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1.  Belonolaimus longicaudatus: An Emerging Pathogen of Peanut in Florida 
Journal of Nematology  2015;47(2):87-96.
Sting nematode (Belonolaimus longicaudatus) is an economically important ectoparasitic nematode that is highly pathogenic on a wide range of agricultural crops in sandy soils of the southeastern United States. Although this species is commonly found in Florida in hardwood forests and as a soilborne pathogen on turfgrasses and numerous agronomic and horticultural crops, it has not been reported infecting peanut. In the summers of 2012 and 2013, sting nematode was found infecting three different peanut cultivars being grown on two separate peanut farms in Levy County, FL. The damage consisted of large irregular patches of stunted, chlorotic plants at both farms. The root systems were severely abbreviated and there were numerous punctate-like isolated lesions observed on pegs and pods of infected plants. Sting nematodes were extracted from soil collected around the roots of diseased peanut over the course of the peanut season at both farm sites. Peanut yield from one of these nematode-infested sites was 64% less than that observed in areas free from sting nematodes. The morphological characters of the nematode populations in these fields were congruous with those of the original and other published descriptions of B. longicaudatus. Moreover, the molecular analyses based on the sequences of D2/D3 expansion fragments of 28S rRNA and internal transcribed spacer (ITS) rRNA genes from the nematodes further collaborates the identification of the sting nematode isolates as B. longicaudatus. The sequences were deposited in GenBank (accession no. KF963097, KF963098 for ITS, and KF96399, KF963100 for D2-D3). The results of the phylogenetic analysis using the sequences of these isolates from peanut compared with those of other isolates from Florida suggests that the sting nematode from both peanut farms are genetically close to B. longicaudatus populations occurring in the state. Peanut plants inoculated with both nematode isolates showed punctate-like isolated lesions on pods and pegs, and an abbreviation of their root systems, whereas those symptoms were not observed on noninoculated peanut plants. To our knowledge, this is the first report of large-scale field damage caused by sting nematode infecting peanut grown under field conditions in Florida.
PMCID: PMC4492293  PMID: 26170470
Arachis hypogaea; Belonolaimus longicaudatus; emerging pathogen; morphology; pathogenicity; peanut; phylogenetics; sting nematode
2.  Effects of the Mi-1, N and Tabasco Genes on Infection and Reproduction of Meloidogyne mayaguensis on Tomato and Pepper Genotypes 
Journal of Nematology  2007;39(4):327-332.
Meloidogyne mayaguensis is a damaging root-knot nematode able to reproduce on root-knot nematode-resistant tomato and other economically important crops. In a growth chamber experiment conducted at 22 and 33°C, isolate 1 of M. mayaguensis reproduced at both temperatures on the Mi-1-carrying tomato lines BHN 543 and BHN 585, whereas M. incognita race 4 failed to reproduce at 22°C, but reproduced well at 33°C. These results were confirmed in another experiment at 26 ± 1.8°C, where minimal or no reproduction of M. incognita race 4 was observed on the Mi-1-carrying tomato genotypes BHN 543, BHN 585, BHN 586 and ‘Sanibel’, whereas heavy infection and reproduction of M. mayaguensis isolate 1 occurred on these four genotypes. Seven additional Florida M. mayaguensis isolates also reproduced on resistant ‘Sanibel’ tomato at 26 ± 1.8°C. Isolate 3 was the most virulent, with reproduction factor (Rf) equal to 8.4, and isolate 8 was the least virulent (Rf = 2.1). At 24°C, isolate 1 of M. mayaguensis also reproduced well (Rf ≥ 1) and induced numerous small galls and large egg masses on the roots of root-knot nematode-resistant bell pepper ‘Charleston Belle’ carrying the N gene and on three root-knot nematode-resistant sweet pepper lines (9913/2, SAIS 97.9001 and SAIS 97.9008) carrying the Tabasco gene. In contrast, M. incognita race 4 failed to reproduce or reproduced poorly on these resistant pepper genotypes. The ability of M. mayaguensis isolates to overcome the resistance of tomato and pepper genotypes carrying the Mi-1, N and Tabasco genes limits the use of resistant cultivars to manage this nematode species in infested tomato and pepper fields in Florida.
PMCID: PMC2586510  PMID: 19259507
Capsicum annuum; bell pepper; resistance; root-knot nematodes; Solanum lycopersicum; sweet pepper
3.  Transfer and Development of Pasteuria penetrans  
Journal of nematology  2007;39(1):55-61.
Pasteuria penetrans isolate P-20 has been attributed as the cause of soil suppressiveness to peanut root-knot nematode in Florida. In this study, P. penetrans was transferred from a suppressive site to a new site and established by growing susceptible hosts to the peanut root-knot nematode during both summer and winter seasons. When two soil fumigants, 1,3-dichloropropene (1,3-D) and chloropicrin, were applied broadcast at the rate of 168 liters/ha and 263 kg/ha, respectively, the bacterium was not adversely affected by 1,3-D but was adversely affected by chloropicrin. In autumn 2005, after the harvest of the second peanut crop, the greatest number of J2 was recorded in the chloropicrin-treated plots, followed by the non-fumigated plots and 1,3-D-fumigated plots. The percentage J2 encumbered with endospores, endospores per J2 and percentage of P. penetrans-infected females were greatest in the non-fumigated plots, followed by 1,3-D- and chloropicrin-fumigated plots. This study demonstrates that P. penetrans can be transferred from a suppressive site to a new site and increased to suppressive densities against the peanut root-knot nematode.
PMCID: PMC2586474  PMID: 19259476
Arachis hypogaea; biological control; chloropicrin; 1,3-dichloropropene; fumigation; management; Meloidogyne arenaria; Pasteuria penetrans; peanut; peanut root-knot nematode; suppressive soil; transferability
4.  Effects of Application Methods and Plastic Covers on Distribution of Cis- and Trans-1,3-Dichloropropene and Chloropicrin in Root Zone 
Journal of Nematology  2005;37(4):483-488.
This study examined the effects of three application methods (chisel injection, Avenger coulter injection, and drip irrigation) and two plastic films (polyethylene film [PE] and virtually impermeable film [VIF]) on distribution of cis- and trans- 1,3-dichloropropene (1,3-D) and chloropicrin (CP) in a Florida sandy soil after application of Telone C35 or Telone In-Line. Regardless of application method, VIF retained greater amounts of cis- and trans-1,3-D and CP in the root zone with longer residential time than PE. There was better retention of the three compounds in the root zone when applied with the Avenger coulter injection rig than chisel injection, especially in combination with VIF. Distribution of the three compounds in the root zone was less predictable when applied by drip irrigation. Following drip irrigation, more than 50% of the three compounds in the PE and VIF-covered beds was found near the end of the drip tapes in one experiment, whereas the distribution was much more uniform in the root zone in a second experiment. Among the three biologically active compounds, CP disappeared from the root zone more rapidly than cis- and trans-1,3-D, especially in the PE-covered beds.
PMCID: PMC2620995  PMID: 19262895
1,3-dichloropropene; chloropicrin; distribution; fumigant; fumigation; methyl bromide alternative; polyethylene mulch; root zone; virtually impermeable film
5.  Effects of Application Strategies of Fumigant and Nonfumigant Nematicides on Cantaloupe Grown in Deep Sand Soils in Florida 
Journal of Nematology  2005;37(3):281-284.
A 2-year study was conducted in which three treatment tactics of oxamyl (at planting application, application every 2 weeks, and rescue applications, as determined by crop symptoms) were compared to fumigant treatments with methyl bromide, 1,3-dichloropropene (1,3-D), and 1,3-D plus chloropicrin for management of Meloidogyne spp. In 2002, treatments that included 1,3-D produced higher yields as determined both by number and weight of marketable fruit. All treatment tactics relying solely on oxamyl, at planting, scheduled treatments, and rescue, were not different from untreated controls for both marketable yield and number of fruit. Gall ratings in 2002 were lowest for 1,3-D at the 112-liters/ha rate, followed by 1,3-D at 84 liters/ha with and without oxamyl. All treatments of oxamyl, except when combined with 1,3-D, had gall ratings not different from untreated plots. In 2004, treatments of methyl bromide and 1,3-D plus chloropicrin had the highest total number of both marketable fruit and highest marketable yields. All treatment strategies relying solely on oxamyl had yields equivalent to the untreated controls. Mean root-gall ratings were lowest for methyl bromide plus chloropicrin and 1,3-D plus chloropicrin treatments. Root-gall ratings for all treatment tactics relying solely on oxamyl were not different from untreated controls.
PMCID: PMC2620971  PMID: 19262874
1,3-dichloropropene; cantaloupe; chloropicrin; Cucumis melo; fumigation; gall rating; management; Meloidogyne; methyl bromide; nonfumigant; oxamyl; rescue; root-knot nematode
6.  Distribution and Downward Movement of Pasteuria penetrans in Field Soil 
Journal of Nematology  2005;37(2):155-160.
Endospores of Pasteuria penetrans were evaluated for their vertical distribution in field soil and their downward movement through soil in the laboratory. In the field trial, the number of endospores attached to second-stage juveniles (J2) of Meloidogyne arenaria race 1 varied greatly in different soil depths. There were higher percentages of J2 with endospores attached in former weed fallow plots during the first 3 years of growing peanut than in former bahiagrass and rhizomal peanut plots (P ≤ 0.05). In weed fallow plots a higher average number of endospores per J2 were maintained in all depths, upper three depths, and upper four depths in 1999, 2000, and 2001, respectively (P ≤ 0.05). However, in 2002, there were no differences in the percentages of J2 with endospores attached and in the average of the numbers of endospores per J2 among the treatments (P > 0.05). In laboratory trials, P. penetrans endospores were observed to move throughout the soil through the percolation of water. After one application of water, some endospores were detected 25 to 37.5 cm deep. Endospores were present at the greatest depth, 37.5 to 50 cm, after the third application of water. These results indicate that rain or water applications by irrigation are likely to move endospores to deeper levels of the soil, but the majority of endospores remain in the upper 0-to-30-cm depth.
PMCID: PMC2620961  PMID: 19262855
bacterium; biological control; distribution; Meloidogyne arenaria; nematode; Pasteuria penetrans; peanut; movement; root-knot nematode; suppressive soil
7.  Persistence and Suppressiveness of Pasteuria penetrans to Meloidogyne arenaria Race 
Journal of Nematology  2004;36(4):540-549.
The long-term persistence and suppressiveness of Pasteuria penetrans against Meloidogyne arenaria race 1 were investigated in a formerly root-knot nematode suppressive site following 9 years of continuous cultivation of three treatments and 4 years of continuous peanut. The three treatments were two M. arenaria race 1 nonhost crops, bahiagrass (Paspalum notatum cv. Pensacola var. Tifton 9), rhizomal peanut (Arachis glabrata cv. Florigraze), and weed fallow. Two root-knot nematode susceptible weeds commonly observed in weed fallow plots were hairy indigo (Indigofera hirsuta) and alyce clover (Alysicarpus vaginalis). The percentage of J2 with endospores attached reached the highest level of 87% in 2000 in weed fallow, and 63% and 53% in 2002 in bahiagrass and rhizomal peanut, respectively. The percentage of endospore-filled females extracted from peanut roots grown in weed fallow plots increased from nondetectable in 1999 to 56% in 2002, whereas the percentages in bahiagrass and rhizomal peanut plots were 41% and 16%, respectively. Over 4 years, however, there was no strong evidence that endospores densities reached suppressive levels because peanut roots, pods, and pegs were heavily galled, and yields were suppressed. This might be attributed to the discovery of M. javanica infecting peanut in this field in early autumn 2001. A laboratory test confirmed that although the P. penetrans isolate specific to M. arenaria attached to M. javanica J2, no development occurred. In summary, P. penetrans increased on M. arenaria over a 4-year period, but apparently because of infection of M. javanica on peanut at the field site root-knot disease was not suppressed. This was confirmed by a suppressive soil test that showed a higher level of soil suppressiveness than occurred in the field (P ≤ 0.01).
PMCID: PMC2620794  PMID: 19262836
Arachis hypogaea; biological control; Meloidogyne arenaria; M. javanica; nematode; Pasteuria penetrans; peanut; persistence; root-knot nematode; suppressive soil
8.  Morphological and Molecular Characterization of Meloidogyne mayaguensis Isolates from Florida 
Journal of Nematology  2004;36(3):232-240.
The discovery of Meloidogyne mayaguensis is confirmed in Florida; this is the first report for the continental United States. Meloidogyne mayaguensis is a virulent species that can reproduce on host cultivars bred for nematode resistance. The perineal patterns of M. mayaguensis isolates from Florida show morphological variability and often are similar to M. incognita. Useful morphological characters for the separation of M. mayaguensis from M. incognita from Florida are the male stylet length values (smaller for M. mayaguensis than M. incognita) and J2 tail length values (greater for M. mayaguensis than M. incognita). Meloidogyne mayaguensis values for these characters overlap with those of M. arenaria and M. javanica from Florida. Enzyme analyses of Florida M. mayaguensis isolates show two major bands (VS1-S1 phenotype) of esterase activity, and one strong malate dehydrogenase band (Rm 1.4) plus two additional weak bands that migrated close together. Their detection requires larger amounts of homogenates from several females. Amplification of two separate regions of mitochondrial DNA resulted in products of a unique size. PCR primers embedded in the COII and 16S genes produced a product size of 705 bp, and amplification of the 63-bp repeat region resulted in a single product of 322 bp. Nucleotide sequence comparison of these mitochondrial products together with sequence from 18S rDNA and ITS1 from the nuclear genome were nearly identical with the corresponding regions from a M. mayaguensis isolate from Mayaguez, Puerto Rico, the type locality of the species. Meloidogyne mayaguensis reproduced on cotton, pepper, tobacco, and watermelon but not on peanut. Preliminary results indicate the M. mayaguensis isolates from Florida can reproduce on tomato containing the Mi gene. Molecular techniques for the identification of M. mayaguensis will be particularly useful in cases of M. mayaguensis populations mixed with M. arenaria, M. incognita, and M. javanica, which are the most economically important root-knot nematode species in Florida, and especially when low (<25) numbers of specimens of these species are recovered from the soil.
PMCID: PMC2620774  PMID: 19262811
isozyme; Meloidogyne arenaria; M. incognita; M. javanica; M. mayaguensis; mitochondrial DNA; molecular diagnosis; morphology; nematode; root-knot nematode; taxonomy
9.  Meloidogyne javanica on Peanut in Florida 
Journal of Nematology  2003;35(4):433-436.
A mixed population of Meloidogyne arenaria race 1 and M. javanica race 3 is reported on peanut from a field in Levy County, Florida. Confirmation of M. javanica on peanut is based on esterase and malate dehydrogenase isozyme patterns resolved on polyacrylamide slab gels following electrophoresis, and perineal patterns. Up to 29% of 290 individual females collected from peanut roots in the field in autumn 2002 showed a typical esterase J3 phenotype for M. javanica. This is the third report of M. javanica infecting peanut in the United States.
PMCID: PMC2620691  PMID: 19262776
Arachis hypogaea; electrophoresis; esterase phenotype; host race; malate dehydrogenase phenotype; Meloidogyne arenaria; Meloidogyne javanica; nematode; peanut; root-knot nematode
10.  Temporal Formation and Immunolocalization of an Endospore Surface Epitope During Pasteuria penetrans Sporogenesis 
Journal of Nematology  2003;35(3):278-288.
The synthesis and localization of an endospore surface epitope associated with the development of Pasteuria penetrans was determined using a monoclonal antibody (MAb) as a probe. Nematodes, uninfected or infected with P. penetrans, were harvested at 12, 16, 24, and 38 days after inoculation (DAI) and then examined to determine the developmental stage of the bacterium. Vegetative growth of P. penetrans was observed only in infected nematodes harvested at 12 and 16 DAI, whereas cells at different stages of sporulation and mature endospores were observed at 24 and 38 DAI. ELISA and immunoblot analysis revealed that the adhesin-associated epitope was first detected at 24 DAI, and increased in the later stages of sporogenesis. These results indicate that the synthesis of adhesin-related proteins occurred at a certain developmental stage relative to the sporulation process, and was associated with endospore maturation. Immunofluorescence microscopy indicated that the distribution of the epitope is nearly uniform on the periphery of each spore, as defined by parasporal fibers. Immunocytochemistry at the ultrastructural level indicated a distribution of the epitope over the parasporal fibers. The epitope also was detected over other structures such as sporangium and exosporium during the sporogenesis process, but it was not observed over the cortex, inner-spore coat, outer-spore coat, or protoplasm. The appearance of the adhesin epitope first at stage III of sporogenesis and its presence on the parasporal fibers are consistent with an adhesin-related role in the attachment of the mature endospore to the cuticle of the nematode host.
PMCID: PMC2620646  PMID: 19262762
biological control; ELISA; immunoblot; immunofluorescence; immunogold labeling; Meloidogyne arenaria; monoclonal antibody; nematode; Pasteuria penetrans; root-knot nematode; SDS-PAGE
11.  Pasteuria spp.: Systematics and Phylogeny of These Bacterial Parasites of Phytopathogenic Nematodes 
Journal of Nematology  2003;35(2):198-207.
Pasteuria spp. include endospore-forming bacterial pathogens of cladoceran crustaceans and plant-parasitic nematodes. Propagation of these nematode pathogens requires attachment of soilborne endospores to nematode hosts, infection, growth, sporulation, and release of endospores to repeat the cycle of infection and propagation. The ability of these bacteria to suppress the levels of plant-parasitic nematodes in the field has made them particularly promising candidates for biocontrol of nematode diseases of plants. Genes encoding 16S ribosomal RNA have been sequenced for the cladoceran (water flea) parasite and type species, Pasteuria ramosa, and for Pasteuria spp. isolated from root-knot (Meloidogyne arenaria race 1 and Meloidogyne sp.), soybean cyst (Heterodera glycines), and sting (Belonolaimus longicaudatus) nematodes. These have provided a phylogenetic basis for their designation to a distinct clade within the family Alicyclobacillaceae of the gram-positive endospore-forming bacteria. Two apparent biotypes of P. penetrans demonstrating a host preference for different Meloidogyne spp. showed identical 16S rDNA sequences, suggesting host-recognition evolves within a given species. The sequences of genes encoding sporulation transcription factors, sigE and sigF, from P. penetrans biotype P-20 show different phylogenetic relationships to other endospore-forming bacteria, supporting their application to further discriminate Pasteuria spp. and biotypes. Distribution of an adhesin-associated epitope on polypeptides from different Pasteuria isolates provides an immunochemical approach to differentiate species and biotypes with specific host preferences. Application of bioinformatics to genomic data, as well as further characterization of the biochemical basis for host recognition, will facilitate development of Pasteuria spp. as benign alternatives to chemical nematicides.
PMCID: PMC2620627  PMID: 19265995
biological control; cladocerans; endospore; Meloidogyne arenaria; nematode; Pasteuria penetrans; Pasteuria ramosa; phylogeny
12.  Ultrastructure and Development of Pasteuria sp. (S-1 strain), an Obligate Endoparasite of Belonolaimus longicaudatus (Nemata: Tylenchida) 
Journal of Nematology  2001;33(4):227-238.
Pasteuria sp., strain S-1, is a gram-positive, obligate endoparasitic bacterium that uses the phytoparasitic sting nematode, Belonolaimus longicaudatus, as its host in Florida. The host attachment of S-1 appears to be specific to the genus Belonolaimus with development occurring only in juveniles and adults of B. longicaudatus. This bacterium is characterized from other described species of Pasteuria using ultrastructure of the mature endospore. Penetration, development, and sporogenesis were elucidated with TEM, LTSEM, and SEM and are similar to other nematode-specific Pasteuria. Recent analysis of 16S rDNA sequence homology confirms its congeneric ranking with other Pasteuria species and strains from nematodes and cladocerans, and corroborates ultrastructural, morphological, morphometric, and host-range evidence suggesting separate species status.
PMCID: PMC2620501  PMID: 19265886
Belonolaimus longicaudatus; development; obligate nematode endoparasitic bacterium; Pasteuria sp. (s-1 strain); sporogenesis; sting nematode; ultrastructure
13.  Efficacy of 1,3-Dichloropropene in Soil Amended with Compost and Unamended Soil 
Journal of Nematology  2001;33(4S):289-293.
1,3-Dichloropropene (1,3-D) is a likely alternative soil fumigant for methyl bromide. The objective was to determine root-knot nematode, Meloidogyne incognita, survival in microplots after exposure to 1,3-D for various periods of time in soil that have previously been amended with compost. The treatments were 1,3-D applied broadcast at 112 liters/ha and untreated controls in both compost-amended and unamended soil. Soil samples were collected from each microplot at 6, 24, 48, 72, and 96 hours after fumigation at three depths (0-15, 15-30, and 30-45 cm). One week after fumigation, six tomato seedlings were transplanted into each microplot and root galling was recorded 6 weeks later. Plants grown in fumigated compost-amended soil had more galls than plants from fumigated unamended soil at P ≤ 0.1. Gall indices from roots in fumigated soil amended with compost were not different from nonfumigated controls. Based on soil bioassays, the number of galls decreased with increasing time after fumigation in both compost-amended and unamended soil at 0-to-15 and 15-to-30 cm depths, but not at 30 to 45 cm deep. Higher soil water content due to the elevated levels of organic matter in the soil at these depths may have interfered with 1,3-D movement, thus reducing its efficacy.
PMCID: PMC2620517  PMID: 19265889
compost-amended soil; deep sand soil; 1,3-dichloropropene; fumigation; Lycopersicon esculentum; Meloidogyne incognita; nematicide; nematode; root-knot nematode; tomato
14.  Effect of Sorghum-Sudangrass and Velvetbean Cover Crops on Plant-Parasitic Nematodes Associated with Potato Production in Florida 
Journal of Nematology  2001;33(4S):285-288.
In a 3-year field study, population densities of Belonolaimus longicaudatus and other plant-parasitic nematodes and crop yields were compared between potato (Solanum tuberosum) cropping systems where either sorghum-sudangrass (Sorghum bicolor × S. arundinaceum) or velvetbean (Mucuna pruriens) was grown as a summer cover crop. Population densities of B. longicaudatus, Paratrichodorus minor, Tylenchorhynchus sp., and Mesocriconema sp. increased on sorghum-sudangrass. Population densities of P. minor and Mesocriconema sp. increased on velvetbean. Sorghum-sudangrass increased population densities of B. longicaudatus and Mesocriconema sp. on a subsequent potato crop compared to velvetbean. Potato yields following velvetbean were not greater than following sorghum-sudangrass despite reductions in population densities of B. longicaudatus.
PMCID: PMC2620518  PMID: 19265888
Belonolaimus longicaudatus; cover crop; cropping sequence; Dolichodorus heterocephalus; Hemicycliophora sp.; management; Meloidogyne incognita; Mesocriconema sp.; modeling; Mucuna pruriens; nematode; Paratrichodorus minor; population dynamics; potato; root-knot nematode; Solanum tuberosum; Sorghum bicolor × S. arundinaceum; sorghum-sudangrass; sting nematode; stubby root nematode; Tlenchorhynchus sp.; velvetbean
15.  Rate Response of 1,3-Dichloropropene for Nematode Control in Spring Squash in Deep Sand Soils 
Journal of Nematology  2000;32(4S):524-530.
The soil fumigant 1,3-dichloropropene (1,3-D) formulated with chloropicrin is viewed as a likely alternative for replacing methyl bromide in Florida when the latter is phased out in 2005. Therefore, it behooves us to learn more about using 1,3-D in deep, sand soils. Two trials were conducted on spring squash to determine the most effective rate of 1,3-D for the control of Meloidogyne spp. Rates tested included 0, 56, 84, 112, and 168 liters/ha of 1,3-D applied broadcast with conventional chisels 30 cm deep. The chisel traces were sealed by disking immediately after fumigant application. Cucurbita pepo cv. Sunex 9602 was sown 7 days after fumigation. The population density of plant-parasitic nematodes in soil and root-knot nematode galling severity was determined at 34 and 65 days after planting (DAP), and the number of marketable fruit and yield were determined. The number of fruit and yield were higher in all plots that received 1,3-D than in untreated controls. The number of Meloidogyne spp. second-stage juveniles was lower in all fumigated plots in trial 1 at both 34 and 65 DAP, and in trial 2 at 65 DAP, than in the untreated control. The severity of root galling was decreased with all treatments in both trials, with broadcast rates of 84, 112, and 168 liters/ha providing the best control of root-knot nematodes in spring squash grown in sandy soil. Satisfactory management of root knot on squash grown in early spring months in north Florida can be achieved with low rates of 1,3-D.
PMCID: PMC2620479  PMID: 19271005
Cucurbita pepo; 1,3-dichloropropene; efficacy; fumigation; management; Meloidogyne arenaria; Meloidogyne incognita; nematode; rate; root-knot nematode; squash
16.  Effects of Potato-Cotton Cropping Systems and Nematicides on Plant-Parasitic Nematodes and Crop Yields 
Journal of Nematology  2000;32(3):297-302.
Belonolaimus longicaudatus has been reported as damaging both potato (Solanum tuberosum) and cotton (Gossypium hirsutum). These crops are not normally grown in cropping systems together in areas where the soil is infested with B. longicaudatus. During the 1990s cotton was grown in a potato production region that was a suitable habitat for B. longicaudatus. It was not known how integrating the production of these two crops by rotation or double-cropping would affect the population densities of B. longicaudatus, other plant-parasitic nematodes common in the region, or crop yields. A 3-year field study evaluated the viability of both crops in monocropping, rotation, and double-cropping systems. Viability was evaluated using effects on population densities of plant-parasitic nematodes and yields. Rotation of cotton with potato was found to decrease population densities of B. longicaudatus and Meloidogyne incognita in comparison with continuous potato. Population densities of B. longicaudatus following double-cropping were greater than following continuous cotton. Yields of both potato and cotton in rotation were equivalent to either crop in monocropping. Yields of both crops were lower following double-cropping when nematicides were not used.
PMCID: PMC2620462  PMID: 19270980
Belonolaimus longicaudatus; cotton; crop rotation; cropping system; double-cropping; Gossypium hirsutum; Meloidogyne incognita; nematode; potato; root-knot nematode; Solanum tuberosum; sting nematode
17.  Damage Function and Economic Threshold for Belonolaimus longicaudatus on Potato 
Journal of Nematology  2000;32(3):318-322.
Belonolaimus longicaudatus has long been recognized as a pathogen of potato (Solanum tuberosum). However, a damage function relating expected yield of potato to population densities of B. longicaudatus at planting has not been derived, and the economic threshold for nematicide application is unknown. The objectives of this study were to derive the damage function of B. longicaudatus on potato and to calculate the economic threshold population density. The damage function data for B. longicaudatus on potato were obtained from an ongoing field study to evaluate cropping systems and nematode management practices. Soil samples were collected from experimental field plots, and nematodes were extracted from a 130-cm³ subsample with a centrifugal-flotation method. A damage function was derived by linear regression of potato yield on nematode population density at planting. Based on this derived damage function and published potato prices, the economic threshold for nematicide application was calculated at 2 to 3 B. longicaudatus/130 cm³ of soil, which was near the detection threshold based on methodology used in this study.
PMCID: PMC2620459  PMID: 19270983
Belonolaimus longicaudatus; damage function; economic threshold; nematode; plant disease loss; potato; Solanum tuberosum; sting nematode
18.  Viability of Heterodera glycines Exposed to Fungal Filtrates 
Journal of Nematology  2000;32(2):190-197.
Filtrates from nematode-parasitic fungi have been reported to be toxic to plant-parasitic nematodes. Our objective was to determine the effects of fungal filtrates on second-stage juveniles and eggs of Heterodera glycines. Eleven fungal species that were isolated from cysts extracted from a soybean field in Florida were tested on J2, and five species were tested on eggs in vitro. Each fungal species was grown in Czapek-Dox broth and malt extract broth. No toxic activity was observed for fungi grown in Czapek-Dox broth. Filtrates from Paecilomyces lilacinus, Stagonospora heteroderae, Neocosmospora vasinfecta, and Fusarium solani grown in malt extract broth were toxic to J2, whereas filtrates from Exophiala pisciphila, Fusarium oxysporum, Gliocladium catenulatum, Pyrenochaeta terrestris, Verticillium chlamydosporium, and sterile fungi 1 and 2 were not toxic to J2. Filtrates of P. lilacinus, S. heteroderae, and N. vasinfecta grown in malt extract broth reduced egg viability, whereas F. oxysporum and P. terrestris filtrates had no effect on egg viability.
PMCID: PMC2620443  PMID: 19270965
biological control; egg; Exophiala pisciphila; fungus; Fusarium oxysporum; Fusarium solani; Gliocladium catenulatum; hatching; Heterodera glycines; juvenile; nematode; Neocosmospora vasinfecta; Paecilomyces lilacinus; Pyrenochaeta terrestris; soybean cyst nematode; Stagonospora heteroderae; toxicity; toxin; Verticillium chlamydosporium; viability
19.  Population Dynamics of Belonolaimus longicaudatusin a Cotton Production System 
Journal of Nematology  2000;32(2):210-214.
Belonolaimus longicaudatus is a recognized pathogen of cotton (Gossypium hirsutum), but insufficient information is available on the population dynamics and economic thresholds of B. longicaudatus in cotton production. In this study, data collected from a field in Florida were used to develop models predicting population increases of B. longicaudatus on cotton and population declines under clean fallow. Population densities of B. longicaudatus increased on cotton, reaching a carrying capacity of 139 nematodes/130 cm³ of soil, but decreased exponentially during periods of bare fallow. The model indicated that population densities should decrease each year of monocropped cotton, if an alternate host is not present between sequential cotton crops. Economic thresholds derived from published damage functions and current prices for cotton and nematicides varied from 2 to 5 B. longicaudatus/130 cm³ of soil, depending on the nematicide used.
PMCID: PMC2620437  PMID: 19270968
Belonolaimus longicaudatus; cotton; economic threshold; fallow; Gossypium hirsutum; modeling; nematode; population decline; population dynamics; population increase; sting nematode
20.  Yield Reduction and Root Damage to Cotton Induced by Belonolaimus longicaudatus 
Journal of Nematology  2000;32(2):205-209.
Sting nematode (Belonolaimus longicaudatus) is recognized as a pathogen of cotton (Gossypium hirsutum), but the expected damage from a given population density of this nematode has not been determined. The objective of this study was to quantify the effects of increasing initial population densities (Pi) of B. longicaudatus on cotton yield and root mass. In a field plot study, nematicide application and cropping history were used to obtain a wide range of Pi values. Cotton yields were regressed on Pi density of B. longicaudatus to quantify yield losses in the field. In controlled environmental chambers, cotton was grown in soil infested with increasing Pi's of B. longicaudatus. After 40 days, root systems were collected, scanned on a desktop scanner, and root lengths were measured. Root lengths were regressed on inoculation density of B. longicaudatus to quantify reductions in the root systems. In the field, high Pi's (>100 nematodes/130 cm³ of soil) reduced yields to near zero. In controlled environmental chamber studies, as few as 10 B. longicaudatus/130 cm³ of soil caused a 39% reduction in fine cotton roots, and 60 B. longicaudatus/130 cm³ of soil caused a 70% reduction. These results suggest that B. longicaudatus can cause significant damage to cotton at low population densities, whereas at higher densities crop failure can result.
PMCID: PMC2620440  PMID: 19270967
Belonolaimus longicaudatus; cotton; crop loss; damage function; damage threshold; Gossypium hirsutum; nematode; plant disease loss; root scanning; sting nematode
21.  A Technique for Determining Live Second-stage Juveniles of Heterodera glycines 
Journal of Nematology  2000;32(1):117-121.
We developed a quick and reliable technique to distinguish live and immobile (presumed dead) Heterodera glycines second-stage juveniles (J2) following their treatment with microbial culture filtrates. About 50 J2 in 1 ml of culture filtrate or water were placed in wells of a 24-well tissue-culture plate. After incubation, the nematodes in the wells were observed with the aid of an inverted microscope. The J2 lay straight and their viability could not be determined by direct microscopic observation. With the addition of one or two drops (50 to 100 µl) of 1 N NaOH into the well, the live nematodes changed their body shape from straight to curled or hook-shaped after about 30 seconds. The nematodes that responded to NaOH by changing their body shape within 3 minutes were considered a live, while those nematodes that failed to respond within 3 minutes and were immobile were presumed to be dead. The technique is simple, fast, and useful for the examination of a large number of samples in which one wants to determine the effects of microbial cultural filtrates on nematodes, or in similar tests.
PMCID: PMC2620428  PMID: 19270956
culture filtrate; Heterodera glycines; juvenile; mortality; nematode; second-stage juvenile; technique
22.  Extraction and Purification of Pasteuria spp. Endospores 
Journal of Nematology  2000;32(1):78-84.
Pasteuria penetrans is an endospore-forming bacterial parasite of root-knot nematodes that has potential as a biological control agent. Biochemical investigations of P. penetrans are limited because of difficulty in obtaining large quantities of endospores free of plant debris and contaminating microorganisms. Our objective was to develop a technique for extraction and purification of P. penetrans endospores from root-knot nematodes. Tomato roots infected with Meloidogyne arenaria that was parasitized by P. penetrans were digested with cytolase. The nematode females along with plant debris were washed with a jet stream of water onto an 800-µm-pore sieve nested on a 250-µm-pore sieve. The materials retained on the 250-µm-pore sieve were centrifuged through a 20% sucrose solution. The resulting loose pellet fraction was collected on a 250-µm-pore sieve and then centrifuged through a 47% sucrose solution. Endospore-filled females were handpicked from the 47% sucrose pellicle fraction. Endospores were released by grinding the females with a glass tissue grinder. The endospores were then filtered through a nylon filter with 8-µm openings, collected by centrifugation, and subjected to buoyant density centrifugation in different media. Further purification by buoyant density centrifugation in a linear gradient of sodium diatrizoate resulted in a preparation of endospores free of debris. This additional step may be desirable for the further characterization of components unique to the endospores.
PMCID: PMC2620432  PMID: 19270952
bacterium; biological control; endospore; extraction; Meloidogyne spp.; nematode; Pasteuria penetrans; purification; root-knot nematode; sodium diatrizoate
23.  Phylogenetic Analysis of Pasteuria penetrans by 16S rRNA Gene Cloning and Sequencing 
Journal of Nematology  1999;31(3):319-325.
Pasteuria penetrans is an endospore-forming bacterial parasite of Meloidogyne spp. This organism is among the most promising agents for the biological control of root-knot nematodes. In order to establish the phylogenetic position of this species relative to other endospore-forming bacteria, the 16S ribosomal genes from two isolates of P. penetrans, P-20, which preferentially infects M. arenaria race 1, and P-100, which preferentially infects M. incognita and M. javanica, were PCR-amplified from a purified endospore extraction. Universal primers for the 16S rRNA gene were used to amplify DNA which was cloned, and a nucleotide sequence was obtained for 92% of the gene (1,390 base pairs) encoding the 16S rDNA from each isolate. Comparison of both isolates showed identical sequences that were compared to 16S rDNA sequences of 30 other endospore-forming bacteria obtained from GenBank. Parsimony analyses indicated that P. penetrans is a species within a clade that includes Alicyclobacillus acidocaldarius, A. cycloheptanicus, Sulfobacillus sp., Bacillus tusciae, B. schlegelii, and P. ramosa. Its closest neighbor is P. ramosa, a parasite of Daphnia spp. (water fleas). This study provided a genomic basis for the relationship of species assigned to the genus Pasteuria, and for comparison of species that are parasites of different phytopathogenic nematodes.
PMCID: PMC2620377  PMID: 19270903
bacterium; biological control; Meloidogyne spp.; nematode; Pasteuria penetrans; Pasteuria ramosa; phylogeny; ribosomal RNA; root-knot nematode; 16S rRNA
24.  Review of Pasteuria penetrans: Biology, Ecology, and Biological Control Potential 
Journal of Nematology  1998;30(3):313-340.
Pasteuria penetrans is a mycelial, endospore-forming, bacterial parasite that has shown great potential as a biological control agent of root-knot nematodes. Considerable progress has been made during the last 10 years in understanding its biology and importance as an agent capable of effectively suppressing root-knot nematodes in field soil. The objective of this review is to summarize the current knowledge of the biology, ecology, and biological control potential of P. penetrans and other Pasteuria members. Pasteuria spp. are distributed worldwide and have been reported from 323 nematode species belonging to 116 genera of free-living, predatory, plant-parasitic, and entomopathogenic nematodes. Artificial cultivation of P. penetrans has met with limited success; large-scale production of endospores depends on in vivo cultivation. Temperature affects endospore attachment, germination, pathogenesis, and completion of the life cycle in the nematode pseudocoelom. The biological control potential of Pasteuria spp. have been demonstrated on 20 crops; host nematodes include Belonolaimus longicaudatus, Heterodera spp., Meloidogyne spp., and Xiphinema diversicaudatum. Pasteuria penetrans plays an important role in some suppressive soils. The efficacy of the bacterium as a biological control agent has been examined. Approximately 100,000 endospores/g of soil provided immediate control of the peanut root-knot nematode, whereas 1,000 and 5,000 endospores/g of soil each amplified in the host nematode and became suppressive after 3 years.
PMCID: PMC2620303  PMID: 19274225
bacterium; Belonolaimus longicaudatus; biological control; biology; cyst nematode; dagger nematode; ecology; endospore; Heterodera spp.; Meloidogyne spp.; nematode; Pasteuria penetrans; review; root-knot nematode; sting nematode; Xiphinema diversicaudatum
25.  Response of Meloidogyne spp., Heterodera glycines, and Radopholus similis to Tannic Acid 
Journal of Nematology  1997;29(4S):737-741.
Tannins, which are water-soluble polyphenols, are toxic to numerous fungi, bacteria, and yeasts. Our objectives were to study the efficacy of tannic acid in control of Meloidogyne arenaria on tomato and its effects on the behavior of M. arenaria, M. incognita, Heterodera glycines, and Radopholus similis. Three concentrations of tannic acid, 0.1, 1.0, and 10 g/500 cm³ of soil, were applied preplant (powder) and at-plant (powder and drench) into soil infested with M. arenaria. Tannic acid at the 1.0-g rate reduced galling compared with the untreated control, regardless of methods of application. The 0.1-g rate resulted in no reduction in galling when applied preplant but reduced galling when applied as a drench and in one of two experiments when applied at-plant. The 10-g rate was phytotoxic to tomato seedlings except when applied 7 days preplant. In the latter case, root galling was suppressed to very low numbers. In behavior studies on water agar, Meloidogyne second-stage juveniles were attracted to areas with an increasing tannic acid gradient. Radopholus similis was repelled from the tannic acid gradient in one of two experiments. There was no effect on H. glycines. The response of M. arenaria second-stage juveniles to different concentrations of tannic acid dissolved in alginate was tested. Movement behavior of the second-stage juveniles were observed at 1,000 and 10,000 μg/ml of tannic acid, but not at 10 and 100 μg/ml.
PMCID: PMC2619828  PMID: 19274278
alginate; attractant; burrowing nematode; Heterodera glycines; Meloidogyne arenaria; Meloidogyne incognita; nematode; polyphenol; Radopholus similis; repellent; root-knot nematode; soybean cyst nematode; tannic acid

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