bluuebunny/crossref_metadata_2025 huggingface dataset:
A Rapid Polymerase Chain Reaction-Based Assay Characterizing Rhizosphere Populations of 2,4-Diacetylphloroglucinol-Producing Bacteria
Authors: McSpadden Gardener Brian B. , Mavrodi Dmitri V. , Thomashow Linda S. , Weller David M.
Published: May 2007

Pseudomonas species that produce 2,4-diacetylphloroglucinol (2,4-DAPG) play a significant role in the suppression of fungal root pathogens in the rhizosphere of crop plants. To characterize the abundance and diversity of these functionally important bacterial populations, we developed a rapid polymerase chain reaction (PCR)-based assay targeting phlD, an essential gene in the phloroglucinol biosynthetic pathway. The phlDgene is predicted to encode a polyketide synthase that synthesizes mono-acetylphloroglucinol, the immediate precursor to 2,4-DAPG. A major portion of the phlD open reading frame was cloned and sequenced from five genotypically distinct strains, and the sequences were screened for conserved regions that could be used as gene-specific priming sites for PCR amplification. Several new phlD-specific primers were designed and evaluated. Using the primers B2BF and BPR4, we developed a PCR-based assay that was robust enough to amplify the target gene from a diverse set of 2,4-DAPG producers and sensitive enough to detect as few as log 2.4 cells per sample when combined with enrichment from a selective medium. Restriction fragment length polymorphism analysis of the amplified phlD sequence allows for the direct determination of the genotype of the most abundant 2,4-DAPG producers in a sample. The method described was useful for characterizing both inoculant and indigenous phlD+ pseudomonads inhabiting the rhizosphere of crop plants. The ability to rapidly characterize populations of 2,4-DAPG-producers will greatly enhance our understanding of their role in the suppression of root diseases.
Mutations that Affect Agrobacterium vitis-Induced Grape Necrosis also Alter Its Ability to Cause a Hypersensitive Response on Tobacco
Authors: Herlache T. C. , Zhang H. S. , Ried C. L. , Carle S. A. , Zheng D. , Basaran P. , Thaker M. , Burr A. T. , Burr T. J.
Published: May 2007

Tn5-induced mutations in Agrobacterium vitis F2/5 resulted in both altered grape necrosis and tobacco leaf panel collapse phenotypes, suggesting that the underlying mechanisms of the reactions are related. The reaction on tobacco resembles the classical hypersensitive response (HR) caused by several plant pathogenic bacteria in that it is observable within 14 h, is inhibited by treatment of plants with metabolic inhibitors, and results in the inability to recover the pathogen from the necrotic zone. Strains of A. vitis differ with regard to their efficiency of causing the reaction on tobacco. An EcoRI fragment from one mutant, M6, which is necrosis-altered and HR-minus, was cloned and sequenced. Sequence analysis revealed that the Tn5 insertion occurred in a region that shares significant homology with genes involved in long chain fatty acid production by the marine bacteria Shewanella spp. and Moritella marina. Complementation of M6 with a cosmid clone from an F2/5 DNA library restored the tobacco HR and grape necrosis phenotypes.
Effect of Potting Mix Microbial Carrying Capacity on Biological Control of Rhizoctonia Damping-Off of Radish and Rhizoctonia Crown and Root Rot of Poinsettia
Authors: Krause Matthew S. , Madden Laurence V. , Hoitink Harry A. J.
Published: May 2007

Potting mixes prepared with dark, highly decomposed Sphagnum peat, with light, less decomposed Sphagnum peat, or with composted pine bark, all three of which were colonized by indigenous microorganisms, failed to consistently suppress Rhizoctonia damping-off of radish or Rhizoctonia crown and root rot of poinsettia. Inoculation of these mixes with Chryseobacterium gleum (C299R2) and Trichoderma hamatum 382 (T382) significantly reduced the severity of both diseases in the composted pine bark mix in which both biocontrol agents maintained high populations over 90 days. These microorganisms were less effective against damping-off in the light and dark peat mixes, respectively, in which populations of C299R2declined. In contrast, crown and root rot, a disease that is severe late in the crop, was suppressed in all three types of mixes. High populations of T382in all three mixes late during the cropping cycle may have contributed to control of this disease.
Antagonism of Nutrient-Activated Conidia of Trichoderma harzianum (atroviride) P1 Against Botrytis cinerea
Authors: Hjeljord Linda Gordon , Stensvand Arne , Tronsmo Arne
Published: May 2007

The effect of preliminary nutrient activation on the ability of conidia of the antagonist Trichoderma harzianum (atroviride) P1 to suppress Botrytis cinerea was investigated in laboratory, greenhouse, and field trials. Preliminary nutrient activation at 21°C accelerated subsequent germination of the antagonist at temperatures from 9 to 21°C; at ≥18°C, the germination time of preactivated T. harzianum P1 conidia did not differ significantly from that of B. cinerea. When coinoculated with B. cinerea, concentrated inocula of preactivated but ungerminated T. harzianum P1 conidia reduced in vitro germination of the pathogen by ≥87% at 12 to 25°C; initially quiescent conidia achieved this level of suppression only at 25°C. Application of quiescent T. harzianum P1 conidia to detached strawberry flowers in moist chambers reduced infection by B. cinerea by ≥85% at 24°C, but only by 35% at 12°C. Preactivated conidia reduced infection by ≥60% at 12°C. Both quiescent and preactivated conidia significantly reduced latent infection in greenhouse-grown strawberries at a mean temperature of 19°C, whereas only preactivated conidia were effective in the field at a mean temperature of 14°C on the day of treatment application. An antagonistic mechanism based on initiation of germination in sufficiently concentrated inocula suggests that at suboptimal temperatures the efficacy of Trichoderma antagonists might be improved by conidia activation prior to application.
Ultrastructure of the Infection ofSorghum bicolorbyColletotrichum sublineolum
Authors: Wharton P. S. , Julian A. M. , O'Connell R. J.
Published: May 2007

Ultrastructural studies of the infection of susceptible and resistant cultivars of Sorghum bicolor by Colletotrichum sublineolum were conducted. Initial penetration events were the same on both susceptible and resistant cultivars. Germ tubes originating from germinated conidia formed globose, melanized appressoria, that penetrated host epidermal cells directly. Appressoria did not produce appressorial cones, but each penetration pore was surrounded by an annular wall thickening. Inward deformation of the cuticle and localized changes in staining properties of the host cell wall around the infection peg suggests that penetration involves both mechanical force and enzymic dissolution. In compatible interactions, penetration was followed by formation of biotrophic globular infection vesicles in epidermal cells. Filamentous primary hyphae developed from the vesicles and went on to colonize many other host cells as an intracellular mycelium. Host cells initially survived penetration. The host plasma membrane invaginated around infection vesicles and primary hyphae and was appressed tightly to the fungal cell wall, with no detectable matrix layer at the interface. Necrotrophic secondary hyphae appeared after 66 h and ramified through host tissue both intercellularly and intracellularly, forming hypostromatic acervuli by 114 h. Production of secondary hyphae was accompanied by the appearance of electron-opaque material within infected cells. This was thought to represent the host phytoalexin response. In incompatible interactions, infection vesicles and primary hyphae were formed in epidermal cells by 42 h. However, they were encrusted with electron-opaque material and appeared dead. These observations are discussed in relation to the infection processes of other Colletotrichum spp. and the host phytoalexin response.
Partial Characterization of a Closterovirus Associated with Apple Mealybug-Transmitted Little Cherry Disease in North America
Authors: Eastwell K. C. , Bernardy M. G.
Published: May 2007

Little cherry disease (LChD) is a serious economic problem of sweet cherry production in western North America where apple mealybug is the principle vector. LChD is associated with a distinct species of double-stranded (ds) RNA. In this study, filamentous virus particles were purified from LChD-infected trees and shown to contain single-stranded RNA corresponding to the previously reported dsRNA isolated from infected trees. The virus particles were characterized and were similar to monopartite members of the genus Closterovirus. A portion of the genome was sequenced and found to be most closely related to the RNA-dependent RNA polymerase of Grapevine leafroll-associated virus-3, a mealybug-transmitted closterovirus. The characteristics of the mealybug-transmitted Little cherry virus in North America are very different from those of a closterovirus associated with a similar disease in Europe.
Quantitative Trait Loci in Sweet Corn Associated with Partial Resistance to Stewart's Wilt, Northern Corn Leaf Blight, and Common Rust
Authors: Brown A. F. , Juvik J. A. , Pataky J. K.
Published: May 2007

Partial resistance to Stewart's wilt (Erwina stewartii, syn. Pantoea stewartii), northern corn leaf blight (NCLB) (Exserohilum turcicum), and common rust (Puccinia sorghi) was observed in an F2:3population developed from a cross between the inbred sweet corn lines IL731a and W6786. The objective of this study was to identify quantitative trait loci (QTL) associated with partial resistance using restriction fragment length polymorphic markers. Phenotypic data were collected for 2 years for Stewart's wilt, NCLB, and common rust but, due to significant family-environment interaction, analysis was conducted individually on data from each year. In 2 years of evaluation for the three diseases, a total of 33 regions in the maize genome were associated with partial resistance describing from 5.9 to 18% of the total phenotypic variability. Of six regions common in both years, three were associated with partial resistance to Stewart's wilt (chromosomes 4:07, 5:03, and 6:04), one was associated with NCLB (chromosome 9:05), and two were associated with common rust (chromosomes 2:04 and 3:04). The rust QTL on 3S mapped to within 20 cM of the rp3 locus and explained 17.7% of the phenotypic variability. Some of the QTL associated with partial resistance to the three diseases have been reported previously, and some are described here for the first time. Results suggest it may be possible to consolidate QTL from various elite backgrounds in a manner analogous to the pyramiding of major resistance genes. We also report here on two QTL associated with anthocyanin production on chromosomes 10:6 and 5:03 in the general location of the a2 gene.
Soil pH and the Activity of a Pelletized Nematophagous Fungus
Authors: Jaffee B. A. , Zasoski R. J.
Published: May 2007

When formulated as assimilative hyphae in alginate pellets, the nematophagous fungus Hirsutella rhossiliensis was more active (i.e., parasitized more assay nematodes) in an acidic vineyard soil than in a neutral vineyard soil. To determine whether soil pH explained the difference, fungus activity was measured in soil from the neutral site that had been acidified (by adding sulfuric acid) and in soil from the acidic site that had been neutralized (by adding calcium hydroxide). As hypothesized, the activity of pelletized Hirsutella rhossiliensis was negatively correlated with soil pH. Maximum activity occurred at pHcalcium chloride4.5, and activity gradually declined to near zero as the pH increased to 6.5 and rapidly declined to near zero as the pH dropped below 4.0. Assays performed on leached soil samples indicated that the effects of sulfuric acid and calcium hydroxide were largely due to pH rather than to specific ions or osmotic potential. The effect of pH, however, was indirect. Heating the neutral soil to 60°C for 2 h did not alter soil pH or electrical conductivity but increased fungus activity to levels equivalent to those in acidified soil. We conclude that, in these two soils, heat treatment or low soil pH suppresses soil organisms that otherwise interfere with growth of Hirsutella rhossiliensis from alginate pellets.
Sequential Sampling for Incidence of Phomopsis Leaf Blight of Strawberry
Authors: Turechek W. W. , Ellis M. A. , Madden L. V.
Published: May 2007

Sequential sampling models for estimation and classification were developed for the incidence of strawberry leaflets infected by Phomopsis obscurans. Sampling protocols were based on a binary power law analysis of the spatial heterogeneity of Phomopsis leaf blight in commercial fields in Ohio. For sequential estimation, samples were collected until mean disease incidence could be estimated with a preselected coefficient of variation of the mean (C). For sequential classification, samples were collected until there was sufficient evidence to classify mean incidence as being below or above a threshold (pt) based on the sequential probability ratio test. Monte-Carlo simulations were used to determine the theoretical average sample number (ASN) and probability of classifying mean incidence as less than pt (operating characteristic) for any true value of incidence. Estimation and classification sampling models were both tested with bootstrap simulations of randomly selected data sets and validated by data sets from another year that were not utilized in developing the models. In general, achieved (or calculated) C after sequentially sampling for estimation was close to the preselected C of 0.2, and mean incidence was estimated with little bias. Achieving a C of 0.1 with less than 75 sampling units (the nominal value for many original data sets) was more problematical, especially with true incidence less than 0.2. ASN for classification was only 9 to 18 at disease incidence values near pt, and approximately five or less at incidence values far from pt. Correct classification decisions were made in over 88% of the validation data sets. Results indicated that it is possible to estimate Phomopsis leaf blight with high precision and with high correct classification probabilities.
Effects of Environmental Factors on Development of Pyrenopeziza brassicae (Light Leaf Spot) Apothecia on Oilseed Rape Debris
Authors: Gilles T. , Fitt B. D. L. , Jeger M. J.
Published: May 2007

The development of Pyrenopeziza brassicae (light leaf spot) apothecia was studied on petiole debris from artificially infected oilseed rape leaves incubated at temperatures from 6 to 22°C under different wetness regimes and in 16 h light/8 h dark or continuous darkness. There was no significant difference between light treatments in numbers of apothecia that developed. Mature apothecia developed at temperatures from 5 to 18°C but not at 22°C. The rate of apothecial development decreased as temperature decreased from 18 to 5°C; mature apothecia were first observed after 5 days at 18°C and after 15 days at 6°C. Models were fitted to estimates of the time (days) for 50% of the maximum number of apothecia to develop (t1; model 1, t1 = 7.6 + 55.8(0.839)T) and the time for 50% of the maximum number of apothecia to decay (t2; model 2, t2 = 24.2 + 387(0.730)T) at temperatures (T) from 6 to 18°C. An interruption in wetness of the petiole debris for 4 days after 4, 7, or 10 days of wetness delayed the time to observation of the first mature apothecia for ≈4 days and decreased the number of apothecia produced (by comparison with continuous wetness). A relationship was found between water content of pod debris and electrical resistance measured by a debris-wetness sensor. The differences between values of t1 predicted by model 1 and observed values of t1 were 1 to 9 days. Model 2 did not predict t2; apothecia decayed more quickly under natural conditions than predicted by model 2.
Species-Specific Detection of Monilinia fructicola from California Stone Fruits and Flowers
Authors: Boehm E. W. A. , Ma Z. , Michailides T. J.
Published: May 2007

A set of molecular diagnostics was developed for Monilinia fructicola, causal agent of brown rot of stone fruits, capable of sensitive detection of the pathogen in planta. Species-specific repetitive sequences were identified from a partial library of 312 recombinant clones hybridized with total DNA, followed by subsequent screening for specificity. One hundred isolates, comprising 12 fungal species common to California stone fruits, were surveyed for specificity. Three clones hybridized to 60 geographically diverse M. fructicola isolates (California, Michigan, Georgia, Oregon, and Australia) to the exclusion of all other fungi surveyed, including the closely related M. laxa (n = 12). Two clones were identical and of extrachromosomal origin (pMF73 and pMF150), whereas the third (pMF210) migrated with uncut DNA. The sensitivity of all three was comparable and capable of detecting 50 pg of fungal DNA in dot blot hybridizations. Six species-specific primer pair sets were designed. They maintained the same specificity patterns observed in the initial hybridization surveys and were sensitive enough to detect 50 fg of fungal DNA template, approximately equivalent to 10 spores. The species-specific clones were capable of detecting the pathogen in planta, specifically from infected plum flowers and nectarine fruit tissue, using both hybridization- and polymerase chain reaction-based methodologies.
Effect of Water Potential on Conidial Germination and Antagonism of Ulocladium atrum Against Botrytis cinerea
Authors: Köhl J. , Molhoek W. M. L.
Published: May 2007

The saprophytic fungus Ulocladium atrum was selected for its ability to competitively exclude Botrytis spp. from aboveground necrotic plant tissues which can play a crucial role in the epidemiology of diseases caused by necrotrophic Botrytis spp. Fungal growth in necrotic aboveground tissues can be hampered by fluctuating water availability. Adaptation to these adverse conditions is a key factor for the successful establishment of an antagonist population in this niche. Conidia of U. atrum germinated at water potentials between -1 and -7 MPa on water agar. Germinated conidia survived one to three interruptions of moist incubation by periods at -10 or -42 MPa. The speed of germination was significantly slower with interruption of the moist period as compared with the control, which had continuously moist incubation. However, the maximum germination percentage was the same for conidia incubated with or without interruption of the moist incubation. In bioassays with necrotic cyclamen leaves at -1, -3, and -7 MPa, U. atrum significantly reduced the sporulation of B. cinerea by more than 80%. The results demonstrate that U. atrum tolerates water stress during competitive substrate colonization with B. cinerea. The antagonist is, therefore, an attractive candidate for field applications on aboveground tissues.
The Role of Ethylene Production in Virulence of Pseudomonas syringae pvs. glycinea and phaseolicola
Authors: Weingart Helge , Ullrich Henriette , Geider Klaus , Völksch Beate
Published: May 2007

The importance of ethylene production for virulence of Pseudomonas syringae pvs. glycinea and phaseolicola was assayed by comparing bacterial multiplication and symptom development in bean and soybean plants inoculated with ethylene-negative (efe) mutants and wild-type strains. The efe mutants of Pseudomonas syringae pv. glycinea were significantly reduced in their ability to grow in planta. However, the degree of reduction was strain-dependent. Population sizes of efe mutant 16/83-E1 that did not produce the phototoxin coronatine were 10- and 15-fold lower than those of the wild-type strain on soybean and on bean, and 16/83-E1 produced very weak symptoms compared with the wild-type strain. The coronatine-producing efe mutant 7a/90-E1 reached fourfold and twofold lower population sizes compared with the wild-type strain on soybean and bean, respectively, and caused disease symptoms typical of the wild-type strain. Experiments with ethylene-insensitive soybeans confirmed these results. The virulence of the wild-type strains was reduced to the same extent in ethylene-insensitive soybean plants as the virulence of the efe mutants in ethylene-susceptible soybeans. In contrast, the virulence of Pseudomonas syringae pv. phaseolicola was not affected by disruption of the efe gene.
Surface Hydrophobicity and Surface Rigidity Induce Spore Germination in Colletotrichum graminicola
Authors: Chaky J. , Anderson K. , Moss M. , Vaillancourt L.
Published: May 2007

We investigated the relationship between physical characteristics of artificial surfaces, spore attachment, and spore germination in Colletotrichum graminicola. Surface hydrophobicity and surface rigidity were both signals for breaking dormancy and initiating spore germination, but spore attachment alone was not an important inducing signal. The presence of a carbon source overrode the necessity for a rigid, hydrophobic substrate for spore germination. Spore attachment was typically stronger to more hydrophobic surfaces, but certain hydrophilic surfaces also proved to be good substrates for spore attachment. In contrast to spore germination, appressorial induction was more dependent on attachment to a rigid substrate than it was on surface hydrophobicity. Appressoria were induced efficiently on hydrophilic surfaces, as long as there was significant conidial attachment to those surfaces.
Persistence of Complex Virulences in Populations of Phytophthora infestans in Western Washington
Authors: Derie M. L. , Inglis D. A.
Published: May 2007

Isolates of Phytophthora infestans, collected from bittersweet, hairy nightshade, petunia, potato, potato vine, and tomato in western Washington, 1998 to 1999, were evaluated for virulence complexity as well as mating type, metalaxyl insensitivity, allozymes of glucose-6-phosphate isomerase and peptidase, and DNA fingerprint with the RG57 probe. Results were compared with those from similar collections made in the same region during the 1990s. Generally, virulence complexity was high for most of the isolates regardless of year, genotype, or host. No marked shift in virulence complexity was evident for the populations studied, and unnecessary virulences were maintained. During 1998 and 1999, isolates of the US-8 and US-11 genotypes had 4 or more virulence factors. US-8 isolates averaged 8.2 and 9.3, whereas US-11 isolates averaged 5.4 and 6.3 virulence factors. The frequency of US-8 isolates that were sensitive to metalaxyl ranged from 5% in 1998 to 72% in 1999. All of the US-11 isolates tested in 1998 and 1999 were insensitive to metalaxyl. From 1996 to 1999 on potato, the recovery of US-8 increased, whereas the recovery of US-11 decreased. No evidence of new genotypes or sexual recombination was found. Western Washington was a desirable location for screening germ plasm for durable resistance to late blight due to the high frequency and persistence of complex virulences.
Combining Biocontrol Agents to Reduce the Variability of Biological Control
Authors: Guetsky Ruth , Shtienberg D. , Elad Y. , Dinoor A.
Published: May 2007

Two biocontrol agents, a yeast (Pichia guilermondii) and a bacterium (Bacillus mycoides), were tested separately and together for suppression of Botrytis cinerea on strawberry leaves. The aims of the research were to determine whether the use of their combination would broaden the environmental conditions under which biological control is effective, and to test the hypothesis that it would reduce the variability of control efficacy under diverse conditions. Applied separately, the biocontrol agents significantly inhibited spore germination, lesion formation, and lesion development at most temperatures, relative humidities, and spray-timing combinations (temperatures: 10, 15, 20, 23, 25, and 30°C; relative humidities: 78, 85, 96, and 100%; and spray-timings: 0, 4, and 7 days before inoculation). However, control efficacy was highly variable, and under certain combinations it was not adequate. Control efficacy achieved by the biocontrol agents applied separately ranged between 38 and 98% (mean 74%) and the coefficient of variation ranged from 9.7 to 75%. The mixture of Bacillus mycoides and Pichia guilermondii suppressed Botrytis cinerea effectively (80 to 99.8% control) under all conditions, and the coefficients of variation were as low as 0.4 to 9% in all cases. Thus, application of both biocontrol agents resulted in better suppression of Botrytis cinerea, and also reduced the variability of disease control. Application of more than one biocontrol agent is suggested as a reliable means of reducing the variability and increasing the reliability of biological control.
Characterization of South African Cryphonectria cubensis Isolates Infected with a C. parasitica Hypovirus
Authors: van Heerden S. W. , Geletka L. M. , Preisig O. , Nuss D. L. , Wingfield B. D. , Wingfield M. J.
Published: May 2007

Cryphonectria cubensis is the causal agent of a serious canker disease of Eucalyptus spp. in tropical and subtropical parts of the world. In this study, a South African C. cubensis isolate was transfected by electroporation with a synthetic RNA transcript corresponding to the full-length coding strand of the C. parasitica hypovirus (CHV1-EP713). Hypovirus infection resulted in pronounced morphological changes that included a striking increase in bright yellow-orange pigment production, a reduction in mycelial growth rate, and reduced sporulation. Greenhouse studies revealed that the virus-containing strain was significantly less virulent than the original virulent C. cubensis isolate. Although the hypovirus was not transmitted through conidia produced by infected C. cubensis, the virus was readily transmitted via hyphal anastomosis to C. cubensis isolates representing a broad range of vegetative compatibility groups. These results suggest that vegetative incompatibility may not pose a strong barrier against virus transmission in South African isolates of C. cubensis and that hypovirus-mediated biological control could provide opportunities to reduce the impact of Cryphonectria canker in South Africa.
Influence of Take-All Epidemics on Winter Wheat Yield Formation and Yield Loss
Authors: Schoeny Alexandra , Jeuffroy Marie-Hélène , Lucas Philippe
Published: May 2007

The effects of take-all epidemics on winter wheat yield formation were determined, and disease-yield relationships were established to assess the agronomic efficacy and economic benefits of control methods. Epidemics were generated in naturally infested fields by varying cropping season, crop order in the rotation, and experimental fungicide seed treatment. Disease incidence and severity were assessed from tillering to flowering. Yield components were measured at harvest. Models simulating the formation of the yield components in the absence of limiting factors were used to estimate the losses caused by take-all. Losses were predicted by the disease level at a specific time or the area under the disease progress curve, reflecting accumulation during a specific period. Losses of grain number per square meter and 1,000-grain weight were linked to cumulative disease incidence between the beginning of stem elongation and flowering, and disease incidence at midstem elongation, respectively. Yield losses were accounted for by both cumulative disease incidence between sowing and flowering, and disease incidence at midstem elongation. Results confirm the importance of nitrogen fertilization in reducing the impact of take-all on wheat.
Evidence for the Predisposition of Fungicide-Resistant Isolates of Venturia inaequalis to a Preferential Selection for Resistance to Other Fungicides
Authors: Köller Wolfram , Wilcox W. F.
Published: May 2007

In the United States, populations of the apple scab pathogen Venturia inaequalis have progressed through three consecutive rounds of fungicide resistance development, first to dodine, then to the benzimidazoles, and most recently to the sterol demethylation inhibitors (DMIs). Analysis of extensive monitoring data have to date provided no indication of detectable cross-resistance or partial cross-resistance of V. inaequalis populations to the three unrelated classes of fungicides prior to the selection of resistant subpopulations. However, in this study, resistance to both benomyl and DMIs developed to significantly higher frequencies within the previously established dodine-resistant population than in the population sensitive to dodine. Accelerated selection of phenotypes double resistant to dodine and the DMI fenarimol was apparent over the course of distinct seasons of apple scab management with either dodine or fenarimol. The data provide evidence for an accelerated speed of resistance development among phenotypes of V. inaequalis already resistant to an unrelated fungicide. This finding represents a departure from the previous model, which assumed entirely independent rounds of resistance developments. The data indicate that phenotypes of V. inaequalis might not only be selected for the trait of fungicide resistance but also for traits allowing a more flexible response to changes in the environment where they compete.
Characterization of Distinct Tombusviruses that Cause Diseases of Lettuce and Tomato in the Western United States
Authors: Obermeier C. , Sears J. L. , Liu H. Y. , Schlueter K. O. , Ryder E. J. , Duffus J. E. , Koike S. T. , Wisler G. C.
Published: May 2007

A soilborne disease of lettuce, associated with necrosis and dieback, has been found with increasing frequency in California and Arizona over the last 10 years. An isometric virus, serologically related to Tomato bushy stunt virus (TBSV), was consistently isolated from lettuce plants with these disease symptoms. Back-inoculation to healthy lettuce plants and subsequent reisolation of the virus from symptomatic lettuce leaves suggested that this virus was the causal agent of this disease. A tombusvirus was also associated with a necrosis disease of greenhouse-grown tomatoes in Colorado and New Mexico. Complementary DNA representing the 3′ end of viral genomic RNAs recovered from diseased lettuce and tomato plants had identical nucleotide sequences. However, these sequences were divergent (12.2 to 17.1%) from sequences of the previously described strains of TBSV, Petunia asteroid mosaic virus (PAMV), Artichoke mottled crinkle virus, and Carnation Italian ringspot virus. Additional tombusvirus isolates were recovered from diseased lettuce and tomato plants and these were most closely related to the TBSV-cherry strain (synonymous with PAMV) and to Cucumber necrosis virus based on comparison of 3′-end sequences (0.1 to 0.6% and 4.8 to 5.1% divergence, respectively). Western blot analysis revealed that the new tombusvirus isolated from diseased lettuce and tomato plants in the western United States is serologically distinct from previously described tombusvirus species and strains. Based on genomic and serological properties, we propose to classify this virus as a new tombusvirus species and name it Lettuce necrotic stunt virus.
Differentiation of Three Homogeneous Groups of Rhizoctonia solani Anastomosis Group 4 by Analysis of Fatty Acids
Authors: Johnk Janell Stevens , Jones Roger K.
Published: May 2007

Profiles of fatty acids from 70 isolates of Rhizoctonia solani anastomosis group (AG)-4 clustered into three groups, corresponding to homogeneous group (HG)-I, HG-II, and a newly described HG-III. Isolates from Georgia peanuts exhibiting limb rot were characterized as gas chromatography (GC) subgroup 1 (GC-1) and contained HG-I isolates. Isolates from diseased soybean hypocotyls grown in North Dakota and sugar beet seedlings, taproots, and tare soil in Minnesota and North Dakota were characterized as GC subgroup 2 (GC-2) and contained predominantly HG-II isolates but also included three distinct isolates based on fatty acid methyl ester (FAME) analysis and morphological features. Selected isolates from North Carolina cucumbers clustered into three distinct groups that corresponded to HG-I, HG-II, and the newly described HG-III. Distinct isolates from the soybean and sugar beet populations clustered with HG-III. Fatty acid profiles of AG-4 were compared with FAME library profiles of AG-1, AG-2 type 2, and AG-3, which were developed in previous studies and were sufficiently different that they could be used to support speciation of this group from R. solani. It is suggested that binomial R. practicola may be appropriate for the portion of AG-4 identified as HG-II.
Association of Cellulytic Enzyme Activities in Eucalyptus Mulches with Biological Control of Phytophthora cinnamomi
Authors: Downer A. J. , Menge J. A. , Pond E.
Published: May 2007

A series of samples were taken from mulched and unmulched trees starting at the surface of mulch or soil to a 15 cm soil depth, forming a vertical transect. Saprophytic fungi isolated from the soil samples on rose bengal medium and surveyed visually were most abundant in mulches and at the interface of mulch and soil (P < 0.05). Microbial activity as assayed by the hydrolysis of fluorescein diacetate was significantly greater in mulch layers than in soils. Cellulase and laminarinase enzyme activities were greatest in upper mulch layers and rapidly decreased in soil layers (P < 0.05). Enzyme activities against Phytophthora cinnamomi cell walls were significantly greater in mulch than in soil layers. When Phytophthora cinnamomi was incubated in situ at the various transect depths, it was most frequently lysed at the interface between soil and mulch (P < 0.001). Roots that grew in mulch layers were significantly less infected with Phytophthora cinnamomi than roots formed in soil layers. In mulched soil, roots were commonly formed at the mulch-soil interface where Phytophthora populations were reduced, whereas roots in unmulched soil were numerous at the 7.5 cm depth where Phytophthora cinnamomi was prevalent. Enzyme activities were significantly and positively correlated with each other, microbial activity, and saprophytic fungal populations, but significantly and negatively correlated with Phytophthora recovery.
Biotic Factors Affecting Expression of the 2,4-Diacetylphloroglucinol Biosynthesis Gene phlA in Pseudomonas fluorescens Biocontrol Strain CHA0 in the Rhizosphere
Authors: Notz R. , Maurhofer M. , Schnider-Keel U. , Duffy B. , Haas D. , Défago G.
Published: May 2007

Production of the polyketide antimicrobial metabolite 2,4-diacetyl-phloroglucinol (DAPG) is a key factor in the biocontrol activity of Pseudomonas fluorescens CHA0. Strain CHA0 carrying a translational phlA′-′lacZ fusion was used to monitor expression of the phl biosynthetic genes in vitro and in the rhizosphere. Expression of the reporter gene accurately reflected actual production of DAPG in vitro and in planta as determined by direct extraction of the antimicrobial compound. In a gnotobiotic system containing a clay and sand-based artificial soil, reporter gene expression was significantly greater in the rhizospheres of two monocots (maize and wheat) compared with gene expression in the rhizospheres of two dicots (bean and cucumber). We observed this host genotype effect on bacterial gene expression also at the level of cultivars. Significant differences were found among six additional maize cultivars tested under gnotobiotic conditions. There was no difference between transgenic maize expressing the Bacillus thuringiensis insecticidal gene cry1Ab and the near-isogenic parent line. Plant age had a significant impact on gene expression. Using maize as a model, expression of the phlA′-′lacZ reporter gene peaked at 24 h after planting of pregerminated seedlings, and dropped to a fourth of that value within 48 h, remaining at that level throughout 22 days of plant growth. Root infection by Pythium ultimum stimulated bacterial gene expression on both cucumber and maize, and this was independent of differences in rhizosphere colonization on these host plants. To our knowledge, this is the first comprehensive evaluation of how biotic factors that commonly confront bacterial inoculants in agricultural systems (host genotype, host age, and pathogen infection) modulate the expression of key biocontrol genes for disease suppression.
Characterization of Synergy Between Cucumber mosaic virus and Potyviruses in Cucurbit Hosts
Authors: Wang Yongzeng , Gaba Victor , Yang Jie , Palukaitis Peter , Gal-On Amit
Published: May 2007

Mixed infections of cucurbits by Cucumber mosaic virus (CMV) and potyviruses exhibit a synergistic interaction. Zucchini squash and melon plants coinfected by the potyvirus Zucchini yellow mosaic virus (ZYMV) and either Fny-CMV (subgroup IA) or LS-CMV (subgroup II) displayed strong synergistic pathological responses, eventually progressing to vascular wilt and plant death. Accumulation of Fny- or LS-CMV RNAs in a mixed infection with ZYMV in zucchini squash was slightly higher than infection with CMV strains alone. There was an increase in CMV (+) strand RNA levels, but no increase in CMV (-) RNA3 levels during mixed infection with ZYMV. Moreover, only the level of capsid protein from LS-CMV increased in mixed infection. ZYMV accumulated to similar levels in singly and mixed infected zucchini squash and melon plants. Coinfection of squash with the potyvirus Watermelon mosaic virus (WMV) and CMV strains increased both the Fny-CMV RNA levels and the LS-CMV RNA levels. However, CMV (-) strand RNA3 levels were increased little or not at all for CMV on coinfection with WMV. Infection of CMV strains (LS and Fny) containing satellite RNAs (WL47-sat RNA and B5*-sat RNA) reduced the accumulation of the helper virus RNA, except when B5*-sat RNA was mixed with LS- CMV. However, mixed infection containing ZYMV and the CMV strains with satellites reversed the suppression effect of satellite RNAs on helper virus accumulation and increased satellite RNA accumulation. The synergistic interaction between CMV and potyviruses in cucurbits exhibited different features from that documented in tobacco, indicating there are differences in the mechanisms of potyvirus synergistic phenomena.
Partial Nucleotide Sequence and Genome Organization of a Canadian Isolate of Little cherry virus and Development of an Enzyme-Linked Immunosorbent Assay-Based Diagnostic Test
Authors: Theilmann J. , Mozafari J. , Reade R. , Wu Z. , Xie W. , Jesperson G. , Bernardy M. , Eastwell K. C. , Rochon D.
Published: May 2007

Approximately 12.4 kb of the genome of a mealybug-transmissible, North American isolate of Little cherry virus (LChV-3, previously designated LChV-LC5) has been cloned and sequenced. The sequenced portion of the genome contains 10 open reading frames (ORFs) and, based on sequence comparisons, encodes a putative RNA helicase (HEL), RNA-dependent RNA polymerase (POL), two coat proteins (CPs), a homologue of HSP70, a 53K protein (p53) that is similar to an equivalent-size protein in other closteroviruses, and a 22K (p22) protein of unknown function. The genome also potentially encodes two small proteins (p5 and p6), one of which is similar to the small hydrophobic proteins of other closteroviruses. Phylogenetic analyses utilizing sequences of the HEL, POL, and HSP70 homologue suggest that LChV-3 is most similar to other mealybug-transmitted closteroviruses. Further comparisons between LChV-3 and a 4.7-kb region of the recently described Little cherry virus-2 (LChV-2) reveals 77% nucleotide sequence identity. Based on this low sequence identity, we propose that LChV-3 be considered a separate species, designated LChV-3. Unexpectedly, the LChV-3 CP duplicate ORF was found to lie upstream of the HSP70 ORF; therefore, the genome organization of LChV-3 is distinct from that of other closteroviruses. Polyclonal antiserum raised to bacterially expressed LChV-3 CP was useful for detection of LChV-diseased trees in the cherry-growing districts of British Columbia, Canada.
Inheritance of Race-Specific Resistance toXanthomonas campestrispv.campestrisinBrassicaGenomes
Authors: Vicente J. G. , Taylor J. D. , Sharpe A. G. , Parkin I. A. P. , Lydiate D. J. , King G. J.
Published: May 2007

The inheritance of resistance to three Xanthomonas campestris pv. campestris races was studied in crosses between resistant and susceptible lines of Brassica oleracea (C genome), B. carinata (BC genome), and B. napus (AC genome). Resistance to race 3 in the B. oleracea doubled haploid line BOH 85c and in PI 436606 was controlled by a single dominant locus (Xca3). Resistance to races 1 and 3 in the B. oleracea line Badger Inbred-16 was quantitative and recessive. Strong resistance to races 1 and 4 was controlled by a single dominant locus (Xca1) in the B. carinata line PI 199947. This resistance probably originates from the B genome. Resistance to race 4 in three B. napus lines, cv. Cobra, the rapid cycling line CrGC5, and the doubled haploid line N-o-1, was controlled by a single dominant locus (Xca4). A set of doubled haploid lines, selected from a population used previously to develop a restriction fragment length polymorphism map, was used to map this locus. Xca4 was positioned on linkage group N5 of the B. napus A genome, indicating that this resistance originated from B. rapa. Xca4 is the first major locus to be mapped that controls race-specific resistance to X. campestris pv. campestris in Brassica spp.
Delayed Symptom Development in ein2-1, an Arabidopsis Ethylene-Insensitive Mutant, in Response to Bacterial Wilt Caused by Ralstonia solanacearum
Authors: Hirsch Judith , Deslandes Laurent , Feng Dong Xin , Balagué Claudine , Marco Yves
Published: May 2007

Wilt disease caused by the phytopathogenic bacterium Ralstonia solanacearum is poorly understood at the molecular level. The possible roles of salicylic acid, jasmonic acid, and ethylene, compounds commonly associated with the plant response to pathogens, in wilt symptom development were investigated using various Arabidopsis thaliana mutants in a Col-0 background, an ecotype that develops wilt symptoms in response to the virulent GMI1000 strain. Following root inoculation, wilt symptoms were delayed in ein2-1, an ethylene-insensitive mutant, in response to several virulent strains of the pathogen. In ein2-1, bacteria invade the plant and multiply, reaching concentrations slightly lower than those detected in susceptible plants but 1 to 2 logs higher than in Nd-1, an A. thaliana ecotype resistant to strain GMI1000. This delay in disease symptom development of ein2-1 plants suggests that ethylene signaling plays a critical role in wilt disease development. Furthermore, a strong accumulation of transcripts corresponding to PR-3 and PR-4, two ethylene-responsive genes, was observed in susceptible Col-0 plants, but not in ein2-1 and Nd-1 plants, providing additional evidence for a role of ethylene in wilt symptom production. However, this hormone is probably not involved in the establishment of resistance to R. solanacearum, because homozygous ein2-1 plants in a resistant background remain fully resistant to strain GMI1000.
An Allee Effect Reduces the Invasive Potential of Tilletia indica
Authors: Garrett K. A. , Bowden R. L.
Published: May 2007

The Karnal bunt pathogen, Tilletia indica, is heterothallic and depends on encounters on wheat spikes between airborne secondary sporidia of different mating types for successful infection and reproduction. This life history characteristic results in reduced reproductive success for lower population densities. Such destabilizing density dependence at low population levels has been described for a range of animals and plants and is often termed an Allee effect. Our objective was to characterize how the Allee effect might reduce the invasive potential of this economically important pathogen. We developed a simple population model of T. indica that incorporates an Allee effect by calculating the probability of infection for different numbers of secondary sporidia in the infection court. An Allee effect is predicted to be important at the frontier of an invasion, for establishment of new foci by a small population of teliospores, and when the environment is nonconducive for the production of secondary sporidia. Using estimated model parameter values, we demonstrated a theoretical threshold population size below which populations of T. indica were predicted to decline rather than increase. This threshold will vary from season to season as a function of weather variables and their effect on the reproductive potential of T. indica. Deployment of partial resistance or use of fungicides may be more useful if they push population levels below this threshold.
A Gene-for-Gene Relationship Underlying the Species-Specific Parasitism of Avena/Triticum Isolates of Magnaporthe grisea on Wheat Cultivars
Authors: Takabayashi N. , Tosa Y. , Oh H. S. , Mayama S.
Published: May 2007

To elucidate genetic mechanisms of the species-specific parasitism of Magnaporthe grisea, a Triticum isolate (pathogenic on wheat) was crossed with an Avena isolate (pathogenic on oat), and resulting F1 progeny were subjected to segregation analyses on wheat cvs. Norin 4 and Chinese Spring. We found two fungal loci, Pwt3 and Pwt4, which are involved in the specific parasitism on wheat. Pwt3 operated on both cultivars while Pwt4 operated only on ‘Norin 4’. Using the cultivar specificity of Pwt4, its corresponding resistance gene was successfully identified in ‘Norin 4’ and designated as Rmg1 (Rwt4). The presence of the corresponding resistance gene indicated that Pwt4 is an avirulence locus. Pwt3 was assumed to be an avirulence locus because of its temperature sensitivity. We suggest that gene-for-gene interactions underlie the species-specific parasitism of M. grisea.
Genetic Uniformity Among Isolates of Peronospora tabacina, the Tobacco Blue Mold Pathogen
Authors: Sukno Serenella A. , Taylor Amy M. , Farman Mark L.
Published: May 2007

As a first step toward analysis of genetic variation and population structure in Peronospora tabacina, we used a collection of random genomic DNA fragments to survey for restriction fragment length polymorphisms (RFLPs) in DNA from a collection of isolates from Kentucky and other tobacco-growing regions of the United States. Also included in the study were isolates from the wild tobacco species, Nicotiana repanda, and from ornamental tobacco, N. alata. In a preliminary survey using DNA from 10 pathogen isolates, no polymorphisms were detected at six single-copy DNA loci using 22 probe-enzyme combinations. Moderately repetitive and highly repetitive regions of the genome were also remarkably similar between isolates, with only 6 of 15 different probes identifying genetic differences. Some of the polymorphic probes were then used to analyze a larger collection of isolates, most of which were from Kentucky. This resulted in the identification of very few additional polymorphisms, indicating that the population of P. tabacina that infects the Kentucky tobacco crop is genetically very homogeneous. The low level of polymorphism detected in this study overall, suggests that genetic variability may be lacking in P. tabacina populations throughout the United States. Two of the RFLP markers gave hybridization patterns that were consistent with P. tabacina being diploid. Frequencies of alleles at these loci and linkage disequilibrium between different marker loci indicated that genetic recombination does not occur frequently in the pathogen population. DNA polymorphisms that were identified in this study enabled us to differentiate the pathogen population into at least 10 haplotypes. One isolate was analyzed in detail and was shown to be genetically stable through several rounds of single-spore isolation and through several pathogenic cycles.