Plant-parasitic Nematodes Associated with Grasses Grown for Seed in the Willamette Valley of Oregon.

Plant-parasitic nematodes (PPN) are an understudied pathogen group in the Oregon cool-season grass seed cropping system. In this survey, the PPN associated with annual ryegrass, bentgrass, fine fescue, orchardgrass, perennial ryegrass, and tall fescue were determined. Thirty-seven fields were sampled in the 2022 or 2023 growing season by collecting 10 soil cores in each of six 100-m transects for nematode extraction and visual identification. PerMANOVA testing indicated significant differences in PPN community composition across grass host and sampling time. Pratylenchus and Meloidogyne were the most commonly encountered nematodes, with maximum population densities of 1,984 and 2,496 nematodes/100 g soil, respectively. Sequencing of the COX1 gene region indicated the presence of P. crenatus, P. fallax, P. neglectus, P. penetrans, and P. thornei, with some of these species being detected for the first time on these grass hosts. The only Meloidogyne sp. found in these grasses was M. nassi, based upon sequencing of the ITS gene region. This first-of-its-kind survey indicates the need for further assessment of the impact of these PPNs on yield and stand longevity in cool-season grass seed fields in Oregon.

Characterization of Globodera ellingtonae Populations from Chile Utilizing Whole Genome Sequencing.

Globodera ellingtonae was originally described from populations collected in the United States. In the original description, ribosomal DNA loci from Globodera sp. collected in Chile and Argentina were similar to G. ellingtonae, suggesting this nematode originated in this region of South America. In an effort to find additional populations of G. elllingtonae, collection trips were conducted in 2017 and 2020 in the Antofagasta and Arica y Parinacota Regions in Northern Chile, respectively. Globodera sp. were more prevalent in Antofagasta (17 samples collected, 53% positive for Globodera sp.) than in Arica y Parincota (16 samples collected, 13% positive for Globodera sp.). The genomes of single cysts (N ≥ 3) from four fields were sequenced. Additionally, the genomes of the G. ellingtonae population from Oregon and a Globodera sp. population originally collected in Antofagasta Region but maintained in culture in France were also sequenced. Based upon a HSP90 sequenced data mined from WSG data, all of the populations from the Antofagasta Region were G. ellingtonae and grouped in a monophyletic clade. A population collected from the Arica y Parincota Region was identified as G. rostochiensis based upon HSP90 data. Genome-wide SNP patterns of the G. ellingtonae populations showed strong clustering based on geographic location indicating that G. ellingtonae has high genetic diversity within Chile. A phylogenetic tree derived from 168,354 binary SNPs in the nuclear genome showed separate but distinct clustering of the Oregon population and the population from Antofagasta maintained in France. The Oregon G. ellingtonae population subtended the Chilean clades and placed on a long branch representing approximately twice the genetic variation observed among all Chilean G. ellingtonae populations. The possibility remains that G. ellingtonae from Chile may be sufficiently diverged to constitute a new species from G. ellingtonae originally described from a population collected in Oregon.

Effect of the trap crop, Solanum sisymbriifolium, on Globodera pallida, Globodera tabacum, and Globodera ellingtonae.

The effect of the nematode trap crop Solanum sisymbriifolium was assessed against three Globodera spp., the potato cyst nematode Globodera pallida (in Idaho), the recently described Globodera ellingtonae (in Oregon), and the tobacco cyst nematode Globodera tabacum (in Connecticut) in field trials. At all locations the ability of S. sisymbriifolium to reduce Globodera encysted second-stage juveniles (J2) in egg densities compared to fallow was considered. For G. ellingtonae, the impact of planting and termination dates of S. sisymbriifolium on final egg densities was also evaluated; and for G. pallida, the ability of the nematode to reproduce on potato (Solanum tuberosum) after exposure to S. sisymbriifolium was determined. Encysted J2 in egg densities of all three Globodera spp. declined from 25 to 68% after trap cropping with S. sisymbriifolium. For G. pallida, S. sisymbriifolium reduced final encysted J2 in egg density by 23 to 50% compared to the fallow treatment, and significantly decreased G. pallida reproduction on potato after exposure to S. sisymbriifolium by 99 to 100% compared to the fallow treatment (P < 0.0001). For G. ellingtonae, the planting date of S. sisymbriifolium in May or June did not impact final egg densities (P = 0.32). Rather, percentage reduction in G. ellingtonae encysted J2 in egg density was most influenced by the length of time to which nematodes were exposed to S. sisymbriifolium, with 30 and 81% reduction after 6 vs 12 wk of exposure, respectively (P < 0.0001). Similar levels of nematode reduction after S. sisymbriifolium were observed for G. tabacum after 12 to 14 wk of exposure to the trap crop; G. tabacum density changes consisted of a 114% increase after susceptible tobacco, a 65% decrease after resistant tobacco, and an 88% decrease after S. sisymbriifolium compared to bare soil. In conclusion, this research demonstrates the widespread applicability of S. sisymbriifolium in reducing a diversity of Globodera spp. present in the USA.The effect of the nematode trap crop Solanum sisymbriifolium was assessed against three Globodera spp., the potato cyst nematode Globodera pallida (in Idaho), the recently described Globodera ellingtonae (in Oregon), and the tobacco cyst nematode Globodera tabacum (in Connecticut) in field trials. At all locations the ability of S. sisymbriifolium to reduce Globodera encysted second-stage juveniles (J2) in egg densities compared to fallow was considered. For G. ellingtonae, the impact of planting and termination dates of S. sisymbriifolium on final egg densities was also evaluated; and for G. pallida, the ability of the nematode to reproduce on potato (Solanum tuberosum) after exposure to S. sisymbriifolium was determined. Encysted J2 in egg densities of all three Globodera spp. declined from 25 to 68% after trap cropping with S. sisymbriifolium. For G. pallida, S. sisymbriifolium reduced final encysted J2 in egg density by 23 to 50% compared to the fallow treatment, and significantly decreased G. pallida reproduction on potato after exposure to S. sisymbriifolium by 99 to 100% compared to the fallow treatment (P < 0.0001). For G. ellingtonae, the planting date of S. sisymbriifolium in May or June did not impact final egg densities (P = 0.32). Rather, percentage reduction in G. ellingtonae encysted J2 in egg density was most influenced by the length of time to which nematodes were exposed to S. sisymbriifolium, with 30 and 81% reduction after 6 vs 12 wk of exposure, respectively (P < 0.0001). Similar levels of nematode reduction after S. sisymbriifolium were observed for G. tabacum after 12 to 14 wk of exposure to the trap crop; G. tabacum density changes consisted of a 114% increase after susceptible tobacco, a 65% decrease after resistant tobacco, and an 88% decrease after S. sisymbriifolium compared to bare soil. In conclusion, this research demonstrates the widespread applicability of S. sisymbriifolium in reducing a diversity of Globodera spp. present in the USA.

Morphological and molecular characterization of Globodera populations from Oregon and Idaho.

An unusual population of cyst nematode was found in soils collected from a Powell Butte, OR field with a cropping history including potato, wheat, other crops, and significant weed presence. These nematodes could not be placed with certainty into any known species and exhibited some unique morphological features in some specimens. Compared with Globodera pallida, the cyst body length was slightly longer and the second-stage juvenile stylet length was slightly shorter. In some individuals, the J2 stylet knob height was greater and the tail annules were more prominent than in G. pallida, and the tail abruptly narrowed, with a slight constriction near the posterior third of the hyaline terminus. Compared with G. rostochiensis, the hyaline tail terminus had a larger number of refractive bodies, and cysts of this population had a smaller Granek's ratio and fewer cuticular ridges between the anus and vulva. In some individuals, the tail termini of second-stage juveniles were more bluntly pointed, and the stylet knobs were more anteriorly directed with greater height. Unlike G. tabacum, the cyst wall often lacked a network-like pattern and, in some individuals, the juvenile tail terminus distinctly narrowed after a constriction. Molecularly, the population was distinct from G. pallida, G. rostochiensis, and G. tabacum. Multiplex polymerase chain reaction of the internal transcribed spacer (ITS) rDNA region gave results similar to G. tabacum; however, ITS restriction fragment length polymorphism patterns were observed to have individual bands in common with G. rostochiensis and G. pallida. Phylogenetic analysis based on ITS1 and -2 rDNA sequences showed greatest similarity to populations from Argentina and Chile; together, they form a moderately supported clade, distinct from G. rostochiensis, G. tabacum, G. "mexicana," European type G. pallida, and several G. pallida populations from South America.

Brassicaceous seed meals as soil amendments to suppress the plant-parasitic nematodes Pratylenchus penetrans and Meloidogyne incognita.

Brassicaceous seed meals are the residual materials remaining after the extraction of oil from seeds; these seed meals contain glucosinolates that potentially degrade to nematotoxic compounds upon incorporation into soil. This study compared the nematode-suppressive ability of four seed meals obtained from Brassica juncea 'Pacific Gold', B. napus 'Dwarf Essex' and 'Sunrise', and Sinapis alba 'IdaGold', against mixed stages of Pratylenchus penetrans and Meloidogyne incognita second-stage juveniles (J2). The brassicaceous seed meals were applied to soil in laboratory assays at rates ranging from 0.5 to 10.0% dry w/w with a nonamended control included. Nematode mortality was assessed after 3 days of exposure and calculated as percentage reduction compared to a nonamended control. Across seed meals, M. incognita J2 were more sensitive to the brassicaceous seed meals compared to mixed stages of P. penetrans. Brassica juncea was the most nematode-suppressive seed meal with rates as low as 0.06% resulting in > 90% suppression of both plant-parasitic nematodes. In general B. napus 'Sunrise' was the least nematode-suppressive seed meal. Intermediate were the seed meals of S. alba and B. napus 'Dwarf Essex'; 90% suppression was achieved at 1.0% and 5.0% S. alba and 0.25% and 2.5% B. napus 'Dwarf Essex', for M. incognita and P. penetrans, respectively. For B. juncea, seed meal glucosinolate-degradation products appeared to be responsible for nematode suppression; deactivated seed meal (wetted and heated at 70 °C for 48 hr) did not result in similar P. penetrans suppression compared to active seed meal. Sinapis alba seed meal particle size also played a role in nematode suppression with ground meal resulting in 93% suppression of P. penetrans compared with 37 to 46% suppression by pelletized S. alba seed meal. This study demonstrates that all seed meals are not equally suppressive to nematodes and that care should be taken when selecting a source of brassicaceous seed meal for plant-parasitic nematode management.

Activity of Hydroxamic Acids from Secale cereale Against the Plant-Parasitic Nematodes Meloidogyne incognita and Xiphinema americanum.

ABSTRACT Cyclic hydroxamic acids are secondary metabolites found in the family Poaceae and have been implicated in the allelopathy of rye (Secale cereale). The toxicity of these compounds against plant-parasitic nematodes is unknown. DIBOA (2,4-dihydroxy-(2H)-1,4-benzoxazin-3(4H)-one), DIMBOA (2,4-hydroxy-7-methoxy-(2H)-1,4-benzoxazin-3(4H)-one), and their degradation products BOA (benzoxazolin-2(3H)-one) and MBOA (6-methoxy-benzoxazolin-2(3H)-one) were screened in vitro against Meloidogyne incognita second-stage juveniles (J2) and eggs and mixed-stages of Xiphinema americanum. Xiphinema americanum was more sensitive to DIBOA and DIMBOA than M. incognita J2, with a maximum apparent mortality of 96 and 92% compared to 73 and 72% at 90 mug/ml. Eggs of M. incognita were less sensitive to the hydroxamic acids than J2; only DIBOA resulted in a 50% reduction in egg hatch, with a lethal concentration (LC(50)) of 74 mug/ml compared to 21 mug/ml for J2. When M. incognita J2 were exposed to DIBOA for 48 h and the compound was removed and replaced with water, the LC(50) value increased from 21.0 to 40.7 mug/ml. MBOA was not toxic to X. americanum or M. incognita eggs, but was toxic to M. incognita J2, with LC(50) values of 44 and 20 mug/ml before and after the compound was removed and replaced with water. BOA was the least toxic hydroxamic acid tested; it did not reduce M. incognita egg hatch after 1 week of exposure or increase X. americanum mortality after 24 h of exposure. While in vitro studies provide a valuable starting point in determining the toxicity of the chemical component of rye, the relevance of the data to soil remains to be determined.

Factors Affecting the Suppression of Heterodera glycines by N-Viro Soil.

Previous laboratory research demonstrated that N-Viro Soil (NVS), an alkaline-stabilized municipal biosolid, suppressed plant-parasitic nematodes. This study continued to explore the use of NVS as a nematode management tool specifically addressing factors that could influence its use. N-Viro Soil from different locations, the components of NVS (de-watered biosolids and fly ash admixtures), and sterilized NVS were applied to sand microcosms to determine effects on nematode survival sand solution pH and ammonia concentrations. This study confirmed the previous finding that an important mechanism of Heterodera glycines suppression by NVS was the generation of alkaline soil conditions. Only the fly ash admixture that resulted in an increase in pH to 10.0 suppressed H. glycines to the same level as NVS. Alkaline-stabilization of biosolids was necessary to achieve nematode suppression. Biosolids applied at rates <3% dry w/w did not suppress H. glycines to the same level as equivalent amounts of NVS. Sand solution pH levels after biosolid application, regardless of rate, were approximately 8.5 whereas 1% and 4% w/w NVS amendment resulted in pH levels of 10.3 and 11.6, respectively. NVS from different processing facilities were all effective in suppressing H. glycines. The NVS source that produced the highest concentration of ammonia did not reduce H. glycines survival to the same level as those sources generating pH levels above 10.1. Microbes associated with NVS appeared not to be responsible for the nematode suppressiveness of the amendment; there was no difference in nematode suppression between autoclaved and nonautoclaved NVS. The role that ammonia plays in the suppression of H. glycines by NVS is still unclear.

Chemical-Mediated Toxicity of N-Viro Soil to Heterodera glycines and Meloidogyne incognita.

N-Viro Soil (NVS) is an alkaline-stabilized municipal biosolid that has been shown to lower population densities and reduce egg hatch of Heterodera glycines and other plant-parasitic nematodes; but the mechanism(s) of nematode suppression of this soil amendment are unknown. This study sought to identify NVS-mediated changes in soil chemical properties and their impact upon H. glycines and Meloidogyne incognita mortality. N-Viro Soil was applied to sand in laboratory assays at 0.5%, 1.0%, 2.0%, and 3.0% dry w/w with a nonamended treatment as a control. Nematode mortality and changes in sand-assay chemical properties were determined 24 hours after incubation. Calculated lethal concentration (LC(90)) values were 1.4% w/w NVS for second-stage juveniles of both nematode species and 2.6 and >3.0% w/w NVS for eggs of M. incognita and H. glycines, respectively. Increasing rates of NVS were strongly correlated (r(2) = 0.84) with higher sand solution pH levels. Sand solution pH levels and, to a lesser extent, the production of ammonia appeared to be the inorganic chemical-mediated factors responsible for killing plant-parasitic nematodes following amendment with NVS.

Sensitivity of Meloidogyne javanica and Tylenchulus semipenetrans to Isothiocyanates in Laboratory Assays.

ABSTRACT Isothiocyanates are released through enzymatic degradation of glucosinolates produced by plants in the family Brassicaceae. Glucosinolate profiles differ among plant species and the isothiocyanate derivatives differ in their toxicity to nematodes. Control of plant-parasitic nematodes in soil by isothiocyanates released from incorporated brassicaceous plant material has been inconsistent. Success might be improved with knowledge of the relative toxicities of various isothiocyanates against nematodes. Laboratory assays were conducted to determine lethal concentration (LC) values in sand of seven commercially available isothiocyanates against Tylenchulus semipenetrans and Meloidogyne javanica. The LC(90) values were 0.01 and 0.03 mumol/ml for 2-phenylethyl isothiocyanate and 0.48 and 0.35 mumol/ml for phenyl isothiocyanate for T. semipenetrans and M. javanica, respectively. Brassicaceous sources of benzyl or 2-phenylethyl isothiocyanate and, to a lesser extent allyl isothiocyanate, are the most promising candidates for plant-parasitic nematode management. The broader context of this research is the development of approaches for consistent and reliable use of plant-derived chemicals for nematode management. The strategy is to select plants in the family Brassicaceae based on their glucosinolate profiles and the sensitivity of the target nematode species to the associated isothiocyanates.

Plant sources of chinese herbal remedies: laboratory efficacy, suppression of Meloidogyne javanica in soil, and phytotoxicity assays.

Extracts of Chinese herbal medicines from plants representing 13 families were tested for their ability to suppress plant-parasitic nematodes. Effective concentration (EC50 and EC90) levels for 18 of the extracts were determined in laboratory assays with Meloidogyne javanica juveniles and all stages of Pratylenchus vulnus. Efficacy of 17 extracts was tested against M. javanica in soil. Generally, EC50 and EC90 values determined in the laboratory were useful indicators for application rates in the soil. Extracts tested from plants in the Liliaceae reduced galling of tomato by M. javanica and were not phytotoxic. Similarly, isothiocyanate-yielding plants in the Brassicaceae suppressed root galling without phytotoxicity. Other plant extracts, including those from Azadirachta indica, Nerium oleander, and Hedera helix, suppressed root galling but were phytotoxic at the higher concentrations tested. Many of these plant sources have been tested elsewhere. Inconsistency in results across studies points to the need for identification of active components and for determination of concentration levels of these components when plant residues or extracts are applied to soil.