Root-lesion Nematode Pratylenchus Penetrans Research Articles

Nematicidal Activity of Phytochemicals against the Root-Lesion Nematode Pratylenchus penetrans.

Plant-parasitic nematodes (PPNs) are highly damaging pests responsible for heavy losses in worldwide productivity in a significant number of important plant crops. Common pest management strategies rely on the use of synthetic chemical nematicides, which have led to serious concerns regarding their impacts on human health and the environment. Plant natural products, or phytochemicals, can provide a good source of agents for sustainable control of PPNs, due to their intrinsic characteristics such as higher biodegradability, generally low toxicity for mammals, and lower bioaccumulation in the environment. In this work, the nematicidal activity of 39 phytochemicals was determined against the root-lesion nematode (RLN) Pratylenchus penetrans using standard direct and indirect contact methodologies. Overall, the RLN was tolerant to the tested phytochemicals at the highest concentration, 2 mg/mL, seldom reaching full mortality. However, high activities were obtained for benzaldehyde, carvacrol, 3-octanol, and thymol, in comparison to other phytochemicals or the synthetic nematicide oxamyl. These phytochemicals were seen to damage nematode internal tissues but not its cuticle shape. Also, the environmental and (eco)toxicological parameters reported for these compounds suggest lower toxicity and higher safety of use than oxamyl. These compounds appear to be good candidates for the development of biopesticides for a more sustainable pest management strategy.

Distribution of Bacterial Endosymbionts of the Cardinium Clade in Plant-Parasitic Nematodes.

Bacteria of the genus "Candidatus Cardinium" and related organisms composing the Cardinium clade are intracellular endosymbionts frequently occurring in several arthropod groups, freshwater mussels and plant-parasitic nematodes. Phylogenetic analyses based on two gene sequences (16S rRNA and gyrB) showed that the Cardinium clade comprised at least five groups: A, B, C, D and E. In this study, a screening of 142 samples of plant-parasitic nematodes belonging to 93 species from 12 families and two orders using PCR with specific primers and sequencing, revealed bacteria of Cardinium clade in 14 nematode samples belonging to 12 species of cyst nematodes of the family Heteroderidae. Furthermore, in this study, the genome of the Cardinium cHhum from the hop cyst nematode, Heterodera humuli, was also amplified, sequenced and analyzed. The comparisons of the average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values for the strain Cardinium cHhum with regard to related organisms with available genomes, combined with the data on 16S rRNA and gyrB gene sequence identities, showed that this strain represents a new candidate species within the genus "Candidatus Paenicardinium". The phylogenetic position of endosymbionts of the Cardinium clade detected in nematode hosts was also compared to known representatives of this clade from other metazoans. Phylogenetic reconstructions based on analysis of 16S rRNA, gyrB, sufB, gloEL, fusA, infB genes and genomes and estimates of genetic distances both indicate that the endosymbiont of the root-lesion nematode Pratylenchus penetrans represented a separate lineage and is designated herein as a new group F. The phylogenetic analysis also confirmed that endosymbionts of ostracods represent the novel group G. Evolutionary relationships of bacterial endosymbionts of the Cardinium clade within invertebrates are presented and discussed.

Host Suitability of Cover Crops to the Root-Lesion Nematode Pratylenchus penetrans Associated with Potato.

Nonhost or poor host cover crops can provide an alternative method for nematode management. A total of 25 cover crop species/cultivars, along with three controls were evaluated in greenhouse experiments for their host suitability to the root-lesion nematode Pratylenchus penetrans. Trials were conducted in a completely randomized design using nematode-infested soil and terminated 3 months after planting. Nematodes were extracted from the roots and soil of each crop to determine their final population densities, reproductive factor (Rf = final population density/initial population density), and distributions in the soil and root habitats. Reproductive factor was used to categorize the host suitability of crops. Faba bean cv. Petite produced the greatest nematode population density in all trials, whereas only alfalfa cv. Bullseye constantly demonstrated the poor host ability to P. penetrans. Annual ryegrass, winter rye cv. ND Dylan, and white proso millet also showed poor hosts in most trials. Five cover crops consistently maintained the population throughout the experiments, with Rf values less than 2, and the remaining tested cover crops were suitable hosts for P. penetrans. The majority of the tested cover crops had less than or equal to 30% of the final population residing in the roots after three months of growth in all the trials. This research helps us gain the knowledge on cover crops and P. penetrans interaction and will be useful for potato growers to select better cover crops and avoid susceptible hosts to manage P. penetrans in infested fields to minimize potato yield losses.

Commercial Potato Cultivars Exhibit Distinct Susceptibility to the Root Lesion Nematode Pratylenchus penetrans

The root lesion nematode Pratylenchus penetrans is an important plant-parasitic nematode of potato. In this study, the susceptibility of commercial potato cultivars to P. penetrans was assessed. Nematode penetration was evaluated in cultivars Agria, Camel, Kennebec, Laura, Royata, and Stemster at 1, 3, 7, and 15 days after inoculation (DAI) with 750 nematodes/plant, and an egression assay at 3 DAI with 1000 nematodes/plant. Reproduction assays of cultivars Agata, Agria, Camel, Désirée, Dirosso, Kennebec, Laura, Picasso, Royata, and Stemster were performed in 2 L pots inoculated with four P. penetrans/g soil and quantified at 60 DAI. Tenue or moderate root cell browning to advanced necrotic areas were observed after nematode penetration, and the number of nematodes/g of root gradually increased with time of infection. A lower number of deposited eggs and nematodes were observed within the roots of cultivar Laura in all assays comparatively to other cultivars. The susceptibility index (SI) was significantly lower in cultivar Laura (0.4–0.6), followed by cultivars Camel and Picasso (0.8–0.9). All remaining cultivars showed SI values above 1. Although the potato susceptibility to the nematode varied among cultivars, no differences on the average number or weight of tubers produced by each plant of inoculated versus non-inoculated plants were detected. Our data reveals that these cultivars have a distinct ability to support the reproduction of P. penetrans.

Detection of the root lesion nematode Pratylenchus penetrans in potato tubers

AbstractThe root lesion nematode Pratylenchus penetrans parasitizes a wide range of economically important crops, including potato (Solanum tuberosum). Damage by P. penetrans impacts not only the potato yield but can also reduce the tuber quality. Detailed information on tuber infection by P. penetrans is scarce for most cultivars and molecular detection of nematodes from infected tubers is needed. The objective of this study was to assess tuber symptomatology due to P. penetrans infection in 10 potato cultivars and to provide an accurate molecular methodology for nematode detection using tuber peels. Sprouts of certified potato seed from cultivars Agata, Agria, Camel, Désirée, Dirosso, Kennebec, Laura, Picasso, Royata, and Stemster were planted in 2 L pots, and soil was inoculated with 4 P. penetrans/g of soil. Sixty days after inoculation, tubers were harvested, inspected for lesions, and the number of nematodes/g of potato peel assessed. Observations of tubers with symptoms showed the presence of P. penetrans in superficial layers of peels around the lenticels and injured necrotic tissue. Different nematode stages were detected in tubers of all inoculated cultivars, varying from 4 to 46 nematodes/g of potato peel. Species‐specific primers showed suitable sensitivity and reproducibility for the detection of P. penetrans in tuber potato peel samples. The molecular detection of P. penetrans directly from tuber peels can facilitate routine nematode inspections of potato seed tubers or cull potatoes for nematode detection, and prevent further dissemination of this species.

Interaction between culture filtrates of Fusarium culmorum isolates and some root lesion nematodes

This study was conducted with root lesion nematodes Pratylenchus penetrans, P. thornei and P. neglectus and three isolates of Fusarium culmorum culture filtrates (ISP, B4, FC5) in vitro conditions. The culture filtrates were diluted 1/0,1/10, 1/20, 1/30, 1/40, 1/50, 1/60, 1/70, 1/80, 90, 1/100 in 1.5 ml microtubes, 250 µl of root lesion nematodes adult+larvae were placed in each microtubes that containing different concentrations of culture filtrate with 50 μl of purified water together with micropipette and incubated at 25±1°C. Nematode mortality were determined after 24 h, 48 h and 72 h. As a result, The mortality effect of culture filtrate of FC5 isolates was determined to be high on three lesion nematode species. Fusarium culmorum culture filtrates of B4 and ISP isolates caused more deaths on P. penetrans than P.thornei and P. neglectus. It has been found to mortality rate increase over time in vitro. The lowest mortality rates were generally found at concentrations of culture filtrates of 1/100 and 1/90, while the highest mortality rates were found at concentrations of 1/0 and 1/10. No difference on P. thornei mortality was detected between the three isolates of F. culmorum culture filtrate at each diluted concentration. In the study, the antagonistic relationship between culture filtrates from F. culmorum isolates and root lesion nematodes were determined in vitro conditions.

A new esophageal gland transcriptome reveals signatures of large scale de novo effector birth in the root lesion nematode Pratylenchus penetrans

BackgroundThe root lesion nematode Pratylenchus penetrans is a migratory plant-parasitic nematode responsible for economically important losses in a wide number of crops. Despite the importance of P. penetrans, the molecular mechanisms employed by this nematode to promote virulence remain largely unknown.ResultsHere we generated a new and comprehensive esophageal glands-specific transcriptome library for P. penetrans. In-depth analysis of this transcriptome enabled a robust identification of a catalogue of 30 new candidate effector genes, which were experimentally validated in the esophageal glands by in situ hybridization. We further validated the expression of a multifaceted network of candidate effectors during the interaction with different plants. To advance our understanding of the “effectorome” of P. penetrans, we adopted a phylogenetic approach and compared the expanded effector repertoire of P. penetrans to the genome/transcriptome of other nematode species with similar or contrasting parasitism strategies. Our data allowed us to infer plausible evolutionary histories that shaped the effector repertoire of P. penetrans, as well as other close and distant plant-parasitic nematodes. Two remarkable trends were apparent: 1) large scale effector birth in the Pratylenchidae in general and P. penetrans in particular, and 2) large scale effector death in sedentary (endo) plant-parasitic nematodes.ConclusionsOur study doubles the number of validated Pratylenchus penetrans effectors reported in the literature. The dramatic effector gene gain in P. penetrans could be related to the remarkable ability of this nematode to parasitize a large number of plants. Our data provide valuable insights into nematode parasitism and contribute towards basic understating of the adaptation of P. penetrans and other root lesion nematodes to specific host plants.

Additive effect of the root-lesion nematode Pratylenchus penetrans and the fungus Rhizoctonia solani on potato yield and damage

The significance of nematodes for disease development caused by other soil-borne pathogens has been demonstrated in many crops throughout the world and specifically prevalent are interactions between plant-parasitic nematodes and species of plant pathogenic fungi. Here, the interaction between the fungus Rhizoctonia solani (AG2) and the migratory endoparasitic root-lesion nematode Pratylenchus penetrans was investigated on potato. The hypotheses were that the combination of R. solani and P. penetrans would result in more severe canker lesions, reduced quality of the tubers and lower tuber yield, and we also expected higher nematode levels to render more nematode damage. To test this, potato plants were grown in pots in two replicate experiments and the presence and/or abundance of the two pathogens were controlled. The first three hypotheses were rejected as (1) the tuber yield decreased when the fungus and nematode occurred together but not more than the sum of their separate effects, i.e. additive, (2) there was no effect of presence of nematodes on the incidence of stem canker, and (3) the quality of tubers was actually partly improved as the presence of the nematodes reduced the likelihood of elephant hide on the tubers in one of the experiments. As expected, there were more visible nematode damages with addition of more nematodes, but beyond that the different nematode levels rendered in most cases similar responses. To have knowledge about interactions between pathogens, as the one showed here, is essential for disease control through appropriate management methods.

Natural variation of root lesion nematode antagonism in the biocontrol fungus Clonostachys rosea and identification of biocontrol factors through genome-wide association mapping.

Biological control is a promising approach to reduce plant diseases caused by nematodes to ensure high productivity in agricultural production. Large‐scale analyses of genetic variation in fungal species used for biocontrol can generate knowledge regarding interaction mechanisms that can improve efficacy of biocontrol applications. In this study, we performed a genome‐wide association study (GWAS) for in vitro antagonism against the root lesion nematode Pratylenchus penetrans in 53 previously genome re‐sequenced strains of the biocontrol fungus Clonostachys rosea. Nematode mortality in C. rosea potato dextrose broth (PDB) culture filtrates was highly variable and showed continuous variation (p < .001) between strains, indicating a polygenic inheritance. Twenty‐one strains produced culture filtrates with higher (p ≤ .05) nematode mortality compared with the PDB control treatment, while ten strains lowered (p ≤ .05) the mortality. The difference in in vitro antagonism against P. penetrans correlated with antagonism against the soybean cyst nematode Heterodera glycines, indicating lack of host specificity in C. rosea. An empirical Bayesian multiple hypothesis testing approach identified 279 single nucleotide polymorphism markers significantly (local false sign rate < 10–10) associated with the trait. Genes present in the genomic regions associated with nematicidal activity included several membrane transporters, a chitinase and genes encoding proteins predicted to biosynthesize secondary metabolites. Gene deletion strains of the predicted nonribosomal peptide synthetase genes nps4 and nps5 were generated and showed increased (p ≤ .001) fungal growth and conidiation rates compared to the wild type. Deletion strains also exhibited reduced (p < .001) nematicidal activity and reduced (p ≤ .05) biocontrol efficacy against nematode root disease and against fusarium foot rot on wheat. In summary, we show that the GWAS approach can be used to identify biocontrol factors in C. rosea, specifically the putative nonribosomal peptide synthetases NPS4 and NPS5.

Impacts of Manures and Manure-Based Composts on Root Lesion Nematodes and Verticillium dahliae in Michigan Potatoes.

The interaction of the root lesion nematode Pratylenchus penetrans and the fungal plant pathogen Verticillium dahliae causes potato early die (PED) complex, which induces premature vine senescence and dramatically reduces yield in potatoes. Management of PED is often achieved through the use of soil fumigants and nematicides, but their adverse effects on soil, human and environmental health, and strict regulations worldwide require alternative control tactics. In this study, we investigated the effects of multiple composts and manures on nematode mortality and PED. In lab assays, root lesion nematodes were exposed to poultry manure, layer ash blend, Dairy Doo, or wood ash for 7 days at rates of 0, 0.1, 1, 10, and 20% by volume of product and assessed for nematode survivorship. Additionally, these products were evaluated for volatile fatty acid content to determine if fatty acid content affects nematode control. In a field trial, the composts and manures were evaluated at two different rates, high (11.2 t/ha) or low (2.8 t/ha), and populations of P. penetrans and V. dahliae were quantified. Our results show that a 1% application rate of poultry manure and layer ash blend provided the greatest nematode control in lab assays with 24.5 and 38.2% reduction, respectively, with greater control at higher rates. In the field, plots treated with poultry manure had significantly higher potato yields and significantly fewer nematodes than control plots. Taken together, our results suggest that poultry manure could be a promising amendment to control PED.

An Expansin-Like Candidate Effector Protein from Pratylenchus penetrans Modulates Immune Responses in Nicotiana benthamiana.

The root lesion nematode (RLN) Pratylenchus penetrans is a migratory species that attacks a broad range of crops. After the RLN is initially attracted to host roots by root exudates and compounds, it releases secretions that are critical for successful parasitism. Among those secretions are nematode virulence factors or effectors that facilitate the entry and migration of nematodes through the roots and modulate plant immune defenses. The recognition of the effectors by host resistance proteins leads to effector-triggered immunity and incompatible plant-nematode interactions. Although many candidate effectors of the RLN and other plant-parasitic nematodes have been identified, the detailed mechanisms of their functions and particularly, their host targets remain largely unexplored. In this study, we sequenced and annotated genes encoding expansin-like proteins, which are major candidate effectors of P. penetrans. One of the genes, Pp-EXPB1, which was the most highly expressed during nematode infection in different plant species, was further functionally characterized via transient expression in the model plant Nicotiana benthamiana and global transcriptome profiling of gene expression changes triggered by this candidate effector in plants. As a result of this investigation, the biological roles of Pp-EXPB1 in nematode parasitism were proposed, the putative cellular targets of the proteins were identified, and the molecular mechanisms of plant responses to the nematode-secreted proteins were outlined.

A novel species of RNA virus associated with root lesion nematode Pratylenchus penetrans.

The root lesion nematode Pratylenchus penetrans is a migratory species that attacks a broad range of plants. While analysing transcriptomic datasets of P. penetrans, we have identified a full-length genome of an unknown positive-sense single-stranded RNA virus, provisionally named root lesion nematode virus 1 (RLNV1). The 8614-nucleotide genome sequence encodes a single large polyprotein with conserved domains characteristic for the families Picornaviridae, Iflaviridae and Secoviridae of the order Picornavirales. Phylogenetic, BLAST and domain search analyses showed that RLNV1 is a novel species, most closely related to the recently identified sugar beet cyst nematode virus 1 and potato cyst nematode picorna-like virus. In situ hybridization with a DIG-labelled DNA probe confirmed the presence of the virus within the nematodes. A negative-strand-specific RT-PCR assay detected RLNV1 RNA in nematode total RNA samples, thus indicating that viral replication occurs in P. penetrans. To the best of our knowledge, RLNV1 is the first virus identified in Pratylenchus spp.

Identification and characterization of the first pectin methylesterase gene discovered in the root lesion nematode Pratylenchus penetrans.

Similar to other plant-parasitic nematodes, root lesion nematodes possess an array of enzymes that are involved in the degradation of the plant cell wall. Here we report the identification of a gene encoding a cell wall-degrading enzyme, pectin methylesterase PME (EC 3.1.1.11), in the root lesion nematode Pratylenchus penetrans. Both genomic and coding sequences of the gene were cloned for this species, that included the presence of four introns which eliminated a possible contamination from bacteria. Expression of the Pp-pme gene was localized in the esophageal glands of P. penetrans as determined by in situ hybridization. Temporal expression of Pp-pme in planta was validated at early time points of infection. The possible function and activity of the gene were assessed by transient expression of Pp-pme in plants of Nicotiana benthamiana plants via a Potato virus X-based vector. To our knowledge, this is the first report on identification and characterization of a PME gene within the phylum Nematoda.

Rhizosphere Microbiomes Modulated by Pre-crops Assisted Plants in Defense Against Plant-Parasitic Nematodes

Plant-parasitic nematodes cause considerable damage to crop plants. The rhizosphere microbiome can affect invasion and reproductive success of plant-parasitic nematodes, thus affecting plant damage. In this study, we investigated how the transplanted rhizosphere microbiome from different crops affect plant-parasitic nematodes on soybean or tomato, and whether the plant’s own microbiome from the rhizosphere protects it better than the microbiome from fallow soil. Soybean plants growing in sterilized substrate were inoculated with the microbiome extracted from the rhizosphere of soybean, maize, or tomato. Controls were inoculated with extracts from bulk soil, or not inoculated. After the microbiome was established, the root lesion nematode Pratylenchus penetrans was added. Root invasion of P. penetrans was significantly reduced on soybean plants inoculated with the microbiome from maize or soybean compared to tomato or bulk soil, or the uninoculated control. In the analogous experiment with tomato plants inoculated with either P. penetrans or the root knot nematode Meloidogyne incognita, the rhizosphere microbiomes of maize and tomato reduced root invasion by P. penetrans and M. incognita compared to microbiomes from soybean or bulk soil. Reproduction of M. incognita on tomato followed the same trend, and it was best suppressed by the tomato rhizosphere microbiome. In split-root experiments with soybean and tomato plants, a systemic effect of the inoculated rhizosphere microbiomes on root invasion of P. penetrans was shown. Furthermore, some transplanted microbiomes slightly enhanced plant growth compared to uninoculated plants. The microbiomes from maize rhizosphere and bulk soil increased the fresh weights of roots and shoots of soybean plants, and microbiomes from soybean rhizosphere and bulk soil increased the fresh weights of roots and shoots of tomato plants. Nematode invasion did not affect plant growth in these short-term experiments. In conclusion, this study highlights the importance of the rhizosphere microbiome in protecting crops against plant-parasitic nematodes. An effect of pre-crops on the rhizosphere microbiome might be harnessed to enhance the resistance of crops towards plant-parasitic nematodes. However, nematode-suppressive effects of a particular microbiome may not necessarily coincide with improvement of plant growth in the absence of plant-parasitic nematodes.

Developing a Real-Time PCR Assay for Direct Identification and Quantification of Pratylenchus penetrans in Soil.

The root-lesion nematode Pratylenchus penetrans is a major pathogen of potato worldwide. Yield losses may be exacerbated by interaction with the fungus Verticillium dahliae in the potato early dying disease complex. Accurate identification and quantification of P. penetrans prior to planting are essential for developing effective integrated pest control measures. However, distinction between P. penetrans and other Pratylenchus spp. based on morphology is a tedious task. A SYBR Green I-based qPCR assay was developed to discriminate, identify, and quantify P. penetrans in field soil. P. penetrans-specific qPCR primers were designed from the D2-D3 region of the 28S rDNA. The specificity of the assay was evaluated using eight isolates of P. penetrans populations and 31 isolates of other nematode species. A standard curve relating threshold cycle and log values of nematode number was generated from artificially infested soils. There was a high correlation between the P. penetrans numbers artificially added to soil or estimated from naturally infested field soils by conventional methods, and the numbers quantified using the qPCR assay. Grinding the field soil prior to DNA extraction improved P. penetrans detection from soil. The qPCR assay will not only be useful for differentiating P. penetrans from mixed populations of Pratylenchus spp., but also for efficient detection and quantification of P. penetrans from field soil. The assay requires no expertise in nematode taxonomy and morphology, and may serve as a useful diagnostic tool in research, diagnostic labs, and extension services for pest management.

Seed Treatment Effects on Maize Seedlings Coinfected with Rhizoctonia solani and Pratylenchus penetrans.

The roots of maize seedlings typically are attacked by a complex of organisms that includes fungal pathogens and plant-parasitic nematodes but few studies have examined the effects of these organisms in combination. Rhizoctonia solani can be an important component of the seedling disease complex; like other fungi, its effect on the plant may be influenced by the activity of nematodes such as the root-lesion nematode Pratylenchus penetrans. In this study, we assessed the impact of seed treatments, including fungicide-nematicide combinations, on maize seedlings exposed to R. solani and P. penetrans alone or in combination. In growth-chamber and greenhouse experiments, seed treated with various active ingredient combinations were planted in an autoclaved sand-soil mixture with or without inoculum of R. solani. In some treatments, a suspension of P. penetrans adults and juveniles was added to the sand-soil mixture. In the greenhouse experiments, infection by R. solani caused dramatic reductions in root length, volume, surface area, and numbers of root tips and root forks, whereas P. penetrans infestation alone reduced only shoot fresh weight. Statistical interactions between the effects of the two organisms were not significant, although fungal infestation significantly reduced the numbers of nematodes extracted from roots. Seed treatments significantly improved most root development variables, and the combination that included four fungicides, thiamethoxam, and abamectin was the best treatment for most variables. Results were similar in the growth-chamber experiments, where R. solani caused significant reductions in nearly all shoot and root development measurements, and seed treatment with sedaxane, alone or combined with abamectin, consistently provided the best results. R. solani was more damaging to seedlings than P. penetrans, and the combination of the two organisms did not cause more damage than R. solani alone. Seed-treatment active ingredients that specifically targeted R. solani (sedaxane) and P. penetrans (abamectin) had large positive effects on seedling health, causing significant improvements in root and shoot growth and development compared with untreated seedlings exposed to these pathogens.

Free-Living Plant-Parasitic Nematodes do not Affect the Efficiency of Seed Tuber Fungicide Treatment against Rhizoctonia solani

Stem canker on germinating potato sprouts is often caused by seed-borne inoculum of the fungus Rhizoctonia solani. However, high amounts of free-living plant-parasitic nematodes have been found in field patches of potato plants with stem canker. Fungicide treatment of the seed tubers can be used to avoid stem canker caused by seed-borne inoculum but it is unknown if nematodes can affect this. To investigate whether free-living plant-parasitic nematodes, the root-lesion nematode Pratylenchus penetrans or a combination of several plant-parasitic nematode genera in a full nematode community, may have a negative effect on the fungicide seed treatment, a pot experiment with seed tubers inoculated with R. solani, half of which were treated with fungicides, was performed. The seed-borne inoculum caused severe damage to the plants, while no fungal damages were observed on the fungicide treated plants. This shows that the nematodes did not affect the fungicide treatment. The probability of black scurf decreased in treatments with a full nematode community, which may be due to the action of fungal-feeding nematodes.

Distribution and Longevity of Pratylenchus penetrans in the Red Raspberry Production System.

One of the major constraints on the production of red raspberries in the Pacific Northwest is the presence of the root-lesion nematode Pratylenchus penetrans. Current management of this nematode relies heavily on preplant soil fumigation; however, regulations have made the practice more difficult and expensive. Additional issues with soil fumigation include lack of efficacy at deeper soil depths and potential inability to penetrate raspberry root material that remains in the field during fumigation which may harbor P. penetrans. To address these issues, two field experiments were conducted in northwestern Washington. In the first experiment, the residency time of P. penetrans in root material from the previous raspberry crop, which was terminated with or without the use of herbicides, was monitored over time. Pratylenchus penetrans was found in root material from 6 to 8 mon after the crop was terminated, and herbicide application did not reduce P. penetrans residency time compared to untreated root material. In a second experiment, the vertical distribution of P. penetrans at three different times during the field establishment process (pre- and postfumigation, and at planting) was determined at two locations. Both locations had detectable prefumigation P. penetrans populations at all depths. However, postfumigation populations showed a different distribution pattern between locations. The location with coarser soil had populations located mainly at shallower depths with a maximum of 44 P. penetrans/100 g soil at 16 to 30 cm deep, whereas the location with finer soil had populations located mainly at deeper depths with a maximum of 8 P. penetrans/100 g soil at 76 to 90 cm deep. At planting, distribution tended to equilibrate among depths at both locations, but the overall population pattern across depth at each location was similar to that observed at postfumigation. Understanding more about the residency time and distribution of this nematode may provide growers with information that can be used to more effectively target P. penetrans.

Biochars reduce infection rates of the root-lesion nematode Pratylenchus penetrans and associated biomass loss in carrot

Biochars, in addition to carbon sequestration, soil amelioration and improvement of plant performance, can measurably reduce disease severity of different pathogen types and even induce system wide defense responses in host plants. The aim of this study was to further investigate if biochars can provide resistance-enhancing effects to host plants faced with plant parasitic nematodes. We asked if these carbonized materials, as a result, may hold the potential to be used as an effective supplement for chemical nematicides. Four different biochars and zeolite (5% v/v) were tested on a carrot (Daucus carota) and root-lesion nematode (Pratylenchus penetrans) pathogen system. Plant biomass, nematode abundances in soil and root tissue (infection rate), as well as plant-nutrient status were quantified. P. penetrans caused decreased shoot- and fine root biomass in infected plants. All applied materials, except for the pine wood biochar, significantly reduced tap root infection rates of P. penetrans by approximately 80% and spelt husk biochar actually reduced infection rates by more than 96%. Infected plants of these treatments produced two to four times more plant biomass than infected plants of the non-material application treatment. It turned out that biochars produced from different feedstocks affected nematode infection rates and host plant biomasses quite differently. Induced resistance is a possible explanation for the highly reduced infection rates, as direct toxic effects of the biochar and effects of altered pH, water holding capacity, soil structure and plant nutrient status could be largely excluded.

The Pratylenchus penetrans Transcriptome as a Source for the Development of Alternative Control Strategies: Mining for Putative Genes Involved in Parasitism and Evaluation of in planta RNAi.

The root lesion nematode Pratylenchus penetrans is considered one of the most economically important species within the genus. Host range studies have shown that nearly 400 plant species can be parasitized by this species. To obtain insight into the transcriptome of this migratory plant-parasitic nematode, we used Illumina mRNA sequencing analysis of a mixed population, as well as nematode reads detected in infected soybean roots 3 and 7 days after nematode infection. Over 140 million paired end reads were obtained for this species, and de novo assembly resulted in a total of 23,715 transcripts. Homology searches showed significant hit matches to 58% of the total number of transcripts using different protein and EST databases. In general, the transcriptome of P. penetrans follows common features reported for other root lesion nematode species. We also explored the efficacy of RNAi, delivered from the host, as a strategy to control P. penetrans, by targeted knock-down of selected nematode genes. Different comparisons were performed to identify putative nematode genes with a role in parasitism, resulting in the identification of transcripts with similarities to other nematode parasitism genes. Focusing on the predicted nematode secreted proteins found in this transcriptome, we observed specific members to be up-regulated at the early time points of infection. In the present study, we observed an enrichment of predicted secreted proteins along the early time points of parasitism by this species, with a significant number being pioneer candidate genes. A representative set of genes examined using RT-PCR confirms their expression during the host infection. The expression patterns of the different candidate genes raise the possibility that they might be involved in critical steps of P. penetrans parasitism. This analysis sheds light on the transcriptional changes that accompany plant infection by P. penetrans, and will aid in identifying potential gene targets for selection and use to design effective control strategies against root lesion nematodes.