In the Pacific Northwest, alfalfa (Medicago sativa) is host to two species of root-knot nematodes, including race 2 of the Columbia root-knot nematode (Meloidogyne chitwoodi) and the northern root-knot nematode (Meloidogyne hapla). In addition to the damage caused to alfalfa itself by M. hapla, alfalfa’s host status to both species leaves large numbers of nematodes available to damage rotation crops, of which potato is the most important. A nematode-resistant alfalfa germplasm release, W12SR2W1, was challenged with both nematode species, to determine the correlation, if any, of resistance to nematode reproduction. Thirty genotypes were screened in replicated tests with M. chitwoodi race 2 or M. hapla, and the reproductive factor (RF) was calculated. The distribution of natural log-transformed RF values was skewed for both nematode species, but more particularly for M. chitwoodi race 2, where more than half the genotypes screened were non-hosts. Approximately 30 percent of genotypes were non-hosts or very poor hosts of M. hapla, but RF values for M. hapla on susceptible genotypes were generally much higher than RF values for genotypes susceptible to M. chitwoodi race 2. The Spearman rank correlation was positive (0.52) and significant (p-value = 0.003), indicating there is some relationship between resistance to these two species of root-knot nematode in alfalfa. However the relationship is not strong enough to suggest genetic loci for resistance are identical, or closely linked. Breeding for resistance or immunity will require screening with each species separately, or with different DNA markers if marker-assisted breeding is pursued. A number of genotypes were identified which are non-hosts to both species. These plants will be intercrossed to develop a non-host germplasm.
alfalfa; host status; Meloidogyne chitwoodi; Meloidogyne hapla; pest resistance
Desiccation tolerance plays an important role in the overwinter survival of the foliar nematode Aphelenchoides fragariae. Survival rates of A. fragariae were compared with those of the anhydrobiotic soil-dwelling nematode Aphelenchus avenae after desiccation (90% RH), cold (4°C) and osmotic (500 mM sucrose) stress treatments. A. fragariae formed aggregates during desiccation and showed higher survival rates than A. avenae under desiccation and osmotic stress. Analysis of transcripts with Illumina RNA-seq indicated that glutaredoxin and other antioxidant-related genes were up-regulated under desiccation stress. Quantitative RT-PCR demonstrated 2.8 fold and 1.3 fold up-regulation of a glutaredoxin gene under desiccated and osmotic stress, respectively, suggesting the participation of antioxidant mechanisms in desiccation tolerance of A. fragariae.
Aphelenchoides fragariae; desiccation; cold stress; osmotic stress; glutaredoxin; survival rate
One hundred and eleven golf courses from 39 counties in the Carolinas were surveyed for plant-parasitic nematodes. Species diversity within habitats was analyzed with five diversity indices including Diversity index (H’), Evenness (J’), Richness (SR), Dominance (λ) and Diversity (H2). The results revealed a remarkably high diversity of 24 nematode species belonging to 19 genera and 11 families. Of those, 23 species were found in SC, 19 species in NC, and 18 species were detected in both states. Helicotylenchus dihystera, Mesocriconema xenoplax, Hoplolaimus galeatus, Tylenchorhynchus claytoni, Belonolaimus longicaudatus, Meloidogyne graminis and Paratrichodorus minor were the most prevalent and abundant species in golf course turfgrasses in both states. Twelve species were new records of plant parasitic nematodes in turfgrasses in both NC and SC. The results also revealed effects of different habitats on diversity of nematode species in turfgrass ecosystem. H’ and SR values were higher in SC than in NC. H’, J’ and H2 values were significantly higher in sandy than in clay soil in NC, but no significant differences between sand and clay soil were detected in SC or in pooled data from both states. There were no significant differences for all indices among the management zones (putting green, fairway and tee) in NC. However, in SC and pooled data, H’, SR and H2 were significantly higher in putting greens than in fairways and tees. Significant differences from different grass species (bermudagrass, creeping bentgrass and zoysiagrass) were detected only in H’, which was significantly higher in zoysiagrass than in bentgrass or bermudagrass in NC. In pooled data, H’ was significantly higher in zoysiagrass samples than in creeping bentgrass samples but was not significantly different from bermudagrass samples.
North Carolina; South Carolina; detection; distribution; diversity; ecology; golf course; identification; plant-parasitic nematode; turfgrass
Previously we showed in laboratory studies that the fungivorus nematode, Aphelenchoides hylurgi, was attracted to and fed upon the chestnut blight fungus, Cryphonectria parasitica, from American chestnut bark cankers and was a carrier of biocontrol, white hypovirulent C. parasitica strains. In the present field study, we recovered Aphelenchoides spp. in almost all (97.0 %) of 133 blight canker tissue assays (three 5-g samples each) from four eastern states. High mean population densities (227 to 474 nematodes per 5 g tissue) of Aphelenchoides spp. were recovered from cankers in Virginia, West Virginia, and Tennessee but not from New Hampshire (mean = 75 nematodes per 5 g tissue). Overall, most canker assays yielded population densities less than 200 nematodes per 5 g tissue. All of 12 very small or young cankers yielded a few to many Aphelenchoides spp. Regression analysis indicated greatest recovery of Aphelenchoides spp. occurred in the month of May (r = 0.94). The results indicate that Aphelenchoides spp. appear to be widespread in blight cankers on American chestnut trees and could play a role in biocontrol of chestnut blight.
biocontrol; chestnut blight; fungivorus nematodes; hypovirulence
In an outside pot experiment, dry pig manure processed on pine sawdust litter and fermented for seven days by house fly larvae (fermented manure), and pine sawdust applied alone, and in combination with a spring application of inorganic nitrogen fertilizer were used to determine their effects on plant parasitic and free-living soil nematodes on sugar beets (cv. Antek). Non amended soil was used as a control. All treatments with fermented pig manure and sawdust with nitrogen fertilizer decreased number of plant parasitic nematodes and also root-fungal feeding nematodes compared to the untreated control. Sawdust applied alone had no effect on plant parasitic and root-fungal feeding nematode suppression. Free-living nematodes which were mainly bacteriovores and fungivores were significantly more abundant in soil amended with fermented pig manure, while the sawdust had no effect on these nematodes. The effect of all tested treatments on omnivores-predators was rather random, and in general, the number of these nematodes decreased after soil amendment applications compared to the untreated control.
fermented animal manure; nematode trophic groups; nitrogen amendments; phytoparasitic nematodes; sawdust
A survey was conducted to determine the assemblage and abundance of plant-parasitic nematodes and their associations with soil factors in organically farmed fields in Minnesota. A total of 31 soil samples were collected from southeast (SE), 26 samples from southwest (SW), 28 from west-central (WC), and 23 from northwest (NW) Minnesota. The assemblage and abundance of plant-parasitic nematodes varied among the four regions. The soybean cyst nematode, Heterodera glycines, the most destructive pathogen of soybean, was detected in 45.2, 88.5, 10.7, and 0% of organically farmed fields with relative prominence (RP) values of 10.3, 26.5, 0.6, and 0 in the SE, SW, WC, and NW regions, respectively. Across the four regions, other common genera of plant-parasitic nematodes were Helicotylenchus (42.6, RP value, same below), Pratylenchus (26.9), Tylenchorhynchus and related genera (9.4), Xiphinema (5.6), and Paratylenchus (5.3). Aphelenchoides, Meloidogyne, Hoplolaimus, Mesocriconema, and Trichodorus were also detected at low frequencies and/or low population densities. The similarity index of plant-parasitic nematodes between two regions ranged from 0.44 to 0.71 and the similarity increased with decreasing distance between regions. The densities of most plant-parasitic nematodes did not correlate with measured soil factors (organic matter, pH, texture). However, the densities of Pratylenchus correlated negatively with % sand, and Xiphinema was correlated negatively with soil pH.
Aphelenchoides; Helicotylenchus; Heterodera glycines; Hoplolaimus; Meloidogyne; Mesocriconema; nematode community; organic-farming; Paratylenchus; Pratylenchus; soybean cyst nematode; Trichodorus; Tylenchorhynchus; Xiphinema
Populations of Bakernema inaequale, C. petasum, C. sphagni, C. mutabile, Ogma octangulare, Xenocriconemella macrodora and Hemicriconemoides chitwoodi were identified and re-described from different geographical areas in the continental United States and molecularly characterized. Two new species of spine nematodes Criconema arkaense n. sp. from Washington County and Lee County, Arkansas and Criconema warrenense n. sp from Warren, Bradley County, Arkansas are also described and named. Criconema arkaense is characterize by having a conspicuous lip region offset from the body with two annuli, short rounded tail with a thin cuticular sheath and subterminal anus. Criconema warrenense n. sp. has two lip region annuli about the same width, first annulus directed posteriorly, separated by a narrow neck annulus and a short conoid tail, unilobed non-folded annulus. The molecular characterization of Criconema arkaense and Criconema warrenense using ITS1 rDNA gene sequence and the molecular phylogenetic relationships of these new species along with the known spines nematodes are provided.
Bakernema inaequale; Criconematidae; Criconema,Criconema arkaense n.sp.; Criconema mutabile; Criconema petasum; Criconema sphagni; Criconema warrenense n.sp.; Hemicriconemoides; Hemicriconemoides chitwoodi; internal transcribed spacer 1; morphology; molecular biology; Ogma; Ogma octangulare; phylogenesis; taxon; Xenocriconemella; Xenocriconemella macrodora
Microorganisms produce volatile organic compounds (VOCs) which mediate interactions with other organisms and may be the basis for the development of new methods to control plant-parasitic nematodes that damage coffee plants. In the present work, 35 fungal isolates were isolated from coffee plant rhizosphere, Meloidogyne exigua eggs and egg masses. Most of the fungal isolates belonged to the genus Fusarium and presented in vitro antagonism classified as mutual exclusion and parasitism against the nematode-predator fungus Arthrobotrys conoides (isolated from coffee roots). These results and the stronger activity of VOCs against this fungus by 12 endophytic bacteria may account for the failure of A. conoides to reduce plant-parasitic nematodes in coffee fields. VOCs from 13 fungal isolates caused more than 40% immobility to Meloidogyne incognita second stage juveniles (J2), and those of three isolates (two Fusarium oxysporum isolates and an F. solani isolate) also led to 88-96% J2 mortality. M. incognita J2 infectivity decreased as a function of increased exposure time to F. oxysporum isolate 21 VOCs. Gas chromatography-mass spectrometry (GC-MS) analysis lead to the detection of 38 VOCs produced by F. oxysporum is. 21 culture. Only five were present in amounts above 1% of the total: dioctyl disulfide (it may also be 2-propyldecan-1-ol or 1-(2-hydroxyethoxy) tridecane); caryophyllene; 4-methyl-2,6-di-tert-butylphenol; and acoradiene. One of them was not identified. Volatiles toxic to nematodes make a difference among interacting microorganisms in coffee rhizosphere defining an additional attribute of a biocontrol agent against plant-parasitic nematodes.
biological control; Meloidogyne exigua; Fusarium; antagonism
A novel entomopathogenic nematode species, Heterorhabditidoides rugaoensis n. sp. RG081015, collected from Rugao, China, is described. The new species is morphologically very similar to H. chongmingensis but can be distinguished from it on the basis of some morphological characteristics, combined with molecular data and a cross-hybridization test. Males of the new species can be recognized on the basis of body length averaging 1396.2 μm; lateral field with one ridge; metastome isoglottoid with one hemispherical swellings comprised of two to three well-developed warts; asymmetric spicules; peloderan bursa. In IJs, EP = 134.5 μm; ES = 149.3 μm; tail length = 82.5 μm; and a = 20.5. Hermaphroditic females have four to five lateral ridges. The 18S rDNA and ITS sequences of the two nematodes share 99% and 98% identity, respectively. Phylogenetic trees of 18S rDNA and ITS indicate that the new species is most closely related to H. chongmingensis; thus, the two nematodes belong to the same genus. Failure of cross-hybridization between them indicates that nematode strain RG081015 is a novel species and is described herein as H. rugaoensis n. sp. The LC50 of the novel species against Galleria mellonella were 24.35 IJs / ml within 48 hours of infection. Morphological characteristics, genetic similarity analyses, and phylogenetic relationships provide strong evidence that some species of Oscheius/Insectivora-group should be reassigned to the genus Heterorhabditidoides.
Entomopathogenic nematode; Heterorhabditidoides rugaoensis n. sp.; Insectivora-group; new nematode species; pathogenicity; phylogeny; Oscheius; taxonomy
Populations of Mesocriconema curvatum, M. kirjanovae, M. onoense, M. ornatum, M. sphaerocephala, M. surinamense, M. vadense, M. xenoplax, and Criconemoides informis from different geographical areas in the continental United States were characterized morphologically and molecularly. A new ring nematode from Washington County, Arkansas, is also described and named Mesocriconema ozarkiense n. sp., This new species is characterized by females with small flattened submedian lobes, lower than or at the same level as the labial disc, vagina straight, very well developed spermatheca without sperm, no more than one anastomoses, L=379-512 μm, V=89-93, stylet length = 49-61 μm, R=107-119, annuli with slightly crenate margins on tail portion and a simple anterior vulval lip. The molecular characterization of M. ozarkiense n. sp. using the ITS rRNA gene sequence and the phylogenesis relationship of this new species with the ring nematodes included in this study are provided.
Criconematidae; Criconemoides; Criconemoides informis; internal transcribed spacer 1; Mesocriconema; Mesocriconema ozarkiense n. sp. M. crenatum; M. curvatum; M. kirjanovae; M. onoense; M. ornatum; M. sphaerocephala; M. surinamense; M. vadense; M. xenoplax; molecular biology; morphology; phylogenesis; taxon
The foliar nematode Aphelenchoides besseyi causes white tip disease in rice (Oryza sativa L.) and floral malady in tuberose (Polianthes tuberosa L.). This nematode is widely distributed in the rice fields of many states of India, including West Bengal (WB), Andhra Pradesh (AP), Madhya Pradesh (MP) and Gujarat (GT). In order to generate information on intraspecific variations of A. besseyi as well as to confirm the identity of the nematode species infecting these important crops, morphological observation was undertaken of A. besseyi isolated from tuberose and rice from WB and rice from AP, MP and GT. The molecular study was only done for rice and tuberose populations from AP and WB. The variations were observed among the populations in the tail, esophageal and anterior regions, including the occurrence of four as well as six lateral lines in the lateral fields. The morphometrics of observed populations showed variations and those could be regarded as a consequence of host-induced or geographical variations. PCR amplification of the rDNA ITS 1 and 2 region of rice (AP) and tuberose (WB) populations of A. besseyi generated one fragment of approximately 830 bp, and the size of the ITS region was 788 bp and 791 bp for tuberose and rice population, respectively. Alignment of the two sequences showed almost 100% similarity. Blast analysis revealed a very high level of similarity of both the Indian strains to a Russian population. The Indian and Russian strains could be differentiated using restriction enzyme Bccl. Host tests revealed that rice (cv. IET 4094), oat (cv. OS-6) and teosinte (cv. TL-1) showed a typical distortion due to the infection of A. besseyi. Five germplasm lines of oat showed no infection of the nematode under field conditions. Local cultivars of onion, maize, chrysanthemum, gladiolus, and Sorghum halepense were also not infected by A. besseyi.
Aphelenchoides besseyi; diagnosis; distribution; host; India; morphology; rice; tuberose
Candidate grape rootstocks were selected through a rigorous screening program initiated with important sources of resistance to Meloidogyne pathotypes and to Xiphinema index in Muscadinia rotundifolia and Vitis species native to North America. Based on their rooting capability and horticultural characteristics, 200 candidates were selected from 5,000 progeny of multiple crosses between commercial grape rootstocks and wild grape species that exhibited resistance to nematodes. After a 15-year screening process, 13 selections emerged with either almost complete or complete combined resistance to M. incognita Race 3, M. incognita pathotype Harmony C, M. arenaria pathotype Harmony A, and X. index, important nematode pests of grapevines. Durability of this broad resistance was tested by challenging the selections with the target nematodes in combination and with the target nematodes in combinations with species not included in the screening process. Durability of resistance of the candidate rootstocks was also tested by exposure to the nematode communities of infested field soils from different locations. Breadth of resistance was determined on the basis of their host status to non-target nematodes, including Mesocriconema xenoplax, Pratylenchus vulnus, Tylenchulus semipenetrans and Paratylenchus hamatus. After a total of 204 separate trials, the rootstocks were released to the grape industry as UCD GRN1, UCD GRN2, UCD GRN3, UCD GRN4, and UCD GRN5. We provide a compilation of current knowledge of the host status of these five newly released rootstocks and of 27 other rootstock cultivars to plant-parasitic nematodes.
Screening; selection; host status; broad resistance; durable resistance
Multiple images of a whole nematode specimen were taken with a high power oil-immersion objective lens and joined together to form one high-resolution megapixel, mosaic photomicrograph of the entire specimen, with the use of a relatively new mounting technique made with a 4% water agar pad. The agar pad kept the specimen nearly level and lateral, and when amended with 10 mM sodium azide, this mounting technique gradually paralyzed the nematode in a natural pose to enable production of sharp, clear images. The individual photographs were joined together and merged into one very large, seamless image. These montaged images will be useful for teaching because the student has access to a virtual specimen that is mounted in the correct orientation, imaged with a research grade microscope, and preserved in a narcotized, living condition. Such specimen images can also serve as representatives of type and voucher specimens without the deterioration typical of real types. The files can be copied and viewed with a computer almost anywhere and at any time, rather than using a more cumbersome, limiting, and expensive microscope.
agar pad technique; digital micrographs; mounting nematodes; type specimens; voucher specimens
Root-knot nematode (Meloidogyne graminicola), an important and widespread pathogen, causes high yield losses in rice with limited information on wheat and on efficient management. Absence of uniform screening protocols is contributing to slow progress of host resistance development. To develop an efficient screening protocol, several greenhouse studies were conducted, and effects of incubation period, inoculum level, inoculation method, seedling age, and their interactions on root-galling severity (RGS) ratings and reproductive factor (RF) values of M. graminicola were determined. At 2 eggs/cm3 soil, significantly lower RGS but higher RF values were observed at 60 days than at 45 days of incubation. Meloidogyne graminicola reproduced six times more on rice than on wheat where the RGS index in both crops increased steadily with increasing inoculum levels, but RF increased at lower levels and decreased beyond a maximum at medium inoculum levels. Inoculum level, container size, seedling age, inoculation method, and their interactions impacted nematode infection and reproduction. The protocol was verified on eleven rice germplasm lines and seven wheat cultivars using the resistance index (RI) calculated from RGS and RF, to screen rice and wheat germplasm.
Meloidogyne graminicola; screening protocol; rice and wheat
A new genus, one new and two known species belonging to family Rhabdolaimidae are described and illustrated. Mediolaimus n. gen. is characterized by small-sized body; papilliform outer labial and cephalic sensilla; long, tubular stoma with stegostom having relatively larger dorsal and smaller sub ventral denticles at same level; cylindrical pharynx with expanded basal bulb having thickened lumen; didelphic gonad having outstretched ovaries; elongate-conoid tail with hemispherical terminus without mucro or spinneret and caudal glands obscure. Mediolaimus obtusicaudatus n. gen. n. sp. is characterized by stoma with a relatively larger dorsal denticle and two fine subventral denticles at same level; amphidelphic reproductive system with outstretched ovaries; tail elongate conoid with smooth hemispherical terminus and absence of male. Rhabdolaimus terrestris
de Man, 1880 and Udonchus tenuicaudatus
Cobb, 1913 are described with additional details. The inter population variations are discussed in the latter species along with a detailed note on the status of Rhabdolaimidae and its proposed placement under superfamily Rhabdolaimoidea of suborder Campydorina.
India; Mediolaimus obtusicaudatus; Udonchus tenuicaudatus; new genus; new species; Rhabdolaimidae; taxonomy
Soil nematode community response to treatments of three, four-year crop rotations (spring wheat-pea-spring wheat-flax, spring wheat-green manure-spring wheat-flax, and spring wheat-alfalfa-alfalfa-flax) under conventional and organic management, and native tall grass restoration (restored prairie) were assessed in June 2003, and July and August 2004. The research site was the Glenlea Long-term Rotation and Crop Management Study, in the Red River Valley, Manitoba, established in 1992. The nematode community varied more with sample occasion than management and rotation. The restored prairie favored high colonizer-persister (c-p) value omnivores and carnivores, and fungivores but less bacterivores. The restored prairie soil food web was highly structured, mature and low-to-moderately enriched as indicated by structure (SI), maturity (MI) and enrichment (EI) index values, respectively. Higher abundance of fungivores and channel index (CI) values suggested fungal-dominated decomposition. Nematode diversity was low even after more than a decade of restoration. A longer time may be required to attain higher diversity for this restored fragmented prairie site distant from native prairies. No consistent differences were found between organic and conventional management for nematode trophic abundance, with the exception of enrichment opportunists of the c-p 1 group which were favored by conventional management. Although EI was lower and SI was higher for organic than conventional their absolute values suggested decomposition channels to be primarily bacterial, and fewer trophic links with both management scenarios. A high abundance of fungivores in the rotation including the green manure indicates greater fungal decomposition.
nematode diversity; organic management; prairie restoration; soil food web
Soybean cyst nematode (SCN) is an obligate, sedentary parasite that is a major pathogen of soybean and accounts for an estimated 1 billion dollars in production losses annually in the United States of America. This paper describes the development of a real-time PCR method for rapid, sensitive, species-specific and accurate identification of SCN alone or on mixed populations with other nematodes in North Carolina. The 83-bp DNA fragment of PrimeTime-real-time PCR was designed based on a 477-bp-SCN-SCAR marker previously proved to be SCN-specific. A total of 44 populations including cyst forming nematodes (Heterodera glycines, H. fici, H. schachtii, H. trifolii, Cactodera weissi, Globodera tabacum, Meloidodera floridensis and other unidentified cyst nematodes) and non-cyst forming nematodes (Ditylenchus dipsaci, Meloidogyne incognita and Xiphinema chambersi) were tested in this study, all SCN populations are tested positive and non-SCN populations negative. This assay for the detection and identification has been successfully applied for testing a single SCN cyst, a 2nd-stage-SCN juvenile, a single SCN egg, up to ten SCN cysts, a 10-fold dilution of a single 2nd-stage-SCN juvenile and 20-fold dilution of one SCN cyst. The assay is not SCN-race specific. It gave an accurate positive result when SCN is mixed with other cyst species. Also, nematode universal primers/probes for real-time PCR amplification as a nematode endogenous control to detect the presence of 18S ribosomal RNA (rRNA) gene were employed in this assay, so that a SCN-negative sample can be tested to exclude false negative. This method will be very useful for a broad range of research programs as well as the regulatory response and management of SCN in North Carolina and other region of the southeastern U.S.A.
DNA sequencing; molecular diagnosis; SCAR marker
To facilitate improved in vivo culture of entomopathogenic nematodes, production of both insect hosts and nematodes should be optimized for maximum fitness, quality, and cost efficiency. In previous studies, we developed an improved diet for Tenebrio molitor, a host that is used for in vivo nematode production, and we demonstrated that single insect diet components (e.g., lipids and proteins) can have a positive or negative impact on entomopathogenic nematode fitness and quality. In this study, we tested components of our improved T. molitor diet (lipids, cholesterol, and a salt [MnSO4]) alone and in combination for effects on host susceptibility and reproductive capacity of Heterorhabditis indica and Steinernema carpocapsae. Our results indicated that moderate levels of lipids (10%) increased host susceptibility to S. carpocapsae but did not affect H. indica, whereas cholesterol and MnSO4 increased host susceptibility to H. indica but not S. carpocapsae. The combined T. molitor diet (improved for increased insect growth) increased host susceptibility to S. carpocapsae and had a neutral effect on H. indica; interactions among single diet ingredients were observed. No effects of insect host diet were detected on the reproductive capacity of either nematode species in T. molitor. Subsequently, progeny infective juveniles, derived from nematodes grown in T. molitor that were fed diets with varying nutritive components were tested for virulence to and reproduction capacity in the target pest Diaprepes abbreviatus. The progeny nematodes produced from differing T. molitor diet treatments did not differ in virulence except H. indica derived from a diet that lacked cholesterol or MnS04 (but contained lipids) did not cause significant D. abbreviatus suppression relative to the water control. We conclude that the improved insect host diet is compatible with production of H. indica and S. carpocapsae, and increases host susceptibility in S. carpocapsae. Furthermore, in a general sense, our results indicate host diets can be optimized for improved in vivo entomopathogenic nematode production efficiency. This is the first report of an insect diet that was optimized for both host and entomopathogenic nematode production. Additionally, our study indicates that host diet may impact broader aspects of entomopathogenic nematode ecology and pest control efficacy.
entomopathogenic nematode; Heterorhabditis; host diet; in vivo; mass production; Steinernema
Scanning electron microscopy (SEM) was applied to paraffin-embedded wood sections to study the histopathology of pine seedlings inoculated with the pinewood nematode (PWN), Bursaphelenchus xylophilus. The sections, which had been previously prepared and observed by light microscopy (LM) on glass slides, were originally obtained from experiments in which pine seedlings had been inoculated with PWN. The cover glass was removed by soaking the glass slide in xylene for 3 to 5 days. The glass slides were cut into small pieces so that each piece contained one wood section. Each piece of the glass slide was attached with double adhesive tape to an aluminum stub. The specimens were sputter-coated with gold and examined with a scanning electron microscope (JEOL-JSM 5200). Compared to LM (as documented in previous reports) SEM provided greater depth of focus and resolution of the damaged wood tissues, nematodes and associated bacteria. SEM made it possible to observe the relationship between bacterial distribution and nematode distribution in wood tissues. SEM observations also suggested the possibility of documenting the death of ray cells and other parenchyma cells in relation to disease development. Finally, the current study of PWN in pine seedlings demonstrated that glass slides prepared for LM observations more than 25 years earlier could be successfully processed for examination by SEM.
bacterial mass; inoculation experiments; Pinus sp.; scanning electron microscopy; technique
A series of experiments were performed to examine the population dynamics of the sugarbeet cyst nematode, Heterodera schachtii, and the nematophagus fungus Dactylella oviparasitica. After two nematode generations, the population densities of H. schachtii were measured in relation to various initial infestation densities of both D. oviparasitica and H. schachtii. In general, higher initial population densities of D. oviparasitica were associated with lower final population densities of H. schachtii. Regression models showed that the initial densities of D. oviparasitica were only significant when predicting the final densities of H. schachtii J2 and eggs as well as fungal egg parasitism, while the initial densities of J2 were significant for all final H. schachtii population density measurements. We also showed that the densities of H. schachtii-associated D. oviparasitica fluctuate greatly, with rRNA gene numbers going from zero in most field-soil-collected cysts to an average of 4.24 x 108 in mature females isolated directly from root surfaces. Finally, phylogenetic analysis of rRNA genes suggested that D. oviparasitica belongs to a clade of nematophagous fungi that includes Arkansas Fungus strain L (ARF-L) and that these fungi are widely distributed. We anticipate that these findings will provide foundational data facilitating the development of more effective decision models for sugar beet planting.
Dactylella oviparasitica; Heterodera schachtii; nematophagous; sugarbeet cyst nematode; suppressive soil; Arkansas Fungus (ARF)
Root-knot nematodes (Meloidogyne spp.) are polyphagous plant parasites of global importance. Successful host infection depends on the particular interaction between a specific nematode species and race and a specific plant species and cultivar. Accurate diagnosis of nematode species is relevant to effective agricultural management; and benefits further from understanding the variability within a single nematode species. Here, we described a population of M. arenaria race 2 from Mendoza (Argentina). This study represents the first morphometric, morphological, biochemical, reproductive, molecular, and host range characterization of a root-knot nematode species from Argentina. Even after gathering morphological and morphometric data of this population and partially sequencing its rRNA, an unequivocal taxonomic assignment could not be achieved. The most decisive data was provided by esterase phenotyping and molecular methods using SCARs. These results highlight the importance of taking a multidimensional approach for Meloidogyne spp. diagnosis. This study contributes to the understanding of the variability of morphological, reproductive and molecular traits of M. arenaria, and provides data on the identification of root-knot nematodes on tomato cultivars from Argentina.
Meloidogyne arenaria; morphology; root-knot nematode; molecular taxonomy
Experiments were conducted in the greenhouse to assess root galling and egg production of three root-knot nematode species, Meloidogyne arenaria, M. incognita, and M. javanica, on several weeds common to Florida agricultural land. Weeds evaluated were Amaranthus retroflexus (redroot pigweed), Cyperus esculentus (yellow nutsedge), Eleusine indica (goosegrass), Portulaca oleracea (common purslane), and Solanum americanum (American black nightshade). Additionally, although it is recommended as a cover crop in southern regions of the U.S., Aeschynomene americana (American jointvetch) was evaluated as a weed following the detection of root galling in a heavy volunteer infestation of an experimental field in southeastern Florida. Weeds were propagated from seed and inoculated with 1000 nematode eggs when plants reached the two true-leaf stage. Tomato (Solanum lycopersicum ‘Rutgers’) was included as a positive control. Aeschynomene americana and P. oleracea roots supported the highest number of juveniles (J2) and had the highest number of eggs/g of root for all three species of Meloidogyne tested. However, though P. oleracea supported very high root levels of the three nematode species tested, its fleshy roots did not exhibit severe gall symptoms. Low levels of apparent galling, combined with high egg production, increase the potential for P. oleracea to support populations of these three species of root-knot nematodes to a degree that may not be appropriately recognized. This research quantifies the impact of P. oleracea as a host for M. arenaria, M. incognita, and M. javanica compared to several other important weeds commonly found in Florida agricultural production, and the potential for A. americana to serve as an important weed host of the three species of root-knot nematode tested in southern regions of Florida.
Aeschynomene americana; Amaranthus retroflexus; Cyperus esculentus; Eleusine indica; Florida; host status; nematode reproduction; Portulaca oleracea; root-knot nematodes; Solanum americanum
The entomopathogenic nematodes (EPN) Heterorhabditis and Steinernema are widely used for the biological control of insect pests and are gaining importance as model organisms for studying parasitism and symbiosis. In this paper recent advances in the understanding of EPN behavior are reviewed. The “foraging strategy” paradigm (distinction between species with ambush and cruise strategies) as applied to EPN is being challenged and alternative paradigms proposed. Infection decisions are based on condition of the potential host, and it is becoming clear that already-infected and even long-dead hosts may be invaded, as well as healthy live hosts. The state of the infective juvenile (IJ) also influences infection, and evidence for a phased increase in infectivity of EPN species is mounting. The possibility of social behavior - adaptive interactions between IJs outside the host - is discussed. EPNs’ symbiotic bacteria (Photorhabdus and Xenorhabdus) are important for killing the host and rendering it suitable for nematode reproduction, but may reduce survival of IJs, resulting in a trade-off between survival and reproduction. The symbiont also contributes to defence of the cadaver by affecting food-choice decisions of insect and avian scavengers. I review EPN reproductive behavior (including sperm competition, copulation and evidence for attractive and organizational effects of pheromones), and consider the role of endotokia matricida as parental behavior exploited by the symbiont for transmission.
Behavior; biological control; ecology; endotokia matricida; entomopathogenic nematode; fitness trade-off; foraging strategy; Heterorhabditis; phased infectivity; Photorhabdus; reproduction; scavenging; sexual maturation; symbiosis; Steinernema; Xenorhabdus