Individual Nematodes Research Articles

Forest types matter for the community and co-occurrence network patterns of soil bacteria, fungi, and nematodes.

Tree growth influences the biological, physical, and chemical properties of the soil through the input of different types of litter and various root exudates. However, our understanding of tree-mediated effects on the composition and diversity of soil biota remains limited. This study aimed to determine the effects of physically neighboring forest types (i.e., an artificial Japanese cedar (Cryptomeria japonica) plantation vs. a broadleaf (Quercus serrata) secondary forest) on individual bacterial, fungal, and nematode communities and the associations among these inter-kingdoms. Bacterial, fungal, and nematode aggregates were estimated using MiSeq high-throughput sequencing system. The amplicon sequence variant richness of fungi and nematodes was significantly greater in the cedar plantation than in the broadleaf forest, and the three soil biota community structures were significantly clustered among the forest types. Environmental factors such as soil pH, C, N, and C/N ratio significantly influenced the three soil biota community structures. The bacterial–fungal–nematode co-occurrence network of the broadleaf forest had more nodes and edges than that of the cedar plantation. Moreover, Ascomycota and Basidiomycota fungi mainly co-occurred with fungivorous nematodes in the cedar and broadleaf forests, respectively. Our results suggested that unique soil biota communities and characteristic co-occurrence network patterns were established among the tripartite inter-kingdom relationships between adjacent forest types.

Combined matrix-assisted laser desorption/ionisation-mass spectrometry imaging with liquid chromatography-tandem mass spectrometry for observing spatial distribution of lipids in whole Caenorhabditis elegans.

Matrix-assisted laser desorption/ionisation-mass spectrometry imaging (MALDI-MSI) is a powerful label-free technique for biomolecule detection (e.g., lipids), within tissue sections across various biological species. However, despite its utility in many applications, the nematode Caenorhabditis elegans is not routinely used in combination with MALDI-MSI. The lack of studies exploring spatial distribution of biomolecules in nematodes is likely due to challenges with sample preparation. This study developed a sample preparation method for whole intact nematodes, evaluated using cryosectioning of nematodes embedded in a 10% gelatine solution to obtain longitudinal cross sections. The slices were then subjected to MALDI-MSI, using a RapifleX Tissuetyper in positive and negative polarities. Samples were also prepared for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis using an Exploris 480 coupled to a HPLC Vanquish system to confirm the MALDI-MSI results. An optimised embedding method was developed for longitudinal cross-sectioning of individual worms. To obtain longitudinal cross sections, nematodes were frozen at -80°C so that all worms were rod shaped. Then, the samples were defrosted and transferred to a 10% gelatine matrix in a cryomold; the worms aligned, and the whole cryomold submerged in liquid nitrogen. Using MALDI-MSI, we were able to observe the distribution of lipids within C. elegans, with clear differences in their spatial distribution at a resolution of 5μm. To confirm the lipids from MALDI-MSI, age-matched nematodes were subjected to LC-MS/MS. Here, 520 lipids were identified using LC-MS/MS, indicating overlap with MALDI-MSI data. This optimised sample preparation technique enabled (un)targeted analysis of spatially distributed lipids within individual nematodes. The possibility to detect other biomolecules using this method thus laid the basis for prospective preclinical and toxicological studies on C. elegans.

Elucidating nematode diversity and prevalence in moose across a wide latitudinal gradient using DNA metabarcoding

Parasitic nematodes are ubiquitous and can negatively impact their host by reducing fecundity or increasing mortality, yet the driver of variation in the parasite community across a wildlife host's geographic distribution remains elusive for most species. Based on an extensive collection of fecal samples (n = 264) from GPS marked moose (Alces alces), we used DNA metabarcoding to characterize the individual (sex, age class) and seasonal parasitic nematode community in relation to habitat use and migration behavior in five populations distributed across a wide latitudinal gradient (59.6°N to 70.5°N) in Norway. We detected 21 distinct nematode taxa with the six most common being Ostertagia spp., Nematodirella spp., Trichostongylus spp., T. axei, Elaphostrongylus alces, and an unclassified Strongylida. There was higher prevalence of livestock parasites in areas with larger sheep populations indicating a higher risk of spillover events. The individual level nematode richness was mostly consistent across study areas, while the number and type of nematode taxa detected at each study area varied considerably but did not follow a latitudinal gradient. While migration distance affected nematode beta-diversity across all sites, it had a positive effect on richness at only two of the five study areas suggesting population specific effects. Unexpectedly, nematode richness was higher in winter than summer when very few nematodes were detected. Here we provide the first extensive description of the parasitic nematode community of moose across a wide latitudinal range. Overall, the population-specific impact of migration on parasitism across the distribution range and variation in sympatry with other ruminants suggest local characteristics affect host-parasite relationships.

A gene with a thousand alleles: The hyper-variable effectors of plant-parasitic nematodes

Pathogens are engaged in a fierce evolutionary arms race with their host. The genes at the forefront of the engagement between kingdoms are often part of diverse and highly mutable gene families. Even in this context, we discovered unprecedented variation in the hyper-variable (HYP) effectors of plant-parasitic nematodes. HYP effectors are single-gene loci that potentially harbor thousands of alleles. Alleles vary in the organization, as well as the number, of motifs within a central hyper-variable domain (HVD). We dramatically expand the HYP repertoire of two plant-parasitic nematodes and define distinct species-specific "rules" underlying the apparently flawless genetic rearrangements. Finally, by analyzing the HYPs in 68 individual nematodes, we unexpectedly found that despite the huge number of alleles, most individuals are germline homozygous. These data support a mechanism of programmed genetic variation, termed HVD editing, where alterations are locus specific, strictly governed by rules, and theoretically produce thousands of variants without errors.

Depth distribution of plant-parasitic nematodes on bentgrass golf greens in Missouri and Indiana.

Control of plant-parasitic nematodes (PPNs) on golf putting greens with nematicides is dependent on the seasonal occurrence and depth distribution of target PPN populations. This study aimed to determine if plant-parasitic nematode populations on golf course putting greens in Missouri and Indiana peaked at a targetable depth at a specific time in the year, focusing primarily on lance (Hoplolaimus spp.) and root-knot (Meloidogyne spp.) nematodes. To elucidate species diversity in the region, rDNA from a subset of lance and root-knot nematodes was sequenced and analyzed, with additional micromorphology of a lance nematode assessed in scanning electron micrographs (SEM). Soil samples were taken to a depth of 25 cm and stratified into 5 cm increments during April, June, August and October at seven sites across Missouri, three in the Kansas City metro of Kansas in 2021 and in ten sites across Indiana in 2022. Samples were stratified in five-centimeter increments and aggregated for a total of 100 cm3 of soil at each depth for each sampling. Samples were processed using a semi-automatic elutriator followed by the sucrose-flotation method, and populations were counted using a hemocytometer and recorded. For molecular characterization, rDNA was extracted and analyzed from 31 individual lance nematodes from one site in Missouri and eight sites in Indiana, and 13 root-knot nematodes from nine sites across Indiana. A significant interaction occurred between sampling month and depth for lance and ring nematodes Missouri/KS, with both PPN populations peaking at the 0-5 cm depth during October, which is well after most targeted nematicide applications are applied. Ring nematodes in Indiana did not follow this trend and were most abundant in August at a depth of 0-5 cm. No significant interaction between depth and month occurred for lance or root-knot nematodes in Indiana, or root-knot nematodes in Missouri/KS. Hoplolaimus stephanus and H. magnistylus were the lance species identified on golf greens, and Meloidogyne naasi, M. graminicola and M. marylandi were the root-knot species identified. Scanning-electron micrographs confirmed morphological characteristics unique to H. stephanus.

Evaluating Vacuum and Steam Heat to Eliminate Pinewood Nematodes in Naturally Infested Whole Pine Logs.

Pinewood nematodes threaten forest health and continue to interfere with international trade because they can be spread around the globe via nematode-infested wood. International Standards for Phytosanitary Measure (ISPM-15) requires that all pine wood be treated at 56°C for 30 min to ensure that all pests and pathogens are killed within sawn wood, whereas fumigation with methyl bromide is the currently approved practice and widely used in treating whole logs. A method of treatment that uses less energy and time or does not rely on environmentally damaging gases is urgently needed. Because vacuum with steam has shown promise in treating several different commodities, the purpose of this study was to use it to eradicate pinewood nematodes in whole logs. Three protocols were applied: 1) 48°C for 15 min., 2) 56°C for 30 min., and 3) 60°C for 60 min. The third protocol reduced the population to statistically zero; however, some samples contained at least 1 survivor. Unfortunately, these surviving nematodes increased in number one month after treatment, and one year later, they continued to reproduce in the wood. Therefore, this protocol needs to be further refined to remove pinewood nematodes completely. Explanation of the survival of individual nematodes within whole logs remains a matter for conjecture: 1) certain portions of the wood were somehow insulated from the heat and did not achieve the lethal temperature, and 2) survival stages may be able to survive temperatures that are deadly to the normal life stages.

Eryngium foetidum Reported as a New Host of Meloidogyne enterolobii in the State of Pará, in the Eastern Amazon Region, Brazil

Amazon chicory (Eryngium foetidum L. [Apiaceae]), also known as culantro, is native to Tropical America and the West Indies. It belongs to the unconventional food plants (UFPs) group, and in addition to be consumed as a spice herb, it possesses a wide range of ethnomedicinal uses (Paul et al. 2011). In 2019, in the eastern Amazon region of Brazil, state of Pará, producers of E. foetidum in the municipality of Castanhal (01°15'363” S 047°10'232” W) reported the occurrence of underdeveloped plants with leaf yellowing and a large number of galls in the root system, which are typical symptoms of root-knotting nematode. Soil and root samples were collected and sent to the Nematology Laboratory (LabNema) located at the Faculty of Agrarian and Veterinary Sciences, UNESP, Jaboticabal, São Paulo, Brazil. A total of 46 second-stage juveniles (J2s) were extracted per 100 cm3 of soil, and a total of 460 eggs and J2s Meloidogyne spp. were found per gram of root. Morphological and molecular techniques were used to identify the species. The analysis of the perineal patter of ten females revealed thin striations in an oval shape with a high and semi-trapezoidal dorsal arch. No striations were observed in the perivulvar region. The labial region of the ten males analyzed exhibited a non-prominent labial disc, fused and slightly recessed submedian lips, with no apparent annulations. The morphological characteristics observed in the adults were consistent with those originally described for Meloidogyne enterolobii (Yang; Eisenback, 1983), confirming the species purity of the recovered population. Three individual nematodes had their 18S rDNA region sequenced (Holterman et al. 2006) which showed an average identity of 99.7% with other sequences of M. enterolobii available in the GenBank database. A Bayesian phylogenetic tree was constructed, providing insights into the specific relationship of M. enterolobii recovered from E. foetidum with other related nematodes. Each of the three sequenced nematodes represented a unique haplotype, resulting in their separation into distinct clades. Moreover, the obtained sequences presented polymorphisms that differed from the M. enterolobii sequences already available in the database, highlighting the genetic diversity of this species in relation to its original host (Silva et al. 2021). The species M. enterolobii was also confirmed using species-specific primers for M. incognita, M. javanica, and M. enterolobii (Zijlstra et al. 2000; Tigano et al. 2010). To confirm the pathogenicity of M. enterolobii on E. foetidum, a modified Koch Postulate was conducted. Six seedlings of E. foetidum were transplanted individually to 10-liter pots containing autoclaved soil. Each pot was then inoculated with 5 mL of a suspension containing 3,000 eggs and J2s from the original population of M. enterolobii obtained from E. foetidum. After 90 days, the inoculated plants exhibited root galls with a plentiful egg mass, in contrast to the healthy non-inoculated plants. The average number of M. enterolobii nematodes recovered from the roots of the inoculated plants was 42,040 eggs and J2s, resulting in a reproduction factor (RF) of 14.0. The importance of reporting the occurrence of M. enterolobii in E. foetidum is due to the fact that this plant species is cultivated in a crop rotation system with other vegetables such as lettuce and coriander, which are also hosts of M. enterolobii. Consequently, different crop rotation strategies and control alternatives need to be considered in areas where E. foetidum is grown. This is the first report of E. foetidum serving as a host for the root-knot nematode M. enterolobii worldwide.

Abundance and distribution of nematodes in petroleum hydrocarbon polluted soil in Okrika Local Government Area, Rivers State, Nigeria

This study examined the abundance of nematodes in petroleum hydrocarbon polluted soils in Okrika region. A total of 80 soil samples were collected at various depths in several sites of the region during the dry and rainy seasons. Three of the sites (Ekerekana, Ogoloma and Okari) were polluted by petroleum hydrocarbon while the control (Abam) was unpolluted. A total of 15 genera containing 749 individual nematodes were extracted from both polluted and unpolluted sites. The highest abundance was recorded at the control site, Abam with 327 (43.7%), followed by Ekerekana with 272 (36.3%), Ogoloma with 97 (12.9%) and Okari with 53 (7.1%) nematodes. In comparison to the dry season, more nematodes, 561 (74.9% out of total population) individuals were extracted during the wet season. During the wet season, nematode abundance was considerably higher (p<0.05) in the unpolluted site than those in polluted sites, but during the dry season, there was no significant difference (p>0.05) between the seasons. Similarly, more nematodes (51.3%) were extracted from the core depth of 0-5 cm than 6-10 cm (33.2%) and 11-15 cm (15.5%). At the depth of 0-5 cm, the control site had a significant (p<0.05) nematode abundance than the polluted sites while at the depths of 6-10 cm and 11-16 cm, although there was a higher numerical abundance of nematodes in the unpolluted sites than the control site, but these were not significant (p>0.05). This study showed that nematodes at the studied sites were relatively sensitive to petroleum hydrocarbon pollution especially at the depth of 0-5 cm in both dry and rainy seasons.

Two Intra-Individual ITS1 rDNA Sequence Variants Identified in the Female and Male Rotylenchulus reniformis Populations of Alabama.

Around 300 different plant species are infected by the plant-parasitic reniform nematode (Rotylenchulus reniformis), including cotton. This is a devasting nematode with a preference for cotton; it is commonly found in Alabama farms and causes severe reduction in yields. Its first internal transcribed spacer (ITS1) region can be sequenced, and potential mutations can be found in order to study the population dynamics of the reniform nematode. The goal of our study was to sequence the ITS1 rDNA region in male and female RNs that were collected from BelleMina, Hamilton, and Lamons locations in Alabama. After separating the single male and female RNs from the samples collected from the three selected listed sites above, the ITS1 region was amplified selectively using specific primers, and the resulting products were cloned and sequenced. Two distinct bands were observed after DNA amplification of male and female nematodes at 550 bp and 730 bp, respectively. The analysis of sequenced fragments among the three populations showed variation in average nucleotide frequencies of female and male RNs. Singletons within the female and male Hamilton populations ranged from 7.8% to 10%, and the variable sites ranged from 13.4% to 26%. However, female and male BelleMina populations had singletons ranging from 7.1% to 19.7% and variable regions in the range of 13.9% to 49.3%. The female and male Lamons populations had singletons ranging from 2.5% to 8.7% and variable regions in the range of 2.9% to 14.2%. Phylogenetic (neighbor-joining) analysis for the two ITS1 fragments (ITS-550 and ITS-730) showed relatively high intra-nematode variability. Different clone sequences from an individual nematode often had greater similarity with other nematodes than with their own sequences. RNA fold analysis of the ITS1 sequences revealed varied stem and loop structures, suggesting both conserved and variable regions in the variants identified from female and male RNs, thus underscoring the presence of significant intra- and inter-nematodal variation among RN populations in Alabama.

Effects of alpine meadows with different degradation gradients on the stability of the soil micro-foodweb in the Tibetan Plateau

Resource-transfer connections among bacteria, fungi, and microbivorous nematodes play an integral role in the decomposition pathways of soil micro-foodweb. Exploring the response of soil bacteria, fungi, and nematodes to grassland degradation is crucial for predicting the direction and trajectory of degradation. However, the knowledge about soil bacteria-fungi-nematode interactions in ecosystem function and nutrient cycling must be highlighted properly, especially in the Qinghai-Tibet Plateau. Therefore, this study focuses on four alpine meadows at different degenerate gradients (non-degraded grassland, slightly degraded grassland, moderately degraded grassland, and severely degraded grassland) on the Qinghai-Tibet Plateau. The investigation of soil bacteria-fungi-nematode interactions uses high-throughput sequencing and correlation network analysis. The results demonstrate significant variations in the composition of soil nematode genera, and bacterial and fungal phyla, across different degradation stages. Moreover, the diversity of nematodes, bacteria and fungi declined as grassland degradation worsened. Principal component analysis (PCA) revealed that alpine meadows with varying degradation gradients significantly influenced the community structure of bacteria, fungi, and nematodes. Mantel analysis indicated that soil water content (P < 0.05), total phosphorus (P < 0.05), and total nitrogen (P < 0.05) were the primary soil properties affecting the soil bacterial community. Similarly, available soil potassium (P < 0.05), microbial biomass carbon (P < 0.05), and microbial biomass nitrogen (P < 0.05) were the main soil factors influencing the soil fungal community. In contrast, the correlation between individual soil characteristics and soil nematodes did not reach a significant level, suggesting that a blend of physical and chemical properties regulates the soil nematode community. As grassland degradation intensified, the number of links in the interaction network of bacteria, fungi, and nematodes increased. Particularly, the networks between fungi and nematodes expanded, and the key species within each network varied with the different stages of grassland degradation. Concurrently, soil micro-foodweb decomposition pathways shifted. In the ND treatment, bacteria and fungi jointly dominated the degradation channels of the soil micro-food web, while the LD and MD treatments mainly dominated the degradation channels of fungi. On the other hand, SD treatment was dominated by bacterial degradation channels. This research adds to our theoretical understanding of meadow soil micro-foodwebs and the sustainable use of alpine grasslands in the Qinghai-Tibet Plateau.

A community-level sampling method for detection of Meloidogyne enterolobii and other root-knot nematodes in sweetpotato storage roots

Meloidogyne enterolobii is an aggressive root-knot nematode (RKN) species that has emerged as a significant pathogen of sweetpotato in the Southeastern United States. M. enterolobii is spread primarily through the movement of infected ‘seed’ sweetpotatoes used for propagation. The RKN resistance in commercially grown sweetpotato cultivars has proven ineffective against this nematode. Detecting RKN in sweetpotato by eye is unreliable, and further distinguishing M. enterolobii from other RKN species that infect sweetpotato is labor intensive; relying on molecular tests conducted on individual nematodes dissected out of host roots. It is often impractical to collect enough nematodes to gauge the composition of RKN species from a single root or multiple roots from across a field. Here, we have developed a community-level survey method to collect skin samples and extract total DNA from batches of sweetpotato storage roots. Combining this method with species-specific PCR assays allowed for quick and sensitive detection of M. enterolobii and other RKN species infecting sweetpotatoes. We tested this method using batches of infected storage roots at varying levels of M. enterolobii infection. We also inoculated skin samples with varying numbers of individual M. enterolobii eggs to determine the method's detection threshold and used this method to conduct surveys for RKN on fresh market sweetpotatoes. Our results show that this method can consistently and reliably detect M. enterolobii in sweetpotato batches at levels as low as 2 eggs per 10 mL skin sample. This method will be a useful tool to help screen for the presence of M. enterolobii in ‘seed’ sweetpotatoes before they are replanted, thereby helping to slow the spread of this nematode to M. enterolobii-free sweetpotato growing operations.

Pratylenchus zeae a causative agent of Corn Root Rot in Jiangsu Province of China.

Corn (Zea mays L.) plays an important role in China's cash crops, not only as food, but a vital raw material for animal husbandry and industry (Li et al. 2022). Pratylenchus zeae is one of the most damaging root-lesion nematodes (RLN) that can result in decreased yield and quality of crops (Liu et al. 2017). In September 2020, five root/soil samples were collected from the rhizosphere of corn (cv. Zhengdan 958), which had weak growth and root brown lesions in Chenzhou Village, Taolin Town, Donghai County, Lianyungang City, Jiangsu Province of China. Nematodes were extracted from the collected samples using the modified Baermann funnel method (Hooper et al. 2005). RLN were found in all samples, an average of 46 RLN per gram of root and 138 RLN per 100 cm3 of soil. The obtained RLN females were sterilized with 0.3% streptomycin sulfate and then inoculated on each carrot disks individually to obtain the purified population. RLN were examined by morphological and molecular characteristics to confirm the species indentification. The main morphological measurements of adult (n = 15) included body length = 524.7 μm (mean) ± 15.1 (standard deviation) (range = 490.7 to 543.6 μm), stylet = 15.2 μm ± 0.8 (14.2 to 16.8 μm), tail length = 30.3 μm ± 2.5 (26.3 to 35.3 μm), a = 25.6 ± 1.3 (24.4 to 29.3), b = 5.3 ± 0.3 (4.7 to 5.8), c = 17.4 ± 1.4 (14.9 to 19.3), two annules on the lip region. No males were found in the specimens. The morphological characters of this population are consistent with the description of P. zeae (Castillo and Vovlas, 2007). Furthermore, DNA was extracted from individual nematodes. The primers of TW81/AB28 and D2A/D3B (Subbotin et al. 2006) were used to amplified the rDNA-ITS region and rDNA 28S D2-D3 region, respectively. The purified PCR products were ligated into One step ZTOPO-Blunt/TA vector and transformed to Escherichia coli strain DH5α, and then sequenced by Sunya Biotechnology Co., Ltd (Henan, China). The obtained seqences were submitted to NCBI. The rDNA-ITS sequences (669 bp, GenBank Accession No: OP456372 and OP466367) exhibited 95.0% to 97.1% of identity with P. zeae sequences (KU198980 and KU198975). The obtained D2-D3 region of the 28S rDNA sequences (782 bp, OP441397 and OP448675) exhibited 99.7% to 100% identity with P. zeae sequences (EU130893 and KY424269). Consequently, both morphological and molecular data confirmed the identity of P. zeae. To further confirm reproduction on corn, single corn seeds (cv. Zhengdan 958) were sown in eight 2-liter pots filled with 1.8-liter of sterilized soil in greenhouse at 28°C. About 15 days after sowing, each pot with one corn plant with the same growth status was selected to inoculate with 1,000 mixed stage nematodes of P. zeae , Eight pots of uninoculated corn plants were used as controls. After 60 days, the inoculated plants were harvested and brown lesions were observed on roots. No symptoms and nematodes was detected in the control. An average number of RLN per pot was 3,752 in soil and 1,183 in roots were extracted, the reproduction factor (final population/initial population) was 4.94, indicating that P. zeae infects and reproduces well on this corn cultivar. P. zeae has only been reported on corn in Guangxi Province, southern in China(Fang et al. 1994). To our knowledge, this is the fist report of P. zeae infecting corn in Jiangsu Province, eastern in China. As P. zeae can cause great damage to corn, necessary measures should be taken to prevent the spread of P. zeae to other areas.

Checklist of soil nematode diversity from Udupi District, Karnataka, India

Nematodes are plentiful in soil and may be found in practically every habitat. Around 25% of global biodiversity is considered to be supported by terrestrial ecosystem soils. There has been less research on nematode populations in Karnataka than there has been in other states. The scarcity of available literature provides up even more opportunities for studying these faunas in this region. As a result, the following investigation was conducted. The major goal of this research was to investigate the nematode diversity in the Udupi area. The collected nematodes were fixed, dehydrated, and displayed on a glass slide after isolation. Published keys were used to identify the species; there were 2,833 individual nematodes recovered. This collection contained 49 soil nematode species, which were classified into 34 genera and 20 families distributed over seven orders.

Identification, Diversity, and Distribution of Meloidogyne spp. in Vegetable Fields of South Georgia, U.S.A.

Root-knot nematode (RKN; Meloidogyne spp.) is the most prevalent plant-parasitic nematode in vegetable fields of Georgia, with an incidence of 67.3%. Because aggressive RKN species are reported in the southeastern United States, molecular-based identification of RKN species was conducted on soil samples taken from a nematode surveillance study in 2018 from 292 RKN-infested vegetable fields in southern Georgia. The RKN-infested soil was potted with tomato cultivar Rutgers, and individual nematode females were isolated from galled roots and subjected to species-specific PCR and mitochondrial haplotype-based RKN species identification. The incidence (%), mean, and maximum relative abundance (second-stage juveniles per 100 cm3 of soil) of the five RKN species identified consisted of M. incognita (91.9, 486, 14,144), M. arenaria (36.0, 707, 14,144), M. floridensis (2.2, 909, 5,264), M. javanica (5.5, 352, 1,488), and M. haplanaria (0.7, 8, 14). A large proportion of fields (29%) had mixed populations of M. incognita and M. arenaria, which may reflect the region's long history of cotton and peanut cultivation. For unknown reasons, mixed populations of M. incognita and M. arenaria were associated with higher population densities. M. incognita is the most important RKN species in vegetable fields, followed by M. arenaria; therefore, pure or mixed populations of these species should be addressed in nematode management programs. Although at a lower incidence, the newly detected species, M. floridensis and M. haplanaria, have the potential to become a major threat since they reproduce on vegetables with Mi-resistant genes.

Nematode functional traits and community structure change from river to the terrestrial border in Segara Anakan Mangrove, Indonesia

Abstract. Maharning AR, Ardli ER, Prabowo RE. 2023. Nematode functional traits and community structure change from river to the terrestrial border in Segara Anakan Mangrove, Indonesia. Biodiversitas 24: 2434-2446. We intended to explore the spatial distribution of nematode functional traits and community structure in Segara Anakan Mangrove, Indonesia. Samples were retrieved from 14 stations, extending from the river to the area adjacent to terrestrial forest. We used taxonomic and functional trait identification, covering body shape, length, tail shape, feeding type, and life history as independent traits, and combined traits by collecting traits describing individual nematodes. The principal component analysis suggested that predatory or omnivorous or long-size nematodes mainly inhabited stations adjacent to the terrestrial forest but were unclear for the mid-stations. There were 29 combined traits and 38 genera of nematodes observed. The detrended correspondence analysis revealed that nematode compositional changes from the river to the terrestrial adjacent area were clearer for the combined traits and nematode genera. Seven combined traits and 12 genera inhabited only the particular stations of the study site. The common nematodes were slender, 1-2 mm long, elongated filiform tails, non-selective deposit feeders, and very tolerant to pollutants, which lived in almost all stations (92.86%). The genus relative abundance (&gt;10%) included Anoplostoma, Eubostrichus, and Chromadorina. The results suggest that distance from the river might regulate the habitability of nematodes, in which nematode functional traits play roles.

Functional traits of soil nematodes define their response to nitrogen fertilization

Abstract Nitrogen (N) fertilization and warming are two crucial global change factors affecting the soil nematode communities. The effects of N fertilization and warming, however, on nematode communities in soils are inconsistent across ecosystems and maybe be even opposite. One key reason is that the commonly used taxonomic diversity is less sensitive to environmental changes than the seldom‐used trait‐based indicators. To verify this, we performed an 8‐year field experiment with four N fertilization levels with and without soil warming and collected an extensive dataset consisting of (i) six traits related to the nematode performance, that is, body size, maximum body length, maximum body width, stylet length, oesophagus length and intestinal length; (ii) the taxonomic alpha (richness and abundance) and beta diversity (Bray–Curtis dissimilarities) of the whole nematode community and each nematode functional group, (iii) soil food web resources (the total taxonomic richness and abundance of plant, bacterial and fungal communities) and (iv) soil properties (pH, total, ammonium and nitrate N, microbial N and C, total and available P and soil water content). We found that N fertilization altered plant diversity and soil nitrate levels, which in turn decreased taxonomic alpha diversity of two nematode functional groups (phytophagous nematodes and predators), but taxonomic diversity for the whole nematode community remained stable. The decreased taxonomic alpha diversity of phytophagous nematodes resulted in increased maximum body width, but decreased stylet length and oesophageal length. Mild warming (~ 0.7°C) had no effects on soil properties and soil food web resources, and the taxonomic diversity or nematode traits remained unchanged. Our results reveal that nematode functional traits show strong responses to N fertilization as individual nematode groups adapted quickly to changed soil properties and food web resources. Taxonomic diversity indices, however, were more stable under these changes showing that the functional composition of nematode communities may respond in the short term despite little effects on species diversity. Thus, the trait‐based indicators not only reveal how nematodes respond to N fertilization, but also how nematodes mediate the effects of N fertilization on ecosystem functioning (i.e. soil nutrient cycling). Read the free Plain Language Summary for this article on the Journal blog.

Velocity of Meloidogyne ethiopica second‐stage juveniles in sandy soil under several soil moisture conditions

AbstractCalculating the velocity and migration of plant‐parasitic nematodes in soil is important for understanding individual roles in nematode populations. Tracking individual nematodes in the soil is difficult, and any marking system can alter the behavior of second‐stage juvenile (J2) nematodes. This study examined the velocity of Meloidogyne ethiopica J2s under different soil moisture conditions and proposed a maximum migration distance. Evaluations considered 16 velocity categories (distances of 1.5, 4.5, 9.5, and 14.5 cm; experimental periods of 3, 6, 16, and 26 d) and four moisture ranges (40–55, 55–70, 70–85, and 85–100%) in pots with sandy soil under controlled temperature and moisture conditions. The weighted average velocity of J2 specimens that reached the roots was 70–75 μm h−1, and the maximum recorded velocity was 378 μm h−1. The fastest average velocities were 100 μm h−1 (Test 1) and 89 μm h−1 (Test 2). The average velocity at 85–100% soil moisture was very similar to the overall weighted average velocity of M. ethiopica J2s because most movement of the J2 specimens occurred at higher soil moisture ranges. Few J2s moved in the 40–55% soil moisture range. At slow velocities (24, 39, and 72 μm h−1), the number of J2s reaching the roots was highly correlated with higher soil moisture. The majority of M. ethiopica J2s did not move or moved at velocities slower than 24 μm h−1, implying that unless the plant roots were near the nematode, they will not be infested.

CRISPR/Cas12a Coupled With Recombinase Polymerase Amplification for Sensitive and Specific Detection of Aphelenchoides besseyi.

Aphelenchoides besseyi (A. besseyi), a seed-borne parasitic nematode, is the causal agent of rice white tip disease (RWTD), which may result in a drastic loss of rice yield. Seed treatments are currently considered to be the most effective means of preventing the spread of RWTD. Therefore, the rapid, highly specific, and accurate detection of A. besseyi from rice seeds is crucial for the surveillance, prevention, and control of RWTD. Here, we describe a novel detection assay that combines recombinase polymerase amplification (RPA) and CRISPR/Cas12a to detect A. besseyi (termed RPA-Cas12a-Ab), with a low limit of detection (LOD) of 1 copy/μl of plasmid or 1:107 diluted DNA extracted from individual nematodes. To improve the user-friendliness, lateral flow strip assay (LFA) was adopted to visualize the detection result. The LOD of the RPA-Cas12a-Ab LFA assay was 1,000 copies/μl plasmid or 1:10 diluted DNA extracted from individual nematodes. The assay developed in this study was able to identify A. besseyi in 45 min with high accuracy and sensitivity without cross reaction with three closely related non-A. besseyi species. Thus, RPA-Cas12a-Ab is a rapid, sensitive, and specific detection system that requires no sophisticated equipment and shows promise for on-site surveillance of A. besseyi.

Discovery of Root-Lesion Nematode (Pratylenchus scribneri) on Corn in Hainan Province of China.

Corn (Zea mays L.) is a very important cereal crop and serves as food, feed, and industrial material (Liu et al. 2016). The root-lesion nematode (RLN) is considered one of the most important plant-parasitic nematodes and can cause economic losses in agriculture worldwide (Jones et al. 2013). In January 2020, five samples were collected from a corn field in Lingshui Lizu Autonomous County, Hainan Province, China. The collected corn plants (cv. Denghai 685) were growing poorly and roots showed distinct lesions and rot. Corn fields with symptoms of stunted plants, and brown lesions on roots were widespread. This corn disease was severe in Lingshui Lizu Autonomous County. RLN were extracted from soil samples by the modified Baermann funnel (Hooper et al. 2005). All the samples contained RLN ranging from 9 to 82 (average 39) RLN per 100 cm3 of soil and 113 to 257 (average 194) RLN per 5 g roots. The extracted RLN were sterilized and cultured on carrot disks at 25°C for 90 days. Afterwards, seeds of corn (cv. Denghai 685) were sown in pots containing 1.8 liters of sterilized soil. Eight plants, one per pot, were infested with 1,000 RLN, eight pots of noninfested corn plants were used as controls, and plants were kept in a greenhouse at 25°C. At 75 days after inoculation, symptoms were like those initially observed in corn fields, whereas no symptoms were observed in the control plants. Nematodes in the soil and roots were extracted using the same method as previously described (Hooper et al. 2005). The average number of RLN per pot was approximately 4,250 in soil and 820 in roots, the reproduction factor (final number of nematodes/initial number of nematodes) was 5.07, no RLN were found in the control. The experiment was conducted twice. The morphological and molecular studies of RLN were examined to confirm species identification. The main morphological measurements of adult females (n = 15) included body length = 526.0 μm ± 17.1 (standard error) (range = 498.0 to 560.5 μm), stylet = 16.0 μm ± 0.3 (15.5 to 16.5 μm), tail length = 29.0 μm ± 1.5 (26.5 to 31.0 μm), a = 23.6 ± 0.6 (22.6 to 24.4), b = 5.6 ± 0.3 (5.2 to 6.0), c = 18.3 ± 0.9 (16.4 to 19.7), V = 78.2% ± 0.6 (77.4 to 79.2%), lip region with two annules. No males were found in the samples. This population was identified as Pratylenchus scribneri, based on the morphological characters (Castillo and Vovlas, 2007). DNA was isolated from individual nematodes followed by proteinase K-based lysis (Wang et al. 2011). The D2/D3 expansion region of the 28S rRNA gene, rDNA-internal transcribed spacer (ITS) region, and mitochondrial cytochrome oxidase I (mtDNA-COI) gene were amplified with primers D2A/-D3B (De Ley et al. 1999), TW81/ AB28 (Vovlas et al. 2011) and JB3/ JB5 (Liu et al. 2018), respectively. The PCR products were purified and ligated into pJET 1.2/blunt cloning vectors and transformed to Escherichia coli strain DH5α, and then sequenced. The obtained 28S rRNA gene D2/D3 region sequences (785bp), ITS sequences (886 bp) and mtDNA-COI (447bp) in this study were submitted to GenBank. The D2/D3 region of the 28S rRNA sequences of the RLN collected in Lingshui (GenBank accession no. MZ701843) showed 99.75% identity with P. scribneri sequences available in the GenBank (KX842628 and KX842625). The ITS sequences of the RLN collected in this study (MZ701842) showed the highest identity of 97.06% with P. scribneri sequences available in the GenBank (KX842626). The mtDNA-COI sequences of the RLN collected in this study (OK040228) showed 100% identity with P. scribneri (MN366409). Both morphological and molecular data confirmed the identity of P. scribneri. P. scribneri has been reported on corn in Inner Mongolia, Hebei, Shanxi, Shandong, Henan, Jiangsu, and Liaoning provinces of China (Li et al. 2019). As far as we know, this is the first report of P. scribneri on corn in Hainan Province, China. Since the RLN can cause considerable damage to corn, strategic measures should be taken to prevent the spread of P. scribneri to other regions in China.

FECUNDITY OF INTESTINAL STRONGYLATA IN HORSES IN CRITICAL CONDITIONS OF CENTRAL YAKUTIA

Horse herd farming based on the year-round pasture management of horses has become one of the main branches of animal husbandry and the basis of the lifestyle and economy of the Yakutia population. Herds are formed according to their sex and age: stallions, mares, and foals are kept in the herd. Helminth infections are widespread among herd horses and are chronic in most cases without pronounced clinical manifestations. Horses seem to be completely healthy from the point of view of the usual visions of the disease. Therefore, no preventive or treatment measures are often applied. In cases of asymptomatic helminth infections (subclinical forms), the huge economic damage caused by them is determined not so much by the death of animals, but it is very difficult to endure wintering in Yakutia with a high invasion degree, and loss of fatness and weight. The Authors of the Article studied the interspecific relationships of equine intestinal nematodes in mixed invasion, and detected changes in the population density of individual nematode species in horses of different age and in different seasons of the year. The Strongylata incidence and infection rate in herd horses was studied based on the results of quantitative coproovoscopic and larvoscopic studies of faeces from herd horses, and on the results of incomplete helminthological dissections on horse ranches of Central Yakutia.