Cyst Nematode Research Articles

The nucleotide-binding domain of NRC-dependent disease resistance proteins is sufficient to activate downstream helper NLR oligomerization and immune signaling.

Nucleotide-binding domain and leucine-rich repeat (NLR) proteins with pathogen sensor activities have evolved to initiate immune signaling by activating helper NLRs. However, the mechanisms underpinning helper NLR activation by sensor NLRs remain poorly understood. Although coiled coil (CC) type sensor NLRs such as the Potato virus X disease resistance protein Rx have been shown to activate the oligomerization of their downstream helpers NRC2, NRC3 and NRC4, the domains involved in sensor-helper signaling are not known. Here, we used Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana to show that the nucleotide-binding (NB) domain within the NB-ARC of Rx is necessary and sufficient for oligomerization and immune signaling of downstream helper NLRs. In addition, the NB domains of the disease resistance proteins Gpa2 (cyst nematode resistance), Rpi-amr1, Rpi-amr3 (oomycete resistance) and Sw-5b (virus resistance) are also sufficient to activate their respective downstream NRC helpers. Using transient expression in the lettuce (Lactuca sativa), we show that Rx (both as full length or as NB domain truncation) and its helper NRC2 form a minimal functional unit that can be transferred from solanaceous plants (lamiids) to Campanulid species. Our results challenge the prevailing paradigm that NLR proteins exclusively signal via their N-terminal domains and reveal a signaling activity for the NB domain of NRC-dependent sensor NLRs. We propose a model in which helper NLRs can perceive the status of the NB domain of their upstream sensors.

GmSABP2-1 encodes methyl salicylate esterase and functions in soybean defense against soybean cyst nematode.

The soybean gene GmSABP2-1 encodes methyl salicylate esterase and its overexpression led to significant reduction in development of pathogenic soybean cyst nematode. Soybean cyst nematode (SCN, Heterodera glycines) is one of the most devastating pests of soybean (Glycine max L. Merr.). In searching for SCN-defense genes, a soybean gene of the methylesterase (MES) family was found to be upregulated in an SCN-resistant soybean line and downregulated in an SCN-susceptible line upon SCN infection. This gene was designated as GmSABP2-1. Here, we report on biochemical and overexpression studies of GmSABP2-1 to examine its possible function in SCN resistance. The protein encoded by GmSABP2-1 is closely related to known methyl salicylate esterases. To determine the biochemical function of GmSABP2-1, a full-length cDNA of GmSABP2-1 was cloned into a protein expression vector and expressed in Escherichia coli. The resulting recombinant GmSABP2-1 was demonstrated to catalyze the demethylation of methyl salicylate. The biochemical properties of GmSABP2-1 were determined. Its apparent Km value was 46.2 ± 2.2μM for methyl salicylate, comparable to those of the known methyl salicylate esterases. To explore the biological significance of GmSABP2-1 in soybean defense against SCN, we first overexpressed GmSABP2-1 in transgenic hairy roots of an SCN-susceptible soybean line. When infected with SCN, GmSABP2-1-overexpressing hairy roots showed 84.5% reduction in the development of SCN beyond J2 stage. To provide further genetic evidence for the role of GmSABP2-1 in SCN resistance, stable transgenic soybean plants overexpressing GmSABP2-1 were produced. Analysis of the GmSABP2-1-overexpressing lines showed a significant reduction in SCN development compared to non-transgenic plants. In conclusion, we demonstrated that GmSABP2-1 encodes methyl salicylate esterase and functions as a resistance-related gene against SCN.

Fungal communities shift with soybean cyst nematode abundance in soils

Plant parasitic nematodes pose a significant threat to food security. Management strategies for these pathogens are limited, and additional tools for their control are needed. Some fungi have shown promise for biocontrol, however, their success in the field has varied. In contrast, some fungal plant pathogens form synergistic associations with nematodes resulting in increased plant disease severity. However, how both groups of fungi change with different plant parasitic nematode abundances in the soil is underexplored. In this study, we used the soybean cyst nematode as a model to understand these changes. We sampled soil from 171 Ohio soybean fields in 2019 and 2021 and determined soybean cyst nematode abundance. We identified the fungi in the samples through amplicon sequencing of the 18S-ITS rDNA regions. Edaphoclimatic factors were used to classify samples into geographic regions to account for environmental differences between sampling locations. We hypothesized that fungal communities would be influenced by both region and soybean cyst nematode abundance. K-near neighbor analysis revealed that fungal communities follow regional patterns. We also found that soybean cyst nematode abundance was associated with changes in these communities regardless of the region. Two potential nematophagous fungi were found to be prevalent in Ohio through core community analysis, although they were enriched when soybean cyst nematode abundance was high. Lastly, differential network analysis showed that interactions among fungal community members change when soybean cyst nematode is present in the soil. Together these results suggest that this nematode significantly shifts the fungal community composition in field soils.

Development and Validation of a Resistance Management Model for the Soybean Cyst Nematode, Heterodera glycines.

Plant-parasitic nematodes are a key yield-limiting pest of crops around the world. Deployment of plant resistance genes are an important management tactic for many economically important plant-parasitic nematodes. The selection for virulence in nematode populations is a major threat to the effectiveness of resistance gene-based management. Little research has gone into resistance management modelling despite the importance of both plant-parasitic nematodes and resistance genes for their management. In this paper, we report on a cyst nematode resistance management model created to explore the factors which are most important for determining the durability of resistance genes to this important family of plant-parasitic nematodes. The relative dominance of virulence expression, the level of inbreeding, and the number of generations per cropping season were the most important factors in predicting resistance gene durability. Aspects of cyst nematode biology that reduce the number of generations per season for a portion of the population had a much smaller effect on the durability of resistance genes. These factors included delayed hatching within a season and early dormancy. The accuracy and utility of the model was tested using the soybean cyst nematode (SCN) rhg1-mediated resistance system. The model accurately predicted the rate at which virulence to the rhg1b resistance gene developed in Iowa over a two-decade period. The model suggested resistance gene pyramids as the most durable management solution for SCN with multiple possible avenues to obtain acceptable efficacy and durability.

Laburnicotides A-F: Acyclic N-Acetyl Oligopeptides from the Nematode-Cyst-Associated Fungus Laburnicola nematophila.

Nematode-associated fungi revealed the potential to produce a broad spectrum of chemical scaffolds. In this study, a mycelial extract of Laburnicola nematophila, a fungal strain derived from the cereal cyst nematode Heterodera filipjevi, was chemically explored and afforded six unprecedentedly reported acylic N-acetyl oligopeptides, laburnicotides A-F (1-6). Structure elucidation of the isolated compounds was established based on comprehensive 1D and 2D NMR spectroscopic analyses together with the acquired HR-ESI-MS spectrometric data. The absolute configuration of amino acid residues in 1-6 was established by performing advanced Marfey's derivatization method. All isolated compounds were assessed for their cytotoxic, antimicrobial, antiviral, and nematicidal activities with no potential activity observed.

Soybean MKK2 establishes intricate signalling pathways to regulate soybean response to cyst nematode infection.

Mitogen-activated protein kinase (MPK) cascades play central signalling roles in plant immunity and stress response. The soybean orthologue of MPK kinase2 (GmMKK2) was recently identified as a potential signalling node whose expression is upregulated in the feeding site induced by soybean cyst nematode (SCN, Heterodera glycines). To investigate the role of GmMKK2 in soybean-SCN interactions, we overexpressed a catabolically inactive variant referred to as kinase-dead variant (KD-GmMKK2) using transgenic hairy roots. KD-GmMKK2 overexpression caused significant reduction in soybean susceptibility to SCN, while overexpression of the wild-type variant (WT-GmMKK2) exhibited no effect on susceptibility. Transcriptome analysis indicated that KD-GmMKK2 overexpressing plants are primed for SCN resistance via constitutive activation of defence signalling, particularly those related to chitin, respiratory burst, hydrogen peroxide and salicylic acid. Phosphoproteomic profiling of the WT-GmMKK2 and KD-GmMKK2 root samples upon SCN infection resulted in the identification of 391 potential targets of GmMKK2. These targets are involved in a broad range of biological processes, including defence signalling, vesicle fusion, chromatin remodelling and nuclear organization among others. Furthermore, GmMKK2 mediates phosphorylation of numerous transcriptional and translational regulators, pointing to the presence of signalling shortcuts besides the canonical MAPK cascades to initiate downstream signalling that eventually regulates gene expression and translation initiation. Finally, the functional requirement of specific phosphorylation sites for soybean response to SCN infection was validated by overexpressing phospho-mimic and phospho-dead variants of two differentially phosphorylated proteins SUN1 and IDD4. Together, our analyses identify GmMKK2 impacts on signalling modules that regulate soybean response to SCN infection.

Fortifying nematode resistance through susceptibility gene inactivation

The predominant genetic defense mechanism against soybean cyst nematode (SCN) in 95% of the North America market is under threat by virulent SCN populations. Usovsky et al. identified GmSNAP02 as an SCN susceptibility gene through fine-mapping of unique bi-parental populations. Loss-of-function of GmSNAP02 confers enhanced resistance to more virulent SCN.

Morphological and molecular characterization of Heterodera ripae, a new record cyst nematode in the rhizosphere soil of Fagopyrum esculentum

Numerous plant parasitic nematodes (PPNs) have the potential to inflict considerable damage on agricultural crops. Through a comprehensive survey aimed at identifying PPNs affecting crops, cyst nematodes were isolated from the rhizosphere soil of buckwheat (Fagopyrum esculentum). Employing both molecular and morphological techniques, this cyst nematode was conclusively identified as Heterodera ripae. Notably, this represents the first documented occurrence of this particular cyst nematode species within the rhizosphere soil of F. esculentum.

Redox signalling in plant-nematode interactions: Insights into molecular crosstalk and defense mechanisms.

Plant-parasitic nematodes, specifically cyst nematodes (CNs) and root-knot nematodes (RKNs), pose significant threats to global agriculture, leading to substantial crop losses. Both CNs and RKNs induce permanent feeding sites in the root of their host plants, which then serve as their only source of nutrients throughout their lifecycle. Plants deploy reactive oxygen species (ROS) as a primary defense mechanism against nematode invasion. Notably, both CNs and RKNs have evolved sophisticated strategies to manipulate the host's redox environment to their advantage, with each employing distinct tactics to combat ROS. In this review, we have focused on the role of ROS and its scavenging network in interactions between host plants and CNs and RKNs. Overall, this review emphasizes the complex interplay between plant defense mechanism, redox signallingand nematode survival tactics, suggesting potential avenues for developing innovative nematode management strategies in agriculture.

Occurrence and distribution of cyst nematodes, Heterodera spp. (Tylenchida: Heteroderidae) associated with black cabbage, Brassica oleracea var. acephala L. (Brassicales: Brassicaceae) in the Eastern Black Sea Region of Türkiye

This study was conducted during 2021-2022 to detect and determine distribution and population of cyst nematodes, Heterodera spp. (Tylenchida: Heteroderidae) in black cabbage Brassica oleracea var. acephala L. (Brassicales: Brassicaceae) production areas of the Eastern Black Sea Region of Türkiye. For it, a total of 77 samples were taken from 53 districts belonging to the Artvin, Giresun, Ordu, Rize, and Trabzon provinces in the region. Soil samples were taken from around the root of the kale plants. Nematodes were extracted by using the centrifugal flotation technique. The nematodes were identified using morphological features and molecular analysis based on Polymerase Chain Reaction (PCR) method. For molecular analysis, the ribosomal DNA region including the gene region of 28S ribosomal RNA (rRNA) (ITS1, 5.8S, ITS2) was amplified using primer sets TW81/AB28. Additionally, a species-specific primer set (Car-F/Car-R) covering the Cytochrome Oxidase I (cox1) region of mitochondrial DNA (mtDNA) was used. As a result of the analysis, cyst nematodes Heterodera cruciferae Franklin, 1945, Heterodera carotae Jones, 1950 and Heterodera fici Kirjanova, 1954 species were identified in the kale production areas in the region. Heterodera carotae is the first record of the cyst nematode species in Türkiye. Heterodera cruciferae, H. carotae, and H. fici were detected from the total collected soil samples at 16.9%, 3.9%, and 1.3% relative frequency, respectively. Among all, Giresun was the most infected province with 35.3% infection rate, followed by Trabzon with 26.3%, Ordu with 21.1% and Rize with 13.3%.

Comparing the effectiveness of real-time PCRs to simultaneously detect and identify viable Globodera pallida and G. rostochiensis

AbstractRecently, the Commission Implementation Regulation (CIR) 2022/1192 was published by the European Union, prescribing regulations on eradication measures for quarantine potato cyst nematodes, including mandated assays for species identification. The CIR 2022/1192 provides a derogation on the use of Beniers et al. (2014) real-time PCR as the validation process was still ongoing. In this study, the validation of a modified version of the Beniers real-time PCR was carried out, following EPPO Standard PM 7/98. Furthermore, the performance was compared to the conventional PCR of Bulman & Marshall (1997) and the Gamel et al. (2017) real-time PCR, both DNA based, which are mandated by CIR 2022/1192. Additionally, the effect of cyst viability was compared with the two real-time PCR assays. The modified Beniers real-time PCR met all validation performance criteria for both target Globodera pallida and G. rostochiensis. The viability assays showed that the DNA based Gamel real-time PCR provided positive results for non-viable, non-infectious cysts, in contrast to the modified RNA based Beniers real-time PCR. Based on the results of this study, the modified Beniers qualifies for the detection and identification of viable Globodera pallida and G. rostochiensis and performs at least equally as well as the other two PCR assays. To encourage the development and improvement of diagnostic protocols, it is suggested that diagnostic laboratories performing official surveys are allowed to use internationally recognized diagnostic protocols when validated in-house according to EPPO standards and accredited by the EU bodies (EPPO, 2021).

Registration of ‘S16‐16814R’: A glyphosate‐tolerant, high‐oleic soybean cultivar

AbstractThere is a demand for soybean [Glycine max (L.) Merr.] cultivars with more functional soybean oil for food and industrial applications. Soybean breeding programs have devoted more efforts to developing high‐yielding soybean cultivars with elevated oil content and improved fatty acid profiles. The University of Missouri – Fisher Delta Research, Extension, and Education Center soybean breeding program has developed and released the high‐oleic soybean cultivar ‘S16‐16814R’ (Reg. no. CV‐561, PI 704911). It is a maturity group 4‐late (relative maturity 4.9), glyphosate‐tolerant (Roundup Ready 1) soybean cultivar with high oleic (84.0%) acid content and a reduced linolenic (3.5%) acid content. S16‐16814R is resistant to multiple nematode species, including soybean cyst nematode races 2 (HG Type 1.2.5.7), 3 (HG Type 5.7), and 5 (HG Type 2.5.7); southern root‐knot nematode; and reniform nematode. S16‐16814R was evaluated from 2017 to 2020 in 61 environments across eight southern US states. It averaged 92% of the check mean for yield, indicating competitive yield performance across multiple environments.

Characterisation and mapping of a Globodera pallida resistance derived from the wild potato species Solanum spegazzinii

Key messageA new resistance locus acting against the potato cyst nematode Globodera pallida was mapped to chromosome VI in the diploid wild potato species Solanum spegazzinii CPC 7195.The potato cyst nematodes (PCN) Globodera pallida and Globodera rostochiensis are economically important potato pests in almost all regions where potato is grown. One important management strategy involves deployment through introgression breeding into modern cultivars of new sources of naturally occurring resistance from wild potato species. We describe a new source of resistance to G. pallida from wild potato germplasm. The diploid species Solanum spegazzinii Bitter accession CPC 7195 shows resistance to G. pallida pathotypes Pa1 and Pa2/3. A cross and first backcross of S. spegazzinii with Solanum tuberosum Group Phureja cultivar Mayan Gold were performed, and the level of resistance to G. pallida Pa2/3 was determined in progeny clones. Bulk-segregant analysis (BSA) using generic mapping enrichment sequencing (GenSeq) and genotyping-by-sequencing were performed to identify single-nucleotide polymorphisms (SNPs) that are genetically linked to the resistance, using S. tuberosum Group Phureja clone DM1-3 516 R44 as a reference genome. These SNPs were converted into allele-specific PCR assays, and the resistance was mapped to an interval of roughly 118 kb on chromosome VI. This newly identified resistance, which we call Gpa VIlspg, can be used in future efforts to produce modern cultivars with enhanced and broad-spectrum resistances to the major pests and pathogens of potato.

The other side of the coin: systemic effects of Serendipita indica root colonization on development of sedentary plant–parasitic nematodes in Arabidopsis thaliana

Main conclusionUpon systemic S. indica colonization in split-root system cyst and root-knot nematodes benefit from endophyte-triggered carbon allocation and altered defense responses what significantly facilitates their development in A. thaliana.Serendipita indica is an endophytic fungus that establishes mutualistic relationships with different plants including Arabidopsis thaliana. It enhances host’s growth and resistance to different abiotic and biotic stresses such as infestation by the cyst nematode Heterodera schachtii (CN). In this work, we show that S. indica also triggers similar direct reduction in development of the root-knot nematode Meloidogyne javanica (RKN) in A. thaliana. Further, to mimick the natural situation occurring frequently in soil where roots are unequally colonized by endophytes we used an in vitro split-root system with one half of A. thaliana root inoculated with S. indica and the other half infected with CN or RKN, respectively. Interestingly, in contrast to direct effects, systemic effects led to an increase in number of both nematodes. To elucidate this phenomenon, we focused on sugar metabolism and defense responses in systemic non-colonized roots of plants colonized by S. indica. We analyzed the expression of several SUSs and INVs as well as defense-related genes and measured sugar pools. The results show a significant downregulation of PDF1.2 as well as slightly increased sucrose levels in the non-colonized half of the root in three-chamber dish. Thus, we speculate that, in contrast to direct effects, both nematode species benefit from endophyte-triggered carbon allocation and altered defense responses in the systemic part of the root, which promotes their development. With this work, we highlight the complexity of this multilayered tripartite relationship and deliver new insights into sugar metabolism and plant defense responses during S. indica–nematode–plant interaction.

Fortify the defense frontline: MAPKs phosphorylate receptor-like cytoplasmic kinase to maintain plant resistance in soybean

Mitogen-activated protein kinase (MAPK) activation is one of the significant immune events that respond to pathogens in plants. A MAPK cascade often contains a MAPK kinase kinase (MAPKKK), a MAPK kinase (MAPKK/MKK), and a MAPK. The well-characterized MAPK cascade, to date, is the MAPKKK3/4/5-MKK4/5-MPK3/6 module. Soybean cyst nematodes (SCN) is one of the most devastating soybean pathogens. However, the early immune components contributing to soybean resistance to SCN and the role of the MAPK cascade in the soybean–SCN interaction remain unclear. A recent study published in Plant Cell discovered that GmMPK3/6 phosphorylates a receptor-like cytoplasmic kinase (RLCK), CDG1-LIKE1 (GmCDL1), and maintains the stability of GmCDL1 in soybean. Remarkably, GmCDL1 enhances GmMPK3/6 activation and resistance to SCN by phosphorylating GmMAPKKK5 and activating the GmMAPKKK5-GmMKK4-GmMPK3/6 cascade. In addition, two L-type lectin receptor kinases (LecRKs), GmLecRK02g and GmLecRK08g, are involved in the GmCDL1 function after the perception of SCN. taken together, this study not only discovers a complete early immune pathway that responds to SCN infection in soybean, but also reveals a molecular mechanism by which plants maintain the activation of the MAPK cascade and resistance.

Breeding for potato cyst nematode resistance in Solanum tuberosum

AbstractTwo species of potato cyst nematodes (PCNs), Globodera rostochiensis (Woll.) and Globodera pallida (Stone), threaten potato (Solanum tuberosum L.) production worldwide. PCNs are remarkably persistent in soil and can reduce potato yield significantly. The most sustainable approach to PCN control is deploying resistant cultivars. This review focuses on the knowledge needed to conduct a successful PCN resistance breeding program, including identifying resistance sources in wild species, introgressing them into cultivated potato, and sifting through clones to identify a marketable potato.

Heterodera glycines HG type 1.2.5.7 causes a decrease in soybean (Glycine max [L.] Merr.) nitrogen fixation and growth variables

ABSTRACT Soybean cyst nematode (SCN), Heterodera glycines, HG type 1.2.5.7 is the most virulent and widespread SCN biotype in the southern states of the USA, but its effect on soybean (Glycine max [L.] Merr.) nitrogen fixation (NF) activity and growth is not well documented. The objectives of this study were to: (1) measure the impact of HG type 1.2.5.7 on NF activity during the early vegetative stage of three soybean genotypes and (2) compare the impact on soybean leaf and root development, and plant growth. The NF activity of two susceptible (S) genotypes (“Ellis” and “Williams 82”) and one resistant (R) genotype (PI 88,788), subjected to a 10,000-egg treatment, was measured using an in-situ flow-through acetylene reduction assay. The average NF activity of the soybean genotypes was decreased by SCN activity. Time points assessment showed that the NF activity was differently impacted among the genotypes. Based on the measured values, the NF activity was decreased by 30% in SCN infected plants of Williams 82 but that of PI 88,788 and Ellis was affected. Likewise, soybean leaf color and leaf area were affected by SCN infection, but there was no impact on root length and root area. These results showed that SCN HG type 1.2.5.7 can exhibit varying negative impact on NF and shoot growth of S soybean genotypes, while R genotypes may be unaffected. Thus, indicating genetic resistance continues to be a critical tool for the management of the SCN.

Haplotype diversity of Heterodera koreana (Tylenchida: Heteroderidae), affecting bamboo in Korea

AbstractIn a survey of plant-parasitic nematodes in agricultural fields, cyst-forming nematodes were found in soil planted bamboo in Korea. The aim of this study was to identify the cyst nematodes based on morphological and molecular characteristics. As the results, the morphology and morphometrics of cysts and second-stage juveniles (J2s) were consistent with those of previous descriptions of Heterodera koreana. In phylogenetic analyses based on DNA sequences, these cyst nematodes were clustered together with clade of H. koreana in internal transcribed spacer (ITS) region, and large subunit D2-D3 segments (LSU D2-D3). These nematodes were clustered together with clade of H. koreana in cytochrome c oxidase subunit I (COI) gene, but a haplotype was different when compared with previous reported haplotypes (haplotype A-C) in Japan. This study showed these cyst nematodes were identified as H. koreana, and a new haplotype of H. koreana is distributed in Korea. We suggest that the new haplotype of H. koreana name as haplotype D.

Analyses of fungal communities from culture-dependent and -independent studies reveal novel mycobiomes associated with Globodera and Heterodera species

Plant-parasitic nematodes are important pathogens of agricultural crops. Of particular importance are cyst nematode species of Globodera and Heterodera which remain a major constraint to global production of crops such as potato, soybean, and wheat. Recent restrictions on the use of some synthetic nematicides has created an urgent need for alternative methods of controlling cyst nematodes. This project aimed to: (i) document fungi found associated cyst nematodes, (ii) determine if the fungi associated with cyst nematodes are dependent on growth conditions or nematode species, (iii) identify core mycobiomes for each of the eight cyst nematode species, (iv) identify the core mycobiome across all cyst nematode species studied, and (v) determine if the fungi associated with Globodera species are structured geographically. Sixty cyst nematode populations from eight species – Globodera sp., G. ellingtonae, G. pallida, G. rostochiensis, H. avenae, H. carotae, H. glycines, and H. schachtii – were collected from several geographic locations globally and screened for fungi using both culture-dependent and -independent methods. The core mycobiome was determined for each species. In total, 744 species of fungi were found associated with cyst nematodes. Fungal species shared across all cyst nematode species included Pochonia sp., Sporothrix sp., Neocosmospora solani, Exophiala equina, Mortierella sp., and Sarocladium kiliense. The core mycobiomes were conserved between cyst nematode species irrespective of their geographic origin. Cyst nematodes collected from the field or from greenhouse cultures also recruited similar core mycobiomes. These findings could be relevant when choosing fungi to evaluate as biocontrol agents against cyst nematodes.

N6-methyladenosine (m6A) sequencing reveals Heterodera glycines-induced dynamic methylation promoting soybean defense.

Unraveling the intricacies of soybean cyst nematode (Heterodera glycines) race 4 resistance and susceptibility in soybean breeding lines-11-452 (highly resistant) and Dongsheng1 (DS1, highly susceptible)-was the focal point of this study. Employing cutting-edge N6-methyladenosine (m6A)-seq and RNA-seq techniques, we delved into the impact of m6A modification on gene expression and plant defense responses. Through the evaluation of nematode development in both resistant and susceptible roots, a pivotal time point (3 days post inoculation) for m6A methylation sequencing was identified. Our sequencing data exhibited robust statistics, successful soybean genome mapping, and prevalent m6A peak distributions, primarily in 3'UTR (Untranslated region) and stop codon regions. Analysis of differentially expressed m6A peaks (DMPs) and expressed genes (DEGs) revealed distinctive patterns between resistant and susceptible genotypes. In the highly resistant line (11-452), key resistance and defense-associated genes displayed increased expression coupled with inhibited methylation, encompassing crucial players like R genes, receptor kinases, and transcription factors. Conversely, the highly susceptible DS1 line exhibited heightened expression correlated with decreased methylation in genes linked to susceptibility pathways, including Mildew Locus O (MLO)-like proteins and regulatory elements affecting defense mechanisms. Genome-wide assessments, GO/KEGG analyses, and DMP/DEG overlap emphasized the intricate interplay of m6A modifications, alternative splicing, microRNA and gene regulation in plant defense. Protein-protein interaction networks illuminated defense-pivotal genes, delineating divergent mechanisms in resistant and susceptible responses. This study sheds light on the dynamic correlation between methylation, splicing, and gene expression, providing profound insights into plant responses to nematode infection.