peptide and are most likely attached to the cell membrane via the GPI anchor. Several of the LTPg genes across the four groups are downregulated in syncytia induced by the beet cyst nematode Heterodera schachtii. We have here studied At3g22600 encoding LTPg5, which is the most strongly downregulated LTPg gene. It is mainly expressed in roots, and a promoter::GUS line was used to confirm the downregulation in syncytia and also showed downregulation in galls of the root knot nematode Meloidogyne incognita. In contrast, infection with bacteria (Pseudomonas syringae) and fungi (Botrytis cinerea) led to the induction of the gene in leaves. This diverse regulation of LTPg5 indicated a role in resistance, which we confirmed with overexpression lines and a T-DNA mutant. The overexpression lines were more resistant to both nematode species and to P. syringae and B. cinerea, while a knock-out mutant was more susceptible to H. schachtii and P. syringae. Thus, LTPg5 encoded by At3g22600 is part of the Arabidopsis resistance mechanism against pathogens. LTPg5 has probably no direct antimicrobial activity but could perhaps act by associating with a receptor-like kinase, leading to the induction of defense genes such as PR1. Full article

Nematode-resistant cover crops can suppress populations of plant-parasitic nematodes. Samples of a loamy and a sandy loam soil were collected from two sugar beet fields in Lower Saxonia, northern Germany (“Jeinsen” and “Gross Munzel”) to measure the effects of cover cropping on the population genetic structure and infectivity of Heterodera schachtii as well as the composition of soil microbial communities. These fields allowed for a comparison of cover cropping with Brassica species resistant to Heterodera schachtii to fallow. In a series of radish bioassays with H. schachtii populations from Jeinsen and Gross Munzel, ratios of second-stage juveniles in roots per eggs in soil were higher in soil from under Brassica cropping than from under fallow. In denaturing gradient gelelectrophoresis, profiles of the parasitism gene vap1 differed between Brassica and fallow treatments in both populations. At Gross Munzel, microbes of soils and within nematode cysts differed between Brassica and fallow areas. Specifically, the frequency and occurrence of isolates of Pochonia chlamydosporia and Exophiala salmonis were lower within the cysts from Brassica than from fallow treatments. Overall, cover cropping with resistant Brassica species affected the bacteria and fungi infecting the cysts and subsequently, the infectivity of the H. schachtii population. Cover crop effects on nematode virulence (vap1 gene) and microbial colonization of the cysts could affect long-term nematode population dynamics. Full article