Ultrastructural analysis of the early development of nematodes is hampered by the impermeability of the eggshell to most commonly used fixatives. High-pressure freezing (HPF), a physical cryo-fixation method, facilitates a fast rate of fixation, and by using this method the issue of the uneven delivery of fixative is circumvented. Although HPF results in a superior preservation of the fine structure, the equipment costs impede a wider application of this method. Self-pressurised rapid freezing (SPRF) is an alternative low-cost cryo-fixation method, and its usefulness was evaluated in an ultrastructural study of the eggshell and the cuticle of unhatched second-stage juveniles (J2) of Globodera rostochiensis and Heterodera schachtii. A comparison with conventional (chemical) fixation demonstrates that SPRF fixation results in a remarkably well-preserved ultrastructure of the entire egg including both the eggshell and the cellular details of the unhatched J2. Therefore, SPRF fixation is proposed as an affordable, relatively easy-to-use and time-efficient technique to study the ultrastructure of unhatched J2 and eggs of nematodes.
Saffron (Crocus sativus) fields in Morocco’s Taliouine and Taznakht regions were surveyed between January and April 2018 to study the diversity and incidence of plant-parasitic nematodes and assess the effects of soil physicochemical properties on the nematodes. Fourteen nematode genera were identified in soil and root samples collected from 66 saffron fields. The most common plant-parasitic nematodes in the Taliouine region were Pratylenchus spp. and Helicotylenchus spp. In the Taznakht region, the most common nematodes were Pratylenchus spp., Tylenchorhynchus spp. and Ditylenchus dipsaci. Nematodes, particularly Pratylenchus spp. and Ditylenchus spp., were abundant and frequent throughout the region. Several nematode genera were significantly associated with soil texture and mineral content, indicating that soil properties play an important role in plant-parasitic nematode communities. This description of plant-parasitic nematode assemblages associated with saffron fields in Morocco and their relationship with soil physicochemical properties provides a starting point from which appropriate nematode management strategies can be implemented.
In certain soils populations of plant-parasitic nematodes (PPN) decline. Understanding this effect may open up environmentally friendly management options. We identified such a suppressive soil containing virtually no PPN. Inoculated Meloidogyne hapla declined in this soil more than in a control soil and reproduction on tomato was reduced. The extracted soil microbiome alone decreased root invasion of second-stage juveniles (J2) and progeny as well as the native soil. We tested the antagonistic potential against PPN that differ in life strategies. The microbiome was most suppressive against two populations of M. hapla and one population of Pratylenchus neglectus, and least suppressive against M. incognita and the ectoparasite Hemicycliophora conida. In a split-root system with M. hapla, plant-mediated but not direct effects of the microbiome significantly reduced root invasion of J2, while direct exposure of M. hapla to the microbiome significantly affected reproduction. Overall, both plant-mediated and direct effects of the microbiome were responsible for the soil suppressiveness against M. hapla.
The lateral field pattern of infective juveniles of the nematode family Steinernematidae is an important taxonomic character. Scanning electron microscopy (SEM) shows the number of ridges and lines or incisures clearly, but does not provide other details. In the present study, ten species from six clades of Steinernematidae have been studied for their lateral field morphology using SEM and high pressure freezing (HPF) with transmission electron microscopy (TEM). Both methods indicated the same number of ridges and lines, although HPF/TEM resulted in a more detailed morphology with differences between the species. The tips of the ridges are either finely rounded or pointed and the lines are V-shaped or have a broadened bottom. These characters represent an additional pattern that may be characteristic for some species within the phylogenetic clades. Further studies of the lateral field morphology of other species is needed to ascertain whether each pattern is clade specific and phylogenetically valuable.
The cyst nematode, Heterodera glycines, is a major pathogen of soybean in tropical regions, which demands novel sustainable management practices. In this work, the use of ethanol against H. glycines was evaluated as both a solution and a fumigant. On second-stage juveniles (J2) of H. glycines, ethanol at low concentration was more effective by direct dipping than by only fumigating the J2. Hatching was significantly reduced by direct dipping in ethanol solutions. Fumigation of H. glycines-infested soil with ethanol reduced infectivity by almost 100% and the number of eggs by about 67% at ethanol concentrations of 48% and 72%, respectively. Only the ethanol at 48% concentration significantly reduced the J2 lipid content, while J2 infectivity and the number of eggs were reduced by dipping at 6% ethanol. The J2 were internally altered by the ethanol solutions. Therefore, ethanol is toxic to H. glycines at low concentrations and affects its pathogenic behaviour rather than simply reducing the lipids.
Watermelon is increasingly produced and consumed in Nigeria and sub-Saharan Africa. However, limited information exists regarding nematode pests and beneficial/free-living nematodes associated with the crop. The present study recorded the abundance and diversity of free-living nematodes from 50 watermelon fields across south-west Nigeria during 2016/2017. Of the 30 genera identified from soil samples, Cephalobus, followed by Rhabditis, Aphelenchus and Aporcelaimus, were predominant. Variation in nematode community structures across the 50 fields was apparent for mean maturity indices, metabolic footprints, feeding-type composition and coloniser-persister (c-p) structure. Faunal analyses characterised 52% of the fields as having stable and enriched soil food webs, which is beneficial for crop production. Significant correlations were apparent between some nematode genera and selected soil properties, and rainfall. This study provides the first information of free-living nematodes associated with watermelon from sub-Saharan Africa, offering novel and baseline information on their abundance and diversity in south-west Nigeria.
The stubby root nematodes are world-wide distributed polyphagous root ectoparasites and can cause damage to a wide range of crops and natural vegetation. In this study, 22 valid and putative species of stubby root nematodes were identified in 37 samples collected in Malaysia, Mexico, Russia, and the USA (California, Florida, Minnesota and Nevada). The analysis of 58 new sequences of the D2-D3 expansion fragments of 28S rRNA gene revealed the following species: Nanidorus minor from California, Florida and Mexico, N. renifer from Florida, two unidentified Nanidorus species from California and Malaysia, respectively, Paratrichodorus allius from Florida and Minnesota, P. pachydermus and Paratrichodorus sp. from Russia, Trichodorus californicus, T. intermedius, and T. obscurus from California, T. obtusus from Florida, eight unidentified Trichodorus species from California, one unidentified Trichodorus species from Nevada, and two unidentified species of stubby root nematodes, one of Trichodorus and another undetermined species from Mexico. Molecular characterisation of T. californicus, T. intermedius and T. obscurus is given for the first time. The phylogenetic tree reconstructed from the analysis of 108 D2-D3 of 28S rRNA gene sequences of 58 valid and putative species of the stubby root nematodes contained five major clades: i) Trichodorus from Europe, Asia and North America; ii) Nanidorus and Trichodorus from Asia; iii) Trichodorus from California; iv) Paratrichodorus from several distant geographical regions; and v) Monotrichodorus from Central and South America. It has been hypothesised that the California Floristic Province is one of the centres of origin and diversification of stubby root nematodes. There is no information on the economic importance of the stubby root nematode species found in California and Mexico. The confirmation of the occurrence of N. renifer and P. allius in Florida should be of concern for the blueberry and potato industries in the state.
There is a demand for novel products for the control of plant-parasitic nematodes, so we characterised the effectiveness of some plant essential oils against Meloidogyne incognita and verified the efficiency of the major component from the most toxic oils and their analogues using in vitro and in vivo assays. In this study, the essential oils from Piptadenia viridiflora, Hyptis suaveolens and Astronium graveolens against M. incognita were evaluated, but only P. viridiflora oil showed toxicity toward M. incognita. Benzaldehyde was its main component according to GC-MS analysis. In in vitro assays, benzaldehyde (100 and 200 μg ml−1) and its oxime (400 μg ml−1) caused a higher rate of M. incognita second-stage juvenile (J2) mortality than the nematicide carbofuran (170 μg ml−1). Reductions of more than 90% in the number of galls and eggs, even greater than that observed with carbofuran, were observed in the assay where the J2 were placed in solutions of benzaldehyde and its oxime 48 h prior to tomato plant inoculation. Application of benzaldehyde together with M. incognita J2 to the substrate resulted only in a reduction in the number of eggs (42-65%); however, its oxime reduced both the number of galls (43-84%) and eggs (23-89%). Therefore, the P. viridiflora oil, its major component benzaldehyde, and the analogue benzaldehyde oxime are toxic to M. incognita. In two different in vivo assays, benzaldehyde oxime was confirmed as toxic to M. incognita with a greater efficacy than benzaldehyde.
Incorporation of castor bean cake into the soil results in the emission of four main compounds: skatole, γ-decalactone, 4-methylphenol and phenol. The toxic effects of these compounds on the life cycle of Meloidogyne incognita were studied here. Only phenol did not reduce hatching of M. incognita second-stage juveniles (J2) at any of the concentrations tested. The other compounds reduced J2 hatching by 42.1 to 59.3% at the highest concentrations. When J2 were immersed in Tween 80® solutions of each compound, at their respective LC50 values and inoculated into soil with tomato plants, reductions in the numbers of galls (48.5-69.3%) and eggs (42.8-62.5%) were observed in comparison with the control; this result was similar to that of carbofuran. The highest reductions in the numbers of galls (21.8-88.5%) and eggs (49.6-98.9%) occurred with all compounds when the highest concentration (500 mg l−1) was applied to the substrate at the moment of J2 inoculation. Volatile compounds of γ-decalactone and skatole failed to cause J2 immobility or mortality. However, phenol caused 95.8% and 4-methylphenol caused 100% mortality. With fumigation, the highest concentration (1000 mg (l substrate)−1) of any tested compound reduced gall numbers at a similar rate to the nematicide dazomet (98.7-100%) and reduced egg numbers (87.1-99.7%). The volatile compounds skatole, γ-decalactone, 4-methylphenol and phenol have a nematicidal effect on the life cycle of M. incognita and alter its pathogenic behaviour on the plant.
The second population of Pratylenchoides riparius, including females and males, is described and illustrated based upon morphological, morphometric and molecular data. The present population from Iran is characterised by some differences with the type population of the species from Hungary in stylet length (24-26 vs 21-22 μm), slightly longer body (1002-1230 vs 830-960 μm), pharynx (202-211 vs 182-190 μm) and tail (64-85 vs 48-57 μm), areolated outer bands of the lateral field (vs non-areolated), widening of the lateral field near tail terminus (vs lateral incisures connecting each other) and presence of males (vs absent). The taxonomic status of the species with regarding the data from the type and presently recovered population, as well as the closely similar species is discussed. The newly recovered population was studied based upon its molecular phylogenetic charactes using the D2-D3 of 28S rRNA and the partial 18S rRNA gene sequences and the results revealed that it forms a clade with P. magnicauda in 28S, but occupies a distant placement from it in 18S phylogeny.