Effect of Heterodera schachtii female age on susceptibility to three fungal hyperparasites in the genus Hyalorbilia.

Three closely related nematophagous fungi in the genus Hyalorbilia were compared for their ability to parasitize females and eggs of Heterodera schachtii at different developmental stages. DoUCR50, StM, and ARF were originally isolated from Heterodera schachtii, Meloidogyne incognita, and Heterodera glycines, respectively. Phylogenetic analysis and pairwise sequence analysis showed that DoUCR50 and StM are more closely related to each other than they are to ARF. DoUCR50 parasitism suppressed 100% of the J2 hatch from 3-week-old H. schachtii females and 75% of the hatch from 4-week-old females. Eggs within 5-week-old females were resistant to parasitism, and hatch of J2 was unaffected by exposure to DoUCR50. StM and ARF did not reduce the hatch of J2 from H. schachtii females of any age. Eggs removed from females and spread onto water agar cultures of the fungi were mostly resistant to parasitism. DoUCR50 parasitized only 16% of such eggs from 3-week-old females. Extracellular hydrolytic enzyme production by the three fungal strains grown on PDA or parasitized H. schachtii females was evaluated using API ZYM (bioMérieux) test strips. All three fungi produced extracellular hydrolytic enzymes when grown on PDA or H. schachtii females. Trypsin-like protease activity was uniquely detected in DoUCR50 grown on PDA and H. schachtii females, with the highest activity associated with the fungus grown on parasitized females.

Multi-Year Field Evaluation of Fluorinated Nematicides Against Meloidogyne incognita in Carrots.

California grows approximately 80% of the U.S. carrot production. The primary production challenges derive from root-knot nematodes (Meloidogyne spp.). Between 2013 and 2016, we evaluated three novel fluorinated nonfumigant nematicides in five field trials. Fluensulfone, fluopyram, and fluazaindolizine were applied as product-ready formulations at various rates, dates, and formulations. They were rated for their efficacy against the Southern root-knot nematode (M. incognita), their ability to mitigate nematode-caused crop damage, and potential to produce marketable carrot yield under high disease pressure. All trials were conducted in randomized complete block designs in M. incognita-infested, sandy-loam fields. Soil population of M. incognita at seeding and harvest, midseason plant vigor and fibrous root galling, harvest taproot galling, and marketable carrot yield were analyzed by ANOVA. Midseason gall ratings were indicative of disease ratings at harvest. All fluazaindolizine and fluensulfone treatments reduced at-harvest galling compared with the untreated controls. Fluopyram resulted in less galling but did not sufficiently protect the lower part of the taproot. Overall, fluazaindolizine at 2.24 kg/ha resulted in the most consistent and highest marketable carrot yield, followed by fluensulfone at 2.95 kg/ha. Both fluazaindolizine and fluensulfone will likely provide effective and target-selective crop protection against root-knot nematodes in fresh carrot production.

Occurrence of Belonolaimus in Sinaloa, Northwestern Mexico: A New Report on Distribution and Host Range.

The present study reports the occurrence of the genus Belonolaimus in the state of Sinaloa, Mexico, associated with native plants (i.e., Ziziphus amole and Stenocereus alamosensis) in a natural coastal ecosystem. Both morphological and molecular approaches were employed to characterize the Sinaloa population. Notwithstanding of some morphological and morphometric variation between Belonolaimus from Sinaloa and other valid species, the characterization indicates that this population might belong to the Belonolaimus longicaudatus species complex. Molecular analyses based on the 28S gene and ITS1-5.8S-ITS2 regions of the ribosomal RNA (rRNA) identified four major clades within Belonolaimus; however, none of the species including B. longicaudatus, B. gracilis, and B. euthychilus were supported as monophyletic; yet monophyly is argued to be a basic requirement of species status. Sequence divergence among different Belonolaimus populations and species varied according to the rRNA dataset (i.e., ITS1-5.8S-ITS2 > 28S > 18S) used, thus showing the importance of using genes with different rates of evolution to estimate species relationships. The fact that Belonolaimus has not been found in other cultivated (including on suitable hosts) areas in Sinaloa and that this population is relatively distant from the common B. longicaudatus groups (i.e., clades A and B) suggests that its appearance was not due to a recent introduction associated with the local agriculture.

Survival of plant pathogens in static piles of ground green waste.

Ground green waste is used as mulch in ornamental landscapes and for tree crops such as avocados. Survival of Armillaria mellea, Phytophthora cinnamomi, Sclerotinia sclerotiorum, and Tylenchulus semipenetrans was assessed for 8 weeks within unturned piles of either recently ground or partially composted green waste. S. sclerotiorum survived at the pile surface and at 10, 30, and 100 cm within the pile for the entire 8 weeks in both fresh green waste (FGW) and aged green waste (AGW). A. mellea and T. semipenetrans did not survive more than 2 days in FGW, while P. cinnamomi persisted for over 21 days in FGW. AGW was less effective in reducing pathogen viability than FGW, most likely because temperatures in AGW peaked at 45 degrees C compared with 70 degrees C in FGW. Survival modeling curves based on pile temperatures indicate the time to inactivate 10 propagules of pathogens was 11, 30, 363, and 50 days for A. mellea, P. cinnamomi, S. sclerotiorum, and T. semipenetrans, respectively. Sclerotia-forming pathogens pose the greatest risk for escape; to ensure eradication of persistent fungi, green waste stockpiles should be turned intermittently to mix pile contents and move pathogen propagules to a location within the pile where they are more likely to be killed by heat, microbial attack, or chemical degradation.

Detection and Investigation of Soil Biological Activity against Meloidogyne incognita.

Greenhouse experiments with two susceptible hosts of Meloidogyne incognita, a dwarf tomato and wheat, led to the identification of a soil in which the root-knot nematode population was reduced 5- to 16-fold compared to identical but pasteurized soil two months after infestation with 280 M. incognita J2/100 cm(3) soil. This suppressive soil was subjected to various temperature, fumigation and dilution treatments, planted with tomato, and infested with 1,000 eggs of M. incognita/100 cm(3) soil. Eight weeks after nematode infestation, distinct differences in nematode population densities were observed among the soil treatments, suggesting the suppressiveness had a biological nature. A fungal rRNA gene analysis (OFRG) performed on M. incognita egg masses collected at the end of the greenhouse experiments identified 11 fungal phylotypes, several of which exhibited associations with one or more of the nematode population density measurements (egg masses, eggs or J2). The phylotype containing rRNA genes with high sequence identity to Pochonia chlamydosporia exhibited the strongest negative associations. The negative correlation between the densities of the P. chlamydosporia genes and the nematodes was corroborated by an analysis using a P. chlamydosporia-selective qPCR assay.

'Candidatus pasteuria usgae' sp. nov., an obligate endoparasite of the phytoparasitic nematode Belonolaimus longicaudatus.

Taxonomically relevant characteristics of a fastidiously Gram-positive, obligately endoparasitic prokaryote (strain S-1) that uses the phytoparasitic sting nematode Belonolaimus longicaudatus as its host are reviewed. 16S rDNA sequence similarity (> or = 93%) confirms its congeneric ranking with other Pasteuria species and strains from nematodes and cladocerans and corroborates morphological, morphometric and host range evidence suggesting a novel taxon. The 16S rDNA sequence of strain S-1 has greatest similarity (96%) to the 16S rDNA sequences of both Pasteuria penetrans from root-knot nematodes (Meloidogyne species) and the recently reported strain of Pasteuria isolated from the soybean cyst nematode Heterodera glycines. Because the obligately endoparasitic nature of prokaryotes in the genus Pasteuria prevents isolation of definitive type strains, strain S-1 is proposed as 'Candidatus Pasteuria usgae' sp. nov.

Evaluation of propargyl bromide for control of barnyardgrass and Fusarium oxysporum in three soils.

With the scheduled phasing out of methyl bromide, there is an urgent need for alternatives. We evaluated the efficacy of propargyl bromide as a potential replacement for methyl bromide for the control of barnyardgrass (Echinochloa crus-galli) and Fusarium oxysporum in an Arlington sandy loam, a Carsitas loamy sand and a Florida muck soil. Soil was mixed with barnyardgrass seeds or F oxysporum colonized on millet seeds, and treated with propargyl bromide at a range of concentrations. The mortality of the fungi and weed seeds was determined after 24 h of exposure at 30 degrees C. The concentrations required to inhibit 50% barnyard seed germination (LC50) were 2.8, 2.4 and 48.5 micrograms g-1 in the sandy loam, loamy sand and muck soil, respectively. In contrast, the LC50 values for F oxysporum were 11.2, 10.8 and 182.1 micrograms g-1 in the sandy loam, loamy sand and muck soil, respectively. The low efficacy of propargyl bromide in the muck soil was a result of the rapid degradation and high adsorption of the compound in the soil. The degradation half-life (t1/2) was only 7 h in the muck soil at an initial concentration of 6.8 micrograms g-1, compared to 60 and 67 h in the sandy loam and loamy sand, respectively. The adsorption coefficients (Kd) were 0.96, 0.87 and 5.6 cm3 g-1 in the sandy loam, loamy sand and muck soil, respectively. These results suggest that registration agencies should consider site-specific properties in recommending application rates for propargyl bromide.

Degradation of soil fumigants as affected by initial concentration and temperature.

Soil fumigation using shank injection creates high fumigant concentration gradients in soil from the injection point to the soil surface. A temperature gradient also exists along the soil profile. We studied the degradation of methyl isothiocyanate (MITC) and 1,3-dichloropropene (1,3-D) in an Arlington sandy loam (coarse-loamy, mixed, thermic Haplic Durixeralf) at four temperatures and four initial concentrations. We then tested the applicability of first-order, half-order, and second-order kinetics, and the Michaelis-Menten model for describing fumigant degradation as affected by temperature and initial concentration. Overall, none of the models adequately described the degradation of MITC and 1,3-D isomers over the range of the initial concentrations. First-order and half-order kinetics adequately described the degradation of MITC and 1,3-D isomers at each initial concentration, with the correlation coefficients greater than 0.78 (r2> 0.78). However, the derived rate constant was dependent on the initial concentration. The first-order rate constants varied between 6 and 10x for MITC for the concentration range of 3 to 140 mg kg(-1), and between 1.5 and 4x for 1,3-D isomers for the concentration range of 0.6 to 60 mg kg(-1), depending on temperature. For the same initial concentration range, the variation in the half-order rate constants was between 1.4 and 1.7x for MITC and between 3.1 and 6.1x for 1,3-D isomers, depending on temperature. Second-order kinetics and the Michaelis-Menten model did not satisfactorily describe the degradation at all initial concentrations. The degradation of MITC and 1,3-D was primarily biodegradation, which was affected by temperature between 20 and 40 degrees C, following the Arrhenius equation (r2 > 0.74).

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.

Phenotypic and molecular analysis of a pasteuria strain parasitic to the sting nematode.

Pasteuria strain S-1 was found to parasitize the sting nematode Belonolaimus longicaudatus. S-1 spores attached to several strains of B. longicaudatus from different geographical locations within the United States. However, they did not adhere to any of the following species: Heterodera schachtii, Longidorus africanus, Meloidogyne hapla, M. incognita, M. javanica, Pratylenchus brachyurus, P. scribneri, P. neglectus, P. penetrans, P. thornei, P. vulnus, and Xiphinema spp. The 16S rRNA genes from Pasteuria strain S-1 and P. penetrans strain Pp from Senegal were obtained by PCR amplification. A DNA sequence analysis showed that the S-1 16S rRNA had 96% or less similarity to the 16S rRNA genes from all previously reported Pasteuria species. Diverse phylogenetic methods all provided robust support for an association of Pasteuria strain S-1, Pasteuria strain NA parasitic to H. glycines, and P. penetrans strain Pp, to the exclusion of P. ramosa. In addition, our study showed intraspecific variation within P. penetrans as inferred by its 98% similarity to P. penetrans strain Pp.

Transfer of Biological Soil Suppressiveness Against Heterodera schachtii.

ABSTRACT Heterodera schachtii-suppressive soil at a rate of either 1 or 10% (dry wt/wt) transferred suppressiveness against the beet cyst nematode to fumigated field plots when mixed into the upper 10-cm soil layer. Soil suppressiveness was established after 1 month of moist fallow and 77 days of Swiss chard cropping in the 10% transfer treatment and after 230 days in the 1% transfer treatment. The number of infective second-stage juveniles (J2) of H. schachtii, monitored initially at 150 degree-day intervals and later at 300 degree-day intervals, indicated the status of suppressiveness in the different treatments during the cropping period. In a greenhouse experiment, amending fumigated field soil with 0.1, 1.0, or 10% suppressive soil, suppressed multiplication of H. schachtii when soils were infested with an additional 5,000 J2. In a second greenhouse experiment, a fumigated sandy loam amended with 10 or 25% suppressive soil and a fumigated loam amended with 25% suppressive soil had significantly fewer eggs per cyst than the nonamended fumigated treatments when 1,000 J2 were added.

Biological Suppression and Natural Population Decline of Heterodera schachtii in a California Field.

ABSTRACT Soil suppressiveness to Heterodera schachtii was demonstrated in a field at the research station of the University of California, Riverside. In two field trials planted to Swiss chard (Beta vulgaris), introduced H. schachtii multiplied 2.7 and 1.7 times more in preplant metam sodium-fumigated plots than in nontreated plots in 1994 and 1995, respectively. In greenhouse experiments, preplant treatments with metam sodium, methyl bromide, methyl iodide, formaldehyde, and aerated steam reduced suppressiveness of soil against H. schachtii to undetectable levels. H. schachtii multiplied significantly less in nontreated soil than in treated soil on Swiss chard. At harvest, the number of infective second-stage juveniles in suppressive soil was close to the lowest detection level, whereas high numbers were encountered in soils initially treated. In a crop rotation trial with host crops of H. schachtii, introduced H. schachtii populations were monitored for five cropping periods over 30 months in initially fumigated versus nontreated suppressive field plots. In fumigated plots, H. schachtii population levels increased in the first and second cropping periods and then declined in the third cropping period. In the fourth and fifth cropping periods, the nematode reproduction factor in the initially fumigated plots was not significantly different from that in suppressive plots.

Evaluation of Biocontrol Activity of Rhizobacteria from Beta vulgaris against Heterodera schachtii.

One hundred fifty rhizobacteria isolated from roots of Swiss chard grown in a soil suppressive to the sugar beet cyst nematode, Heterodera schachtii, were tested for their influence on the nematode's ability to hatch and infect roots. Two screening systems were used that focused on the ability of bacteria to inhibit either nematode hatching or root infection. Most of the bacterial strains reduced hatching in vitro compared to the control, while with 5% of the strains there were 0% hatch. Seven percent of all strains significantly reduced second-stage juvenile (J2) infection of mustard roots raised in soil-less growth pouches. Eighteen strains from four genera (Bacillus, Pseudomonas, Variovorax, and Arthrobacter) were selected for greenhouse trials. In the greenhouse trials four bacterial strains, including two strains of Bacillus megaterium, reduced nematode infection of sugar beet when eggs were used as inoculum. Seven bacterial strains reduced nematode infection of sugar beet in one of two trials when plants were inoculated with J2. Most of the effective strains were Bacillus spp., primarily B. megaterium. Colonization of sugar beets roots by two B. megaterium strains was found to be stable over 30 days in the greenhouse.

Life Cycle and Mating Behavior of Belonolaimus longicaudatus in Gnotobiotic Culture.

The life cycle of Belonolaimus longicaudatus was observed in vitro on excised roots of Zea mays. Roots were cultured on Gamborg's B5 medium in petri dishes with 1.5% agar adjusted to pH 5.8 and incubated at 28 degrees C in darkness. Second-stage juveniles (J2) fed on the roots and started the second molt (M2) to the third-stage juveniles 2 days after inoculation (DAI). The third molt (M3) to the fourth-stage juveniles occurred 7 DAI, followed by the fourth molt (M4) to males 13 DAI or to females 14 DAI. Nematode gender differences were observed by the end of the fourth molt. The first male appeared 15 DAI and the first female 17 DAI, after which mating occurred. Males were attracted to females, and mating was observed. Mating was required for reproduction. Fertilized females began to lay eggs 19 DAI and continued egg laying without the further presence of males during a 90-day observation. All of the eggs hatched. Unfertilized females rarely laid eggs, and none of the eggs were able to hatch. Feeding took place between each molt and before egg deposition occurred. The first-stage juveniles molted in the eggs 4 days after deposition, and J2 hatched from eggs 5 days after egg deposition. The life cycle from J2 to J2 was completed in 24 days.

Survey of crop losses in response to phytoparasitic nematodes in the United States for 1994.

Previous reports of crop losses to plant-parasitic nematodes have relied on published results of survey data based on certain commodities, including tobacco, peanuts, cotton, and soybean. Reports on crop-loss assessment by land-grant universities and many commodity groups generally are no longer available, with the exception of the University of Georgia, the Beltwide Cotton Conference, and selected groups concerned with soybean. The Society of Nematologists Extension Committee contacted extension personnel in 49 U.S. states for information on estimated crop losses caused by plant-parasitic nematodes in major crops for the year 1994. Included in this paper are survey results from 35 states on various crops including corn, cotton, soybean, peanut, wheat, rice, sugarcane, sorghum, tobacco, numerous vegetable crops, fruit and nut crops, and golf greens. The data are reported systematically by state and include the estimated loss, hectarage of production, source of information, nematode species or taxon when available, and crop value. The major genera of phytoparasitic nematodes reported to cause crop losses were Heterodera, Hoplolaimus, Meloidogyne, Pratylenchus, Rotylenchulus, and Xiphinema.

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 degrees 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.

Cloning of Genes Involved in the Synthesis of Pyrrolnitrin from Pseudomonas fluorescens and Role of Pyrrolnitrin Synthesis in Biological Control of Plant Disease.

A soil isolate of Pseudomonas fluorescens (BL915) was shown to be an effective antagonist of Rhizoctonia solani-induced damping-off of cotton. Investigation of the biological basis of this antagonism revealed that the strain produces pyrrolnitrin, a secondary metabolite known to inhibit R. solani and other fungi. Mutants of strain BL915 that did not produce pyrrolnitrin and did not suppress damping-off of cotton by R. solani were generated by exposure to N-methyl-N' -nitro-N-nitrosoguanidine. A gene region that was capable of restoring pyrrolnitrin production to the non-pyrrolnitrin-producing mutants and of conferring this ability upon two other P. fluorescens strains not otherwise known to produce this compound or to be capable of suppressing damping-off caused by R. solani was isolated from strain BL915. The non-pyrrolnitrin-producing strains (mutants of BL915 and the other two P. fluorescens strains) which synthesized pyrrolnitrin after the introduction of the gene region from strain BL915 were also shown to be equal to strain BL915 in their ability to suppress R. solani-induced damping-off of cotton. These results indicate that we have isolated from P. fluorescens BL915 a gene(s) that has a role in the synthesis of pyrrolnitrin and that the production of this compound has a role in the ability of this strain to control damping-off of cotton by R. solani.

The challenge of research and extension to define and implement alternatives to methyl bromide.

Over the past 30 years, methyl bromide (MBr), a broad spectrum fumigant, has been used extensively for soilborne disease and pest control in the production of many fruit, vegetable, turf, and nursery crops. Recently, agricultural emissions of MBr were implicated as a potentially significant contributor to stratospheric ozone depletion. As a precautionary measure for global ozone protection, the U.S. Environmental Protection Agency has enforced federal legislation which mandates a complete phase-out of MBr use within the United States by 1 January 2001. Thus, new cost effective, environmentally compatible strategies for control of nematodes and other soilborne pests and pathogens must be developed and tested in a relatively short time to avoid significant losses in crop productivity. The extent to which certain agricultural industries that are now heavily reliant on MBr are affected will depend on the development of sustainable, integrated tactics to pest control, such as combinations of cultural, chemical, and biological tactics. New muhidisciplinary research and extension programs must be developed to address and overcome major constraints and incompatibilities that have prevented such tactics from being widely adopted.

Drip irrigation as a delivery system for infestation of field plots with nematodes.

A drip irrigation delivery system was used to infest field sites with the plant-parasitic root-knot nematodes, Meloidogyne incognita. Juvenile or egg inocula passed through the system without blockage of emitters or harm to the nematodes. Field sites so infested were available for experimentation. Delivery of approximately 5 x 10 to 10 juveniles or 10 to 3 x 10 eggs per emitter through the drip system resulted in heavy root galling of tomatoes planted next to the drip emitters. Nematodes feeding on bacteria (Acrobeloides sp.) and on fungi (Deladenus durus) also were successfully applied through the drip system. This method has potential for uniformly infesting experimental sites with plant-parasitic or entomogenous nematodes and for manipulation of nematode community structure for soil ecological studies.

[Complications of arteriography--details at the time of the occurrence of side effects]. / Komplikationen der Arteriographie--Angaben zum Zeitpunkt des Auftretens der Zwischenfälle.

Own observations and the review of the literature show that complications of arteriography may appear after some hours or some days. The causal context is given in general for complications that may be due to the puncture. This is valid, too, for renal insufficiency induced by contrast substance. Neurological symptoms appearing hours or days after cerebral angiography are more difficult to evaluate. Different influences may contribute. The information of the patient before arteriography may in some cases include complications of possible follow-up treatment.