Identification of Meloidogyne species on traditional Chinese medicine plants in the Qinling mountain area of China and their aggressiveness to different medicinal herbs.

Root-knot nematodes (Meloidogyne spp.) are plant-parasitic nematodes that cause serious damage on a worldwide basis. There are many species of traditional Chinese medicine (TCM) plants, but only a few have been reported to be infected by Meloidogyne species. From 2020 to 2022, a survey was conducted in the Qinling mountain area, which is the main producing region of TCM plants in China. Obvious galling symptoms were observed on the root systems of fifteen species of TCM plants. Females were collected from diverse diseased TCM plants and subsequently identified at morphological and molecular level. Among the twenty diseased root samples collected, Meloidogyne hapla populations were identified in twelve samples (60%) and Meloidogyne incognita populations were identified in eight samples (40%). Among the fifteen species of diseased TCM plants, eight of them, namely Scutellaria baicalensis, Leonurus japonicus, Dioscorea zingiberensis, Cornus officinalis, Viola philippica, Achyranthes bidentata, Senecio scandens, and Plantago depressa were reported to be infected by Meloidogyne species for the first time. The host status of five species of TCM plants for two M. hapla isolates and one M. incognita isolate from TCM plants in this study was then evaluated. Differences in TCM plants' response to nematode infection were apparent when susceptibility was evaluated by the egg counts per gram fresh weight of root and the reproduction factor of the nematodes. Among the five species of TCM plants tested, Salvia miltiorrhiza and Gynostemma pentaphyllum were the most susceptible, while S. baicalensis and V. philippica were not considered suitable hosts for M. hapla or M. incognita.

Potential of nicotinamide adenine dinucleotide (NAD) for management of root-knot nematode in tomato.

Nicotinamide adenine dinucleotide (NAD) has been shown to induce plant defense responses to different plant pathogens, including reducing northern root-knot nematode, Meloidogyne hapla, penetration and increasing plant mass in tomato. We wanted to further evaluate NAD that are effective against the more economically important species, M. incognita and whether NAD treatments of tomato seedlings in transplant trays can protect plants in the field. Different NAD concentrations (1 mM, 0.1 mM and 0.01 mM) and three application timings (pre; post; pre and post inoculation) were evaluated in growth room and greenhouse trials. The highest tested NAD concentration (1 mM) suppressed second-stage juveniles (J2) infection for all three application methods. Root gall ratings at 30 days after inoculation (DAI) were also suppressed by 1 mM NAD compared to the other two concentrations, and egg mass number was significantly suppressed for all concentrations and application timings compared to the non-treated control. The rate of 1 mM NAD for all three application timings also improved plant growth at 30 DAI. Long-term effects of 1 mM NAD (pre, pre + post, or post applications) on nematode infection, growth and yield of tomato were evaluated in two additional experiments. All NAD applications suppressed root galls after 60 days, but only the pre + post 1 mM NAD application suppressed gall severity at 105 days, as well as suppressed egg counts by 50% at 60 DAT. No significant difference in plant biomass and fruit yield after 105 days was observed among the treatments. Two field trials were conducted in spring and fall 2020 using tomato seedlings (cv. HM 1823) treated with two different NAD concentrations (1 mM and 5 mM in spring; 5 mM and 10 mM in fall) and transplanting seedlings in fumigated (chloropicrin ± 1,3-dichloropropene) and non-fumigated plastic-mulch beds. No significant impact of NAD in terms of reducing RKN severity or overall tomato growth and production was seen in fumigated beds, but in non-fumigated beds 5 mM NAD slightly increased early fruit yield in spring, and 10 mM NAD reduced root-knot soil populations in fall.

Genome-wide analysis of a putative lipid transfer protein LTP_2 gene family reveals CsLTP_2 genes involved in response of cucumber against root-knot nematode (Meloidogyne incognita).

Plant lipid transfer proteins (LTPs) are small basic proteins that play important roles in the regulation of various plant biological processes as well as the response to biotic and abiotic stresses. However, knowledge is limited on how this family of proteins is regulated in response to nematode infection in cucumber. In the present study, a total of 39 CsLTP_2 genes were identified by querying databases for cucumber-specific LTP_2 using a Hidden Markov Model approach and manual curation. The family has a five-cysteine motif (5CM) with the basic form CC-Xn-CXC-Xn-C, which differentiates it from typical nsLTPs. The members of CsLTP_2 were grouped into six families according to their structure and their phylogenetic relationships. Expression data of CsLTP_2 genes in 10 cucumber tissues indicated that they were tissue-specific genes. Two genes showed significant expression change in roots of resistant and susceptible lines during nematode infection, indicating their involvement in response to Meloidogyne incognita. This systematic analysis provides a foundation of knowledge for future studies of the biological roles of CsLTP_2 genes in cucumber in response to nematode infection and may help in the efforts to improve M. incognita-resistance breeding in cucumber.

Nematicidal activity of 5-iodoindole against root-knot nematodes.

Multi-drug resistance in nematodes is a serious problem as lately several resistant phenotypes have emerged following the intermittent usage of synthetic nematicides. Contemporary research continues to focus on developing and/or repurposing small molecule inhibitors that are eco-friendly. Here, we describe the repurposing of the indole derivative, 5-iodoindole, as a nematicide for the root-knot nematode, Meloidogyne incognita. 5-Iodoindole effectively killed juveniles and freshly hatched juveniles by inducing multiple vacuole formation. Notably, at higher dosage (50 µg/mL), 5-iodoindole induced rapid juvenile death within 6 h. Microscopic analysis confirmed that the rapid death was due to the generation of reactive oxygen species (ROS). Computational docking attributed this ROS production to the antagonistic effect of 5-iodoindole on glutathione S-transferase (GST), which is known to play a critical role in the suppression of ROS in nematode models. Furthermore, 5-iodoindole also effectively reduced the gall formations and eggs masses of M. incognita on Solanum lycopersicum roots in pot experiments, and importantly it did not harm the physiological properties of the plant. Overall, the study provides valuable insights on the use of 5-iodoindole as an alternate measure to control root-knot nematodes. Overall, our findings suggest the efficacy of 5-iodoindole should be studied under field conditions.

Synthesis and nematicidal activities of 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea against Meloidogyne incognita.

Two series of novel 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea were designed and synthesized. The bioassay results showed that most of the test compounds showed good nematicidal activity against M. incognita at the concentration of 10.0mgL-1in vivo. The compounds A13, A17 and B3 showed excellent nematicidal activity on the second stage juveniles of the root-knot nematode with the inhibition rate of 51.3%, 58.3% and 51.3% at the concentration of 1.0mgL-1 respectively. It suggested that the structure of 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea could be optimized further.

Reproduction of Meloidogyne incognita and M. graminis on Several Grain Sorghum Hybrids.

A total of 27 grain sorghum hybrids were evaluated in a series of greenhouse experiments to determine their susceptibility to Meloidogyne incognita race 3 and M. graminis. Each hybrid was inoculated with 2,000 nematode eggs/pot. Reproduction by M. incognita was numerically greater than M. graminis on 93% of the hybrids tested, indicating that grain sorghum is a better host for M. incognita than M. graminis. A wide variation in host suitability was observed on these hybrids in a second experiment as reproduction by M. incognita ranged from 395 to 3,818 eggs/g of root. Only two hybrids, Terral RV9782 and RV9823, consistently supported <20% reproduction by M. incognita compared to the most susceptible hybrid, Golden Acres 5556. Reproduction of four isolates of M. incognita was evaluated on six selected hybrids in a third greenhouse experiment. Hybrid susceptibility was similar to that observed in the previous experiment for all isolates. A difference in isolate aggressiveness was observed between two of the four isolates across all hybrids. In fields where damaging populations of M. incognita are present, most grain sorghum hybrids will likely maintain or increase the nematode population for the subsequent crop.

Diversity of Root-knot Nematodes Associated with Tubers of Yam (Dioscorea spp.) Established Using Isozyme Analysis and Mitochondrial DNA-based Identification.

The root-knot nematodes (RKN), Meloidogyne spp., represent an important threat to yam (Dioscorea spp.) production in West Africa. With the aim to establish the diversity of RKN species affecting yam tubers, for control and resistance screening purposes, surveys were conducted in the main yam producing areas of Nigeria. Galled tubers (N = 48) were collected from farmers' stores and markets in nine states in Nigeria and in one district in Ghana. RKN isolated from yam tubers were identified using enzyme phenotyping (esterase and malate dehydrogenase) and mitochondrial DNA (mtDNA) NADH dehydrogenase subunit 5 (Nad5) barcoding. Examination of 48 populations revealed that yam tubers were infested by Meloidogyne incognita (69%), followed by M. javanica (13%), M. enterolobii (2%), and M. arenaria (2%). Most of the tubers sampled (86%) were infected by a single species, and multiple species of RKN were detected in 14% of the samples. Results of both identification methods revealed the same species, confirming their accuracy for the identification of these tropical RKN species. In addition to M. incognita, M. javanica, and M. enterolobii, we report for the first time M. arenaria infecting yam tubers in Nigeria. This finding extends the list of yam pests and calls for caution when developing practices for yam pest management.

Broad Meloidogyne Resistance in Potato Based on RNA Interference of Effector Gene 16D10.

Root-knot nematodes (Meloidogyne spp.) are a significant problem in potato (Solanum tuberosum) production. There is no potato cultivar with Meloidogyne resistance, even though resistance genes have been identified in wild potato species and were introgressed into breeding lines. The objectives of this study were to generate stable transgenic potato lines in a cv. Russet Burbank background that carry an RNA interference (RNAi) transgene capable of silencing the 16D10 Meloidogyne effector gene, and test for resistance against some of the most important root-knot nematode species affecting potato, i.e., M. arenaria, M. chitwoodi, M. hapla, M. incognita, and M. javanica. At 35 days after inoculation (DAI), the number of egg masses per plant was significantly reduced by 65% to 97% (P < 0.05) in the RNAi line compared to wild type and empty vector controls. The largest reduction was observed in M. hapla, whereas the smallest reduction occurred in M. javanica. Likewise, the number of eggs per plant was significantly reduced by 66% to 87% in M. arenaria and M. hapla, respectively, compared to wild type and empty vector controls (P < 0.05). Plant-mediated RNAi silencing of the 16D10 effector gene resulted in significant resistance against all of the root-knot nematode species tested, whereas R Mc1(blb) , the only known Meloidogyne resistance gene in potato, did not have a broad resistance effect. Silencing of 16D10 did not interfere with the attraction of M. incognita second-stage juveniles to roots, nor did it reduce root invasion.

Tulipaline A: structure-activity aspects as a nematicide and V-ATPase inhibitor.

Carbonyl groups are known to form covalent adducts with endogenous proteins, but so far, their nematicidal mechanism of action of has been overlooked. The nematicidal activity of ten lactones was tested in vitro against the root knot nematodes Meloidogyne incognita and Meloidogynearenaria. In particular, the saturated lactones α-methylene-γ-butyrolactone or tulipaline A (1) and γ-butyrolactone (3) were active against M. incognita with an EC50/48h of 19.3±10.0 and 40.0±16.2mg/L respectively. Moreover the α, ß-unsaturated lactone 5,6-dihydro-2H-pyran-2-one (2) exhibited the strongest nematicidal activity against the two species with EC50/48h 14.5±5.3 and 21.2±9.7mg/L respectively. Here we propose that the toxic effects of lactones and aldehydes on M.incognita and M. arenaria might be a consequence of their vacuolar-type H(+)-ATPase (V-ATPase) inhibition activity; in fact α-methylene-γ-butyrolactone (1) and salicylaldehyde (12) produced an increased pH in lysosomal-like organelles on HeLa human cell line and this alteration was most likely related to a V-ATPase impairment.

Optimization and utilization of emerging waste (fly ash) for growth performance of chickpea (Cicer arietinum L.) plant and mitigation of root-knot nematode (Meloidogyne incognita) stress.

The sustainable management of large amounts of fly ash (FA) is a concern for researchers, and we aim to determine the FA application in plant development and nematicidal activity in the current study. A pot study is therefore performed to assess the effects of adding different, FA-concentrations to soil (w/w) on the infection of chickpea plants with the root-knot nematode Meloidogyne incognita. Sequence characteristic amplified region (SCAR) and internal transcribed spacer (ITS) region-based-markers were used to molecularly confirm M. incognita. With better plant growth and chickpea yield performance, FA enhanced the nutritious components of the soil. When compared with untreated, uninoculated control (UUC) plants, the inoculation of M. incognita dramatically reduced chickpea plant growth, yield biomass, and metabolism. The findings showed that the potential of FA to lessen the root-knot nematode illness in respect of galls, egg-masses, and reproductive attributes may be used to explain the mitigating effect of FA. Fascinatingly, compared with the untreated, inoculated control (UIC) plants, the FA treatment, primarily at 20%, considerably (p ≤ 0.05) boosted plant growth, yield biomass, and pigment content. Additionally, when the amounts of FA rose, the activity of antioxidants like superoxide dismutase-SOD, catalase-CAT, and peroxidase-POX as well as osmo-protectants like proline gradually increased. Therefore, our findings imply that 20% FA can be successfully applied as a potential strategy to increase biomass yield and plant growth while simultaneously reducing M. incognita infection in chickpea plants.

Morphological and Morphometrical Characterization of Meloidogyne incognita from Different Host Plants in Four Districts of Punjab, India.

The population of M. incognita, the root knot nematode (RKN) was found infesting five different host plants (okra, banana, sunflower, bottle gourd, and brinjal) out of 24 examined from four districts of Punjab, India (Gurdaspur, Ludhiana, Patiala, and Hoshiarpur). Morphological and morphometrical characterization indicated that in the case of mature female, the characters of body length and width, neck length, ratio 'a', anus to tail terminus (ATT), interphasmid distance (IPD), and perineal pattern were recorded as stable characters. These taxonomic characters can be reliable for identification. All characters of second-stage juvenile (J2) such as body length, stylet length, head to median bulb length (H-MB), distance from median bulb to excretory pore (MB-EP), tail length, anal body width (ABW), and ratios C and C' were highly variable. Analysis of interpopulation morphometric characters of mature female of M. incognita, namely, body length, width, and ratio 'a' were moderately variable characters (CV 0.26% to 20%) and stylet length, neck length, length of median bulb (LMB), and width of median bulb (WMB) were highly variable (CV 1.0% to 36.1%). In the perineal pattern, the two characters ATT and IPD were moderately variable (CV 8.8% to 17.6%) and two characters, anus to vulval slit (AVS) and length of vulval slit (LVS), were highly variable (CV 2.1% to 40.5%). In J2, body length, stylet length, H-MB, MB-EP, ABW, tail length, ratios C, and C' were highly variable characters (CV > 12%).

Evaluation of Cover Crops with Potential for Use in Anaerobic Soil Disinfestation (ASD) for Susceptibility to Three Species of Meloidogyne.

Several cover crops with potential for use in tropical and subtropical regions were assessed for susceptibility to three common species of root-knot nematode, Meloidogyne arenaria, M. incognita, and M. javanica. Crops were selected based on potential use as organic amendments in anaerobic soil disinfestation (ASD) applications. Nematode juvenile (J2) numbers in soil and roots, egg production, and host plant root galling were evaluated on arugula (Eruca sativa, cv. Nemat), cowpea (Vigna unguiculata, cv. Iron & Clay), jack bean (Canavalia ensiformis, cv. Comum), two commercial mixtures of Indian mustard and white mustard (Brassica juncea & Sinapis alba, mixtures Caliente 61 and Caliente 99), pearl millet (Pennisetum glaucum, cv. Tifleaf III), sorghum-sudangrass hybrid (Sorghum bicolor × S. bicolor var. sudanense, cv. Sugar Grazer II), and three cultivars of sunflower (Helianthus annuus, cvs. 545A, Nusun 660CL, and Nusun 5672). Tomato (Solanum lycopersicum, cv. Rutgers) was included in all trials as a susceptible host to all three nematode species. The majority of cover crops tested were less susceptible than tomato to M. arenaria, with the exception of jack bean. Sunflower cv. Nusun 5672 had fewer M. arenaria J2 isolated from roots than the other sunflower cultivars, less galling than tomato, and fewer eggs than tomato and sunflower cv. 545A. Several cover crops did not support high populations of M. incognita in roots or exhibit significant galling, although high numbers of M. incognita J2 were isolated from the soil. Arugula, cowpea, and mustard mixture Caliente 99 did not support M. incognita in soil or roots. Jack bean and all three cultivars of sunflower were highly susceptible to M. javanica, and all sunflower cultivars had high numbers of eggs isolated from roots. Sunflower, jack bean, and both mustard mixtures exhibited significant galling in response to M. javanica. Arugula, cowpea, and sorghum-sudangrass consistently had low numbers of all three Meloidogyne species associated with roots and are good selections for use in ASD for root-knot nematode control. The remainder of crops tested had significant levels of galling, J2, and eggs associated with roots, which varied among the Meloidogyne species tested.

Antagonistic Bacteria Bacillus velezensis VB7 Possess Nematicidal Action and Induce an Immune Response to Suppress the Infection of Root-Knot Nematode (RKN) in Tomato.

Meloidogyne incognita, the root-knot nematode (RKN), a devastating plant parasitic nematode, causes considerable damage to agricultural crops worldwide. As a sedentary root parasite, it alters the root's physiology and influences the host's phytohormonal signaling to evade defense. The sustainable management of RKN remains a challenging task. Hence, we made an attempt to investigate the nematicide activity of Bacillus velezensis VB7 to trigger the innate immune response against the infection of RKN. In vitro assay, B. velezensis VB7 inhibited the hatchability of root-knot nematode eggs and juvenile mortality of M. incognita by 87.95% and 96.66%, respectively at 96 hrs. The application of B. velezensis VB7 challenged against RKN induced MAMP-triggered immunity via the expression of transcription factors/defense genes by several folds pertaining to WRKY, LOX, PAL, MYB, and PR in comparison to those RKN-inoculated and healthy control through RT-PCR. Additionally, Cytoscape analysis of defense genes indicated the coordinated expression of various other genes linked to immune response. Thus, the current study clearly demonstrated the effectiveness of B. velezensis VB7 as a potential nematicide and inducer of immune responses against RKN infestation in tomato.

Chemo-Ecological Insights into the Use of the Non-Host Plant Vegetable Black-Jack to Protect Two Susceptible Solanaceous Crops from Root-Knot Nematode Parasitism.

Plant parasitic nematodes (PPNs) develop through three major stages in their life cycle: hatching, infection, and reproduction. Interruption of any of these stages can affect their growth and survival. We used screenhouse pot experiments, laboratory in vitro hatching and mortality assays, and chemical analysis to test the hypothesis that the non-host Asteraceae plant vegetable black-jack (Bidens pilosa) suppresses infection of the PPN Meloidogyne incognita in two susceptible Solanaceae host plants, tomato (Solanum lycopersicum) and black nightshade (S. nigrum). In intercrop and drip pot experiments, B. pilosa significantly reduced the number of galls and egg masses in root-knot nematode (RKN)-susceptible host plants by 3-9-fold compared to controls. Chemical analysis of the most bioactive fraction from the root exudates of B. pilosa identified several classes of compounds, including vitamins, a dicarboxylic acid, amino acids, aromatic acids, and a flavonoid. In in vitro assays, the vitamins and aromatic acids elicited the highest inhibition in egg hatching, whereas ascorbic acid (vitamin) and 2-hydroxybenzoic acid (aromatic acid) elicited strong nematicidal activity against M. incognita, with LC50/48 h values of 12 and 300 ng/µL, respectively. Our results provide insights into how certain non-host plants can be used as companion crops to disrupt PPN infestation.

Combined effects of vermicompost and vermicompost leachate on the early growth of Meloidogyne incognita stressed Withania somnifera (L.) Dunal.

Roots of Withania somnifera, an important medicinal herb, are prone to the infection of Meloidogyne incognita (a root parasitic nematode). The infection lowers the quality and quantity of plant material and poses a challenge in crop cultivation and obtaining desirable yield. In the present study, in vitro inhibitory activity of vermicompost leachate (Vcom-L) was assessed against % hatch and survival of M. incognita in a 96 h assay. Then, Vcom-L was used as soil supplement in combination with vermicompost (Vcom) to evaluate their nematode inhibitory and stress alleviating effect in W. somnifera, in a pot experiment. Root galling intensity and growth performance of nematode-stressed W. somnifera raised from seeds pre-soaked in distilled water (DW), Vcom-L, vermicompost tea (Vcom-T) and different dilutions of Vcom-L were assessed. We observed 79% suppression of egg hatching and 89% juvenile (J2) mortality after 96 h compared to control, at 100% concentration of Vcom-L. Significant reduction in gall formation with increase in growth parameters of seedlings was observed after combined application of Vcom (60% or 100%) + Vcom-L and was evident as enhancement in seedling biomass and contents of chlorophyll and protein. However, proline, total phenolics and malondialdehyde (MDA) content declined significantly in these combinations compared to the control (0% Vcom). Activity of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidise (APX), guaiacol peroxidise (GPX) and glutathione reductase (GR) declined with Vcom as well as Vcom + Vcom-L and corresponded with decline in the accumulation of reactive oxygen species in leaves. Further, 1:5 and 1:10 dilutions of Vcom-L in combination with Vcom (60%) proved beneficial in mitigating the nematode-induced stress in W. somnifera. Present results showed the potential of Vcom and Vcom-L in standardised combination as an effective strategy in controlling the pathogenicity of M. incognita in medicinal plants such as W. somnifera.

Bacterial strains integrated with surfactin molecules of Bacillus subtilis MTCC441 enrich nematocidal activity against Meloidogyne incognita.

Bacteria secrete lipopeptide (surfactin) molecules, which are known to act as natural antibiotics. Recently, research on lipopeptide molecules has grown because of their role in increasing resistance to plant pathogens. Isolated surfactin molecules at several concentrations, viz. 35, 25, 15 and 5 ppm, were used in an in vitro study for analysis of egg hatching inhibition and second-stage juvenile (J2) mortality of the nematode Meloidogyne incognita. Cell suspensions (1.2 × 108  cfu·ml-1 ) of both Bacillus subtilis (MTCC-441) and Pseudomonas putida (MTCC-102) were used in J2 inoculated tomato plants. Root-dip treatment of tomato seedlings with the crude lipopeptide (35 ppm) was also performed to analyse biocontrol potential. MALDI-TOF MS analysis was conducted to determine specific lipopeptide molecules. Data showed nematode egg mortality of 85% in the 35 ppm surfactin concentration 96 h after exposure. The maximum ovicidal activity was 83.97% after incubation with 35 ppm surfactin for the same exposure period. Plant growth attributes and biochemical parameters were significantly improved when bacterial cultures were applied before J2 inoculation of tomato seedlings. We also recorded a reduction in egg masses, nematode population and root galling. The J2 penetration into tomato roots was effective in the root-dip experiments. Surfactin mass peak was determined at m/z 1058 [M+Na]+ using MALDI-TOF MS. These results indicate that bacterial cell suspensions can be used as a potent and versatile source to deal with nematode infection and provide a rich source of bioactive compounds with antinematode activity.

Selection and characterization of two Bacillus thuringiensis strains showing nematicidal activity against Caenorhabditis elegans and Meloidogyne incognita.

Bacillus thuringiensis has been widely used as a biological control agent against insect pests. Additionally, nematicidal strains have been under investigation. In this report, 310 native strains of B. thuringiensis against Caenorhabditis elegans were tested. Only the LBIT-596 and LBIT-107 strains showed significant mortality. LC50s of spore-crystal complexes were estimated at 37.18 and 31.89 µg/mL for LBIT-596 and LBIT-107 strains, respectively, while LC50s of partially purified crystals was estimated at 23.76 and 20.25 µg/mL for LBIT-596 and LBIT-107, respectively. The flagellin gene sequence and plasmid patterns indicated that LBIT-596 and LBIT-107 are not related to each other. Sequences from internal regions of a cry5B and a cyt1A genes were found in the LBIT-596 strain, while a cry21A, a cry14A and a cyt1A genes were found in the LBIT-107 strain. Genome sequence of the LBIT-107 strain showed new cry genes, along with other virulence factors, hence, total nematicidal activity of the LBIT-107 strain may be the result of a multifactorial effect. The highlight of this contribution is that translocation of spore-crystal suspensions of LBIT-107 into tomato plants inoculated at their rhizosphere decreased up to 90% the number of galls of Meloidogyne incognita, perhaps the most important nematode pest in the world.

A method for evaluating root-knot nematode infection in rice using a transparent paper pouch.

The root-knot nematode (RKN) Meloidogyne incognita is one of the most economically damaging plant-parasitic nematodes. Molecular studies of the plant-RKN interaction have been vigorously carried out in dicotyledonous model plants, while the host range of M. incognita is wide including monocotyledonous plants. As M. incognita causes quality and yield losses in rice (Oryza sativa L.) cultivated in both upland and irrigated systems, we developed a method to examine the plant-RKN interaction in this model monocotyledonous crop plant. Here, we show that a transparent paper pouch could be used to evaluate nematode infection rates in rice with similar results to that of the traditional soil method. The system using a transparent paper pouch can be used to observe the spatial and temporal distribution of developing galls and can save the space of growth chamber.

Reproduction of Meloidogyne marylandi and M. incognita on several Poaceae.

The susceptibility of 22 plant species to Meloidogyne marylandi and M. incognita was examined in three greenhouse experiments. Inoculum of M. marylandi was eggs from cultures maintained on Zoysia matrella "Cavalier" or Cynodon dactylon x C. trasvaalensis "Tifdwarf". Inoculum of M. incognita was eggs from cultures maintained on Solanum lycopersicum 'Rutgers'. In each host test the inoculum density was 2,000 nematode eggs/pot. None of the three dicot species tested (Gossypium hirsutum, Arachis hypogaea, and S. lycopersicum) were hosts for M. marylandi but, as expected, M. incognita had high levels of reproduction on G. hirsutum and S. lycopersicum. Meloidogyne marylandi reproduced on all of the 19 grass species (Poaceae) tested but reproduction varied greatly (P = 0.05) among these hosts. The following grasses were identified for the first time as hosts for M. marylandi: Buchloe dactyloides (buffalograss), Echinochloa colona (jungle rice), Eragostis curvula (weeping lovegrass), Paspalum dilatatum (dallisgrass), P. notatum (bahiagrass), Sorghastrum, nutans (indiangrass), Tripsacum dactyloides (eastern gamagrass), and Zoysia matrella (zoysiagrass). No reproduction of M. incognita was observed on B. dactyloides, Cyndon dactylon (common bermudagrass), E. curvula, P. vaginatum (seashore paspalum), S. nutans, T. dactyloides, Z. matrella or Z. japonica. Reproduction of M. incognita was less than reproduction of M. marylandi on the other grass species, except for the Zea mays inbred line B73 on which M. incognita had greater reproduction than did M. marylandi (P = 0.05) and Stenotaphrum secundatum (St. Augustinegrass) on which M. incognita and M. marylandi had similar levels of reproduction.

Silencing of a Meloidogyne incognita selenium-binding protein alters the cuticular adhesion of Pasteuria penetrans endospores.

Pasteuria penetrans is an endospore forming hyperparasitic bacterium of the plant-pathogenic root-knot nematode, Meloidogyne incognita. For successful parasitization, the first step is adherence of bacterial endospores onto the cuticle surface of nematode juveniles. The knowledge of molecular intricacies involved during this adherence is sparse. Here, we identified a M. incognita selenium-binding protein (Mi-SeBP-1) differentially expressed during the initial interaction of M. incognita and P. penetrans, and show that it is involved in modulating parasitic adhesion of bacterial endospores onto nematode cuticle. Selenium-binding proteins (SeBPs) are selenium associated proteins important for growth regulation, tumor prevention and modulation of oxidation/reduction in cells. Although reported to be present in several nematodes, the function of SeBPs is not known in Phylum Nematoda. In situ hybridization assay localized the Mi-SeBP-1 mRNA to the hypodermal cells. RNAi-mediated silencing of Mi-SeBP-1 significantly increased the adherence of P. penetrans endospores to the nematode juvenile cuticle. Silencing of Mi-SeBP-1 did not change the nematode's ability to parasitize plants and reproduction potential within the host. These results suggest that M. incognita Mi-SeBP-1 might be involved in altering the attachment of microbial pathogens on the nematode cuticle, but is not involved in nematode-host plant interaction. This is the first report for a function of SeBP in Phylum Nematoda.