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1.  RKN Lethal DB: A database for the identification of Root Knot Nematode (Meloidogyne spp.) candidate lethal genes 
Bioinformation  2012;8(19):950-952.
Root Knot nematode (RKN; Meloidogyne spp.) is one of the most devastating parasites that infect the roots of hundreds of plant species. RKN cannot live independently from their hosts and are the biggest contributors to the loss of the world's primary foods. RNAi gene silencing studies have demonstrated that there are fewer galls and galls are smaller when RNAi constructs targeted to silence certain RKN genes are expressed in plant roots. We conducted a comparative genomics analysis, comparing RKN genes of six species: Meloidogyne Arenaria, Meloidogyne Chitwoodi, Meloidogyne Hapla, Meloidogyne Incognita, Meloidogyne Javanica, and Meloidogyne Paranaensis to that of the free living nematode Caenorhabditis elegans, to identify candidate genes that will be lethal to RKN when silenced or mutated. Our analysis yielded a number of such candidate lethal genes in RKN, some of which have been tested and proven to be effective in soybean roots. A web based database was built to house and allow scientists to search the data. This database will be useful to scientists seeking to identify candidate genes as targets for gene silencing to confer resistance in plants to RKN.
Availability
The database can be accessed from http://bioinformatics.towson.edu/RKN/
doi:10.6026/97320630008950
PMCID: PMC3488838  PMID: 23144556
RKN; Meloidogyne; web based database; RNAi; C. elegans
2.  Suppression of Meloidogyne incognita and M. javanica by Pasteuria penetrans in Field Soil 
Journal of Nematology  1996;28(1):43-49.
The role of Pasteuria penetrans in suppressing numbers of root-knot nematodes was investigated in a 7-year monocuhure of tobacco in a field naturally infested with a mixed population of Meloidogyne incognita race 1 and M. javanica. The suppressiveness of the soil was tested using four treatments: autoclaving (AC), microwaving (MW), air drying (DR), and untreated. The treated soil bioassays consisted of tobacco cv. Northrup King 326 (resistant to M. incognita but susceptible to M. javanica) and cv. Coker 371 Gold (susceptible to M. incognita and M. javanica) in pots inoculated with 0 or 2,000 second-stage juveniles of M. incognita race 1. Endospores of P. penetrans were killed by AC but were only slightly affected by MW, whereas most fungal propagules were destroyed or inhibited in both treatments. Root galls, egg masses, and numbers of eggs were fewer on Coker 371 Gold in MW, DR, and untreated soil than in AC-treated soil. There were fewer egg masses than root galls on both tobacco cultivars in MW, DR, and untreated soil than in the AC treatment. Because both Meloidogyne spp. were suppressed in MW soil (with few fungi present) as well as in DR and untreated soil, the reduction in root galling, as well as numbers of egg masses and eggs appeared to have resulted from infection of both nematode species by P. penetrans.
PMCID: PMC2619665  PMID: 19277344
bacterium; biological control; Meloidogyne incognita; M. javanica; nematode; Nicotiana tabacum; Pasteuria penetrans; root-knot nematode; suppressive soil; tobacco
3.  Host suitability of Ixora spp. for the Root-knot Nematodes Meloidogyne incognita Race 1 and M. javanica 
Journal of Nematology  1992;24(4S):722-728.
Eight commonly cultivated Ixora species or cultivars were tested for their suitability as hosts and their level of tolerance to Meloidogyne incognita race 1 and M. javanica in a greenhouse study. Twenty weeks postinoculation with 5,000 eggs per pot, M. incognita race 1 and M. javanica produced galls and formed egg masses on roots of all eight Ixora species or cultivars tested. However, only M. javanica-infected 'Petite Yellow' and 'Maui' had decreases (P ≤ 0.05) in root wet weights, suggesting that the other cultivars were more tolerant to these root-knot nematode species. Differential host suitability to each Meloidogyne species was based on the relative number of galls, galls per gram root weight, egg masses, and second-stage juveniles produced per plant. 'Bonnie Lynn,' 'Maui,' and 'Petite Red' were good to excellent hosts for both Meloidogyne spp. Ixora coccinea was a good host for M. incognita race 1 but less suitable for M. javanica. 'Singapore' and 'Petite Yellow' were poor hosts for M. incognita race 1 but excellent hosts for M. javanica. 'Nora Grant' and I. casei 'Super King' were poor hosts for both species of root-knot nematodes.
PMCID: PMC2629859  PMID: 19283052
host-parasite relationship; Ixora spp.; Meloidogyne incognita race 1; M. javanica; nematode; ornamental; root-knot nematode; woody ornamental
4.  Efficacy Evaluation of Fungus Syncephalastrum racemosum and Nematicide Avermectin against the Root-Knot Nematode Meloidogyne incognita on Cucumber 
PLoS ONE  2014;9(2):e89717.
The root-knot nematode (RKN) is one of the most damaging agricultural pests.Effective biological control is need for controlling this destructive pathogen in organic farming system. During October 2010 to 2011, the nematicidal effects of the Syncephalastrum racemosum fungus and the nematicide, avermectin, alone or combined were tested against the RKN (Meloidogyne incognita) on cucumber under pot and field condition in China. Under pot conditions, the application of S. racemosum alone or combined with avermectin significantly increased the plant vigor index by 31.4% and 10.9%, respectively compared to the M. incognita-inoculated control. However, treatment with avermectin alone did not significantly affect the plant vigor index. All treatments reduced the number of root galls and juvenile nematodes compared to the untreated control. Under greenhouse conditions, all treatments reduced the disease severity and enhanced fruit yield compared to the untreated control. Fewer nematodes infecting plant roots were observed after treatment with avermectin alone, S. racemosum alone or their combination compared to the M. incognita-inoculated control. Among all the treatments, application of avermectin or S. racemosum combined with avermectin was more effective than the S. racemosum treatment. Our results showed that application of S. racemosum combined with avermectin not only reduced the nematode number and plant disease severity but also enhanced plant vigor and yield. The results indicated that the combination of S. racemosum with avermectin could be an effective biological component in integrated management of RKN on cucumber.
doi:10.1371/journal.pone.0089717
PMCID: PMC3933638  PMID: 24586982
5.  Sensitive PCR Detection of Meloidogyne arenaria, M. incognita, and M. javanica Extracted from Soil 
Journal of nematology  2006;38(4):434-441.
We have developed a simple PCR assay protocol for detection of the root-knot nematode (RKN) species Meloidogyne arenaria, M. incognita, and M. javanica extracted from soil. Nematodes are extracted from soil using Baermann funnels and centrifugal flotation. The nematode-containing fraction is then digested with proteinase K, and a PCR assay is carried out with primers specific for this group of RKN and with universal primers spanning the ITS of rRNA genes. The presence of RKN J2 can be detected among large numbers of other plant-parasitic and free-living nematodes. The procedure was tested with several soil types and crops from different locations and was found to be sensitive and accurate. Analysis of unknowns and spiked soil samples indicated that detection sensitivity was the same as or higher than by microscopic examination.
PMCID: PMC2586468  PMID: 19259460
Detection; diagnosis; Meloidogyne arenaria; Meloidogyne incognita; Meloidogyne javanica; PCR; root-knot nematode; soil
6.  Phenotypic Expression of rkn1-Mediated Meloidogyne incognita Resistance in Gossypium hirsutum Populations 
Journal of nematology  2006;38(2):250-257.
The root-knot nematode Meloidogyne incognita is a damaging pest of cotton (Gossypium hirsutum) worldwide. A major gene (rkn1) conferring resistance to M. incognita was previously identified on linkage group A03 in G. hirsutum cv. Acala NemX. To determine the patterns of segregation and phenotypic expression of rkn1, F1, F2, F2:3, BC1F1 and F2:7 recombinant inbred lines (RIL) from intraspecific crosses between Acala NemX and a closely related susceptible cultivar Acala SJ-2 were inoculated in greenhouse tests with M. incognita race 3. The resistance phenotype was determined by the extent of nematode-induced root galling and nematode egg production on roots. Suppression of root galling and egg production was highly correlated among individuals in all tests. Root galling and egg production on heterozygous plants did not differ from the susceptible parent phenotype 125 d or more after inoculation, but were slightly suppressed with shorter screening (60 d), indicating that rkn1 behaved as a recessive gene or an incompletely recessive gene, depending on the screening condition. In the RIL, rkn1 segregated in an expected 1 resistant: 1 susceptible ratio for a major resistance gene. However, within the resistant class, 21 out of 34 RIL were more resistant than the resistant parent Acala NemX, indicating transgressive segregation. These results suggest that rkn1-based resistance in G. hirsutum can be enhanced in progenies of crosses with susceptible genotypes. Allelism tests and molecular genetic analysis are needed to determine the relationship of rkn1 to other M. incognita resistance sources in cotton.
PMCID: PMC2586458  PMID: 19259455
cotton; Gossypium hirsutum; Meloidogyne incognita; resistance; rkn1; root-knot nematode; phenotypic expression; transgressive segregation
7.  Optimum Initial Inoculum Levels for Evaluation of Resistance in Tomato to Meloidogyne spp. at Two Different Soil Temperatures 
Journal of Nematology  1982;14(4):536-539.
The effects of Meloidogyne incognita or M. javanica at five initial inoculum levels of 20, 100, 200, 1,000, and 2,000 eggs and infective juveniles per seedling on 'Floradade,' 'Nemarex,' 'Patriot,' and 'PI 129149-2(sib)-5' tomatoes maintained at 25 or 32.5 C were studied. The number of egg masses on roots of the susceptible cultivar Floradade was similar for both species of root-knot nematodes at either 2.5 or 32.5 C soil temperatures. At 25 C, very low numbers of egg masses were produced by both species of root-knot nematodes on Nematex, Patriot, and Lycopersicon peruvianum PI 129149-2(sib)-5. At 32.5 C, the best inoculum level for assessing resistance in these tomato genotypes was 200 eggs and infective juveniles per seedling. With 28 days of incubation, this temperature and inoculum level produced quantitative differences in resistance for both species of Meloidogyne.
PMCID: PMC2618214  PMID: 19295749
Lycopersicon; root-knot nematode
8.  Nematicidal activity of fervenulin isolated from a nematicidal actinomycete, Streptomyces sp. CMU-MH021, on Meloidogyne incognita 
An isolate of the actinomycete, Streptomyces sp. CMU-MH021 produced secondary metabolites that inhibited egg hatch and increased juvenile mortality of the root-knot nematode Meloidogyne incognita in vitro. 16S rDNA gene sequencing showed that the isolate sequence was 99% identical to Streptomyces roseoverticillatus. The culture filtrates form different culture media were tested for nematocidal activity. The maximal activity against M. incognita was obtained by using modified basal (MB) medium. The nematicidal assay-directed fractionation of the culture broth delivered fervenulin (1) and isocoumarin (2). Fervenulin, a low molecular weight compound, shows a broad range of biological activities. However, nematicidal activity of fervenulin was not previously reported. The nematicidal activity of fervenulin (1) was assessed using the broth microdilution technique. The lowest minimum inhibitory concentrations (MICs) of the compound against egg hatch of M. incognita was 30 μg/ml and juvenile mortality of M. incognita increasing was observed at 120 μg/ml. Moreover, at the concentration of 250 μg/ml fervenulin (1) showed killing effect on second-stage nematode juveniles of M. incognita up to 100% after incubation for 96 h. Isocoumarin (2), another bioactive compound produced by Streptomyces sp. CMU-MH021, showed weak nematicidal activity with M. incognita.
doi:10.1007/s11274-010-0588-z
PMCID: PMC3128717  PMID: 21841897
Streptomyces sp. CMU-MH021; Fervenulin; Nematicidal activity; Root-knot nematode
9.  Effect of Ammonium Ions on Egg Hatching and Second-Stage Juveniles of Meloidogyne incognita in Axenic Tomato Root Culture 
Journal of Nematology  1995;27(3):346-352.
Eggs, either dispersed or in masses, and second-stage juveniles (J2) of Meloidogyne incognita were exposed to different concentrations of ammonium ions in a nutrient agar medium upon which excised tomato roots were growing. Egg hatch and J2 penetration of the roots was slowed or inhibited at high (54 and 324 mg/liter) but not at low (1.5 and 9 mg/liter) concentrations of ammonium nitrate. The effect of ammonium on J2 appeared to be temporary and reversible. High potassium nitrate concentration (1,116 mg/liter) did not modify egg hatch or J2 penetration. There was no adverse effect from the high ammonium nitrate concentrations or an equivalent potassium nitrate concentration on root dry weight. Ammonium ions influence nematodes both directly and via plant roots and do so independently of microbial organisms.
PMCID: PMC2619617  PMID: 19277298
ammonium ion; axenic culture; hatching; Meloidogyne incognita; root-knot nematode
10.  Geostatistical modeling of the spatial variability and risk areas of southern root-knot nematodes in relation to soil properties 
Geoderma  2010;156(3-4):243-252.
Identifying the spatial variability and risk areas for southern root-knot nematode [Meloidogyne incognita (Kofoid & White) Chitwood] (RKN) is key for site-specific management (SSM) of cotton (Gossypium hirsutum L.) fields. The objectives of this study were to: (i) determine the soil properties that influence RKN occurrence at different scales; and (ii) delineate risk areas of RKN by indicator kriging. The study site was a cotton field located in the southeastern coastal plain region of the USA. Nested semivariograms indicated that RKN samples, collected from a 50×50 m grid, exhibited a local and regional scale of variation describing small and large clusters of RKN population density. Factorial kriging decomposed RKN and soil properties variability into different spatial components. Scale dependent correlations between RKN data showed that the areas with high RKN population remained stable though the growing season. RKN data were strongly correlated with slope (SL) at local scale and with apparent soil electrical conductivity deep (ECa-d) at both local and regional scales, which illustrate the potential of these soil physical properties as surrogate data for RKN population. The correlation between RKN data and soil chemical properties was soil texture mediated. Indicator kriging (IK) maps developed using either RKN, the relation between RKN and soil electrical conductivity or a combination of both, depicted the probability for RKN population to exceed the threshold of 100 second stage juveniles/100 cm3 of soil. Incorporating ECa-d as soft data improved predictions favoring the reduction of the number of RKN observations required to map areas at risk for high RKN population.
doi:10.1016/j.geoderma.2010.02.024
PMCID: PMC2921668  PMID: 20717481
Cotton; Factorial kriging; Indicator kriging; Logistic regression; Nematodes; Risk map; Semivariogram; Soil properties; Southern root-knot nematode; Spatial variability
11.  Effects of Root Decay on the Relationship between Meloidogyne spp. Gall Index and Egg Mass Number in Cucumber and Horned Cucumber 
Journal of Nematology  1992;24(4S):707-711.
A greenhouse study was conducted to determine if root necrosis had an effect on the relationship between root-knot nematode gall index and egg mass number. Thirty-four cultigens of Cucumis (14 accessions, 12 cultivars, and six breeding lines of C. sativus, and two accessions of C. metuliferus) were evaluated against four root-knot species (Meloidogyne arenaria race 2, M. incognita race 1, M. incognita race 3, and M. javanica) measuring gall index, root necrosis, and egg mass number. Root necrosis affected the gall index-egg mass relationship. At lower root necrosis values, a stronger relationship existed between gall index and egg mass number than at higher root necrosis values. Root tissue was destroyed by root necrosis, and normal root-knot nematode reproduction would not occur, even though root galling was still observed. The races of M. incognita tested had a greater effect in predisposing C. sativus and C. metuliferus to root necrosis than did M. arenaria race 2 or M. javanica. This study showed that root necrosis had an adverse affect on the relationship between gall index and egg mass number in cucumber.
PMCID: PMC2629864  PMID: 19283049
African horned cucumber; cucumber; Cucumis sativus; Cucumis metuliferus; Meloidogyne arenaria; Meloidogyne incognita; Meloidogyne javanica; nematode; resistance; root-knot nematode
12.  In-vitro Assays of Meloidogyne incognita and Heterodera glycines for Detection of Nematode-antagonistic Fungal Compounds 
Journal of Nematology  1999;31(2):172-183.
In-vitro methods were developed to test fungi for production of metabolites affecting nematode egg hatch and mobility of second-stage juveniles. Separate assays were developed for two nematodes: root-knot nematode (Meloidogyne incognita) and soybean cyst nematode (Heterodera glycines). For egg hatch to be successfully assayed, eggs must first be surface-disinfested to avoid the confounding effects of incidental microbial growth facilitated by the fungal culture medium. Sodium hypochlorite was more effective than chlorhexidine diacetate or formaldehyde solutions at surface-disinfesting soybean cyst nematode eggs from greenhouse cultures. Subsequent rinsing with sodium thiosulfate to remove residual chlorine from disinfested eggs did not improve either soybean cyst nematode hatch or juvenile mobility. Soybean cyst nematode hatch in all culture media was lower than in water. Sodium hypochlorite was also used to surface-disinfest root-knot nematode eggs. In contrast to soybean cyst nematode hatch, root-knot nematode hatch was higher in potato dextrose broth medium than in water. Broth of the fungus Fusarium equiseti inhibited root-knot nematode egg hatch and was investigated in more detail. Broth extract and its chemical fractions not only inhibited egg hatch but also immobilized second-stage juveniles that did hatch, confirming that the fungus secretes nematode-antagonistic metabolites.
PMCID: PMC2620357  PMID: 19270887
bioassay technique; biological control; culture broth; egg hatch; fungus; Fusarium equiseti; Heterodera glycines; in-vitro assay; Meloidogyne incognita; microbial secondary metabolites; nematicide; nematode
13.  QTL Analysis for Transgressive Resistance to Root-Knot Nematode in Interspecific Cotton (Gossypium spp.) Progeny Derived from Susceptible Parents 
PLoS ONE  2012;7(4):e34874.
The southern root-knot nematode (RKN, Meloidogyne incognita) is a major soil-inhabiting plant parasite that causes significant yield losses in cotton (Gossypium spp.). Progeny from crosses between cotton genotypes susceptible to RKN produced segregants in subsequent populations which were highly resistant to this parasite. A recombinant inbred line (RIL) population of 138 lines developed from a cross between Upland cotton TM-1 (G. hirsutum L.) and Pima 3–79 (G. barbadense L.), both susceptible to RKN, was used to identify quantitative trait loci (QTLs) determining responses to RKN in greenhouse infection assays with simple sequence repeat (SSR) markers. Compared to both parents, 53.6% and 52.1% of RILs showed less (P<0.05) root-galling index (GI) and had lower (P<0.05) nematode egg production (eggs per gram root, EGR). Highly resistant lines (transgressive segregants) were identified in this RIL population for GI and/or EGR in two greenhouse experiments. QTLs were identified using the single-marker analysis nonparametric mapping Kruskal-Wallis test. Four major QTLs located on chromosomes 3, 4, 11, and 17 were identified to account for 8.0 to 12.3% of the phenotypic variance (R2) in root-galling. Two major QTLs accounting for 9.7% and 10.6% of EGR variance were identified on chromosomes 14 and 23 (P<0.005), respectively. In addition, 19 putative QTLs (P<0.05) accounted for 4.5–7.7% of phenotypic variance (R2) in GI, and 15 QTLs accounted for 4.2–7.3% of phenotypic variance in EGR. In lines with alleles positive for resistance contributed by both parents in combinations of two to four QTLs, dramatic reductions of >50% in both GI and EGR were observed. The transgressive segregants with epistatic effects derived from susceptible parents indicate that high levels of nematode resistance in cotton may be attained by pyramiding positive alleles using a QTL mapping approach.
doi:10.1371/journal.pone.0034874
PMCID: PMC3325951  PMID: 22514682
14.  Effect of Gamma-irradiation and Heat on Root-knot Nematode, Meloidogyne javanica 
Journal of Nematology  1997;29(1):30-34.
Effects of gamma-irradiation on the root-knot nematode Meloidogyne javanica were investigated. A dose of 7.5 kGy killed all second-stage juveniles (J2) within 1 day after treatment. Egg hatch was completely inhibited at 6.25 kGy. A bioassay on tomato measuring galling and egg production was used to determine the infectivity of irradiated J2 and J2 hatched from irradiated eggs. The J2 and eggs irradiated with a dose of 4.25 kGy did not induce galls or reproduce on tomato plants. When nematodes were exposed to combined irradiation and heat treatment, no synergistic effect on J2 or eggs was measured. Heat treatment at 49° C for 10 minutes or 20 minutes without irradiation immobilized J2 and prevented egg development. Irradiation rates needed to kill or incapacitate M. javanica were high and may be impractical as a quarantine measure.
PMCID: PMC2619752  PMID: 19274131
Cobalt-60; control; dose; gamma-irradiation; heat treatment; kGy; Meloidogyne javanica; quarantine; root-knot nematode
15.  Comparison of Sequences from the Ribosomal DNA Intergenic Region of Meloidogyne mayaguensis and Other Major Tropical Root-Knot Nematodes 
Journal of Nematology  1997;29(1):16-22.
The unusual arrangement of the 5S ribosomal gene within the intergenic sequence (IGS) of the ribosomal cistron, previously reported for Meloidogyne arenaria, was also found in the ribosomal DNA of two other economically important species of tropical root-knot nematodes, M, incognita and M. javanica. This arrangement also was found in M. hapla, which is important in temperate regions, and M. mayaguensis, a virulent species of concern in West Africa. Amplification of the region between the 5S and 18S genes by PCR yielded products of three different sizes such that M. mayaguensis could be readily differentiated from the other species in this study. This product can be amplified from single juveniles, females, or egg masses. The sequences obtained in this region for one line of each of M. incognita, M. arenaria, and M. javanica were very similar, reflecting the close relationships of these lineages. The M. mayaguensis sequence for this region had a number of small deletions and insertions of various sizes, including possible sequence duplications.
PMCID: PMC2619761  PMID: 19274129
Meloidogyne arenaria; Mdoidogyne incognita; Meloidogyne javanica; Meloidogyne mayaguensis; PCR; rDNA; root-knot nematode
16.  Reproduction and Development of Meloidogyne incognita and M. javanica on Guardian Peach Rootstock 
Journal of Nematology  1999;31(3):334-340.
Guardian peach rootstock was evaluated for susceptibility to Meloidogyne incognita race 3 (Georgia-peach isolate) and M. javanica in the greenhouse. Both commercial Guardian seed sources produced plants that were poor hosts of M. incognita and M. javanica. Reproduction as measured by number of egg masses and eggs per plant, eggs per egg mass, and eggs per gram of root were a better measure of host resistance than number of root galls per plant. Penetration, development, and reproduction of M. incognita in Guardian (resistant) and Lovell (susceptible) peach were also studied in the greenhouse. Differences in susceptibility were not attributed to differential penetration by the infectivestage juveniles (J2) or the number of root galls per plant. Results indicated that M. incognita J2 penetrated Guardian roots and formed galls, but that the majority of the nematodes failed to mature and reproduce.
PMCID: PMC2620375  PMID: 19270905
host-parasitic relationship; Meloidogyne incognita; Meloidogyne javanica; nematode; peach; Prunus persica; resistance; root-knot nematode; rootstock
17.  Effect of Meloidogyne incognita and Importance of the Inoculum on the Yield of Eggplant 
Journal of Nematology  1986;18(4):487-490.
The relationship between population densities of race 1 of Meloidogyne incognita and yield of eggplant was studied. Microplots were infested with finely chopped nematode-infected pepper roots to give population densities of 0, 0.062, 0.125, 0.25, 0.50, 1, 2, 4, 8, 16, 32, 64, and 128 eggs and juveniles/cm³ soil. Both plant growth and yield were suppressed by the nematode. A tolerance limit of 0.054 eggs and juveniles/cm³ soil and a minimum relative yield of 0.05 at four or more eggs and juveniles/cm³ soil were derived by fitting the data with the equation y = m + (1 - m)zP⁻T. Maximum nematode reproduction rate was 12,300. Hatch of eggs from egg masses in water or from sodium hypochlorite dissolved egg masses was similar (41% and 39%), but egg viability was significantly greater from egg masses in water (58%) than from sodium hypochlorite dissolved egg masses (12%) after 4 weeks. Greater numbers of nematodes were collected from roots of tomatoes from soil infested with entire egg masses than from tomato roots from soil infested with egg masses dissolved by sodium hypochlorite.
PMCID: PMC2618571  PMID: 19294216
root-knot nematode; tolerance limit; eggplant; inoculum; Solanum melongena; Meloidogyne incognita
18.  The complex hybrid origins of the root knot nematodes revealed through comparative genomics 
PeerJ  2014;2:e356.
Root knot nematodes (RKN) can infect most of the world’s agricultural crop species and are among the most important of all plant pathogens. As yet however we have little understanding of their origins or the genomic basis of their extreme polyphagy. The most damaging pathogens reproduce by obligatory mitotic parthenogenesis and it has been suggested that these species originated from interspecific hybridizations between unknown parental taxa. We have sequenced the genome of the diploid meiotic parthenogen Meloidogyne floridensis, and use a comparative genomic approach to test the hypothesis that this species was involved in the hybrid origin of the tropical mitotic parthenogen Meloidogyne incognita. Phylogenomic analysis of gene families from M. floridensis, M. incognita and an outgroup species Meloidogyne hapla was carried out to trace the evolutionary history of these species’ genomes, and we demonstrate that M. floridensis was one of the parental species in the hybrid origins of M. incognita. Analysis of the M. floridensis genome itself revealed many gene loci present in divergent copies, as they are in M. incognita, indicating that it too had a hybrid origin. The triploid M. incognita is shown to be a complex double-hybrid between M. floridensis and a third, unidentified, parent. The agriculturally important RKN have very complex origins involving the mixing of several parental genomes by hybridization and their extreme polyphagy and success in agricultural environments may be related to this hybridization, producing transgressive variation on which natural selection can act. It is now clear that studying RKN variation via individual marker loci may fail due to the species’ convoluted origins, and multi-species population genomics is essential to understand the hybrid diversity and adaptive variation of this important species complex. This comparative genomic analysis provides a compelling example of the importance and complexity of hybridization in generating animal species diversity more generally.
doi:10.7717/peerj.356
PMCID: PMC4017819  PMID: 24860695
Genome sequencing; Phylogenomics; Meloidogyne incognita; Meloidogyne hapla; Meloidogyne floridensis; Comparative genomics; Hybrid speciation
19.  Effects of Nematicides and Herbicides Alone or Combined on Meloidogyne incognita Egg Hatch and Development 
Journal of Nematology  1987;19(Annals 1):67-70.
The effects of nematicides carbofuran (C) and fenamiphos (F) and herbicides metribuzin (M) and trifluralin (T), alone and in combination, on hatching, penetration, development, and reproduction of Meloidogyne incognita race 3 were determined under laboratory conditions. To study hatching, entire egg masses were exposed to nematicides (6 μg/ml), herbicides (0.5 μg/ml), and their combinations over a period of 16 days; the hatched juveniles were extracted and counted every 48 hours. Second-stage juveniles that hatched from day 6 to day 8 were used as inoculum to determine the effects of the chemicals on penetration, development, and reproduction of M. incognita on tomato 4, 16, and 32 days after inoculation. F, F + T, and F + M inhibited hatching; whereas, C, T, M, C + T, and C + M did not affect hatching, penetration, development of females, or reproduction. Since so few juveniles hatched from the fenamiphos treatments, we were not able to use them for the postinfection development study. There was no apparent reduction in the effect of the nematicides by the herbicides.
PMCID: PMC2618687  PMID: 19290279
carbofuran; fenamiphos; hatching; herbicide; life cycle; Meloidogyne incognita; metribuzin; nematicide; pesticide interaction; root-knot nematode; survival; trifluralin
20.  Fitness of Virulent Meloidogyne incognita Isolates on Susceptible and Resistant Cowpea 
Journal of Nematology  2005;37(4):457-466.
A study of life-history traits was made to determine factors associated with the fitness of Meloidogyne incognita isolates virulent to resistance gene Rk in cowpea. Egg hatch, root penetration, egg mass production, and fecundity (eggs per egg mass) of avirulent and virulent phenotypes were compared among M. incognita isolates, isofemale lines, and single descent lines over multiple generations on resistant and susceptible cowpea. Variation (P ≤ 0.05) in both hatch and root penetration rates was found among isolates at a given generation. However, this variation was not consistent within nematode lines among generations, and there was no correlation with level of virulence, except for penetration and virulence on resistant cowpea at generation 20. Resistant and susceptible cowpea roots were penetrated at similar levels. Differences in reproductive factors on resistant plants were correlated with levels of virulence expression. In some isofemale lines, single descent lines, and isolates, lower (P ≤ 0.05) rates of egg mass production and fecundity on susceptible cowpea were associated with virulence to Rk, indicating a trade-off between reproductive fitness and virulence. Other virulent nematode lines from the same isolates did not have reduced reproductive ability on susceptible cowpea over 27 generations. Thus, virulent lineages varied in reproductive ability on susceptible cowpea, contributing to adaptation and maintenance of virulence within M. incognita populations under stabilizing selection.
PMCID: PMC2620988  PMID: 19262891
cowpea; fitness; genetic variation; Meloidogyne incognita; resistance; root-knot nematode; selection; Vigna unguiculata; virulence
21.  Effects of Irrigation, Nitrogen, and a Nematicide on Pearl Millet 
Journal of Nematology  1995;27(4S):571-574.
Pearl millet is used mainly as a temporary forage crop in the southern United States. A new pearl millet hybrid has potential as a major grain crop in the United States. The effects of nematodes, irrigation, a nematicide, and nitrogen rates on a new pearl millet grain hybrid, HGM-100, and nematode population changes were determined in a 2-year study. Root-knot nematodes (Meloidogyne incognita race 1) entered the roots of pearl millet and caused minimal galling, but produced large numbers of eggs that hatched into second-stage juveniles. Root-gall indices ranged from 1.00 to 1.07 on a 1-5 scale and were not affected by irrigation or rates of nitrogen. Yield of pearl millet was up to 31% higher under no supplemental irrigation than under irrigation, 16% higher in fenamiphos-treated plots than untreated plots, and 56% higher in plots treated with 38 kg nitrogen/ha than plots treated with 85 kg nitrogen/ha. In southern Georgia, pearl millet appears to be resistant to ring nematode (Criconemella ornata) but favors development and reproduction of M. incognita.
PMCID: PMC2619661  PMID: 19277324
chemical control; Criconemella ornata; irrigation; Meloidogyne incognita; millet; nematode; nitrogen; Pennisetum glaucum; ring nematode; root-knot nematode
22.  Transcription profile of soybean-root-knot nematode interaction reveals a key role of phythormones in the resistance reaction 
BMC Genomics  2013;14:322.
Background
Root-knot nematodes (RKN– Meloidogyne genus) present extensive challenges to soybean crop. The soybean line (PI 595099) is known to be resistant against specific strains and races of nematode species, thus its differential gene expression analysis can lead to a comprehensive gene expression profiling in the incompatible soybean-RKN interaction. Even though many disease resistance genes have been studied, little has been reported about phytohormone crosstalk on modulation of ROS signaling during soybean-RKN interaction.
Results
Using 454 technology to explore the common aspects of resistance reaction during both parasitism and resistance phases it was verified that hormone, carbohydrate metabolism and stress related genes were consistently expressed at high levels in infected roots as compared to mock control. Most noteworthy genes include those encoding glycosyltransferases, peroxidases, auxin-responsive proteins and gibberellin-regulated genes. Our data analysis suggests the key role of glycosyltransferases, auxins and components of gibberellin signal transduction, biosynthesis and deactivation pathways in the resistance reaction and their participation in jasmonate signaling and redox homeostasis in mediating aspects of plant growth and responses to biotic stress.
Conclusions
Based on this study we suggest a reasonable model regarding to the complex mechanisms of crosstalk between plant hormones, mainly gibberellins and auxins, which can be crucial to modulate the levels of ROS in the resistance reaction to nematode invasion. The model also includes recent findings concerning to the participation of DELLA-like proteins and ROS signaling controlling plant immune or stress responses. Furthermore, this study provides a dataset of potential candidate genes involved in both nematode parasitism and resistance, which can be tested further for their role in this biological process using functional genomics approaches.
doi:10.1186/1471-2164-14-322
PMCID: PMC3701510  PMID: 23663436
Root–knot nematode; Glycine max; Transcriptome; Pyrosequencing; Plant–pathogen interaction; Hormone
23.  Histopathology of Root-knot Nematode (Meloidogyne incognita) Infection on White Yam (Dioscorea rotundata) Tubers 
Journal of Nematology  1988;20(1):23-28.
White yam tissues naturally and artificially infected with root-knot nematodes were fixed, sectioned, and examined with a microscope. Infective second-stage juveniles of Meloidogyne incognita penetrated and moved intercellularly within the tuber. Feeding sites were always in the ground tissue layer where the vascular tissues are distributed in the tubers. Giant cells were always associated with xylem tissue. They were thin walled with dense cytoplasm and multinucleated. The nuclei of the giant cells were only half the size of those found in roots of infected tomato plants. Normal nematode growth and development followed giant cell formation. Females deposited eggs into a gelatinous egg mass within the tuber, and a necrotic ring formed around the female after eggs had been produced. Second-stage juveniles hatched, migrated, and re-infected other areas of the tuber. No males were observed from the tuber.
PMCID: PMC2618787  PMID: 19290181
giant cell; histopathology; Meloidogyne incognita; root-knot nematode; yam
24.  Host Suitability of Potential Cover Crops for Root-knot Nematodes 
Journal of Nematology  1999;31(4S):619-623.
Several potential cover crops were evaluated for their susceptibility to Meloidogyne arenaria race 1, M. incognita race 1, and M. javanica in a series of five greenhouse experiments. No galls or egg masses were observed on roots of castor (Ricinus communis), cowpea (Vigna unguiculata cv. Iron Clay), crotalaria (Crotalaria spectabilis), or American jointvetch (Aeschynomene americana). Occasional egg masses (rating ≤1.0 on 0-5 scale) were observed on marigold (Tagetes minuta) in one test with M. incognita, on sesame (Sesamum indicum cv. Paloma) in a test with M. arenaria, and on sunn hemp (Crotalaria juncea cv. Tropic Sun) in 1 of 2 tests with M. incognita; otherwise, these crops were free of egg masses. Numbers of second-stage juveniles (J2) hatched from eggs per root system were low (≤10/pot) for the abovementioned crops. Egg-mass levels and numbers of hatched J2 of M. incognita on pearl millet (Pennisetum typhoides, Tifleaf II hybrid) were comparable to those on a susceptible tomato (Lycopersicon esculentum cv. Rutgers). In a test with M. arenaria, egg mass levels and numbers of J2 on Japanese millet (Echinochloa frumentacea) were similar to those on tomato. Japanese millet was susceptible to each of the nematode isolates tested. However, several of the crops evaluated were very poor hosts or non-hosts of the nematode isolates, including several legumes (cowpea, crotalaria, jointvetch, sunn hemp) that have potential use in both nematode and nitrogen management.
PMCID: PMC2620418  PMID: 19270926
Aeschynomene americana; castor; cowpea; Crotalaria juncea; Crotalaria spectabilis; Echinochloa frumentacea; host-plant resistance; jointvetch; marigold; Meloidogyne arenaria; Meloidogyne incognita; Meloidogyne javanica; millet; nematode; nematode management; Pennisetum glaucum; Pennisetum typhoides; Ricinus communis; sesame; Sesamum indicum; sunn hemp; sustainable agriculture; Tagetes minuta; Vigna unguiculata
25.  Effect of Meloidogyne incognita, M. hapla, and M. javanica on the Severity of Fusarium Wilt of Chrysanthemum 
Journal of Nematology  1969;1(2):122-125.
Rooted cuttings of Chrysanthemum morifolium 'Yellow Delaware' (Fusarium-susceptible) and 'White Iceberg' (Fusarium-resistant) were greenhouse-grown in: (i) non-infested soil; (ii) soil infested with Fusarium oxysporum alone; (iii) soil infested with Meloidogyne incognita, M. javanica or M. hapla; and (iv) each nematode separately plus the fungus. All nematode species infected roots of both cultivars and caused characteristic root-knot symptoms but did not appreciably affect growth meassured by plant weight. Nematodes did not break Fusarium wilt resistance of 'White Iceberg'; however, wilt symptoms appeared earlier and were more severe among 'Yellow Delaware' plants inoculated with Meloidogyne javanica and F. oxysporum than with similar combinations of the fungus and M. incognita or M. hapla or with the fungus alone.
PMCID: PMC2617816  PMID: 19325666

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