Parasitism of Hoplolaimus galeatus on Diploid and Polyploid St. Augustine grasses

''Floratam'' and ''FX-313'' St. Augusfinegrasses (Stenotaphrum secundatum) were compared in a time-course experiment for host suitability and susceptibility to the lance nematode, Hoplolaimus galeatus. Nematode densities were determined in the soil and acid-fuchsin stained roots 42, 84, 126, 168, and 210 days after pots containing 230 cm³ of autoclaved native Margate fine sand/pot were infested with 104 ± 9 nematodes and maintained at 25 ± 2 C in the laboratory. ''FX-313'' was a more suitable host for H. galeatus. Numbers of H. galeatus reached a maximum at 210 days after inoculation, with 5,550 and 4,120 nematodes (adults plus juveniles)/pot for ''FX-313'' and ''Floratam,'' respectively. Root and shoot dry weights of both grasses were not affected by H. galeatus throughout the experiment. Three polyploid, 2n = 30 to 32 (''Floratam,'' ''FX-10,'' and ''Bitterblue'') and three diploid, 2n = 18 (''FX-313,'' ''Florida Common,'' and ''Seville'') S. secundatum genotypes were inoculated with H. galeatus (99 ± 9/pot) and compared with uninoculated controls 210 days after inoculation. St. Augustinegrass genotypes differed as hosts of H. galeatus. ''FX-313'' and ''Florida Common'' represented the high and low extremes, respectively, for nematode reproduction (9,750 and 5,490 nematodes/pot or 4,239 and 2,387 nematodes/100 cm³ of soil). However, differences in root and shoot growth were not detected 210 days after inoculation with H. galeatus.     

In Vitro Culture and Feeding Behavior of Belonolaimus longicaudatus on Excised Zea mays Roots

A greenhouse population of the sting nematode, Belonolaimus longicaudatus, obtained from an infested golf course in California''s Coachella Valley, was surface-decontaminated and cuhured on excised roots of Zea mays supported by Gamborg''s B5 medium. At 26-27 °C the females laid eggs, and newly emerged juveniles of the second generation completed three molts within 29 days after egg deposition. Sixty days after inoculation with 60 females and 40 males, an average of 529 nematodes and 83 eggs were recovered from the culture. The feeding process consisted of probing, stylet penetration, ingestion, and stylet retraction. Feeding seemed to be necessary before egg deposition or molting occurred. The sting nematode was observed feeding exclusively as an ectoparasite and preferably at the region of cell division and elongation. Vigorous feeding by many nematodes usually caused discoloration of root tips and termination of growth.     

Host Status of 'SeaIsle 1' Seashore Paspalum (Paspalum vaginatum) to Belonolaimus longicaudatus and Hoplolaimus galeatus

Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging pathogens of turfgrasses in Florida. However, the host status of seashore paspalum (Paspalum vaginatum) is unknown. Glasshouse experiments were performed in 2002 and 2003 to determine the tolerance of ''SeaIsle 1'' seashore paspalum to a population of B. longicaudatus and a population of H. galeatus, and to compare to ''Tifdwarf'' bermudagrass for differences. Both nematode species reproduced well on either grass, but only B. longicaudatus consistently reduced root growth as measured by root length. Belonolaimus longicaudatus reduced root growth (P ≤ 0.05) by 35% to 45% at 120 days after inoculation on both grasses. In 2003, higher inoculum levels of H. galeatus reduced root growth (P ≤ 0.05) by 19.4% in seashore paspalum and by 14% in bermudagrass after 60 and 120 days of exposure, respectively. Percentage reductions in root length caused by H. galeatus and B. longicaudatus indicated no differences between grass species, although Tifdwarf bermudagrass supported higher soil population densities of both nematodes than SeaIsle 1 seashore paspalum.     

Yield Reduction and Root Damage to Cotton Induced by Belonolaimus longicaudatus

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.     

Slit Injection of 1,3-Dichloropropene for Management of Belonolaimus longicaudatus on Established Bermudagrass

Belonolaimus longicaudatus is a serious problem on bermudagrass, a common warm-season turfgrass, in Florida. The cancellation of organophosphate nematicides necessitates that new management tools be identified for use on sports turf. Postplant application of 1,3-dichloropropene (1,3-D) on bermudagrass was evaluated for management of B. longicaudatus on golf course fairways and driving ranges. A series of 10 experiments were conducted to evaluate the effectiveness of 1,3-D in reducing population densities of B. longicaudatus and enhancing bermudagrass recovery from nematode damage. In 5 of 10 experiments, 1,3-D injected at 46.8 liters/ha was effective in reducing population densities of B. longicaudatus (P < 0.05) compared to untreated plots 2 to 4 weeks after treatment. One month after treatment, population densities of B. longicaudatus ranged from 59% to 97% of those in untreated plots. Nematode suppression generally lasted 2 months or less. Turf visual performance was improved following injection with 1,3-D (P < 0.05) over untreated plots when other factors were not limiting. Turf root development also was enhanced following injection with 1,3-D. Postplant injection of 1,3-D could be a useful nematode management tool for certain sports turf applications.     

Alternatives to Fenamiphos for Management of Plant-Parasitic Nematodes on Bermudagrass

Plant-parasitic nematodes can be very damaging to turfgrasses. The projected cancellation of the registration for fenamiphos in the near future has generated a great deal of interest in identifying acceptable alternative nematode management tactics for use on turfgrasses. Two field experiments were conducted to evaluate the effectiveness of repeated applications of several commercially available nematicides and root biostimulants for reducing population densities of plant-parasitic nematodes and (or) promoting health of bermudagrass in nematode-infested soil. One experimental site was infested with Hoplolaimus galeatus and Trichodorus obtusus, the second with Belonolaimus longicaudatus. In both trials, none of the experimental treatments reduced population densities (P ≤ 0.1) of plant-parasitic nematodes, or consistently promoted turf visual performance or turf root production. Nematologists with responsibility to advise turf managers regarding nematode management should thoroughly investigate the validity of product claims before advising clientele in their use.     

Effects of Pratylenchus zeae and Quinisulcius acutus Alone and in Combination on Sorghum

Host-parasite relationships of Pratylenchus zeae and Quinisulcius acutus, alone or in combination, were studied on sorghum in the greenhouse and laboratory. Q. acutus at 1,000 or 5,000 nematodes per 15-cm-d pot and P. zeae at 500 nematodes per pot significantly suppressed plant height and fresh and oven dry shoot and root weights. A mixture of 1,000 Q. acutus and 500 P. zeae per pot resulted in greatest suppression of growth. Roots of plants inoculated with Q. acutus alone were reduced in number and size and showed lesions and discoloration. Reproduction of this nematode 42 days after inoculation was much greater in treatments of 100 or 1,000 than 5,000 nematodes. The population density of the two species at 6 weeks after inoculation was significantly less when combined than for each species alone. When the two species were combined, reproduction of P. zeae was greater than that of Q. acutus, but the final populations per gram of root weight were the same. Q. acutus fed ectoparasitically on epidermal cells of sorghum roots in the zone of elongation and differentiation when observed under in vitro conditions.     

Spatial Patterns of Belonolaimus spp. Among and Within Citrus Orchards on Florida's Central Ridge

A survey was initiated to determine the incidence of Belonolaimus spp. (sting nematodes) in citrus orchards in the central ridge region of Florida, following widespread damage by these nematodes to young trees replanted after freezing weather in 1989-90. Sting nematodes were detected in 50% of 210 samples and in 64% of 84 orchards surveyed. More orchards in Polk County were infested with sting nematodes (82%) than in counties to the north (36%) or south (48%). Principal component analysis of morphometric data separated six of seven sting nematode populations in northeastern Polk County from six populations in adjacent regions. Stylet:tail ratio for nematodes in northeastern Polk County tend to be > 1.0 and were ≪ 1.0 for all other populations. Patchiness of nematodes within an orchard was associated with stunted trees (23% smaller), reduced root mass density (25% lower), and low fruit yield (57% reduction). Soil texture did not vary among trees of different size in the orchard, but soil water potential between irrigation events was highest beneath small trees with low root mass density. Results of the survey indicate that the incidence of sting nematodes in orchards on the central ridge is much higher than previously estimated and that sting nematodes can cause substantial damage in replanted orchards. Further research is needed to evaluate the significance of sting nematode population variability and its relationship to citrus crop loss in Florida.     

Influence of Nematodes and Light Sources on Growth and Nodulation of Soybean

The influence of nematodes on nodulation of soybean varied according to their modes of parasitism. In the greenhouse, nodule formation was stimulated by the endoparasites, Meloidogyne hapla and Pratylenchus penetrans, but was inhibited slightly by the ectoparasite, Belonolaimus longicaudatus. In an experiment under controlled conditions in a phytotron, Heterodera glycines severely inhibited nodule formation, whereas plants inoculated with B. longicaudatus and P. penetrans had more nodules per g root than nematode-free plants. Nitrogen-fixing capacity, however, was inhibited by all three nematode species. Different light sources used in the phytotron experiment also influenced growth and nodulation of soybean. A fluorescent plus incandescent light regime resulted in plants with the greatest shoot weight, pod number, and nodules per g root. Plants grown under Lucalox lamps had excessive stem elongation.     

Effects and Dynamics of a Nematode Community on Soybean

The relationships between densities of all members of a plant-parasitic nematode community and yield of ''Davis'' soybean and between final and preplant population levels were examined in small plots on sandy soils in north-central Florida. Plant-parasitic nematodes present in the community included Belonolaimus longicaudatus, Criconemella sphaerocephala, Meloidogyne incognita, Paratrichodorus minor, Pratylenchus brachyurus, and Xiphinema sp. Plant growth, including stand count, soybean yield (kg/ha), and size of young plants, was occasionally inversely correlated (P ≤ 0.05) with densities of B. longicaudatus or P. brachyurus, but not with densities of other species or with a range of soil variables. The nature of this relationship varied with season, with more severe stand losses noted during 1987 than in 1988. Final population densities (Pf) of most nematode species showed significant (P ≤ 0.05) linear relationships to densities measured at planting or earlier (Pi). These relationships were stronger (higher r²) with the ectoparasite B. longicaudatus than with the endoparasites M. incognita and P. brachyurus. Criconemella sphaerocephala declined under soybean cultivation, reaching levels near zero after two seasons. A quadratic model showed an improvement (P ≤ 0.05) over the linear model in describing the relationship between Pf and Pi measured at planting for B. longicaudatus, and gave a better indication of the leveling off of Pf at high values of Pi.     

Effects of a Commercial Formulation of Bacillus firmus I-1582 on Golf Course Bermudagrass Infested with Belonolaimus longicaudatus

One of the primary pests of bermudagrass (Cynodon spp.) on golf courses in the southeastern United States is Belonolaimus longicaudatus (sting nematode). In 2011, a commercial formulation of Bacillus firmus I-1582, Nortica 5WG, was launched in the United States for management of plant-parasitic nematodes on turfgrasses. To test the efficacy of late winter/early spring application of this biopesticide on B. longicaudatus, two field trials in 2009 compared B. firmus with fenamiphos and untreated control treatments. In 2011, two additional field trials compared treatment with B. firmus with untreated control only. These trials measured treatment effects on the population density of B. longicaudatus, turf root length, and turf percent green cover. In all four trials, treatment with B. firmus improved root length and decreased numbers of B. longicaudatus in contrast to the untreated. These results indicate that late winter/early spring application of B. firmus is an effective biopesticide treatment for management of B. longicaudatus on golf course bermudagrass.     

Effects and Dynamics of a Nematode Community on Maize

Relationships between nematode density and yield and between final and preplant population levels were examined in small maize plots on sandy soils in north-central Florida. Plant-parasitic nematodes present in the community included Belonolaimus longicaudatus, Criconemella sphaerocephala, Meloidogyne incognita, Paratrichodorus minor, Pratylenchus brachyurus, and a Xiphinema sp. Plant growth--including stand count, grain yield, stalk weight, and size of young plants--often was inversely correlated (P ≤ 0.05) with densities of B. longicaudatus and occasionally with P. brachyurus, but not with densities of other species or with a range of soil variables. More severe losses in grain yields from B. longicaudatus occurred in 1987 than in 1988, although mean preplant nematode densities in February were similar in both years (4.4 vs. 3.9/100 cm³ soil). Final population densities of most nematode species were linearly related (P ≤ 0.05) to densities measured at planting or earlier. These relationships were stronger (higher r²) with the ectoparasites B. longicaudatus and C. sphaerocephala than with the endoparasites M. incognita and P. brachyurus. No significant correlations were found between population densities of different nematode species.     

Effect of Soybean Tip Removal on Penetration and Development of Heterodera glycines

On a few occasions, soybeans with broken root tips were included in tests to evaluate resistance to Heterodera glycines. Although females developed on these plants, the numbers tended to be lower than on similarly treated intact roots. To test the possibility that removal of the root meristem affected nematode development, a culture system using pruned soybeans was devised that permitted access to the roots without disturbing the plants. Treatments included removal of 2 mm of root tip at various times ranging from 24 hours before to 10 days after inoculation, or roots left intact. In each experiment, all roots were inoculated at the same time with equal numbers of freshly hatched second-stage juveniles of Heterodera glycines. No differences in nematode development were detected in plants with root tips removed after inoculation compared to the control. When tips were removed at or before inoculation, fewer juveniles entered roots and relatively fewer nematodes developed. Penetration levels and development correlated with root tip removal such that progressively fewer nematodes entered roots and relatively greater numbers of nematodes remained undeveloped as the time interval between root tip removal and inoculation was increased.     

Saline Irrigation Affects Belonolaimus longicaudatus and Hoplolaimus galeatus on Seashore Paspalum

Seashore paspalum (Paspalum vaginatum) has great potential for use in salt-affected turfgrass sites. Use of this grass on golf courses, athletic fields, and lawns in subtropical coastal areas may aid in conservation of freshwater resources. Belonolaimus longicaudatus and Hoplolaimus galeatus are considered among the most damaging root pathogens of turfgrasses in Florida. Glasshouse experiments were performed in 2002 and 2003 to examine the effects of increasing levels of irrigation salinity on B. longicaudatus and H. galeatus. Irrigation treatments were formulated by concentrating deionized water to six salinity levels (0, 5, 10, 15, 20, and 25 dS/m). Final population densities of H. galeatus followed a negative linear regression (r² = 0.92 and 0.83; P <= 0.01) with increasing salinity levels. Final population densities of B. longicaudatus were quadratically (r² = 0.72 and 0.78; P <= 0.01) related to increasing salinity levels from 0 to 25 dS/m. An increase in population densities of B. longicaudatus was observed at moderate salinity levels (10 and 15 dS/m) compared to 0 dS/m. Root-length comparisons revealed that B. longicaudatus caused root stunting at low salinity levels, 0 to 10 dS/m, but roots were not affected at 15 to 25 dS/m. These results indicate that the ability of B. longicaudatus to feed and stunt root growth was negatively affected at salinity levels of 15 dS/m and above.     

Ultrastructure and Development of Pasteuria sp. (S-1 strain), an Obligate Endoparasite of Belonolaimus longicaudatus (Nemata: Tylenchida)

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.     

Damage Function and Economic Threshold for Belonolaimus longicaudatus on Potato

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.     

Effects of a Mutant Strain and a Wild Type Strain of Verticillium lecanii on Heterodera glycines Populations in the Greenhouse

A wild type strain ofVerticillium lecanii and a mutant strain with increased tolerance to the fungicide benomyl were evaluated in greenhouse experiments for effects on Heterodera glycines populations. Nematodes were applied at 300 eggs and juveniles per 4,550-cm³ pot (two soybean plants in 4,990 g loamy sand per pot) and at both 300 and 10,000 eggs and juveniles per 1,720-cm³ pot (one soybean plant in 2,060 g sand per pot). With 300 nematodes added per pot, both V. lecanii strains significantly reduced nematode populations in loamy sand (fungus applied at 0.02% dry weight per dry weight loamy sand) and sand (0.006% and 0.06% fungus application rates). The mutant strain applied at 0.002% to sand also significantly reduced cyst numbers. When 10,000 nematodes were added per pot, only the mutant strain at 0.06% significantly decreased population. Various media were tested for isolation of the fungus strains from prills, loamy sand, and sand, but the fungi were recovered from few of the greenhouse pots.     

Effects of Belonolaimus longicaudatus Management and Nitrogen Fertility on Turf Quality of Golf Course Fairways

Field experiments evaluated the effects of nematicide and fertility on performance of ‘Tifway 419’ bermudagrass parasitized by the sting nematode (Belonolaimus longicaudatus). Plot treatments were nontreated or nematicide (1,3-dichloropropene) treated combined with different nitrogen (N) fertilizer levels. Effects of treatments on numbers of B. longicaudatus and turf performance were compared. Nematicide consistently reduced numbers of B. longicaudatus, but fertilizer level had no effect on B. longicaudatus. Turf performance of nematicide-treated plots was improved compared with nontreated plots during both experiments. Increasing N fertilizer level improved turf performance in nematicide-treated plots in some cases, but had no effect on turf performance in nontreated plots in either experiment. Results suggest that increasing N fertilizer levels may not improve turf performance at sites infested with B. longicaudatus unless nematode management tactics are effective in reducing nematode densities.     

Induced Resistance to Meloidogyne hapla by other Meloidogyne species on Tomato and Pyrethrum Plants

Advance inoculation of the tomato cv. Celebrity or the pyrethrum clone 223 with host-incompatible Meloidogyne incognita or M. javanica elicited induced resistance to host-compatible M. hapla in pot and field experiments. Induced resistance increased with the length of the time between inoculations and with the population density of the induction inoculum. Optimum interval before challenge inoculation, or population density of inoculum for inducing resistance, was 10 days, or 5,000 infective nematodes per 500-cm³ pot. The induced resistance suppressed population increase of M. hapla by 84% on potted tomato, 72% on potted pyrethrum, and 55% on field-grown pyrethrum seedlings, relative to unprotected treatments. Pyrethrum seedlings inoculated with M. javanica 10 days before infection with M. hapla were not stunted, whereas those that did not receive the advance inoculum were stunted 33% in pots and 36% in field plots. The results indicated that advance infection of plants with incompatible or mildly virulent nematode species induced resistance to normally compatible nematodes and that the induced resistance response may have potential as a biological control method for plant nematodes.