Search

Soil suppressiveness can reduce the damage by plant parasitic nematodes (PPN) in agricultural soils and is conveyed by the activity of soil microorganisms. While natural suppressiveness has been reported, it is still poorly understood if soil suppressiveness can be elicited by manipulating the soil microbial community. In the present study we assessed the number of the Pratylenchus penetrans (Pp) and the bacterial and fungal community composition over 7 years in a long-term soil health experiment. The field experiment consisted of an organic and conventional agricultural land management system and three soil health treatments (SHT): an untreated control (CT), anaerobic disinfestation (AD) and a combination of marigold cover cropping, compost and chitin amendment (CB). The land management systems were kept continuously, while the soil health treatments were applied only twice in seven years. The microbial community significantly differed between the organic and conventional system, but there was no significant difference in Pp numbers between the two systems. However, both the CB treatment and to a lesser extent the AD treatment reduced Pp numbers and increased yield with the effect being the strongest in the years immediately after the treatment. Accordingly, both the bacterial and fungal community differed significantly between the treatments, the differences being largest in the years after the treatments. Notably, the CB treatment elicited both long-term changes in the microbial community and a reduction of Pp numbers lasting for at least three years. These results indicated that a combination of treatments can lead to an altered soil microbial community in combination with persisting suppressiveness of Pp. [ABSTRACT FROM AUTHOR]

Easter lilies, Lilium longiflorum cv. Nellie White are a staple of the floral industry. In the U.S. most of the Easter lilies are grown in Oregon and California along the coast where there is a micro climate that is favorable to growth of lilies. The main pest when growing lilies in the field is Pratylenchus penetrans, the root lesion nematode. Easter lilies are one of the most expensive crops to produce because of the cost of chemicals used to control P. penetrans and other pathogens that infect the lilies. Our previous study had shown that transgenic Easter lilies containing a rice cystatin gene (Oc-IΔD86 that has a deleted Asp86) were resistant to P. penetrans in vitro. This study examined growth characteristics of five independently transformed lines of the cystatin Easter lilies compared to non-transformed Nellie White for three seasons in the field in Brookings, Oregon. Liles grown in three soil chemical treatments 1) preplant fumigation, 2) preplant fumigation plus at plant organophosphate, and 3) at plant organophosphate were compared to those grown in nontreated soil. Growth characteristics evaluated included: time of shoot emergence, survival of plants, size of plants, visual ratings of plant health, basal roots and stem roots, weight of foliage and roots, and number and size of bulblets that developed on stems. Nematodes were counted following their extraction from the roots. While not totally resistant, when planted in the field, transformed lines demonstrated and maintained a degree of resistance to lesion nematode over two growing seasons and displayed desirable growth and quality characteristics similar to non-transformed lilies.

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. [ABSTRACT FROM AUTHOR]

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.

Easter lily bulbs for greenhouse forcing are produced in Del Norte County, California and Curry County, Oregon, USA. Pratylenchus penetrans infestation seriously affects growth of field grown bulbs. During two consecutive years of field trials containing 22 treatments, commercially prepared formulations of essential oils (EOs) were compared to an untreated control and to a standard chemical fumigant treatment (FU) (1,3-dichloropropene and metam sodium) applied preplant followed by phorate (PH) at planting to determine their value in the management of lesion nematode, and in improving plant health. The EO products Duogard, EF400, EF300, and Cinnamite were tested as preplant dips to bulblet planting stock. The treated bulblets were tested either alone, in combination with PH at-planting, at planting following FU or in combination with PH at planting following FU. The organophosphates ethoprop and fosthiazate were also tested either alone, or at a reduced rate in combination with a reduced rate of PH. With respect to bulb circumference, ten treatments consistently outperformed the control. In consecutive years, three treatments had healthier looking roots than the control. At harvest, levels of lesion nematode within roots were consistently lower in nine treatments. EOs were beneficial in mitigating nematode damage.Easter lily bulbs for greenhouse forcing are produced in Del Norte County, California and Curry County, Oregon, USA. Pratylenchus penetrans infestation seriously affects growth of field grown bulbs. During two consecutive years of field trials containing 22 treatments, commercially prepared formulations of essential oils (EOs) were compared to an untreated control and to a standard chemical fumigant treatment (FU) (1,3-dichloropropene and metam sodium) applied preplant followed by phorate (PH) at planting to determine their value in the management of lesion nematode, and in improving plant health. The EO products Duogard, EF400, EF300, and Cinnamite were tested as preplant dips to bulblet planting stock. The treated bulblets were tested either alone, in combination with PH at-planting, at planting following FU or in combination with PH at planting following FU. The organophosphates ethoprop and fosthiazate were also tested either alone, or at a reduced rate in combination with a reduced rate of PH. With respect to bulb circumference, ten treatments consistently outperformed the control. In consecutive years, three treatments had healthier looking roots than the control. At harvest, levels of lesion nematode within roots were consistently lower in nine treatments. EOs were beneficial in mitigating nematode damage.

Easter lilies, Lilium longiflorum cv. Nellie White are a staple of the floral industry. In the U.S. most of the Easter lilies are grown in Oregon and California along the coast where there is a micro climate that is favorable to growth of lilies. The main pest when growing lilies in the field is Pratylenchus penetrans, the root lesion nematode. Easter lilies are one of the most expensive crops to produce because of the cost of chemicals used to control P. penetrans and other pathogens that infect the lilies. Our previous study had shown that transgenic Easter lilies containing a rice cystatin gene (Oc-IΔD86 that has a deleted Asp86) were resistant to P. penetrans in vitro. This study examined growth characteristics of five independently transformed lines of the cystatin Easter lilies compared to non-transformed Nellie White for three seasons in the field in Brookings, Oregon. Liles grown in three soil chemical treatments 1) preplant fumigation, 2) preplant fumigation plus at plant organophosphate, and 3) at plant organophosphate were compared to those grown in nontreated soil. Growth characteristics evaluated included: time of shoot emergence, survival of plants, size of plants, visual ratings of plant health, basal roots and stem roots, weight of foliage and roots, and number and size of bulblets that developed on stems. Nematodes were counted following their extraction from the roots. While not totally resistant, when planted in the field, transformed lines demonstrated and maintained a degree of resistance to lesion nematode over two growing seasons and displayed desirable growth and quality characteristics similar to non-transformed lilies.

Root-knot (Meloidogyne incognita (Kofoid & White) Chitwood), reniform (Rotylenchulus reniformis Lindford & Oliveira), and lesion nematodes (Pratylenchus penetrans (Cobb) Filipjev & Schuurmans Stekhoven) are plant-parasitic nematodes that feed on soybean (Glycine max (L.) Merr.) roots, limiting seed production. The availability of resistance in soybeans to these nematodes is limited. However, new sources of resistance can be discovered in wild relatives of agronomic crops. Perennial Glycine species, wild relatives to soybean, are a source of valuable genetic resources with the potential to improve disease resistance in soybean. To determine if these perennials have resistance against nematodes, 18 accessions of 10 perennial Glycine species were evaluated for their response to M. incognita and R. reniformis, and eight accessions of six perennial Glycine species were evaluated for their response to P. penetrans. Pot experiments were conducted for M. incognita and R. reniformis in a growth chamber and in vitro experiments were conducted for P. penetrans. We found both shared and distinct interactions along the resistance-susceptible continuum in response to the three plant-parasitic nematode species. Ten and 15 accessions were classified as resistant to M. incognita based on eggs per gram of root and gall index, respectively. Among them, G. tomentella plant introductions (PIs) 446983 and 339655 had a significantly lower gall index than the resistant soybean check cv. Forrest. Of three R. reniformis resistant accessions identified in this study, G. tomentella PI 441001 showed significantly greater resistance to R. reniformis than the resistant check cv. Forrest based on nematodes per gram of root. In contrast, no resistance to P. penetrans was recorded in any perennial Glycine species.

Root-knot (Meloidogyne incognita (Kofoid & White) Chitwood), reniform (Rotylenchulus reniformis Lindford & Oliveira), and lesion nematodes (Pratylenchus penetrans (Cobb) Filipjev & Schuurmans Stekhoven) are plant-parasitic nematodes that feed on soybean (Glycine max (L.) Merr.) roots, limiting seed production. The availability of resistance in soybeans to these nematodes is limited. However, new sources of resistance can be discovered in wild relatives of agronomic crops. Perennial Glycine species, wild relatives to soybean, are a source of valuable genetic resources with the potential to improve disease resistance in soybean. To determine if these perennials have resistance against nematodes, 18 accessions of 10 perennial Glycine species were evaluated for their response to M. incognita and R. reniformis, and eight accessions of six perennial Glycine species were evaluated for their response to P. penetrans. Pot experiments were conducted for M. incognita and R. reniformis in a growth chamber and in vitro experiments were conducted for P. penetrans. We found both shared and distinct interactions along the resistance-susceptible continuum in response to the three plantparasitic nematode species. Ten and 15 accessions were classified as resistant to M. incognita based on eggs per gram of root and gall index, respectively. Among them, G. tomentella plant introductions (PIs) 446983 and 339655 had a significantly lower gall index than the resistant soybean check cv. Forrest. Of three R. reniformis resistant accessions identified in this study, G. tomentella PI 441001 showed significantly greater resistance to R. reniformis than the resistant check cv. Forrest based on nematodes per gram of root. In contrast, no resistance to P. penetrans was recorded in any perennial Glycine species. [ABSTRACT FROM AUTHOR]

The root-lesion nematode, Pratylenchus penetrans , is a ubiquitous parasite of roots of temperate fruit trees. It affects early growth of trees replanted into former orchard sites where populations have built up and may contribute to decline complexes of older trees. Most British Columbia, Canada, apple acreage is planted with M.9 rootstock, but growers are increasingly considering Geneva-series rootstocks such as G.41 and G.935. Among these rootstocks, responses to P. penetrans , specifically, are poorly known. To compare the resistance and tolerance to P. penetrans of G.41, G.935, and M.9 rootstocks ('Ambrosia' scion), a field microplot experiment was established in spring of 2020 at the Summerland Research and Development Centre. The experimental design was a two by three factorial combination of: P. penetrans inoculation (+/-) and rootstock (G.41, G.935, and M.9), with 20 replicate microplots of each of the six treatment combinations arranged in a randomized complete block design. The P. penetrans inoculum was 5,400 nematodes per microplot (54 P. penetrans liter -1 soil), which is below commonly accepted damage thresholds. Though P. penetrans population densities were lower for the G.41 rootstock by the end of the 2021 growing season, the effects of P. penetrans were similar among rootstocks. In the establishment year (2020), P. penetrans caused significant reductions in aboveground growth. In 2021, shoot growth and root weight were reduced by P. penetrans . The nematode also reduced rates of leaf gas exchange and stem water potential. These data suggest that while G.41 and G.935 may have other horticultural benefits over M.9, they are equally susceptible to P. penetrans at the early stages of tree growth., Competing Interests: The author(s) declare no conflict of interest.

Annual rotation green manure crops of selected brassicas, buckwheat, forage pearl millet, forage radish, black-eyed Susan, sesame, sudangrass, and velvetbean were evaluated to determine impacts on lesion nematode Pratylenchus penetrans and dagger nematode Xiphinema americanum populations densities. Canadian forage pearl millet ' 101'and 'Tifgrain 102' millet effectively controlled P. penetrans but increased population densities ofX. americanum. Black-eyed Susan (Rudbeckia hirta) and sudangrass 'Trudan 8' also reduced P. penetrans densites but notX. americanum. Rapeseed and other brassicas as a green manure reducedX. americanum densities but did not suppress P. penetrans. Brassica juncea 'Pacific Gold', B. napus 'Dwarf Essex', but not mustard 'Caliente' resulted in low densities ofX. americanum in soil. Velvetbean and sesame increased both P. penetrans and X. americanum population densities. A moderately suitable host plant such as buckwheat was ineffective in managing both P. penetrans and X. americanum populations. These results emphasize that suppressive effects of any given cover crop are nematode-specific, and that within a group of cover crops, e.g., the Brassicaceae, species and varieties can vary substantially in their effectiveness. If the rotation crop chosen is a good host for the nematodes present, it may exacerbate the problem instead of controlling it. [ABSTRACT FROM AUTHOR]

A survey was conducted in Washington State in 2015 and 2016 to gauge grower perceptions, understanding, and current practices regarding soil quality. Soil quality has been defined as the ability of the soil to sustain plants, animals, and humans over time. Many current practices of modern agriculture can be detrimental to soil quality, including soil tillage and soil fumigation, both of which are commonly used for the Washington red raspberry (Rubus idaeus) production system. The area between red raspberry beds, known as the alleyway, is frequently tilled and kept bare, without groundcover, to manage weeds. Growers commonly fumigate the soil before planting red raspberry to manage soilborne pathogens and plant-parasitic nematodes. The majority of red raspberry growers surveyed consider soil quality quite often in relation to the management of their fields. The majority of growers during both years considered cover crops to have a positive impact on soil quality. However, growers also perceived soil fumigation to have a positive impact on soil quality. The majority of growers responded that they were willing to adopt alleyway cover crops for a variety of reasons, including improving red raspberry production, physical soil quality, and beneficial soil microorganism populations. This survey demonstrated that there is interest in soil quality among growers; however, there is a difference in perceptions between growers and researchers regarding how management practices impact soil quality.

Sweet cherry growers are increasingly using semi-dwarfing rootstocks, including the Gisela® series, when replanting orchards. Little is known of the susceptibility of these new cherry rootstocks to Pratylenchus penetrans, a recognized pest of temperate fruit trees worldwide. Two field experiments were planted in 2010, one in the Okanagan Valley of British Columbia and one in the Annapolis Valley of Nova Scotia. Each experiment was a factorial combination of three rootstocks (Gi.3, Gi.5, and Gi.6) × three training systems, with six replicate four-tree plots of each of the nine combinations. Both sites were fumigated prior to planting and population densities of P. penetrans in roots and root-zone soil were subsequently monitored from 2013 through 2017. None of the P. penetrans population parameters (nematodes/kg soil, nematodes/g fine root, and nematodes/kg soil including roots) differed among rootstocks at either site, suggesting that the rootstocks did not differ in their ability to host P. penetrans. At the British Columbia site only there was an inverse relationship between P. penetrans population densities and tree size for Gi.3 trees in four years and for Gi.6 in 2017, suggesting that Gi.3 rootstock is less tolerant than Gi.5 and Gi.6 rootstocks.

Summary: Seven species belonging to Suborder Hoplolaimina are characterised using integrative taxonomy, considering both morphological and molecular phylogenetic analyses of the 28S-rRNA, ITS-rRNA and COI gene sequences. It is evident that, as more populations of Pratylenchus zeae are continuously characterised, the species continues to display an ever-increasing intraspecific genetic variation within the 28S-rRNA and ITS-rRNA genes. However, the COI gene sequences exhibit minimum intraspecific variation and thus might be the most powerful DNA barcoding marker for the precise identification of P. zeae and should therefore be recommended as a complementary technique in the identification process of the species. Pratylenchus zeae , Meloidogyne graminicola and Heterodera pratensis are characterised herein for the first time in Korea, while the presence in Korea of P. penetrans , P. scribneri , H. avenae , and M. marylandi , is molecularly confirmed. [ABSTRACT FROM AUTHOR]

Summary: Robust and accurate identification of root-lesion nematodes (Pratylenchus spp.) is an essential step for determining their potential threat to crop yields and, consequently, development of an efficient agronomic management strategy. It is recognised that DNA-based techniques provide rapid identification of a range of plant-parasitic nematodes including Pratylenchus spp. Efficient and repeatable DNA extraction is central to molecular methodologies. Here, six common DNA extraction protocols were compared to evaluate their efficiency to obtain quality DNA samples for Pratylenchus penetrans. Samples with five and ten individuals of P. penetrans were successfully extracted and amplified by all extraction methods tested, whereas samples with a single nematode presented challenges for DNA amplification. Among all methods tested, the DNA extraction protocol with glass beads proved to be efficient for P. penetrans and all other species tested (P. crenatus , P. neglectus and P. thornei), generating high quality DNA at comparatively low cost and with a rapid sample throughput. [ABSTRACT FROM AUTHOR]

Continuous cropping often leads yield losses via deteriorated soil biological and chemical properties. In this mini review, the importance of organic matter amendment is focused in relation to the management of soilborne plant pathogens and plant parasitic nematodes. A wide range of examples are shown for organic matter amendment including composts, cover crops and simple organic compound to suppress the densities of pathogens and nematodes. A novel diagnostic method using real-time PCR is also shown for plant parasitic nematodes. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Root‐lesion nematodes of the genus Pratylenchus are migratory endoparasites with worldwide economic impact on several important crops including potato, where certain species like P. penetrans, P. neglectus, and P. scribneri reduce the yield and quality of potato tubers. Morphological identification of Pratylenchus spp. is challenging, and recent advancements in molecular techniques provide robust and rapid diagnostics to differentiate species without the need of specialist skills. However, the fact that molecular diagnostics are not available for all Pratylenchus species means that there are limitations in worldwide application. In general, root‐lesion nematodes are difficult to manage once introduced into agricultural land and damage can be related to pathogenicity and population densities. In addition, root‐lesion nematodes interact with fungi such as Verticillium dahliae, resulting in disease complexes that enhance the damage inflicted on the potato crop. Management interventions are often focused on limiting nematode reproduction before planting crops and include the application of nematicides, and cultural practices such as crop rotation, cover crops, biofumigation, and biological control. Understanding the limitations of the available crop protection strategies is important and there are many gaps for further study. This review discusses the status of the diagnosis, distribution, pathogenicity, and management of the main species of root‐lesion nematodes, reported to infect potatoes worldwide, and highlights areas for potential future research. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Early maturing varieties of soybean have a high yield potential in Europe, where the main biotic threat to soybean cultivation are root lesion nematodes (Pratylenchus spp.). Nitrogen fixation in root nodules by highly efficient inoculants of Bradyrhizobium japonicum is an incentive to grow soybean in low-input rotation systems. We investigated density-dependent effects of Pratylenchus penetrans on nitrogen fixation by co-inoculated B. japonicum. Less than 130 inoculated nematodes affected the number and weight of nodules, the density of viable bacteroids in nodules, and nitrogen fixation measured as concentration of ureides in leaves. With more inoculated nematodes, the percentage that invaded the roots increased, and adverse effects on the symbiosis accelerated, leading to non-functional nodules at 4,000 and more nematodes. When P. penetrans invaded roots that had fully established nodules, growth of nodules, density of bacteroids, and nitrogen fixation were affected but not the number of nodules. In contrast, nodulation of already infested roots resulted in a high number of small nodules with decreased densities of bacteroids and nitrogen fixation. P. penetrans invaded and damaged the nodules locally, but they also significantly affected the nodule symbiosis by a plant-mediated mechanism, as shown in an experiment with split-root systems. [ABSTRACT FROM AUTHOR]

The bacterial endosymbiont Wolbachia interacts with different invertebrate hosts, engaging in diverse symbiotic relationships. Wolbachia is often a reproductive parasite in arthropods, but an obligate mutualist in filarial nematodes. Wolbachia was recently discovered in plant-parasitic nematodes, and, is thus far known in just two genera Pratylenchus and Radopholus, yet the symbiont’s function remains unknown. The occurrence of Wolbachia in these economically important plant pests offers an unexplored biocontrol strategy. However, development of Wolbachia-based biocontrol requires an improved understanding of symbiont-host functional interactions and the symbiont’s prevalence among nematode field populations. This study used a molecular-genetic approach to assess the prevalence of a Wolbachia lineage (wPpe) in 32 field populations of Pratylenchus penetrans. Populations were examined from eight different plant species in Washington, Oregon, and California. Nematodes were also screened for the endosymbiotic bacterium Cardinium (cPpe) that was recently shown to co-infect P. penetrans. Results identified wPpe in 9/32 and cPpe in 1/32 of P. penetrans field populations analyzed. No co-infection was observed in field populations. Wolbachia was detected in nematodes from 4/8 plant-hosts examined (raspberry, strawberry, clover, and lily), and in all three states surveyed. Cardinium was detected in nematodes from mint in Washington. In the wPpe-infected P. penetrans populations collected from raspberry, the prevalence of wPpe infection ranged from 11 to 58%. This pattern is unlike that in filarial nematodes where Wolbachia is an obligate mutualist and occurs in 100% of the host. Further analysis of wPpe-infected populations revealed female-skewed sex ratios (up to 96%), with the degree of skew positively correlating with wPpe prevalence. Uninfected nematode populations had approximately equal numbers of males and females. Comparisons of 54 wPpe 16S ribosomal RNA sequences revealed high similarity across the geographic isolates, with 45 of 54 isolates being identical at this locus. The complete absence of wPpe among some populations and low prevalence in others suggest that this endosymbiont is not an obligate mutualist of P. penetrans. The observed sex ratio bias in wPpe-infected nematode populations is similar to that observed in arthropods where Wolbachia acts as a reproductive manipulator, raising the question of a similar role in plant-parasitic nematodes.

Since the discovery of Wolbachia in plant-parasitic nematodes (PPNs), there has been increased interest in this earliest branching clade that may hold important clues to early transitions in Wolbachia function in the Ecdysozoa. However, due to the specialized skills and equipment of nematology and the difficulty in culturing most PPNs, these PPN-type Wolbachia remain undersampled and poorly understood. To date, there are few established laboratory methods for working with PPN-type Wolbachia strains, and most research has relied on chance discovery and comparative genomics. Here, we address this challenge by providing detailed methods to assist researchers with more efficiently collecting PPNs and screen these communities, populations, or single nematodes with a newly developed PPN-type Wolbachia-specific PCR assay. We provide an overview of the typical yields and outcomes of these methods, to facilitate further targeted cultivation or experimental methods, and finally we provide a short introduction to some of the specific challenges and solutions in following through with comparative or population genomics on PPN-type Wolbachia strains., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Pratylenchus penetrans is an economically important root-lesion nematode species that affects agronomic and ornamental plants. Understanding its diversity is of paramount importance to develop effective control and management strategies. This study aimed to characterize the morphological and genetic diversity among seven European isolates. An isolate from the USA was included in the molecular analyses for comparative purposes. Morphometrics of the European P. penetrans isolates generally were within the range of the original descriptions for this species. However, multiple morphometric characteristics, including body length, maximum body width, tail length and length of the post-vulval uterine sac showed discrepancies when compared to other populations. Nucleotide sequence-based analyses revealed a high level of intraspecific diversity among the isolates. We observed no correlation between D2-D3 rDNA- and COXI-based phylogenetic similarities and geographic origin. Our phylogenetic analyses including selected GenBank sequences also suggest that the controversy surrounding the distinction between P. penetrans and P. fallax remains.

Abstract In Florida, strawberry is an economically important horticultural crop. Plant-parasitic nematodes present a significant obstacle for strawberry growers in the sandy soils found in this state. Historically, nematodes have been controlled through the application of soil fumigants; however, there is interest in exploring the use of non-fumigant nematicides that can provide similar levels of control and at the same time be more environmentally friendly than fumigants. Using two field experiments, the effects of non-fumigant chemical and biological nematicides on plant vigour and fruit yield, populations of plant-parasitic nematodes, and populations of free-living nematode feeding groups were evaluated. In experiment 1, soil fumigation with metam potassium, as well as all non-fumigant chemical nematicides (fluopyram, fluensulfone, fluazaindolizine) failed to increase plant vigour, fruit yield, or suppress populations of plant-parasitic nematodes. In experiment 2, fluopyram increased fruit yield and suppressed Belonolaimus longicaudatus populations by the end of the growing season. Biological nematicides (Dazitol®, Majestene®, and Nemakill®) did not enhance crop establishment or suppress plant-parasitic nematode populations. Soil fumigation with 1,3-dichloropropene + chloropicrin provided consistent enhancement of plant vigour and fruit yield, as well as suppressed populations of B. longicaudatus ; however, populations of free-living nematodes were also reduced. Overall, soil application of fluopyram shows potential to be a component of an integrated pest management strategy for the control of B. longicaudatus populations on strawberry in Florida. Highlights • Chemical and biological nematicides were evaluated on strawberry in Florida. • In experiment 1, metam potassium and non-fumigant nematicides did not improve yield. • In experiment 2, fluopyram increased yield and suppressed Belonolaimus longicaudatus. • Fumigation with 1,3D + chloropicrin improved yield, vigor, and suppressed nematodes. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Abstract A decline in red raspberry (Rubus idaeus) crop longevity has been partially attributed to parasitism by the plant-parasitic nematode Pratylenchus penetrans. Soil fumigation with 1,3-dichloropropene (1,3-D) and chloropicrin is the most common management practice for P. penetrans in this production system, but the effects are variable and there are strict regulations regarding application; alternatives are needed. A three-year study was conducted in a replanted commercial red raspberry field with a history of P. penetrans to evaluate brassicaceous seed meal (BSM) soil amendment combined with root inoculum removal (RR) as preplant alternatives to traditional soil fumigation with 1,3-D and chloropicrin. Additionally, the fumigant metam sodium at full and half rates with root removal was evaluated. Treatments were applied once prior to planting and included: BSM at 3.4 t ha−1 with RR, full rate metam sodium (692 L ha−1; Max Fum) with RR, half rate metam sodium with RR (Min Fum), and full rate metam sodium without RR (Max Fum–RR; control). Data collected included: P. penetrans population dynamics in soil and raspberry roots, microbial (bacterial and fungal) communities in soil, raspberry vegetative growth, estimated yield, and fruit total soluble solids (TSS) concentration. Population densities of P. penetrans in soil and roots were significantly higher in BSM (983–4801 P. penetrans g−1 of root) than in Max Fum and Max Fum–RR (32–802 and 40–1509 P. penetrans g−1 of root, respectively) during the first two years of the study. By the end of the study there were no differences in P. penetrans population densities among the treatments. Root removal did not affect P. penetrans densities as there were no significant differences between Max Fum and Max Fum–RR. Min Fum was more effective than BSM at reducing P. penetrans population densities, but not as effective as Max Fum or Max Fum-RR. The only difference among treatments regarding the soil microbial community was observed in the first spring after treatment application, when BSM had a soil bacterial community that differed from the other treatments; this difference did not persist into the next sampling date. The most noticeable differences in bacterial and fungal soil communities were due to season, not treatment. There were no significant differences in raspberry vegetative growth the first summer after planting. Yield or TSS did not differ in the second and third summer after planting. The current data show that Min Fum, BSM amended to soil at this experimental rate, and root removal are not effective in reducing P. penetrans population densities, but yield and fruit quality are not compromised under the conditions of the experiment. [ABSTRACT FROM AUTHOR]

Wolbachia and Cardinium are among the most important and widespread of all endosymbionts, occurring in nematodes and more than half of insect and arachnid species, sometimes as coinfections. These symbionts are of significant interest as potential biocontrol agents due to their abilities to cause major effects on host biology and reproduction through cytoplasmic incompatibility, sex ratio distortion, or obligate mutualism. The ecological and metabolic effects of coinfections are not well understood. This study examined a Wolbachia–Cardinium coinfection in the plant-parasitic nematode (PPN), Pratylenchus penetrans, producing the first detailed study of such a coinfection using fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic analysis. Results from FISH and single-nematode PCR showed 123/127 individuals in a focal population carried Cardinium (denoted strain cPpe), and 48% were coinfected with Wolbachia strain wPpe. Both endosymbionts showed dispersed tissue distribution with highest densities in the anterior intestinal walls and gonads. Phylogenomic analyses confirmed an early place of cPpe and long distance from a sister strain in another PPN, Heterodera glycines, supporting a long history of both Cardinium and Wolbachia in PPNs. The genome of cPpe was 1.36 Mbp with 35.8% GC content, 1,131 predicted genes, 41% having no known function, and missing biotin and lipoate synthetic capacity and a plasmid present in other strains, despite having a slightly larger genome compared to other sequenced Cardinium. The larger genome revealed expansions of gene families likely involved in host–cellular interactions. More than 2% of the genes of cPpe and wPpe were identified as candidate horizontally transferred genes, with some of these from eukaryotes, including nematodes. A model of the possible Wolbachia–Cardinium interaction is proposed with possible complementation in function for pathways such as methionine and fatty acid biosynthesis and biotin transport.

Inoculation with antagonistic soil microorganisms has shown potential to suppress replant disease of apple in orchard soils. Pseudomonas spp. may have the potential to reduce Pratylenchus penetrans populations on apple. Pseudomonas spp. were isolated from the rhizosphere of sweet cherry and screened for antagonistic characteristics. Two highly antagonistic Pseudomonas isolates, P10-32 and P10-42, were evaluated for growth promotion of apple seedlings, suppression of P. penetrans populations, and root colonization in soil from three orchards. During the isolate screening, Pseudomonas fluorescens P10-32 reduced in vitro growth of fungal pathogens, had protease activity, had capacity to produce pyrrolnitrin, suppressed P. penetrans populations, and increased plant biomass. Pseudomonas fluorescens P10-42 reduced in vitro growth of fungal pathogens, had protease activity, suppressed P. penetrans populations, and increased plant biomass. In potted orchard soil, inoculating apple with P. fluorescens P10-32 suppressed P. penetrans populations in one of the three soils examined. Inoculation with P. fluorescens P10-42 improved plant growth in two of the soils and suppressed P. penetrans abundance in one soil. In one of the soils, P. fluorescens P10-42 was detected on the roots 56 days postinoculation. Overall, we conclude that Pseudomonas spp. play a role in suppressing P. penetrans on apple in orchard soil. [ABSTRACT FROM AUTHOR]

Wolbachia and Cardinium are among the most important and widespread of all endosymbionts, occurring in nematodes and more than half of insect and arachnid species, sometimes as coinfections. These symbionts are of significant interest as potential biocontrol agents due to their abilities to cause major effects on host biology and reproduction through cytoplasmic incompatibility, sex ratio distortion, or obligate mutualism. The ecological and metabolic effects of coinfections are not well understood. This study examined a Wolbachia – Cardinium coinfection in the plant-parasitic nematode (PPN), Pratylenchus penetrans , producing the first detailed study of such a coinfection using fluorescence in situ hybridization (FISH), polymerase chain reaction (PCR), and comparative genomic analysis. Results from FISH and single-nematode PCR showed 123/127 individuals in a focal population carried Cardinium (denoted strain cPpe), and 48% were coinfected with Wolbachia strain wPpe. Both endosymbionts showed dispersed tissue distribution with highest densities in the anterior intestinal walls and gonads. Phylogenomic analyses confirmed an early place of cPpe and long distance from a sister strain in another PPN, Heterodera glycines , supporting a long history of both Cardinium and Wolbachia in PPNs. The genome of cPpe was 1.36 Mbp with 35.8% GC content, 1,131 predicted genes, 41% having no known function, and missing biotin and lipoate synthetic capacity and a plasmid present in other strains, despite having a slightly larger genome compared to other sequenced Cardinium. The larger genome revealed expansions of gene families likely involved in host–cellular interactions. More than 2% of the genes of cPpe and wPpe were identified as candidate horizontally transferred genes, with some of these from eukaryotes, including nematodes. A model of the possible Wolbachia – Cardinium interaction is proposed with possible complementation in function for pathways such as methionine and fatty acid biosynthesis and biotin transport. [ABSTRACT FROM AUTHOR]

Preplant soil incorporation of compost has previously shown potential to suppress Pratylenchus penetrans populations and improve replant establishment of newly planted fruit trees at old orchard sites. Using a greenhouse experiment, we evaluated the effects of preplant incorporation of three compost amendments (agricultural waste compost, yard trimmings compost, and municipal waste compost) on growth of apple seedlings, P. penetrans populations, microbial populations associated with soil suppressiveness, and microbial activity using soil collected from three orchard sites. We also aimed to identify the biological factors that were most strongly associated with enhanced plant growth in compost-amended soil. In four out of nine compost type × soil combinations, compost improved apple seedling growth relative to the control. In four out of nine compost type × soil combinations, compost reduced P. penetrans root infestation relative to the control, and in one compost type × soil combination, compost amendment increased P. penetrans root infestation. Composts stimulated soil microbial activity and abundance, resulting in greater populations of Pseudomonas spp. in soil in seven of the nine compost type × soil combinations. In a step-wise multiple linear regression, P. penetrans root infestation and the total abundance of fungi in soil accounted for the most variance in plant biomass, with increases in both variables correlated with reduced seedling biomass. Overall, composts show variability in the capacity to promote the establishment of fruit trees in orchard soil. Future research should be directed toward understanding variability in compost-induced plant growth promotion among different orchard soils and compost types. [ABSTRACT FROM AUTHOR]

A four-year study was conducted to establish the effects of drip and microsprinkler irrigation systems on populations of Pratylenchus penetrans , early plant growth, and fruit yield of sweet cherry trees planted into soil previously used for apple production. The effects of irrigation type on root colonization by arbuscular mycorrhizal fungi, the abundance of soil microbial antagonists, soil biological suppressiveness, plant nutrition, and soil water content were also evaluated. Trees that received drip irrigation had larger trunk diameters than trees irrigated with microsprinklers throughout the first four years of orchard establishment, with greater total fruit yield also observed with drip irrigation in the fourth growing season. Populations of P. penetrans in soil and roots were smaller under drip irrigation than microsprinkler. Trees irrigated using drip emitters had higher per cent root colonization by arbuscular mycorrhizal fungi than those irrigated by microsprinklers, and had a greater density of fine roots in soil. Drip irrigation also increased soil volumetric water content in the root zone, with soil moisture content often approaching or exceeding field water-holding capacity relative to that of microsprinklers. Overall, drip irrigation shows potential as a component of an integrated pest management strategy for reducing the effect of P. penetrans on newly planted sweet cherry. [ABSTRACT FROM AUTHOR]

Pratylenchus penetrans is one of the most important species of root lesion nematodes (RLNs) because of its detrimental and economic impact in a wide range of crops. Similar to other plantparasitic nematodes (PPNs), P. penetrans harbours a significant number of secreted proteins that play key roles during parasitism. Here, we combined spatially and temporally resolved nextgeneration sequencing datasets of P. penetrans to select a list of candidate genes aimed at the identification of a panel of effector genes for this species. We determined the spatial expression of transcripts of 22 candidate effectors within the oesophageal glands of P. penetrans by in situ hybridization. These comprised homologues of known effectors of other PPNs with diverse putative functions, as well as novel pioneer effectors specific to RLNs. It is noteworthy that five of the pioneer effectors encode extremely proline-rich proteins. We then combined in situ localization of effectors with available genomic data to identify a non-coding motif enriched in promoter regions of a subset of P. penetrans effectors, and thus a putative hallmark of spatial expression. Expression profiling analyses of a subset of candidate effectors confirmed their expression during plant infection. Our current results provide the most comprehensive panel of effectors found for RLNs. Considering the damage caused by P. penetrans, this information provides valuable data to elucidate the mode of parasitism of this nematode and offers useful suggestions regarding the potential use of P. penetrans-specific target effector genes to control this important pathogen. [ABSTRACT FROM AUTHOR]

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 precrops 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. [ABSTRACT FROM AUTHOR]

Cover crops can provide many benefits to agroecosystems, such as lessening soil erosion and increasing water infiltration. However, cover crop use is not common in established red raspberry (Rubus idaeus) fields in the Pacific Northwest. Raspberry growers are concerned about resource competition between the cover crop and raspberry crop, as well as increasing population densities of the plant-parasitic nematode Pratylenchus penetrans, which has a wide host range and has been shown to reduce raspberry plant vigor and yield. A 2-yr study was conducted in an established 'Meeker' raspberry field in northwest Washington to evaluate the effects of nine alleyway cover crops, mowed weed cover, and the industry standard of bare cultivated soil on P. penetrans population dynamics, raspberry yield, and fruit quality. The host status for P. penetrans of cover crops included in the field experiment, as well as Brassica juncea 'Pacific Gold' and Sinapis alba 'Ida Gold', was also evaluated in greenhouse experiments. In the field experiment, P. penetrans population densities did not increase in alleyway cover crop roots over time or in alleyway soil surrounding cover crop roots (means range from 0 to 116 P. penetrans/100 g of soil) compared with the bare cultivated control (means range from 2 to 55 P. penetrans/100 g of soil). Pratylenchus penetrans populations did not increase over time in raspberry grown adjacent to alleyways with cover crops (means range from 1,081 to 6,120 P. penetrans/g of root) compared with those grown adjacent to bare cultivated soil alleyways (means range from 2,391 to 5,536 P. penetrans/g of root). Raspberry grown adjacent to bare cultivated soil did not have significantly higher yield or fruit quality than raspberry grown adjacent to cover crops in either year of the experiment. In the greenhouse assays, 'Norwest 553' wheat and a perennial ryegrass mix were poor hosts for P. penetrans, whereas 'Nora' and 'TAM 606' oat and 'Pacific Gold' and 'Ida Gold' mustard were good hosts. These results support the idea that the potential benefits of alleyway cover crops outweigh the potential risk of increasing P. penetrans population densities and do not compromise raspberry yield or fruit quality. [ABSTRACT FROM AUTHOR]

Replant disease (RD) presents a significant barrier to establishing productive orchards on old orchard soil. Using a field and greenhouse experiment, this study evaluated the influence of agricultural waste compost (AWC) soil amendments, and bark chip mulch (BM) on Pratylenchus penetrans populations and new growth of sweet cherry planted into soil previously used for apple production. Impacts of the treatments on the abundance of rhizosphere microorganisms associated with soil suppressiveness were also evaluated. In the field experiment, fumigation, and preplant incorporation of AWC combined with surface application of BM increased trunk cross-sectional area (TCSA) compared to untreated control plots. Fumigation initially decreased P. penetrans populations in soil, however populations recovered by the end of the first growing season. AWC and BM suppressed P. penetrans populations in roots relative to fumigation or the control. AWC increased the abundance of total bacteria, 2,4-diacetylphloroglucinol-producing (DAPG) bacteria, and pyrrolnitrin-producing (PRN) bacteria in the rhizosphere, relative to fumigation and the control. In the greenhouse experiment, fumigation, AWC, and yard trimmings compost (YTC) increased shoot length of apple and sweet cherry seedlings as well as suppressed P. penetrans populations compared to the control. AWC also increased the abundance of total bacteria, Pseudomonas spp., DAPG + bacteria, and PRN + bacteria relative to fumigation and the control. Overall, composts and BM show potential as alternatives to fumigation for suppression of RD on sweet cherry, with promotion of beneficial rhizosphere microorganisms a possible contributing mechanism in compost-induced soil suppressiveness. [ABSTRACT FROM AUTHOR]

Fatty acid- and retinol-binding proteins ( FARs) are unique to nematodes, and are implicated in a wide range of metabolic and parasitic related functions. Along with the in silico analyses performed in this study, three different FAR members of this family were identified in Pratylenchus penetrans. The cDNA corresponding to the fatty acid- and retinol-binding Pp-far-1 gene was cloned and herein characterized at the molecular level for the first time for this genus. The translated 185 amino acid sequence of P. penetrans FAR-1 sequence, with a predicted molecular weight of 20.82 kDa and a pI of 5.49, shares highest sequence identity to FARs of other migratory nematodes of the Pratylenchidae family (90% to P. vulnus and 80% to Radopholus similis). In situ hybridization localizes Pp-far-1 transcripts in the hypodermis of the nematode. RT- qPCR detected Pp-far-1 transcripts for all nematode developmental stages, with highest expression levels found in juveniles, adult females and adult males, respectively. Pp-far-1 is also highly expressed during infection and establishment of P. penetrans in roots of different host plants, such as corn, lily and soybean. The importance of Pp-far-1 was studied by in planta RNA interference ( RNAi) assays using stable soybean hairy root lines. Targeting Pp-far-1 decreased expression of the nematode Pp-far-1 and significantly reduced the reproduction of nematodes, with a total of between 44% and 70% fewer nematodes in comparison to the average number of nematodes counted for control lines. The results indicate that suppressing the expression levels of Pp-far-1 can act as an effective target gene to control P. penetrans. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The success of all plant-parasitic nematodes is dependent on the completion of several complex behaviors. The lesion nematode Pratylenchus penetrans is an economically important parasite of a diverse range of plant hosts. Unlike the cyst and root-knot nematodes, P. penetrans moves both within and outside of the host roots and can feed from both locations. Adult females of P. penetrans require insemination by actively moving males for reproduction and can lay eggs both within and outside of the host roots. We do not have a complete understanding of the molecular basis for these behaviors. One candidate modulator of these behaviors is the neurotransmitter serotonin. Previous research demonstrated an effect of exogenously applied serotonin on the feeding and male mating behaviors of cyst and root-knot nematodes. However, there are no data on the role of exogenous serotonin on lesion nematodes. Similarly, there are no data on the presence and function of endogenous serotonin in any plant-parasitic nematode. Here, we establish that exogenous serotonin applied to P. penetrans regulates both feeding and sex-specific behaviors. Furthermore, using immunohistochemistry and pharmacological assays, our data suggest that P. penetrans utilizes endogenous serotonin to regulate both feeding and sex-specific behaviors. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Aims: Plant damage inflicted by aboveground herbivores can occur at different stages of plant development and can induce plant responses that affect the growth of belowground herbivores. This study explores impacts of aboveground herbivory at different plant development stages on the population dynamics of root-feeding nematodes. Methods: We simulated aboveground herbivory by clipping the foliage of the grass species Holcus lanatus, and tested how plant defoliation at different times (1, 4 or 7 weeks after nematode inoculation) influenced the population of two root-feeding nematode species: the endoparasitic Pratylenchus penetrans and the ectoparasitic Tylenchorhynchus dubius. Results: Defoliation increased the total abundance of P. penetrans and the number per unit root mass (density) of both P. penetrans and T. dubius. Defoliation enhanced the density of P. penetrans, however, only when plants were defoliated early. Timing did not influence the density of T. dubius, although both abundance and density increased over time. Defoliation increased the nitrogen concentration of plant roots, but reduced root biomass. The strongest reduction of root biomass occurred after early defoliation. Conclusions: Our study indicates that plant responses to aboveground herbivory and their effects on belowground herbivores can be influenced by the time when plants are defoliated, as well as by the belowground herbivore species and their interactions. [ABSTRACT FROM AUTHOR]

Eight fungal isolates (ELRF I to 8) were recovered from necrotic roots of Easter lilies, Lilium longiflorum cv. Nellie White, grown in a field in the U.S. Pacific Northwest. The eight fungal isolates were identified by sequencing and molecular phylogenetic analyses based on their ITS i DNA region. Five isolates were identified as Fusarium oxysporum, two as F. tricinctum, and one as Rhizoctonia sp. AG-I. This constitutes the first report of Rhizoctonia sp. AG-I infecting lilies worldwide and the first report of F. tricinctum infecting lilies in the United States. To study and validate their pathogenicity, pure cultures of each isolate were used to infect the roots of Easter lily plants growing in vitro. In addition, Easter lily plants growing in vitro were infected either with or without Pratylenchus penetrans, the root lesion nematode, prior to placing a culture plug of fungus 1 cm from a lily root. Pratylenchus penetrans is a nematode species commonly found in the sampled fields. The presence of both nematode and Rhizoctonia sp. AG-1 isolate ELRF 3 in infected lilies was evaluated by molecular analyses, confirming the infection of roots 3 days after inoculation, prior to development of disease symptoms. Necrosis and root rot developed more rapidly with all eight fungal isolates when there had been prior infection with P. penetrans, the major nematode parasitizing Easter lily roots in the field in Oregon. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cover cropping is a common practice in U.S. Midwest carrot production for soil conservation, and may affect soil ecology and plant-parasitic nematodes--to which carrots are very susceptible. This study assessed the impact of cover crops--oats (Avena sativa), radish (Raphanus sativus) cv. Defender, rape (Brassica napus) cv. Dwarf Essex, and a mixture of oats and radish--on plantparasitic nematodes and soil ecology based on the nematode community in Michigan carrot production systems. Research was conducted at two field sites where cover crops were grown in Fall 2014 preceding Summer 2015 carrot production. At Site 1, rootlesion (Pratylenchus penetrans) and stunt (Tylenchorhynchus sp.) nematodes were present at low population densities (less than 25 nematodes/100 cm³ soil), but were not significantly affected (P > 0.05) by cover crops. At Site 2, P. penetrans population densities were increased (P ≤ 0.05) by 'Defender' radish compared to other cover crops or fallow control during cover crop growth and midseason carrot production. At both sites, there were few short-term impacts of cover cropping on soil ecology based on the nematode community. At Site 1, only at carrot harvest, radish-oats mixture and 'Dwarf Essex' rape alone enriched the soil food web based on the enrichment index (P ≤ 0.05) while rape and radish increased structure index values. At Site 2, bacterivore abundance was increased by oats or radish cover crops compared to control, but only during carrot production. In general, cover crops did not affect the nematode community until nearly a year after cover crop growth suggesting that changes in the soil community following cover cropping may be gradual. [ABSTRACT FROM AUTHOR]

Lilium longiflorum cv. Nellie White, commonly known as Easter lily, is an important floral crop with an annual wholesale value of over $26 million in the United States. The root-lesion nematode, Pratylenchus penetrans, is a major pest of lily due to the significant root damage it causes. In this study, we investigated the cytological aspects of this plant-nematode interaction using bright-field and transmission electron microscopy. We took advantage of an in vitro culture method to multiply lilies and follow the nematode infection over time. Phenotypic reactions of roots inoculated with P. penetrans were evaluated from 0 to 60 d after nematode infection. Symptom development progressed from initial randomly distributed discrete necrotic areas to advanced necrosis along entire roots of each inoculated plant. A major feature characterizing this susceptible host response to nematode infection was the formation of necrosis, browning, and tissue death involving both root epidermis and cortical cells. Degradation of consecutive cell walls resulted in loss of cell pressure, lack of cytoplasmic integrity, followed by cell death along the intracellular path of the nematode's migration. Pratylenchus penetrans was never seen in the vascular cylinder as the layer of collapsed endodermal cells presumably blocked the progression of nematodes into this area of the roots. This study presents the first detailed cytological characterization of P. penetrans infection of Easter lily plants. [ABSTRACT FROM AUTHOR]