Meloidogyne Incognita Research Articles

Effect of Temperature on the Embryogenesis of Three Geographically Distinct Populations of Meloidogyne incognita Is Driven by Intrinsic Thermal Acclimation Reaction.

Research interest in the mechanisms enabling plant-parasitic nematodes to adjust their physiological performance and cope with changing temperatures has intensified in light of global warming. Here, we show that geographically distinct populations of the root-knot nematode Meloidogyne incognita, which is prevalent in the three main pepper-growing regions in Israel-Carmel Valley (Carmel), Jordan Valley (JV), and Arava Rift (Arava)-possess persistent differences in their thermal acclimation capacity, which affect pre- and postembryonic development. The optimal temperature for embryonic growth completion was 25°C for the Carmel population; 25 and 30°C for the JV population; and 30°C for the Arava population. Cumulative hatching percentages showed variations among populations; relative to hatching at 25°C, the Carmel population experienced hatching reduction at the higher studied temperatures 30 and 33°C, while the JV and Arava populations exhibited an increase in hatching at 30 and 33°C, respectively. Juvenile survival indicates that at the lowest temperature (20°C), the Carmel population gained the highest survival rates throughout the experimental duration, while at the same duration at 33°C, the Arava population gained the highest survival rate. Infective juveniles of the Carmel population demonstrated increased penetration of tomato roots at 25°C compared to the JV and Arava populations. Inversely, at 33°C, increased penetration was observed for the Arava compared to the Carmel and JV populations. Altogether, the Arava population's performance at 33°C might incur distinct fitness costs, resulting in consistent attenuation compared to the Carmel population at 25°C. Precisely defining a population's thermal acclimation response might provide essential information for models that predict the impact of future climate change on these populations.

Sensitivity of Meloidogyne incognita and Rotylenchulus reniformis to cyclobutrifluram.

Cyclobutrifluram, a succinate dehydrogenase inhibitor fungicide, is being evaluated as a seed-applied nematicide in cotton and soybean to manage plant-parasitic nematodes. Currently, there is no information on the toxicity, ovicidal activity, nematode recovery, or effects on nematode infection for Meloidogyne incognita or Rotylenchulus reniformis after exposure to low concentrations of cyclobutrifluram. Nematode toxicity assays were performed in aqueous solutions of cyclobutrifluram, while root infection assays were conducted on tomato. Nematode paralysis was observed after 2 h of exposure to 0.5 µg/ml cyclobutrifluram for both nematode species. Based on an assay of nematode motility, the 2-hr EC50 value for M. incognita and R. reniformis was 0.48 and 1.07 µg/ml, respectively. In a comparable assay with a similar nematicide, continuous exposure to 0.5 µg/ml cyclobutrifluram for 24 h resulted in at least 45% more immotile nematodes for both species compared to those treated with 0.5 µg/ml fluopyram. Continuous exposure to concentrations >1.0 µg/ml suppressed hatching for both species compared to the water control. Nematode recovery from paralysis was greater than 80% for M. incognita and R. reniformis 24 h after nematodes were rinsed and removed from a 1-h treatment to their respective 2-hr EC50 concentrations. Nematode infection of tomato roots was reduced following a 1-h treatment with aqueous solutions of cyclobutrifluram, ranging from 0.12 to 0.48 µg/ml for M. incognita and 0.27 to 1.07 µg/ml for R. reniformis. Overall, the toxicity of cyclobutrifluram to these nematode species was greater than that of fluopyram and although the effects of cyclobutrifluram were reversible, low concentrations were effective at reducing the ability of both nematodes to infect tomato roots.

Metabolomics and histopathological analysis of two tomato cultivars after co-infection with soil-borne pathogens (Southern root-knot nematode and Fusarium wilt fungus)

Southern root-knot nematode (Meloidogyne incognita) and Fusarium wilt fungus (Fusarium oxysporum) are one of the most predominant pathogens responsible for substantial agricultural yield reduction of tomato. The current study planned to assess the effects of M. incognita (Mi) and F. oxysporum (Fo) and their co-infection on two tomato cultivars, Zhongza 09 (ZZ09) and Gailing Maofen 802 (GLM802). The present study examined the effects of co-infection on leaf morphology, chlorophyll content, leaf area, and histopathology. The present study used metabolomics to evaluate plant-pathogen interactions. The outcomes of the current study revealed that chlorophyll content and leaf area decreased more in GLM802 during co-infection. In co-infection (Fo + Mi), the chlorophyll content reduction in ZZ09 was 11%, while in GLM802 the reduction reached up to 31% as compared to control. Moreover, the reduction in leaf are in ZZ09 was 31%, however, in the GLM802 reduction was observed 54% as compared to control plants. Similarly, GLM802 stems exhibited larger brown patches on their vascular bundles than ZZ09 stems. The rate of browning of GLM802 stems was 247% more than ZZ09, during co-infection. Moreover, GLM802 roots exhibited a higher abundance of hyphae and larger galls than ZZ09 roots. In metabolic studies, glutathione, succinic acid, and 2-isopropylmalic acid decreased, whereas spermine and fumaric acid increased in GLM802 co-infected stems. It indicates that GLM802 is weakly resistant; therefore, F. oxysporum and other pathogens readily damage tissue. In the co-infected stem of ZZ09, L-asparagine and shikimic acid increased, but pipecolic acid, L-saccharine, and 2-isopropylmalic acid declined. L-asparagine was crucial in preserving the stability of nitrogen metabolism, chlorophyll synthesis, and leaf growth in ZZ09. Shikimic acid's substantial accumulation could explain the limited extent of browning observed in the vascular bundles of ZZ09. Thus, the present study provides insight into M. incognita and F. oxysporum co-infection in two tomato cultivars, which may aid breeding efforts to generate commercially viable resistant cultivars. However, further research on the relationship between M. incognita and F. oxysporum in different host plants is required in the future.

EMERGING NEMATOLOGICAL PROBLEMS IN THE CAMPANIA REGION

During a fifteen-year period of work carried out by the Test facility for the farm acquisition to be used to conduct efficacy trials against nematodes, a survey was carried out on nematological problems occurring in the Campania Region. The investigations has shown that the genus Meloidogyne spp., in particular the species M. incognita, is the most widespread and harmful trophic group. These nematodes are found in particular on horticultural plants in the sandy soils of the coastal areas and especially under protected conditions. Infestations of the cyst nematodes Globodera rostochiensis and G. pallida on potato in the neighbouring Caserta and Neapolitan areas, H. goettingiana on leguminous plants, especially peas, and H. daverti on carnation in the Vesuvian foothills are also common. In the Nocerino-Sarnese countryside, infestations by the stem and bulb nematode, Ditylenchus dipsaci, on onions are still severe. On fruit trees are common Mesocriconema xenoplax on actinidia, Meloidogyne incognita and M. hapla on peach trees and Tylenchulus semipenetrans on citrus fruits. Among the virus transmitting nematodes, Xiphinema diversicaudatum is very harmful to peach trees in Terra di Lavoro, in the Carinola (Caserta) area, and X. index to grapevines in the Benevento and Avellino areas. Key Words: phytoparasitic nematodes, crops, distribution, damage.

Distinct changes in tomato-associated multi-kingdom microbiomes during Meloidogyne incognita parasitism

BackgroundThe interplay between root-knot nematode (RKN) parasitism and the complex web of host-associated microbiota has been recognized as pivotal for effective management of the pest. However, studies assessing this relationship have focussed on the bacterial and fungal communities, neglecting the unicellular eukaryotic members. Here, we employed amplicon sequencing analysis of the bacterial 16S rRNA, fungal ITS and eukaryotic 18S rRNA genes, and comprehensively examined how the microbiome composition, diversity and networking developed with time in the rhizospheres and roots of RKN-inoculated and non-inoculated tomato plants.ResultsAs expected, infection with the RKN Meloidogyne incognita decreased plant growth. At individual timepoints, we found distinct bacterial, fungal and eukaryote community structures in the RKN-inoculated and non-inoculated rhizospheres and roots, and RKN inoculation affected several taxa in the root-associated microbiome differentially. Correlation analysis revealed several bacterial and fungal and few protist taxa that correlated negatively or positively with M. incognita. Moreover, network analysis using bacterial, fungal and eukaryotic data revealed more dynamic networks with higher robustness to disturbances in the RKN-inoculated than in the non-inoculated rhizospheres/roots. Hub taxa displayed a noticeable successional pattern that coincided with different phases of M. incognita parasitism. We found that fungal hubs had strong negative correlations with bacteria and eukaryotes, while positive correlations characterized hub members within individual kingdoms.ConclusionOur results reveal dynamic tomato-associated microbiomes that develop along different trajectories in plants suffering M. incognita infestation and non-infested plants. Overall, the results identify stronger associations between RKN and bacterial and fungal taxa than between eukaryotic taxa and RKN, suggesting that fungal and bacterial communities could play a larger role in the regulation of RKN. The study identifies several putative RKN-antagonistic bacterial and fungal taxa and confirms the antagonistic potential previously identified in other taxa.

Understanding the dynamics of Meloidogyne incognita infestation in pomegranate orchards of Himachal Pradesh, India (year 2018, 2019 and 2021) and its management strategies

This study investigates the prevalence and dynamics of pomegranate wilt disease induced by Meloidogyne incognita across the Kullu, Mandi and Solan districts of Himachal Pradesh (India), revealed notable spatial and temporal variations in nematode populations and galling severity across the regions. The highest average nematode infestation of 9.25 % was observed at Nauni with highest counts of average second-stage juvenile (J2) larvae (449 larvae per 100 cc of soil) followed by Hurla (Kullu) with 7.42 % infestation. Correlation analysis reveals a strong positive relationship between larval population and galling severity suggesting a potential link between nematode levels and plant damage. Common disease symptoms were leaf size reduction, yellowing and gradual decline of pomegranate plants, often observed in patches within orchards. Microscopic identification revealed distinctive pear-shaped body of mature females while J2 larvae displayed vermiform shapes and the associated species of M. incognita was confirmed through examination of the perineal pattern. Pathogenicity test reveals initiation of leaf yellowing symptom after 45 days of inoculation of larval suspension and root galling was observed after 60 days onward followed by plant decline under greenhouse conditions. Results from pot and field experiments demonstrated the efficacy of Fluopyram and Fluensulfone in reducing nematode populations and galling severity. Treatment with drenching of Fluopyram at the rate of 2 ml/L reduced 98.56 % larvae under field and 99.00 % larvae/100 cc soil under pot conditions. Statistical analysis (paired t-test and MANOVA) confirms significant differences in galling severity and larval population before and after drenching. The study also underscores the importance of weed management in disease mitigation as several weed species (Chenopodium album and Solanum nigrum) were identified as potential reservoirs for M. incognita in infested pomegranate plant basin. This investigation contributes to the advancement of management practices for pomegranate cultivation that addresses both nematode and weed infestations ultimately enhancing crop resilience and productivity.

Utilizing endophytic plant growth-promoting bacteria and the nematophagous fungus Purpureocillium lilacinum as biocontrol agents against the root-knot nematode (Meloidogyne incognita) on tomato plants

AbstractThe present investigation was designed to assess how administering biocontrol agents (BCAs) made from the nematophagous endophytic fungus MR20 (Purpureocillium lilacinum) or a mixture of endophytic plant growth-promoting bacteria MR12 (Pseudomonas fluorescens), and MR25 (Serratia marcescens), could enhance tomato growth and resistance to Meloidogyne incognita under in vivo conditions. The three strains’ cell-free culture filtrates showed a strong nematocidal impact (P < 0.05) on M. incognita infective second-stage juveniles (J2s). The highest mortality rates by the three BCAs were at concentrations of 80%, followed by 60%, 40%, and 20%. The in vitro hatching of free eggs was found to be considerably (P < 0.05) reduced as the concentrations of the endophytic bacteria MR12, and MR25 were increased in the order 1 × 105, 1 × 106, 1 × 107, 0.5 × 108, to 1 × 108 colony forming units (CFU)/mL, and the maximum nematicidal activity in killing M. incognita free eggs occurred at 1 × 108 CFU/mL. A statistically significant (P < 0.05) reduction in the percentage of M. incognita-free eggs retrieved was seen in vitro when various concentrations of the endophytic fungus MR20 were applied compared to the negative control (distilled water). The endophytic fungus MR20 had the highest nematicidal activity against M. incognita free eggs at a concentration of 3 × 106 CFU/mL. The application of P. lilacinum or a combination of P. fluorescens and S. marcescens to tomato plants in the presence of M. incognita under greenhouse conditions resulted in a significant increase (P < 0.05) in root and shoot fresh weight, number of leaves, weight of leaves, and stem diameter when compared to the positive control treatment that contained only M. incognita. Treatment with P. lilacinum was more effective (P < 0.05) than P. fluorescens and S. marcescens in reducing egg masses per root, J2s per 100 g of soil, egg masses per 100 g of soil, J2s + eggs of M. incognita per 100 g of soil, reproduction factor, and reduction percentage after 60 days under greenhouse conditions. Nevertheless, the reduction of M. incognita gall size categorization (> 4 mm, 2–4 mm, and < 2 mm) was more effectively achieved by treatment with P. fluorescens and S. marcescens than by treatment with P. lilacinum. Under in vivo conditions, the tested P. lilacinum or a mixture of P. fluorescens and S. marcescens effectively controlled nematode population densities below the economic threshold.

Local and systemic transcriptome and spliceome reprogramming induced by the root-knot nematode Meloidogyne incognita in tomato.

Root-knot nematodes (Meloidogyne spp.) are widely spread root parasites that infect thousands of vascular plant species. These highly polyphagous nematodes engage in sophisticated interactions with host plants that results in the formation of knot-like structures known as galls whose ontogeny remains largely unknown. Here, we determined transcriptome changes and alternative splicing variants induced by Megalaima incognita in galls and neighboring root cells at two distinct infective stages. M. incognita induced substantial transcriptome changes in tomato roots both locally in galls and systemically in neighboring cells. A considerable parallel regulation of gene expression in galls and neighboring cells were detected, indicative of effective intercellular communications exemplified by suppression of basal defense responses particularly during the early stage of infection. The transcriptome analysis also revealed that M. incognita exerts a tight control over the cell cycle process as a whole that results in an increase of ploidy levels in the feeding sites and accelerated mitotic activity of the gall cells. Alternative splicing analysis indicated that M. incognita significantly modulates pre-mRNA splicing as a total of 9064 differentially spliced events from 2898 genes were identified where intron retention and exon skipping events were largely suppressed. Furthermore, a number of differentially spliced events were functionally validated using transgenic hairy root system and found to impact gall formation and nematode egg mass production. Together, our data provide unprecedented insights into the transcriptome and spliceome reprogramming induced by M. incognita in tomato with respect to gall ontogeny and nematode parasitism.

Integrated Analysis of the lncRNA-miRNA-mRNA Expression Profiles in Response to Meloidogyne incognita in Radish (Raphanus sativus L.)

Radish is an important root vegetable that is widely grown in Asia. The root-knot nematode (RKN) Meloidogyne incognita seriously affects the growth and development of radish root and causes poor appearance quality. However, the molecular mechanism of radish response to RKNs remains poorly understood. In this study, a total of 220 lncRNAs, 1144 mRNAs, 20 miRNAs and 153 proteins were differential expressed between the RKN-infected and WT samples. Correlation analysis of all DEPs compared with all DGEs showed that 8 mRNAs-DEPs showed a changed abundance. The results showed that 18 DEmiRNAs have 167 target DEGs in 220 miRNA-target modules and 29 DElncRNAs were predicted as putative targets of 16 DEmiRNAs in 37 miRNA-target modules. In all, 6 DGEs in the ABA pathway and 2 DGEs in the JA pathway were identified under RKN infection, respectively. The four regulatory networks of lncRNA-miRNA-mRNA were constructed in response to RKN infection. qRT-PCR analysis found that the expression pattern of 6 DElncRNAs, 6 DEmRNAs, 6 DEmiRNAs and 6 DEPs were consistent with sequencing results. These results provide a theoretical basis for studying the molecular mechanism of radish in response to M. incognita and breeding resistant varieties to this nematode.

Pathogenicity of Root-knot Nematode, Meloidogyne incognita on Tuberose (Polianthes tuberosa L.)

A pot experiment was carried out in the net house of Department of Nematology, Assam Agricultural University, Jorhat, to study the pathogenicity of root-knot nematode, Meloidogyne incognita, on tuberose. The study found that as the inoculum level of M. incognita increased from 10 to 10,000 second-stage juveniles (J2) per pot (containing 1 kg of sterilized soil), all plant growth parameters of tuberose decreased progressively. At the highest inoculum level, the plants became severely stunted, chlorotic, and produced very few bulblets. It was noted that at the highest inoculum level, the root systems were significantly reduced, with larger and mostly coalescent galls. The treatments with no nematode (check and associated check) were free from galls and eggmasses. An initial inoculum level of 100 J2 of M.incognita per kg of soil caused significant reduction in all growth parameters and proved to be pathogenic to the tuberose plants. The number of galls and eggmasses increased gradually in the plants with an initial inoculum level of 10 to 1000 J2 of M.incognita per kg of soil, but it declined at highest inoculum level of 10,000 J2 per kg of soil. The experiment also recorded the variation in nematode population in the soil as well as their reproductive rates. These findings can be used to develop effective, targeted methods for minimizing the damage caused by root-knot nematode in tuberose crops.

IN SILICO STUDIES OF IDENTIFIED COMPOUNDS FROM ETHANOLIC EXTRACT OF CELOSIA TRIGYNA AGAINST NEMATODE (Meloidogyne incognita)

Plant-parasitic nematodes (PPNs) have been found to be a major cause of considerable economic burden on the horticultural crops industry. Celosia genus have been renown to purpose antimicrobial activity. This study tends to swing the antimicrobial activity of one species of this genus i.e. Celosia trigyna against root-knot nematodes employing molecular docking methods. The identification of compounds in the leaf extract with the potential to inhibit Meloidogyne incognita was carried out in silico. Liquid Chromatography Mass Spectrometry (LCMS) was employed for analysis of the ethanolic leaf extract of this plant, it revealed the presence of cis-9-tetradecanoic acid, vitexin-glucoside, 6”-O-deoxyhexoside, gallic acid, malic acid, sinapic acid, 3-feruloylquinic acid in the extract. Site-directed multiligand docking of the identified compounds was performed on protein 2MIF of Meloidogyne incognita and employing Bifenazate and Boscalid as positive controls. The binding affinity of 3-feruloylquinic (– 4.2 kcal/mol) was significantly higher than the positive control compounds i.e. Bifenazate and Boscalid (– 4.0 kcal/mol and – 3.9 kcal/mol). The interactions of this molecule with the amino acids of the protein showed that the mechanism of its inhibitory action is similar to that of the cocrystallized ligand. This result validates that the antimicrobial activity of the ethanolic extract of the leaves of C. trigyna could be employed against root-knot nematodes for sustainable agriculture.

Botanicals as Phyto-nematicides Against Root-knot Disease Caused by the Root-knot Nematode (Meloidogyne incognita) via Promoting growth, Yield, and Biochemical Performance of Cabbage Crop

Botanicals as Phyto-nematicides Against Root-knot Disease Caused by the Root-knot Nematode (Meloidogyne incognita) via Promoting growth, Yield, and Biochemical Performance of Cabbage Crop

Draft genome sequence of Bacillus thuringiensis strain V-AB8.18, a novel isolate with potential nematicidal activity.

We report the draft genome of Bacillus thuringiensis strain V-AB8.18, comprising 308 contigs totaling 6,182,614 bp, with 35% G + C content. It contains 6,151 putative protein-coding genes, including App6 and Cry5-like crystal proteins, exhibiting 99% pairwise identity to nematicidal proteins App6Aa2 and Cry5Ba2, active against Meloidogyne incognita and Meloidogyne hapla.

Bacillus, Pseudomonas and Serratia control Meloidogyne incognita (Rhabditida: Meloidogynidae) and promote the growth of tomato plants

Plant-parasitic nematodes cause significant losses in agriculture worldwide. Among these parasites, the root-knot nematode (Meloidogyne spp.) stands out. Consequently, control methods are being developed to combat this pest. Among these methods is biological control, the main agents of which are bacteria. This study aimed to evaluate the impact of 102 bacterial isolates on the mortality of Meloidogyne incognita J2 juveniles, hypothesizing that certain isolates could effectively control the nematode and promote tomato plant growth. Five experiments were conducted to test this hypothesis. First, the effect of the 102 bacterial isolates on J2 mortality was assessed under laboratory conditions. Second, five isolates (Bacillus cereus IBCBb130, Bacillus cereus IBCBb116, Pseudomonas aeruginosa IBCBb122, Bacillus proteolyticus IBCBb136, and Serratia sp. IBCBb118), which showed >68% J2 mortality, were evaluated over 72 h. Third, these isolates were tested at seven different concentrations. Fourth, their efficacy in controlling M. incognita on tomato plants was assessed in a greenhouse setting. Fifth, their potential to promote tomato plant growth without nematode inoculation was evaluated. The results revealed that the 102 bacterial isolates caused mortality ranging from 2 to 79.57% for M. incognita J2. At a 10% dilution, five isolates (B. cereus IBCBb130, P. aeruginosa IBCBb122, B. cereus IBCBb116, B. proteolyticus IBCBb136, and Serratia sp. IBCBb118) maintained mortality rates above 68%. Among these strains, B. cereus IBCBb130 and P. aeruginosa IBCBb122 were particularly effective, with B. cereus IBCBb130 showing high mortality rates under laboratory conditions and P. aeruginosa IBCBb122 significantly reducing nematode populations in greenhouse pots. Additionally, Serratia sp. IBCBb118 demonstrated notable potential for promoting tomato plant growth. In conclusion, specific bacterial isolates exhibit strong potential for the biocontrol of M. incognita and enhancement of tomato plant growth, suggesting a viable alternative to chemical nematicides. These findings provide insight into the development of sustainable agricultural practices through targeted manipulation of the soil microbiota. Future research should explore the underlying mechanisms of these interactions and their long-term effects on crop yield and soil health.

Multistress‐tolerant Pseudomonas aeruginosa SSVP3 uses multiple strategies to control Meloidogyne incognita in tomato

AbstractTomato is the most important horticultural crop, and India is its second largest producer; however, the plant‐pathogenic nematode Meloidogyne incognita is a serious pest of tomato, causing detrimental losses in its production. This study is focused on the use of multistress‐tolerant Pseudomonas aeruginosa SSVP3 to control this devastating nematode and attempts to decipher the roles of its metabolites and volatile organic compounds (VOCs) during this interaction. The supernatant of P. aeruginosa SSVP3 caused 58% mortality in M. incognita J2 juveniles. The major nematicidal metabolites, identified using liquid chromatography‐mass spectrometry, were pyocyanin, pyoluteorin, pyochelin, benthocyanin and phenazines. The VOCs secreted by P. aeruginosa SSVP3, which were identified via gas chromatography–mass spectrometry (HS‐SPME‐GC–MS) in a mixture, caused 98% mortality in M. incognita J2 juveniles. In a pot experiment, seedling root treatment with P. aeruginosa SSVP3 activated the induced systemic response in tomato plants to M. incognita by increasing the activity of defence and antioxidant enzymes as well as the proline and phenolic contents. The malondialdehyde content in P. aeruginosa SSVP3‐treated plants decreased as compared to those in the other treatment groups. The number of galls formed and the number of endoparasitic stages of M. incognita in the untreated nematode control plants were much greater (19 and 35) than those in the P. aeruginosa SSVP3‐treated (0.33 and 1) and chemically treated groups (0.67 and 1.33). These results indicate that P. aeruginosa and its secreted metabolites and VOCs have a high potential for controlling nematodes.

Correction: Nematicidal potency of silver nanoparticles and extracts from Borreria verticillata (L.) G. Mey against Meloidogyne incognita on tomato plants

Correction: Nematicidal potency of silver nanoparticles and extracts from Borreria verticillata (L.) G. Mey against Meloidogyne incognita on tomato plants

Exploring the nematicidal mechanisms and control efficiencies of oxalic acid producing Aspergillus tubingensis WF01 against root-knot nematodes.

Root-knot nematodes (RKN; Meloidogyne spp.) are among the highly prevalent and significantly detrimental pathogens that cause severe economic and yield losses in crops. Currently, control of RKN primarily relies on the application of chemical nematicides but it has environmental and public health concerns, which open new doors for alternative methods in the form of biological control. In this study, we investigated the nematicidal and attractive activities of an endophytic strain WF01 against Meloidogyne incognita in concentration-dependent experiments. The active nematicidal metabolite was extracted in the WF01 crude extract through the Sephadex column, and its structure was identified by nuclear magnetic resonance and mass spectrometry data. The strain WF01 was identified as Aspergillus tubingensis based on morphological and molecular characteristics. The nematicidal and attractive metabolite of A. tubingensis WF01 was identified as oxalic acid (OA), which showed solid nematicidal activity against M. incognita, having LC50 of 27.48 μg ml-1. The Nsy-1 of AWC and Odr-7 of AWA were the primary neuron genes for Caenorhabditis elegans to detect OA. Under greenhouse, WF01 broth and 200 μg ml-1 OA could effectively suppress the disease caused by M. incognita on tomatoes respectively with control efficiency (CE) of 62.5% and 70.83%, and promote plant growth. In the field, WF01-WP and 8% OA-WP formulations showed moderate CEs of 51.25%-61.47% against RKN in tomato and tobacco. The combined application of WF01 and OA resulted in excellent CEs of 66.83% and 69.34% toward RKN in tomato and tobacco, respectively. Furthermore, the application of WF01 broth or OA significantly suppressed the infection of J2s in tomatoes by upregulating the expression levels of the genes (PAL, C4H, HCT, and F5H) related to lignin synthesis, and strengthened root lignification. Altogether, our results demonstrated that A. tubingensis WF01 exhibited multiple weapons to control RKN mediated by producing OA to lure and kill RKN in a concentration-dependent manner and strengthen root lignification. This fungus could serve as an environmental bio-nematicide for managing the diseases caused by RKN.

Biocontrol potential of endophytic fungi against phytopathogenic nematodes on potato (Solanum tuberosum L.)

Root-knot nematodes (RKNs) are a vital pest that causes significant yield losses and economic damage to potato plants. The use of chemical pesticides to control these nematodes has led to environmental concerns and the development of resistance in the nematode populations. Endophytic fungi offer an eco-friendly alternative to control these pests and produce secondary metabolites that have nematicidal activity against RKNs. The objective of this study is to assess the efficacy of Aspergillus flavus (ON146363), an entophyte fungus isolated from Trigonella foenum-graecum seeds, against Meloidogyne incognita in filtered culture broth using GC–MS analysis. Among them, various nematicidal secondary metabolites were produced: Gadoleic acid, Oleic acid di-ethanolamide, Oleic acid, and Palmitic acid. In addition, biochemical compounds such as Gallic acid, Catechin, Protocatechuic acid, Esculatin, Vanillic acid, Pyrocatechol, Coumarine, Cinnamic acid, 4, 3-indol butyl acetic acid and Naphthyl acetic acid by HPLC. The fungus was identified through morphological and molecular analysis, including ITS 1–4 regions of ribosomal DNA. In vitro experiments showed that culture filtrate of A. flavus had a variable effect on reducing the number of egg hatchings and larval mortality, with higher concentrations showing greater efficacy than Abamectin. The fungus inhibited the development and multiplication of M. incognita in potato plants, reducing the number of galls and eggs by 90% and 89%, respectively. A. flavus increased the activity of defense-related enzymes Chitinas, Catalyse, and Peroxidase after 15, 45, and 60 days. Leaching of the concentrated culture significantly reduced the second juveniles’ stage to 97% /250 g soil and decreased the penetration of nematodes into the roots. A. flavus cultural filtrates via soil spraying improved seedling growth and reduced nematode propagation, resulting in systemic resistance to nematode infection. Therefore, A. flavus can be an effective biological control agent for root-knot nematodes in potato plants. This approach provides a sustainable solution for farmers and minimizes the environmental impact.

Effect of Silybum marianum (L.) Gaertn. Leaf and Seed Extracts Prepared Using Different Solvents on Root-Knot Nematode

Objective: The nematicidal effect of milk thistle leaves and seeds prepared with different solvents on Meloidogyne incognita was investigated. Materials and Methods: Acetone, ethanol and distilled water were used. The in vitro was carried out in 6 cm petri dishes. The extractions were studied with 500 and 1000 μg/ml (ppm). The in vitro and pot experiments designed random plots with 5 replications for each extraction, solvent and concentration. Four hundred second stage juvenile larvae (J2) were used as inoculum and dead individuals were counted after 48 hours. Five days after transplantation, nematode inoculation was carried out with 500 J2 per pot. After 24 hours, 30 ml of the solution was applied to the soil at 1000 ppm concentration. The experiment was terminated for 50 days. Then, gall and egg mass counts were made. Results: In vitro, the mortality rate at 1000 ppm was found to be similar in acetone (78.0%) and ethanol (80.8%) solvents in leaf extraction, while the highest was detected in ethanol (94.0%) in the seed extract. In distilled water solvent, 68.0% mortality was determined in the leaf extract and 62.2% mortality in the seed extract. There was no statistically significant difference between the leaf and seed extracts in number of galls and egg masses. No statistical difference could be determined between the solvents in the number of egg masses in seed extraction. While the number of galls in the leaf extract was found to be higher than in acetone (8.8 unit/root) and ethanol (8.0 unit/root) in distilled water (18.0 unit/root) and the difference between them was found to be significant, no statistically significant difference in the number of egg mass between the solvents. Conclusion: It was observed that all solvents of the leaf and seed extract suppressed galls and egg masses by more than 80% compared to the control.

Design, Synthesis, Nematicidal Evaluation, and Molecular Docking Study of Pyrano[3,2-c]pyridones against Meloidogyne incognita.

Root-knot nematodes pose a serious threat to crops by affecting production and quality. Over a period of time, substantial work has been done toward the development of effective and environmentally benign nematicidal compounds. However, due to the inefficiencies of previously reported synthetics in achieving the target of safe, selective, and effective treatment, it is necessary to develop new efficacious and safer nematicidal agents considering human health and environment on top priority. This work aims to highlight the efficient and convenient l-proline catalyzed synthesis of pyrano[3,2-c]pyridone and their use as potential nematicidal agents. In vitro results of larval mortality and egg hatching inhibition revealed maximum nematicidal activity against Meloidogyne incognita from compounds 15b, 15m, and 15w with LC50 values of 28.8, 46.8, and 49.18 μg/mL at 48 h, respectively. Under similar conditions, pyrano[3,2-c]pyridones derivatives 15b (LC50 = 28.8 μg/mL) was found at par with LC50 (26.92 μg/mL) of commercial nematicide carbofuran. The in vitro results were further validated with in silico studies with the most active compound 15b nematicidal within the binding to the pocket of acetylcholine esterase (AChE). In docking, binding free energy values for compound 15b were found to be -6.90 kcal/mol. Results indicated that pyrano[3,2-c]pyridone derivatives have the potential to control M. incognita.