Distribution, Diversity, and Soil Associations of Wine Grape Plant-Parasitic Nematodes in Georgia, U.S.A., Vineyards.

Wine grape (Vitis vinifera and V. vinifera hybrids) production in Georgia occurs in three distinct regions (North, West, and South) which can be characterized by sandy, sandy-loam, or sandy clay-loam soils. We studied plant-parasitic nematode (PPN) communities in 15 wine grape vineyards from the three primary growing regions to understand which nematodes are a concern and what soil characteristics are associated with their occurrence and relative abundance. Twelve genera of PPNs were detected throughout the state: Belonolaimus, Helicotylenchus, Hemicycliophora, Heterodera, Hoplolaimus, Meloidogyne, Mesocriconema, Paratrichodorus, Paratylenchus, Pratylenchus, Tylenchorhynchus, and Xiphinema. Nonmetric multidimensional scaling ordination and multirank permutation procedure identified PPN community differences and soil characteristics that were associated by region. Indicator species analysis identified Helicotylenchus, Mesocriconema, Tylenchorhynchus, and Xiphinema as statistically associated with the West while Meloidogyne and Paratrichodorus were associated with the South. Our analyses further suggested that soil texture (percent sand, percent clay, and percent silt) and the lime buffer capacity at equilibrium (LBCEQ) were associated with PPN community structure while pH was not. When focused on a single vineyard in the North, multiple logistic regression analysis suggested a statistically significant association between Meloidogyne spp. and soil characteristics, including percentages of sand, pH, and LBCEQ. Our study supports the association between soil characteristics and specific nematode genera, as well as the emergence of LBCEQ, the soil measurement with the strongest statistical association with nematode community structure and Meloidogyne presence.

Differences in distribution and community structure of plant-parasitic nematodes in pecan orchards between two ecoregions of Georgia.

In Georgia, pecans are commercially grown in the Piedmont and Coastal Plain ecoregions which are characterized by sandy-loam, sandy, and/or clay soils. If well-drained, these soils are suitable for pecan production, but the soil characteristics differ enough between ecoregions in which the plant-parasitic nematode (PPN) communities could differ substantially. We studied PPN communities in pecan orchards to evaluate the potential for ecoregion differences. In total, 11 genera (Helicotylenchus, Hemicycliophora, Heterodera, Hoplolaimus, Meloidogyne, Mesocriconema, Pratylenchus, Paratylenchus, Paratrichodorus, Tylenchorhynchs, Xiphenema) were recovered from pecan orchards in the Piedmont and Coastal Plain ecoregions. However, Non-Metric Multi-Dimensional Scaling ordination, Multi-Rank Permutation Procedure, and Indicator Species Analyses indicated that the pecan PPN communities strongly differed between ecoregions and that different genera were strongly associated with different ecoregions. For 9 of the 11 PPN genera, the maximum counts occurred in Coastal Plain locations, suggesting that the well-drained sandy soils of the Coastal Plain and comparatively ill-drained red clay soils of the Piedmont may be conducive and unfavorable for movement/reproduction of PPNs, respectively.

Rapid detection of pecan root-knot nematode, Meloidogyne partityla, in laboratory and field conditions using loop-mediated isothermal amplification.

Meloidogyne partityla is the dominant root-knot nematode (RKN) species parasitizing pecan in Georgia. This species is known to cause a reduction in root growth and a decline in the yields of mature pecan trees. Rapid and accurate diagnosis of this RKN is required to control this nematode disease and reduce losses in pecan production. In this study, a loop-mediated isothermal amplification (LAMP) method was developed for simple, rapid, and on-site detection of M. partityla in infested plant roots and validated to detect the nematode in laboratory and field conditions. Specific primers were designed based on the sequence distinction of the internal transcribed spacer (ITS)-18S/5.8S ribosomal RNA gene between M. partityla and other Meloidogyne spp. The LAMP detection technique could detect the presence of M. partityla genomic DNA at a concentration as low as 1 pg, and no cross reactivity was found with DNA from other major RKN species such as M. javanica, M. incognita and M. arenaria, and M. hapla. We also conducted a traditional morphology-based diagnostic assay and conventional polymerase chain reaction (PCR) assay to determine which of these techniques was less time consuming, more sensitive, and convenient to use in the field. The LAMP assay provided more rapid results, amplifying the target nematode species in less than 60 min at 70°C, with results 100 times more sensitive than conventional PCR (~2-3 hrs). Morphology-based, traditional diagnosis was highly time-consuming (2 days) and more laborious than conventional PCR and LAMP assays. These features greatly simplified the operating procedure and made the assay a powerful tool for rapid, on-site detection of pecan RKN, M. partityla. The developed LAMP assay will facilitate accurate pecan nematode diagnosis in the field and contribute to the management of the pathogen.

First report of the stubby-root nematode Nanidorus minor infecting Paspalum vaginatum, seashore paspalum grass in Georgia, USA.

We found that Nanidorus spp. was pathogenic to seashore paspalum (Paspalum vaginatum) turfgrass as its population increased from 100 to 2,080 nematodes per pot 180 days after inoculation under greenhouse conditions. Morphological measurements of adult females were similar to those described for N. minor. Molecular analysis also confirmed the morphological identification by targeting three different regions of the genomic DNA. Three primer pairs targeting 18S rDNA (360F/932R), 28S rDNA (D2A/D3B) and ITS1 rDNA (BL18/5818) were used in singleplex PCR. Forward and reverse sequences of each individual primer set were then subjected to multiple alignment and the complimentary sequences were assembled into a consensus sequence. Upon nucleotide blast on the NCBI website, they were all confirmed to be N. minor. A one-step multiplex PCR method using specific primers and a fragment size of 190 bp also confirmed the identity of N. minor. To the best of our knowledge, this is the first report of N. minor infecting seashore paspalum turfgrass in Georgia.

Effect of spirotetramat and fluensulfone on population densities of Mesocriconema xenoplax and Meloidogyne incognita on peach.

Management of plant-parasitic nematodes (PPNs) on peach is needed for a longer period of time than is typically afforded by pre-plant fumigant nematicides. Two post-plant nematicides, spirotetramat and fluensulfone, were evaluated for control of Meloidogyne incognita and Mesocriconema xenoplax under laboratory and greenhouse conditions. In vitro assays were conducted to test the effect of spirotetramat at 0.017 and 0.026 kg a.i./ha and fluensulfone at 3.92 kg a.i./ha on the mobility of both M. incognita and M. xenoplax in 24-well plates for 24, 48, and 72 hr, compared to a water control. Both fluensulfone and spirotetramat reduced mobility of M. xenoplax, but only fluensulfone reduced the mobility of M. incognita, compared to the untreated control. In peach greenhouse trials, both spirotetramat at 0.017 kg a.i./ha and fluensulfone at 3.92 kg a.i./ha reduced M. incognita numbers by 62 and 77% at 40 d after inoculation (DAI), respectively; neither chemical reduced populations at 70 DAI. Fluensulfone reduced M. xenoplax numbers by 84, 94, and 96% at 30, 60, and 90 DAI, respectively. No effects were observed for spirotetramat on M. xenoplax. At 40 DAI, dual applications of spirotetramat 30 d apart reduced M. incognita numbers by 58 and 54% for both 0.017 and 0.026 kg a.i./ha rates, respectively; no reductions were observed at 70 DAI. No effect was seen for a dual application of spirotetramat on M. xenoplax. These post-plant nematicides may provide additional options for management of PPNs on peach.Management of plant-parasitic nematodes (PPNs) on peach is needed for a longer period of time than is typically afforded by pre-plant fumigant nematicides. Two post-plant nematicides, spirotetramat and fluensulfone, were evaluated for control of Meloidogyne incognita and Mesocriconema xenoplax under laboratory and greenhouse conditions. In vitro assays were conducted to test the effect of spirotetramat at 0.017 and 0.026 kg a.i./ha and fluensulfone at 3.92 kg a.i./ha on the mobility of both M. incognita and M. xenoplax in 24-well plates for 24, 48, and 72 hr, compared to a water control. Both fluensulfone and spirotetramat reduced mobility of M. xenoplax, but only fluensulfone reduced the mobility of M. incognita, compared to the untreated control. In peach greenhouse trials, both spirotetramat at 0.017 kg a.i./ha and fluensulfone at 3.92 kg a.i./ha reduced M. incognita numbers by 62 and 77% at 40 d after inoculation (DAI), respectively; neither chemical reduced populations at 70 DAI. Fluensulfone reduced M. xenoplax numbers by 84, 94, and 96% at 30, 60, and 90 DAI, respectively. No effects were observed for spirotetramat on M. xenoplax. At 40 DAI, dual applications of spirotetramat 30 d apart reduced M. incognita numbers by 58 and 54% for both 0.017 and 0.026 kg a.i./ha rates, respectively; no reductions were observed at 70 DAI. No effect was seen for a dual application of spirotetramat on M. xenoplax. These post-plant nematicides may provide additional options for management of PPNs on peach.

Fusarium wilt of cotton may commonly result from the interaction of Fusarium oxysporum f. sp. vasinfectum with Belonolaimus longicaudatus.

The interaction between Fusarium oxysporum f. sp. vasinfectum (Fov) and Meloidogyne incognita (root-knot nematode) resulting in Fusarium wilt (FW) of cotton is well-known. Although Belonolaimus longicaudatus (sting nematode) can also interact with Fov and cause FW, it has long been believed that virtually all of the FW in Georgia is caused by the interaction of Fov with M. incognita. In recent years, FW has been reported more frequently in Georgia, which suggests that something affecting the disease complex may have changed. In 2015 and 2016, a survey of 27 Georgia cotton fields in 10 counties was conducted. At least 10 soil and stem samples per field were collected from individual plants showing symptoms of FW to quantify plant-parasitic nematode levels and identify Fov races. Fov race 1 was identified in all samples in 2015, but one sample also had the LA110 genotype and another sample also had the LA108 genotype. In 2016, all Fov races and genotypes found in 2015 were present, however, MDS-12 and LA127/140 also were found. Meloidogyne incognita was present in 18% of fields in 2015 and 40% in 2016, whereas B. longicaudatus was present in all fields in 2015 and 75% of fields in 2016. Regardless of whether they occurred separately or together, M. incognita and B. longicaudatus were present, respectively, in 18% and 55% of individual samples in 2015 and 40% and 51% in 2016. However, M. incognita without B. longicaudatus was found in 7% of samples in 2015 and 34% in 2016, whereas B. longicaudatus without M. incognita was found in 45% of samples in 2015 and 44% in 2016. We conclude that Fov race 1 continues to be the dominant race in Georgia and many instances of FW in Georgia may be due to Fov interacting with B. longicaudatus and not M. incognita as previously believed.The interaction between Fusarium oxysporum f. sp. vasinfectum (Fov) and Meloidogyne incognita (root-knot nematode) resulting in Fusarium wilt (FW) of cotton is well-known. Although Belonolaimus longicaudatus (sting nematode) can also interact with Fov and cause FW, it has long been believed that virtually all of the FW in Georgia is caused by the interaction of Fov with M. incognita. In recent years, FW has been reported more frequently in Georgia, which suggests that something affecting the disease complex may have changed. In 2015 and 2016, a survey of 27 Georgia cotton fields in 10 counties was conducted. At least 10 soil and stem samples per field were collected from individual plants showing symptoms of FW to quantify plant-parasitic nematode levels and identify Fov races. Fov race 1 was identified in all samples in 2015, but one sample also had the LA110 genotype and another sample also had the LA108 genotype. In 2016, all Fov races and genotypes found in 2015 were present, however, MDS­12 and LA127/140 also were found. Meloidogyne incognita was present in 18% of fields in 2015 and 40% in 2016, whereas B. longicaudatus was present in all fields in 2015 and 75% of fields in 2016. Regardless of whether they occurred separately or together, M. incognita and B. longicaudatus were present, respectively, in 18% and 55% of individual samples in 2015 and 40% and 51% in 2016. However, M. incognita without B. longicaudatus was found in 7% of samples in 2015 and 34% in 2016, whereas B. longicaudatus without M. incognita was found in 45% of samples in 2015 and 44% in 2016. We conclude that Fov race 1 continues to be the dominant race in Georgia and many instances of FW in Georgia may be due to Fov interacting with B. longicaudatus and not M. incognita as previously believed.

First Report of the Yellow Nutsedge Cyst Nematode, Heterodera cyperi, in Georgia, U.S.A.

Soil samples collected during a survey for plant-parasitic nematodes in Tift County GA in summer 2017 were submitted for routine diagnosis of nematodes to the Extension Nematology Lab at the Department of Plant Pathology, University of Georgia, Athens, Georgia. Cyst nematodes recovered by centrifugal flotation technique were discovered in the samples from two research sites in a field with a history of tobacco and vegetable production. Cyst nematodes from tobacco (10 cysts/100 cm 3 of soil) and vegetable (2 cysts/100 cm 3 of soil) sites had similar morphological features. Morphology and morphometric measurements of the cysts and J2 ( Fig. 1A-C ) were in agreement with those of Heterodera cyperi ( Golden et al., 1962 ; Romero and López-Llorca, 1996 ). Measurements of J2 ( n = 12) included the length (range = 443-494 µm, mean = 467.4 µm) and width (18.3-24.4 µm, 20.6 µm) of body, stylet (19.1-20.8 µm, 20.3 µm), tail (61.6.0-66.4 µm, 64.2 µm), body width at anus (11.9-14.1 µm, 12.8 µm), and hyaline tail terminus (22.7-29.2 µm, 26.3 µm). The lateral field of J2 had three lines. Cysts ( n = 10; Fig. 1C ) were lemon-shaped, light to dark brown in color with protruding neck and vulval cone. The cysts had ambifenestrated vulval cone and no bullae was present. Morphometrics included body length excluding neck (370.5-714.4 µm, 555.7 µm); body width (165.6-411.1 µm, 310.9 µm); neck length (36.5-66.3 µm, 49.8 µm); fenestra length (26.3-42.5 µm, 35.8 µm), and fenestra width (19.1-31.5 µm, 23.8 µm). DNA was extracted from single cysts ( n = 3) and internal transcribed spacer (ITS) of rRNA and partial cytochrome oxidase I ( COI ) genes were amplified with primers TW81/AB28 and Het-coxiF/Het-coxiR, respectively ( Subbotin et al., 2001 ; Subbotin, 2015 ) and sequenced. The resulting sequences were deposited into the GenBank database (Accession no. MG825344 and MG857126) and also subjected to BLAST searches in the database. ITS sequence of H. cyperi showed 100% similarity (100% coverage) with that of a H. cyperi population from Spain (AF274388). COI sequence of H. cyperi showed 89% similarity (98% coverage) with that of H. guangdongensis (MF425735), and 88% similarity (83% coverage) with that of H. elachista (KC618473). The pathogenicity of H. cyperi was examined under greenhouse conditions using tobacco cv. K340, tomato cv. Tribute, cucumber cv. Thunder, and yellow nutsedge ( Cyperus esculentus L.). 3-wk-old seedlings of the test plants were transferred into Deepot D25L cell containers (5-cm-diam. × 25.4-cm deep) filled with sterilized sand: sand: soil mixture (1:2) and then inoculated with 1,000 eggs and J2 of H. cyperi . The plants were grown for 90 d in a greenhouse before examination of roots and extraction of cysts from the soil. Results showed that the nematode failed to reproduce on tobacco, tomato, and cucumber whereas white females and mature cysts of H. cyperi were observed on yellow nutsedge roots ( Fig. 1E ). The results confirmed that yellow nutsedeg was a host for the nematode, and tobacco, tomato, or cucumber were non-hosts. In the United States, H. cyperi was reported from Florida, North Carolina, and Arkansas ( Subbotin et al., 2010 ) infecting Cyperus spp. Yellow nutsedge is considered a serious weed problem in many cropping systems including peanut, cotton, tobacco, and vegetable crops in the Southern United States. To our knowledge, this is the first report of H. cyperi infecting yellow nutsedge in Georgia. Figure 1Photomicrographs of Heterodera cyperi from yellow nutsedge in Georgia. Whole body (A), the anterior region (B), and the posterior region (C) of J2. Cysts (D) recovered from the soil and the vulval cone of cyst with the ambifenestrate fenestra (E). A mature cyst (F) on the surface of yellow nutsedge root infected with the nematode.

Incidence and Pathogenicity of Plant-Parasitic Nematodes Associated with Blueberry (Vaccinium spp.) Replant Disease in Georgia and North Carolina.

Blueberry replant disease (BRD) is an emerging threat to continued blueberry (Vaccinium spp.) production in Georgia and North Carolina. Since high populations of ring nematode Mesocriconema ornatum were found to be associated with commercially grown blueberries in Georgia, we hypothesized that M. ornatum may be responsible for predisposing blueberry to BRD. We therefore tested the pathogenicity of M. ornatum on 10-wk-old Rabbiteye blueberries (Vaccinium virgatum) by inoculating with initial populations (Pi) of 0 (water control), 10, 100, 1,000. and 10,000 mixed stages of M. ornatum/pot under both greenhouse (25 ± 2°C) and field microplot conditions. Nematode soil population densities and reproduction rates were assessed 75, 150, 225, and 255, and 75, 150, 225, and 375 d after inoculation (DAI) in both the greenhouse and field experiments, respectively. Plant growth parameters were recorded in the greenhouse and field microplot experiments at 255 and 375 DAI, respectively. The highest M. ornatum population density occurred with the highest Pi level, at 75 and 150 DAI under both greenhouse (P < 0.01) and field (P < 0.01) conditions. However, M. ornatum rate of reproduction increased significantly in pots receiving the lowest Pi level of 10 nematodes/plant compared with the pots receiving Pi levels of 100, 1,000, and 10,000 nematodes 75 DAI. Plant-parasitic nematode populations were determined in commercial blueberry replant sites in Georgia and North Carolina during the 2010 growing season. Mesocriconema ornatum and Dolichodorus spp. were the predominant plant-parasitic nematodes in Georgia and North Carolina, respectively, with M. ornatum occurring in nearly half the blueberry fields sampled in Georgia. Other nematode genera detected in both states included Tylenchorhynchus spp., Hoplolaimus spp., Hemicycliophora spp., and Xiphinema spp. Paratrichodorus spp. was also found only in Georgia. In Georgia, our results indicate that blueberry is a host for M. ornatum and its relationship to BRD warrants further investigation.

Differences in Immune Defense Evasion of Selected Inbred Lines of Heterorhabditis Bacteriophora in Two White Grub Species.

We determined virulence of seven Heterorhabditis bacteriophora strain GPS11 inbred lines possessing superior infective juvenile longevity, and heat and ultra violet radiation tolerance against white grubs Popillia japonica and Cyclocephala borealis. At 1 and 2 weeks after treatment, inbred line A2 was significantly more virulent towards P. japonica compared to the parent strain GPS11 and inbred lines A7, A8, A12 and A21; and line A2 caused significantly higher C. borealis mortality than lines A6 and A12. Penetration, encapsulation and survival of two inbred lines, A2 and A12, that showed the highest and lowest virulence against both grub species were then assessed. There were no differences between the two lines for the total number of nematodes penetrated in either P. japonica or C. borealis within the first 24 h, but a significantly higher percentage of penetrated nematodes were alive in line A2 compared to the line A12 in both grub species. P. japonica immune response over time to hemocoel-injected nematodes of A2, A12 and the parent strain was further investigated. While all injected nematodes were encapsulated at 6 h post injection, non-encapsulated living nematodes were detected at 12 and 24 h post injection, showing the breakage out of encapsulation. A higher percentage of non-encapsulated living nematodes and a lower percentage of dead nematodes were found in line A2 as compared to the line A12 after 12 h post injection. These data suggest that virulence differences in the studied H. bacteriophora inbred lines are not due to differences in nematode penetration or recognition by the grub immune system, but are related to the ability of the infective juveniles to break out of encapsulation.

Genetic variation and relationships between isolates and species of the entomopathogenic nematode genus Heterorhabditis deciphered through isozyme profiles.

We studied variation in isozyme patterns of 8 metabolic enzymes in 5 species of Heterorhabditis (H. bacteriophora, H. indica, H. marelata, H. megidis, and H. zealandica) comprising 18 isolates. Isozyme banding patterns of all the 8 enzymes were species specific; however, 3 enzymes, i.e., arginine kinase, fumarate hydratase, and malate dehydrogenase, displayed distinct patterns among all the 18 isolates. Phylogenetic analysis of the isozyme patterns produced dendrograms depicting a high degree of genetic variation among Heterorhabditis species, with the average pairwise distance of 0.2000. Trees constructed using different phylogenetic methods showed a relatively close genetic relationship between H. megidis and H. zealandica and between H. bacteriophora and H. indica. Also, H. bacteriophora HP88 was the most distant species from H. megidis (UK isolate), H. marelatus (Oregon isolate), and H. zealandica (X1 isolate) with pairwise distance of 0.1957, 0.2228, and 0.2120, respectively. Phylogenetic analysis also revealed genetic variation among H. bacteriophora isolates with the average pairwise distance of 0.1507. GPS2 and GPS3 were the most closely related isolates with the average distance of only 0.0870, followed by GPS1 and GPS2 with average distance of 0.1087. In contrast, KMD19 and HP88, OH25, and HP88, and OH25 and Acows isolates were the most divergent populations with a pairwise distance of 0.2011 and 37 character differences. Pairwise distance analysis also revealed that genetic divergence among populations of H. bacteriophora is relatively independent of geographic distance. Overall, these results demonstrate strong subspecies structuring in H. bacteriophora.

Comparative analysis of the expressed genome of the infective juvenile entomopathogenic nematode, Heterorhabditis bacteriophora.

We report the first cDNA-sequencing project of the entomopathogenic nematode, Heterorhabditis bacteriophora. A total of 1246 expressed sequence tags (ESTs) were generated by random sequencing of clones from a cDNA library of the infective juvenile stage. The ESTs were annotated resulting in 1072 useful ESTs that were categorized into functional categories according to Kyoto Encyclopedia of Genes and Genomes. Approximately 459 of 1072 ESTs (43%) had significant similarities to annotated sequences in GenBank. Of these, 417 had significant similarities to the free-living nematode Caenorhanditis elegans proteins. Most ESTs (18%) belonged to the genetic information processing category followed by metabolism (15% ESTs) and environmental information processing (15%) pathways. Several interesting ESTs were found that may have roles in the infectivity and survival of infective juveniles. These included proteases, dauer pathway genes (akt-1, pdk-1 & daf-7) and aging and stress resistance genes such as superoxide dismutase (sod-4), heat shock genes (hsp-4 & hsp-6), and eat genes, and signaling proteins like G-protein coupled receptors, regulators of G-protein signaling (rgs), and serine/threonine kinases. Other interesting ESTs include systemic RNAi defective protein (sid-1), ribonuclease III family members (rnh-2 &rnc) and transposase gene (Tc3A). About 67% of the ESTs did not find matches in any of the searched databases suggesting potentially novel genes in this enomopathogenic nematode. Note: Sequences described in this paper have been deposited in Genbank under the accessions DN 152655-DN 152999, and DN 153000-DN 153726.

Infection Behavior and Overwintering Survival of Foliar Nematodes, Aphelenchoides fragariae, on Hosta.

We studied the pathogenicity and overwintering survival of the foliar nematode, Aphelenchoides fragariae, infecting Hosta spp. Nematodes applied to either lower or upper sides of noninjured and injured hosta leaves were able to infect and produce typical symptoms on nine cultivars. Leaves of only four cultivars (Borschi, Fragrant Blue, Patomic Pride, and Olive Bailey Langdon) showed no symptoms of nematode infection. The nematodes overwintered as juveniles and adults in soil, dry leaves, and dormant buds, but not in roots. Nematode winter survival was higher in dormant buds and soil from the polyhouse than in an open home garden. Of the nematodes found in the dormant buds, 35% to 79% were located between the first two outside layers of the buds. The nematodes tolerated 8 hr exposure to 40 degrees C and -80 degrees C in leaf tissues. Relative humidity influenced nematode migration from soil to leaves. The presence of nematodes only on the outer surface of foliage (leaves and petioles) confirmed the migration of A. fragariae on the surface of the plants. Of the total number of nematodes found on the foliage, 25% to 46% and 66% to 77% were alive at 90% and 100% relative humidity, respectively, suggesting that high moisture is required for the survival and upward movement of nematodes. We conclude that A. fragariae can overwinter in soil, infected dry leaves, and dormant buds and migrate in films of water on the outer surface of the plant during spring to leaves to initiate infection.

Both heat-shock and cold-shock influence trehalose metabolism in an entomopathogenic nematode.

Heat-shock response is highly conserved in animals and microorganisms, and it results in the synthesis of heat-shock proteins. In yeast, heat-shock response has also been reported to induce trehalose accumulation. We explored the relationship between heat- (35 C) or cold-shock (1 and 10 C) and trehalose metabolism in the entomopathogenic nematode, Heterorhabditis bacteriophora. Because both heat- and cold-shocks may precede desiccation stress in natural soil environments, we hypothesized that nematodes may accumulate a general desiccation protectant, trehalose, under both situations. Indeed, both heat- and cold-shocks influenced trehalose accumulation and activities of enzymes of trehalose metabolism in H. bacteriophora. Trehalose increased by 5- and 6-fold in heat- and cold-shocked infective juveniles, respectively, within 3 hr of exposure, compared with the nematodes maintained at 25 C (culture temperature). The activity of trehalose-6-phosphate synthase (T6PS), an enzyme involved in the synthesis of trehalose, also significantly increased in both heat- and cold-shocked nematodes during the first 3 hr of exposure. Generally, the trehalose levels and activities of T6PS declined to their original levels within 3 hr when nematodes were transferred back to 25 C. In both heat- and cold-shocked nematodes, trehalase activity decreased significantly within the first 3 hr and generally returned to the original levels within 3 hr when these nematodes were transferred back to 25 C. The results demonstrate that the trehalose concentrations in H. bacteriophora are influenced by both heat- and cold-shocks and are regulated by the action of 2 trehalose-metabolizing enzymes, T6PS and trehalase. The accumulated trehalose may enhance survival of nematodes under both cold and warm conditions, but it may also provide simultaneous protection against desiccation that may result from subsequent evaporation or freezing. This is the first report of the relationship between trehalose metabolism and heat-shock for the Nematoda.

Effectiveness of a Hot Water Drench for the Control of Foliar Nematodes Aphelenchoides fragariae in Floriculture.

Effectiveness of a hot water drench for the control of Aphelenchoides fragariae infesting hosta (Hosta sp.) and ferns (Matteuccia pensylvanica) was studied. Drenching with hot water at 70 degrees C and 90 degrees C in October reduced (P < 0.05) A. fragariae in the soil but not in the leaves relative to the control (25 degrees C) 300 days after treatment (DAT). Plants drenched with 90 degrees C water had lower numbers of nematode-infected leaves per plant than those treated with 25 degrees C and 70 degrees C water (P < 0.05). Hot water treatments had no adverse effect on the growth parameters of hosta. Boiling water (100 degrees C) applied once a month for 3 consecutive months (April, May, June) consistently reduced the number of infected leaves and the severity of infection relative to the control 150 DAT in hosta but not in ferns (P < 0.05). Boiling water (100 degrees C) caused a 67% reduction in A. fragariae population in hosta leaves, 50% in fern fronds, and 61% to 98% in the soil over the control 150 DAT. A boiling water drench had no effect on the fern growth but caused 49% and 22% reduction in the number and size of hosta leaves, respectively, over the control in 2002. We conclude that 90 degrees C water soil drench in the autumn or early spring could prove effective in managing foliar nematodes on hosta in nurseries and landscapes.

Identification of alternatives for the management of foliar nematodes in floriculture.

The foliar nematodes, Aphelenchoides spp, have emerged as important pests of ornamentals in North America during the last decade. Due to the ban on the use of potentially toxic pesticides, there are currently no nematicides registered to manage foliar nematodes on ornamentals. Therefore, we have evaluated a biological [Burkholderia cepacia (syn Pseudomonas cepacia)], two plant products [clove (Syzygium aromaticum) extract and Nimbecidine (azadirachtin)] and twelve chemical pesticides registered for the management of insects, mites, slugs or diseases of ornamentals, against Aphelenchoides fragariae on the most popular ornamental, hosta (Hosta spp), for two consecutive years. We found ZeroTol (270 g liter-1 peroxyacetic acid), currently labeled as a broad-spectrum fungicide/algicide, to be a very potent nematicide that killed 100% of the nematodes in water suspension. It also caused over 70% reduction in A fragariae population in soil and in the leaves without any phytotoxicity. B cepacia caused 67-85% reduction in A fragariae population in leaves and 50% reduction in the soil whereas insecticidal soap caused over 72% reduction in leaves and 61% reduction in the soil. Clove extract and Nimbecidine did not show any potential for the control of A fragariae on hosta. Although all twelve chemical pesticides were effective in reducing the population of A fragariae in the soil 45 days after treatment (DAT), only diazinon 475 g liter-1 EC, trichlorfon 800 g kg-1 SP, ethoprophos 100 g kg-1 GR, oxamyl 100 g kg-1 GR and ZeroTol caused over 70% reduction in nematode population compared with the control. In the leaves, only diazinon EC, trichlorfon SP, insecticidal soap, oxamyl GR and ZeroTol consistently caused over 70% nematode population reduction compared with the control at 45 DAT in both years. Thus, only diazinon EC, trichlorfon SP, oxamyl GR and ZeroTol consistently caused over 70% reduction in nematode population both in soil and leaves. Due to the recent ban by the US Environmental Protection Agency on the use of the first three of these formulations, only ZeroTol would serve as an effective tool to manage foliar nematodes in ornamentals. Although not as effective as ZeroTol in the soil, insecticidal soap is the only other alternative for foliar nematode management.