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.
Belonolaimus longicaudatus; development; obligate nematode endoparasitic bacterium; Pasteuria sp. (s-1 strain); sporogenesis; sting nematode; ultrastructure
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.
bacterium; biological control; Meloidogyne spp.; nematode; Pasteuria penetrans; Pasteuria ramosa; phylogeny; ribosomal RNA; root-knot nematode; 16S rRNA
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.
alginate; attractant; burrowing nematode; Heterodera glycines; Meloidogyne arenaria; Meloidogyne incognita; nematode; polyphenol; Radopholus similis; repellent; root-knot nematode; soybean cyst nematode; tannic acid
Effects of tannic acid on hatching of Heterodera glycines eggs were determined in vitro using three batches of eggs obtained from greenhouse cultures in Florida or from naturally infested field soil in Minnesota. A quadratic model fits the percentage egg hatch. Hatch increased with increasing tannic acid concentrations from 0 to about 39 mg/liter, then declined with further increases in concentration. Tannic acid did not induce hatching of dormant eggs obtained from the field.
cyst; dormancy; egg; hatch; Heterodera glycines; soybean cyst nematode; tannic acid
The incidence of adhesion of Pasteuria penetrans endospores to Meloidogyne incognita second-stage juveniles (J2) was studied after pretreatment of the latter with monoclonal antibodies (MAb), cationized ferritin, and other organic molecules in replicated trials. Monoclonal antibodies developed to a cuticular epitope of M. incognita second-stage juveniles gave significant reductions in attachment of P. penetrans endospores to treated nematodes. MAb bound to the entire length of J2 except for the area of the lateral field, where binding was restricted to the incisures. Since reductions in attachment with MAb treatment were modest, it is uncertain if these results implicated a specific surface protein as a factor that interacted in binding of the endospore to the nematode cuticle. Endospore attachment was decreased following treatment of the nematode with the detergents sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB). Endospore attachment to live nematodes was significantly greater than attachment to dead nematodes. Attachment rates of three P. penetrans isolates to M. incognita race 3 varied between isolates. The effects of neuraminidase, pronase, pepsin, trypsin, lipase, and Na periodate on endospore attachment were inconsistent. The cationic dye alcian blue, which binds sulfate and carboxyl groups on acidic glycans, had no consistent effect on endospore attachment. The incidence of endospore attachment was significantly lower but modest, at best, for nematodes that were treated with cationized ferritin alone or cationized ferritin following monoclonal antibody. The lack of consistency or extreme reduction in most experiments suggests that attachment of P. penetrans spores to M. incognita is not specified by only one physico-chemical factor, but may involve a combination of at least two physico-chemical factors (including surface charge and movement of the J2). This points to a need for analysis of combined or factorial treatment effects.
adhesion; bacterium; binding site; biological control; cationized ferritin; cuticle; Meloidogyne incognita; monoclonal antibody; nematode; Pasteuria penetrans; root-knot nematode; surface charge; surface coat
The biological control of Meloidogyne arenaria on peanut (Arachis hypogaea) by Pasteuria penetrans was evaluated using a six x six factorial experiment in field microplots over 2 years. The main factors were six inoculum levels of second-stage juveniles (J2) of M. arenaria race 1 (0, 40, 200, 1,000, 5,000, and 25,000 J2/microplot, except that the highest level was 20,000 J2/microplot in 1995) and six infestation levels of P. penetrans as percentages of J2 with endospores attached (0, 20, 40, 60, 80, and 100%). The results were similar in 1994 and 1995. Numbers of eggs per root system, J2 per 100 cm³ soil at harvest, root galls, and pod galls increased with increasing nematode inoculum levels and decreased with increasing P. penetrans infestation levels (P ≤ 0.05), except that there was no effect of P. penetrans infestation levels on J2 per 100 cm³ soil in 1994 (P> 0.05). There were no statistical interaction effects between the inoculum levels of J2 and the infestation levels of P. penetrans (P > 0.05). When the infestation level was increased by 10%, the number of eggs per root system, root galls, and pod galls decreased 7.8% to 9.4%, 7.0% to 8.5%, and 8.0% to 8.7% in 1994 and 1995, respectively, whereas J2 per 100 cm³ soil decreased 8.8% in 1995 (P ≤ 0.05). The initial infestation level of P. penetrans contributed 81% to 95% of the total suppression of pod galls, whereas the infection of J2 of the subsequent generations contributed only 5% to 19% suppression of pod galls. The major suppressive mechanism of M. arenaria race 1 by P. penetrans on peanut is the initial endospore infestation of J2 at planting.
Arachis hypogaea; bacterium; biological control; endospore; Meloidogyne arenaria; Pasteuria penetrans; peanut; root-knot nematode
The potential of Pasteuria penetrans for suppressing Meloidogyne arenaria race 1 on peanut (Arachis hypogaea) was tested over a 2-year period in a field microplot experiment. Endospores of P. penetrans were mass-produced on M. arenaria race 1 infecting tomato plants. Endospores were inoculated in the first year only at rates of 0, 1,000, 3,000, 10,000, and 100,000 endospores/g of soil, respectively, into the top 20 cm of microplots that were previously infested with M. arenaria race 1. One peanut seedling was planted in each microplot. In the first year, root gall indices and pod galls per microplot were significantly reduced by 60% and 95% for 100,000 endospores/g of soil, and 20% and 65% for 10,000 endospores/g of soil, respectively. Final densities of second-stage juveniles (J2) in soil were not significantly different among the treatments. The number of endospores attached to J2 and percentage of J2 with attached endospores significantly increased with increasing endospore inoculation levels. Pasteuria penetrans significantly reduced the densities of J2 that overwintered. In the second year, root and pod gall indices, respectively, were significantly reduced by 81% and 90% for 100,000 endospores/g of soil, and by 61% and 82% of 10,000 endospores/g of soil. Pod yields were significantly increased by 94% for 100,000 and by 57% for 10,000 endospores/g of soil, respectively. The effect of P. penetrans on final densities of J2 in soil was not significant. Regression analyses verified the role of P. penetrans in the suppression of M. arenaria. The minimum number of endospores required for significantly suppressing M. arenaria race 1 on peanut was 10,000 endospores/g of soil.
Arachis hypogaea; bacterium; biological control; endospore; Meloidogyne arenaria; Pasteuria penetrans; peanut; root-knot nematode
Six methods for quantification of the endospore concentrations of Pasteuria penetrans from tomato roots are described. Mortar disruption and machine disruption methods gave the highest estimations (endospores per gram of root material) of 83.7 and 79.0 million, respectively. These methods were significantly superior to incubation bioassay (47.7 million), enzymatic disruption (32.1 million), and enzymatic disruption + flotation (25.8 million) methods. A centrifugation bioassay method gave the lowest estimation of 12.7 million.
bacterium; biological control; endospore; Lycopersicon esculentum; Meloidogyne arenaria; method; Pasteuria penetrans; root-knot nematode; tomato
Two years of data collected from the Florida Nematode Assay Laboratory of the Florida Cooperative Extension Service and 4 years of data from the Florida Department of Agriculture and Consumer Services, Division of Plant Industry, were compiled to find out the distribution of Pasteuria spp. on nematodes in Florida soils. Information recorded came from 335 samples and included nematode genera with Pasteuria endospores attached, host plants associated with the samples, and the origins of the samples. Pasteuria spp. were detected on 14 different plant-parasitic nematode genera in 41 Florida counties and associated with over 39 different plant species and in seven fallow fields. Pasteuria-infected nematodes were associated with a wide range of plant hosts, although frequency of associations with these hosts reflected the sample bias of the laboratories involved. Meloidogyne and Hoplolaimus spp. were the two nematode genera most frequently associated with Pasteuria. Pasteuria spp. were observed attached to members of these two genera in 176 and 59 soil samples, respectively.
bacterium; biological control; endospore; Florida; lance nematode; Meloidogyne; Hoplolaimus; nematode; Pasteuria penetrans; Pasteuria spp.; root-knot nematode
The effects of 12 summer crop rotation treatments on population densities of Meloidogyne arenaria race 1 and on yields of subsequent spring vegetable crops were determined in microplots. The crop sequence was: (i) rotation crops during summer 1991 ; (ii) cover crop of rye (Secale cereale) during winter 1991-92; (iii) squash (Cucurbita pepo) during spring 1992; (iv) rotation crops during summer 1992; (v) rye during winter 1992-93; (vi) eggplant (Solanum melongena) during spring 1993. The 12 rotation treatments were castor (Ricinus communis), cotton (Gossypium hirsutum), velvetbean (Mucuna deeringiana), crotalaria (Crotalaria spectabilis), fallow, hairy indigo (Indigofera hirsuta), American jointvetch (Aeschynomene americana), sorghum-sudangrass (Sorghum bicolor x S. sudanense), soybean (Glycine max), horsebean (Canavalia ensiformis), sesame (Sesamum indicum), and peanut (Arachis hypogaea). Compared to peanut, the first eight rotation treatments resulted in lower (P ≤ 0.05) numbers of M. arenaria juveniles on most sampling dates. Soybean, horsebean, and sesame rotations were less effective in suppressing nematodes. Yield of squash was greater (P ≤ 0.05) following castor, cotton, velvetbean, and crotalaria than following peanut. Compared to the peanut rotation, yield of eggplant was enhanced (P ≤ 0.10) following castor, crotalaria, hairy indigo, American jointvetch, and sorghum-sudangrass. Several of these rotation crops may provide a means for depressing M. arenaria population densities on a short-term basis to enhance yields in a subsequent susceptible vegetable crop.
Aeschynomene americanum; Arachis hypogaea; Canavalia ensiformis; cropping systems; Crotalaria spectabilis; Cucurbita pepo; fallow; Glycine max; Gossypium hirsutum; Indigofera hirsuta; Meloidogyne arenaria; Mucuna deeringiana; nematode; nematode management; Ricinus communis; Sesamum indicum; Solanum melongena; Sorghum bicolor; sustainable agriculture
Attachment of relatively low numbers of endospores from two isolates of Pasteuria spp. to several species of nematodes was consistently achieved in 2-5 minutes with a centrifugation technique. The rate of attachment of Pasteuria penetrans at 10⁴ endospores/0.1 ml/tube to second-stage juveniles (J2) of Meloidogyne javanica, M. incognita race 1, M. incognita race 3, and M. arenaria races 1 and 2 in two tests averaged 4.4, 5.2, 0.1, 0.3, and 0 endospores per J2, respectively. The rate of attachment Pasteuria sp. at 10³ endospores/0.1 ml/tube to individuals of Hoplolaimus galeatus, Belonolaimus longicaudatus, M. arenaria race 1, M. javanica, and M. incognita race 1 in two tests averaged 0.8, 0.04, 0, 0, and 0 endospores per nematode, respectively. The rate of attachment of P. penetrans to M. javanica at 10³, 10⁴, or 10⁵ endospores/0.1 ml/tube from two tests averaged 1.0, 5.7, and 28.3 endospores per J2, respectively. All of the J2 had endospores attached following centrifugation in tubes with 10⁴ and 10⁵ endospores/0.1 ml/tube.
bacterium; biological control; centrifugation; endospore; Meloidogyne arenaria; M. incognita; M. javanica; method; nematode; Pasteuria penetrans; Pasteuria sp.
Microplot experiments were conducted in 1989 and 1990 to determine the relationship between yield of peanut (Arachis hypogaea) and inoculum density ofMeloidogyne arenaria race 1. Nine inoculum densities were used, ranging from 0-200 eggs/100 cm³ soil (1989) or from 0-100 eggs/100 cm³ (1990), and each density was replicated 10 times. In 1989, higher final densities (mean of 1,171 juveniles [J2]/100 cm³ soil) were obtained in plots inoculated with 0.5 to 50 eggs/100 cm³ soil than in plots inoculated with 100 to 200 eggs/100 cm³ (313 J2/100 cm³ soil). In 1990, final densities of M. arenaria reached high levels (≥ 1,111 J2/100 cm³ soil) in all inoculated plots. Pod yield and dry weight of foliage at harvest were negatively correlated (P ≤ 0.05) with inoculum density in both seasons. In 1989, the relationship between pod weight (y) and initial density (x) was described by Seinhorst's equation, with y = 0.088 + 0.91(0.90)⁽x⁻¹⁾ and r² = 0.826. In 1990, the relationship was y = 0.22 + 0.78(0.97)⁽x⁻¹⁾ and r² = 0.794. These equations suggest tolerance limits of approximately 1 egg/100 cm³ soil, which may require specialized methods, such as bioassay, for detection.
Arachis hypogaea; bioassay; damage function; Meloidogyne arenaria; nematode; peanut; root-knot nematode; Seinhorst's equation; threshold; tolerance limit
Meloidogyne incognita race 1, M. javanica, M. arenaria race 1, M. hapla, and an undescribed Meloidogyne sp. were analyzed by comparing isozyme phenotypes of esterase, malate dehydrogenase, phosphoglucomutase, isocitrate dehydrogenase, and α-glycerophosphate dehydrogenase. Isozyme phenotypes were obtained from single mature females by isoelectric focusing electrophoresis. Of these five isozymes, only esterase and phosphoglucomutase could be used to separate all five Meloidogyne spp.; however, the single esterase electromorphs were similar for M. incognita and M. hapla. Yet when both nematodes were run on the same gel, differences in their esterase phenotypes were detectable. Isozyme phenotypes from the other three isozymes revealed a great deal of similarity among M. incognita, M. javanica, M. arenaria, and the undescribed Meloidogyne sp.
biochemical systematics; electrophoresis; enzyme; esterase; α-glycerophosphate dehydrogenase; isocitrate dehydrogenase; isoelectric focusing; malate dehydrogenase; Meloidogyne arenaria; M. hapla; M. incognita; M. javanica; nematode; phosphoglucomutase
The specific gravity of spores of Pasteuria penetrans collected from Meloidogyne arenaria was found to be around 1.28. Increasing the sucrose concentration used for the extraction of Pratylenchus scribneri from a specific gravity of 1.14 to 1.26 led to the recovery of higher numbers of specimens filled with spores of Pasteuria sp. (P ≤ 0.05). The numbers of spore-filled specimens of Hoplolaimus galeatus recovered from field soil were not affected by the concentration of the sucrose solutions. Belonolaimus longicaudatus was recovered from field soil in greater numbers in sucrose solutions with specific gravities of 1.22 and 1.26 than with a specific gravity of 1.14 (P ≤ 0.05).
Belonolaimus longicaudatus; biological control; centrifugal flotation; extraction; Hoplolaimus galeatus; nematode; Pasteuria spp.; Pratylenchus scribneri; specific gravity
A field infested with Meloidogyne arenaria and with a history of peanut yield losses was divided into two equal parts. One-half of the field (bahia site) was planted to bahiagrass in 1986 and maintained through 1987. The other half (peanut site) was planted to soybean in 1986 and peanut in 1987 with hairy vetch planted each fall as a cover crop. In 1988 identical nematicide treatments including 1,3-dichloropropene (1,3-D), aldicarb, and ethoprop were applied to the two sites, and the sites were planted with the peanut cultivar Florunner. At mid-season, population levels of M. arenaria second-stage juveniles in the bahia site were relatively low, compared with those in the peanut site. At harvest, however, population levels were high in both sites. No nematicide treatment increased yields over the untreated control in either site (P ≤ 0.05). Bahiagrass alone and the combination of bahiagrass and 1,3-D applied broadcast resulted in 6.6-fold and 9.7-fold increases in yield, respectively, over the untreated control in the peanut site. All treatments in the bahia site resulted in increased vegetative growth and yields, compared with the duplicate treatments in the peanut site.
aldicarb; Arachis hypogaea; bahiagrass; crop rotation; 1,3-dichloropropene; ethoprop; Meloidogyne arenaria; nematicide; Paspalum notatum; peanut
Three tests were conducted to evaluate the efficacy of fumigant and nonfumigant nematicides for control of Meloidogyne arenaria race 1 on peanut. Methyl bromide, 1,3-D, methyl isothiocyanate, and methyl isothiocyanate mixtures were applied 7 or 8 days preplant either broadcast or in-the-row. Aldicarb, ethoprop, fenamiphos, and F5145 were applied at different rates and by different methods at-plant or at early flowering. Of the 32 treatments evaluated, only seven resulted in yield increases (P = 0.05), although early season vigor was high in all treated plots. During the latter one-third of the growing season, however, nematode control was not adequate in most treatments resulting in heavy peg, pod, and root infection by M. arenaria.
1,3-D; aldicarb; Arachis hypogaea; chemical control; ethoprop; F5145; fenamiphos; fumigant; Meloidogyne arenaria; methyl bromide; methyl isothiocyanate; nematicide; nonfumigant; peanut; root-knot nematode
The effect of two cropping and tillage systems on the population dynamics of four nematode species was evaluated on a loamy sand. Hairy vetch succeeded by corn or grain sorghum was seeded in split plots randomized within whole plots of no-tillage versus conventional tillage over four growing seasons (1980-83). The vetch-corn cropping system increased the density of Meloidogyne incognita 2.9 x more than the vetch-grain sorghum cropping system. In contrast, the vetch-grain sorghum cropping system increased the density of Criconemella ornata 0.7 x more than the vetch-corn cropping system. Meloidogyne incognita and C. ornata were affected more by these cropping systems than were Pratylenchus brachyurus or Paratrichodorus minor. Multiple cropping systems, vetch varieties, and crop host preference affected nematode population densities, whereas tillage treatments, conventional or no-tillage, had little effect on them.
corn; Criconemella ornata; cultural practice; grain sorghum; lesion nematode; Meloidogyne incognita; multiple cropping; Paratrichodorus minor; phytoparasitic nematode; population dynamics; root-knot nematode; Pratylenchus brachyurus; Sorghum bicolor; stubby-root nematode; tillage; vetch; Vicia spp.; Zea mays
The efficacy of the nematode parasite Paecilomyces lilacinus, alone and in combination with phenamiphos and ethoprop, for controlling the root-knot nematode Meloidogyne javanica on tobacco and the ability of this fungus to colonize in soil under field conditions were evaluated for 2 years in microplots. Combinations and individual treatments of the fungus grown on autoclaved wheat seed, M. javanica eggs (76,000 per plot), and nematicides were applied to specified microplots at the time of transplanting tobacco the first year. Vetch was planted as a winter cover crop, and the fungus and nematicides were applied again the second year to specified plots at transplanting time. The fungus did not control the nematode in either year of these experiments. The average root-gall index (0 = no visible galls and 5 = > 100 galls per root system) ranged from 2.7 to 3.9 the first year and from 4.3 to 5.0 the second in nematode-infested plots treated with nematicides. Plants with M. javanica alone or in combination with P. lilacinus had galling indices of 5.0 both years; the latter produced lower yields than all other treatments during both years of the study. Nevertheless, the average soil population densities of P. lilacinus remained high, ranging from 1.2 to 1.3 × 106 propagules/g soil 1 week after the initial inoculation and from 1.6 to 2.3 × 104 propagules/g soil at harvest the second year. At harvest the second year the density of fungal propagules was greatest at the depth of inoculation, 15 cm, and rapidly decreased below this level.
biocontrol; ethoprop; 1,3-dichloropropene; fenamiphos; fungal egg parasite; Meloidogyne javanica; nematicide; Nicotiana tabacum; Paecilomyces lilacinus; root-knot nematode; tobacco; vetch; Vicia vilIosa
The organo-phosphate experimental nematicide, O-ethyl S,S-di-sec-butyl phosphorodithioate (FMC 67825), provided yield increases of corn comparable to carbofuran. Both the emulsifiable concentrate and granular formulations of FMC 67825 were equally effective. The evaluations, duplicated in two adjacent fields, clearly demonstrated the importance of the type of plant pathogenic nematodes infesting the sites. Where Belonolaimus longicaudatus occurred, yield increases ranged from 73% to 80%, whereas in the adjacent field (without B. longicaudatus) yields increases ranged from - 14% to 28%.
Belonolaimus; carbofuran; corn; Criconemella; FMC 67825; lesion nematode; Meloidogyne; nematicide; Paratrichodorus; Pratylenchus; ring nematode; root-knot nematode; sting nematode; stubby-root nematode; Zea mays