Most Australian vegetable growers apply fumigants or nematicides as a precautionary nematode control measure when crops susceptible to root-knot nematode (RKN, Meloidogyne spp.) are grown in soils and environmental conditions suitable for the nematode. The only way growers can make rational decisions on whether these expensive and environmentally disruptive chemicals are required is to regularly monitor RKN populations and decide whether numbers prior to planting are high enough to cause economic damage. However, such monitoring programs are difficult to implement because nematode quantification methods vary in efficiency and the damage threshold for RKN on highly susceptible vegetable crops is often < 10 root-knot nematodes /200 mL soil. Consequently, five nematode quantification methods were tested to see whether they could reliably detect these very low population densities of RKN. Two novel methods produced consistent results: 1) extracting nematodes from 2 L soil samples using Whitehead trays, quantifying the RKN DNA in the nematode suspension using molecular methods, and generating a standard curve so that the molecular results provided an estimate of the total number of RKN individuals in the sample, and 2) a bioassay in which two tomato seedlings were planted in pots containing 2 L soil and the number of galls produced on roots were counted after 21-25 days. Both methods could be used to quantify low populations of RKN, but bioassays are more practical because expensive equipment and facilities are not required and they can be done at a local level by people lacking nematological or molecular skills.

The root-knot nematode (RKN), Meloidogyne javanica, causes severe damage to a wide variety of crops. These nematodes significantly reduce tomato yield globally, causing symptoms such as stunted growth, galls on roots, chlorosis, and wilting, ultimately leading to host death. Classical nematode control methods, such as the application of chemical nematicides, are very effective; however, their use is limited due to conflicts with sustainable agriculture. Therefore, biological methods, are gaining attention as more environmentally friendly options. In the present study, 47 strains of bacteria were isolated from the rhizosphere of RKN-infected plants. The effect of these strains was studied on egg hatching and second stage infective juveniles (J2s) mortality of M. javanica, in vitro. Then, three holes were made in the soil around the roots of non-inoculated and nematode inoculated tomato plants and a suspension of 15 mL of three isolates with the greatest negative effect on hatching and J2s mortality (107 CFU/ml), was poured into the holes. Stenotrophomonas maltophilia CPHE1, Peribacillus frigoritolerans Rhs-L31 and Bacillus cereus Pt0-RL12 improved the vegetative indices of inoculated plants compared to control plants. These strains significantly reduced nematode hatching and significantly increased mortality of nematode J2s; and in greenhouse pot experiments significantly reduced the number of nematode eggs and egg masses, root galls, and nematode reproduction factor. In each case, inoculation with the bacterial strains significantly increased peroxidase and superoxide dismutase activity, and decreased catalase activity in tomato roots infected with M. javanica. The present study indicates the potential of these bacterial strains for biocontrol of M. javanica on tomato.

Root-knot nematode (RKN) causes severe yield loss in cucumber. Understanding the interactions of biocontrol agent-soil microbiomes and RKNs is essential for enhancing the efficacy of biocontrol agents and nematicides to curb RKN damage to cucumber. The field experiment in this work was conducted to determine the ability of Bacillus velezensis GHt-q6 to colonize cucumber plants, investigate its effect on the control of RKNs, and assess its influence on soil microbiology in the inter-root zone of cucumber plants. After 10 days post-treatment (DPT), GHt-q6-Rif could stably colonize the roots (4.55 x 10(4) cfu center dot g(-1)), stems (3.60 x 10(3) cfu center dot g(-1)), and leaves (3.60 x 10(2) cfu center dot g(-1)) of cucumber. The high-throughput sequencing results suggested that the bacterial community diversity increased at the late development phase (p > 0.05). The strain GHt-q6 increased the relative abundance of beneficial bacteria (Gemmatimonadaceae, Sphingomonadaceae, Pseudomonadaceae). Throughout the complete cucumber growth period, strain GHt-q6 significantly increased soil urease, sucrase, accessible potassium, and phosphorus (p < 0.05). However, strain GHt-q6 had a minimal effect on catalase activity. At the pulling stage, strain GHt-q6 exhibited 43.35% control effect on cucumber RKNs, which was 7.54% higher than that of Bacillus subtilis. The results highlighted the significant potential of the strain GHt-q6 to manage cucumber RKNs and improve soil microecology. Hence, the applications of B. velezensis GHt-q6 can enhance the nematicidal action to curb RKN infecting cucumber.

The Meloidogyne spp., commonly known as root-knot nematodes (RKN), are obligate sedentary endoparasites considered among the most damaging plant-parasitic nematodes globally. They harm crops by using parasitic proteins to alter host cell physiology, which promotes parasitism and reduces crop yield. Traditional RKN management, primarily through chemical control, negatively impacts the nutritional value, soil texture, and vegetable production, and poses risks to human health and the environment. An emerging eco-friendly and costeffective alternative is the use of plant growth-promoting microbes (PGPM)-mediated biological approaches. The PGPM enhances plant growth directly by solubilizing phosphorus and iron, fixing nitrogen, producing phytohormones, siderophores, and ammonia, or indirectly through competition, antibiosis, hydrogen cyanide, 1-aminocyclopropane-1-carboxylate (ACC) deaminase enzyme, and exopolysaccharides (EPS) production. This review explores various RKN management strategies, emphasizing green biological approaches, their benefits and drawbacks, current commercial status and usage, and the underlying genes, challenges, and limitations associated with these methods.

BACKGROUNDRoot-knot nematodes (RKN, Meloidogyne spp.) are economically significant pests that cause immense damage to a wide range of crops. Among them, M. incognita and M. enterolobii are of particular concern, as their high virulence and broad host range. RKN are challenging for detection due to their subterranean lifestyle underground. Also, the mixed infection of nematodes in field crops complicates the need for more accurate diagnostic and quantification technologies.RESULTSTo address this challenge, we developed and optimized a novel duplex droplet digital PCR (ddPCR) method, using primer/probe sets targeting M. incognita and M. enterolobii, to simultaneously identify and quantify both species within a single assay. The innovative ddPCR diagnostic demonstrated excellent performance in terms of sensitivity, precision and reproductivity when quantifying the eggs and soil samples containing juveniles of both species. Moreover, the application of the duplex ddPCR method enables the monitoring of population dynamics of M. incognita and M. enterolobii under competitive environmental conditions. Our results indicated that the reproduction factor of M. incognita possibly inhibited when in mixed populations of M. enterolobii.ConclusionIn this study, we first applied duplex ddPCR technique for differentiating mixed infections of M. incognita and M. enterolobii, offering a valuable tool for species detection and quantification. It enables the monitoring of population dynamics for both species, which is crucial for providing theoretical guidance for the implementation of timely and effective control measures. (c) 2024 Society of Chemical Industry.

The root-knot nematode (Meloidogyne spp.) is an obligate plant parasite and is one of the largest threats to the Australian sweetpotato industry, causing crop losses of up to 57% of marketable yield. In this study, two potential fungal biocontrol agents were encapsulated in alginate granules and their nematophagous activity was assessed in a laboratory-based microcosm experiment. Both species of fungi significantly reduced numbers of root-knot nematodes in red ferrosol soil. A greater reduction was observed in untreated field soil prior to introduction of root-knot nematodes and fungal biocontrol agents compared to soil that had been heat-sterilised. In a ten-week glasshouse experiment, no significant difference in the root-knot nematode populations in sweetpotato roots and soil was found between fungal biocontrol agent and control treatments. There was a trend towards an increase in the sweetpotato storage root weight and reduction in storage root damage in fungal biocontrol agent compared to control treatments, and both yield and damage levels were similar to those achieved from the use of chemical nematicide treatments. These results demonstrate the need for greater understanding of the interactions between soil biological populations and introduced nematophagous fungi if effective biocontrol is to be consistently achieved with these bioagents under field conditions.

Root-knot nematodes (RKN; Meloidogyne spp.) are among the most damaging plant-parasitic nematodes. They parasitize almost every species of higher plant and induce the formation of galls along the plant roots, which are detrimental to plant growth. North Carolina's leading field crops are sweetpotato (Ipomoea batatas (L.) Lam.), soybean (Glycine max L. Merr), cotton (Gossypium hirsutum L.), and tobacco (Nicotiana tabacum L.), which are all hosts to several root-knot nematode species. This pathogen represents a major threat to farmers, obligating them to seek alternative crops that are non-host to root-knot nematodes that will help decrease soil populations and provide economic revenue. We tested seven sesame cultivars for their host status and potential resistance to four Meloidogyne species (M. arenaria, M. incognita, M. enterolobii, and M. hapla). We inoculated sesame seedlings with 1,000 nematode eggs of each species. Sixty days after inoculation, we harvested the plants to evaluate a visual gall severity rating, measure final egg counts, and calculate the reproductive factor (RF). All sesame cultivars had a significantly lower RF than the tomato (Solanum lycopersicum L.) cv. Rutgers control for all species of RKN except M. arenaria. The RF values for sesame cultivars inoculated with M. incognita and M. hapla were not significantly different from one another; however, there were significant differences in RF among sesame cultivars inoculated with M. enterolobii, suggesting that genetic variability of the host may play an important role in host status and conferring resistance.

Meloidogyne spp. are the most devastating plant-parasitic nematodes affecting tomato worldwide. Although resistant cultivars and rootstocks are used, selection for virulence occurs in the pathogen. Consequently, using other resistance sources, such as Solanum torvum, could improve resistance durability. Several experiments in microplots and plastic greenhouses were carried out to determine the potential use of S. torvum as a tomato rootstock to protect against M. incognita and M. javanica. In microplots, the relationship between nematode density at transplanting (Pi) and multiplication rate did not differ between Meloidogyne species in either ungrafted or grafted tomato. However, maximum multiplication rate and maximum density on grafted tomato were 1.27% and 2.93% those on ungrafted, respectively. The grafted tomato plants yielded between 2.9 and 7.5 more times than the ungrafted plants at Pi >= 100 eggs + J2s per 100 cm(3) of soil, but no differences were observed in plastic greenhouse where a large amount of scion-rooting occurred. In microplots, the quality of the tomato fruits of ungrafted and grafted plants was affected by the Pi. In parallel, some pot experiments were conducted on S. torvum and susceptible eggplant to determine the putative selection for nematode virulence to S. torvum and the nematode fitness cost. These showed that the nematode subpopulations infected and reproduced less on S. torvum than on eggplant. However, the female fertility was only reduced after development of three or four subpopulations on S. torvum. Finally, a histopathological study showed that nematode infection and development in S. torvum was delayed compared to eggplant.

This study investigates the efficacy of Trichoderma spp. and Bacillus spp., as well as their gamma radiation-induced mutants, as potential biological control agents against Meloidogyne javanica (Mj) in tomato plants. The research encompasses in vitro assays, greenhouse trials, and molecular identification methodologies to comprehensively evaluate the biocontrol potential of these agents. In vitro assessments reveal significant nematicidal activity, with Bacillus spp. demonstrating notable effectiveness in inhibiting nematode egg hatching (16-45%) and inducing second-stage juvenile (J2) mortality (30-46%). Greenhouse trials further confirm the efficacy of mutant isolates, particularly when combined with chitosan, in reducing nematode-induced damage to tomato plants. The combination of mutant isolates with chitosan reduces the reproduction factor (RF) of root-knot nematodes by 94%. By optimizing soil infection conditions with nematodes and modifying the application of the effective compound, the RF of nematodes decreases by 65-76%. Molecular identification identifies B. velezensis and T. harzianum as promising candidates, exhibiting significant nematicidal activity. Overall, the study underscores the potential of combined biocontrol approaches for nematode management in agricultural settings. However, further research is essential to evaluate practical applications and long-term efficacy. These findings contribute to the development of sustainable alternatives to chemical nematicides, with potential implications for agricultural practices and crop protection strategies.

Yam is an important medicinal and edible dual-purpose plant with high economic value. However, nematode damage severely affects its yield and quality. One of the major effects of nematode infestations is the secondary infection of pathogenic bacteria or fungi through entry wounds made by the nematodes. Understanding the response of the symbiotic microbial community of yam plants to nematodes is crucial for controlling such a disease. In this study, we investigated the rhizosphere and how endophytic microbiomes shift after nematode infection during the tuber expansion stage in the Dioscorea opposita Thunb. cultivar Tiegun. Our results revealed that soil depth affected the abundance of nematodes, and the relative number of Meloidogyne incognita was higher in the diseased soil at a depth of 16 to 40 cm than those at a depth of 0 to 15 and 41 to 70 cm. The abundance of and interactions among soil microbiota members were significantly correlated with root-knot nematode (RKN) parasitism at various soil depths. However, the comparison of the microbial alpha-diversity and composition between healthy and diseased rhizosphere soil showed no difference. Compared with healthy soils, the co-occurrence networks of M. incognita-infested soils included a higher ratio of positive correlations linked to plant health. In addition, we detected a higher abundance of certain taxonomic groups belonging to Chitinophagaceae and Xanthobacteraceae in the rhizosphere of RKN-infested plants. The nematodes, besides causing direct damage to plants, also possess the ability to act synergistically with other pathogens, especially Ramicandelaber and Fusarium, leading to the development of disease complexes. In contrast to soil samples, RKN parasitism specifically had a significant effect on the composition and assembly of the root endophytic microbiota. The RKN colonization impacted a wide variety of endophytic microbiomes, including Pseudomonas, Sphingomonas, Rhizobium, Neocosmospora, and Fusarium. This study revealed the relationship between RKN disease and changes in the rhizosphere and endophytic microbial community, which may provide novel insights that help improve biological management of yam RKNs.