bluuebunny/crossref_metadata_2025 huggingface dataset:
Rubber Tree (Hevea brasiliensis) Bark Necrosis Syndrome I: Still No Evidence of a Biotic Causal Agent
Authors: Pellegrin F. , Nandris D. , Chrestin H. , Duran-Vila N.
Published: May 2007

The main constraint in rubber plantations worldwide is the cessation of latex production because of two syndromes: (i) tapping panel dryness (TPD), a reversible physiological response to overexploitation; and (ii) bark necrosis (BN), an irreversible syndrome spreading from the collar toward the tapping cut. Early BN symptoms develop in the inner phloem tissues but never affect the cambium. Necrotic patches appear in the outer phloem, inducing bark cracking and peeling, but these alterations never lead to tree death. BN spreads gradually to neighboring rubber trees, and evidence of linear disease centers suggest that a pathogen may be involved, possibly transmitted by the tapping knife. Several previous etiological investigations (fungi, phytoplasma, bacteria, and virus) were performed (3) on leaves, bark, and latex using different methods (e.g., isolation, transmission, chemical treatments, and optic and electron microscope studies). Recent works focused on mechanically transmissible pathogens, such as viroid (2) or virus/double strand RNA, using RNA extraction (nonionic cellulose and appropriate ethanol concentrations) and treatment with RNase A, followed with sequential polyacrylamide gel electrophoresis (s-PAGE), reverse transcription-polymerase chain reaction (RT-PCR), degenerate oligonucleotide primer-PCR (DOP-PCR), and cloning and sequence analysis. While numerous viroid-like (between 250 and 400 nucleotides) and double strand virus-like (1,800 bp) low-molecular-weight RNAs were observed, no definite correlation was found with the BN status of trees. Sequencing of the various isolated RNAs only identified plasmids, nonpathogenic bacteria and yeasts, but none of the suspected pathogens. In addition, previous and recent transmission trials (tapping knife disinfection, bud grafting, bark implantation, and etc.) failed to confirm the involvement of a biotic agent. In conclusion, since these etiological investigations were inconclusive, a physiological disease is now suspected that involves exogenous stresses, nonoptimal vascular relations at the rootstock/scion junction and impaired cyanide metabolism (1,4).

References: (1) H. Chrestin et al. Plant Dis. 88:1047, 2004. (2) N. Duran-Vila et al. J. Gen. Virol. 69:3069, 1988. (3) D. Nandris et al. Eur. J. For. Pathol. 21:340, 1991. (4) D. Nandris et al. Plant Dis. 88:1047, 2004.
An Outbreak of Bacterial Speck Caused by Pseudomonas syringae pv. tomato on Tomato Transplants Grown in Commercial Seedling Companies Located in the Western Mediterranean Region of Turkey
Authors: Basim H. , Basim E. , Yilmaz S. , Dickstein E. R. , Jones J. B.
Published: May 2007

A serious outbreak of a leaf spot disease was observed on tomato transplants grown in commercial seedling companies in southwestern Turkey (Antalya) during the springs of 2002 and 2003. Disease incidence was more severe in the western Mediterranean Region of Turkey. Occurrence of the outbreak resulted in approximately 20 and 25% seedling losses in the springs of 2002 and 2003, respectively. The initial symptoms consisted of pronounced water-soaked, dark brown-to-black spots on young expanding leaves that were 1 to 2 mm in diameter. Later, a number of leaf spots on older leaves enlarged and coalesced, causing leaf desiccation and finally, seedling death. In addition, in 2003 the disease incidence was approximately 5% in 142 commercial greenhouses. Tomato production was unaffected since significant outbreaks did not occur on greenhouse plants. No fruit symptoms were observed. Twenty-six strains were isolated from diseased tomato seedlings and plants from different greenhouses located in different places and all were gram negative and fluorescent on King's B medium. All strains were levan and gelatin liquefaction positive and oxidase and arginine dihydrolase negative. None of the 26 strains utilized erythritol and l-lactate as the sole carbon source (1,2). Fatty acid analysis identified the strains as Pseudomonas syringae pv. tomato with similarity indices ranging from 0.876 to 0.932%. Pathogenicity of the isolates was confirmed on 4-week-old tomato seedlings (cv. Biotek Selin) sprayed with the bacterial suspensions containing 108 CFU/ml of sterile water. Later, a number of leaf spots on the leaves enlarged and coalesced, causing leaf desiccation. Inoculated and control tomato seedlings were covered with polyethylene bags and placed in a growth chamber at 25°C for 48 h and then the bags were removed. Small (1 to 2 mm), water-soaked, dark brown-to-black spots similar to those observed in the greenhouses of commercial seedling companies and commercial greenhouses that produce tomato developed on the young expanding leaves of inoculated plants within 7 to 10 days. No symptoms developed on control plants. The bacterium was reisolated from inoculated plants and identified as a strain of Pseudomonas syringae pv. tomato. Koch's postulates were fulfilled. To our knowledge, this is the first report for the occurrence and outbreak of the bacterial speck disease on tomato transplants in greenhouses of commercial tomato seedling production companies in Turkey.

References: (1) D. C. Hildebrand et al. Pages 60–80 in: Laboratory Guide for Identification of Plant Pathogenic Bacteria. N. D. Schaad, ed. The American Phytopathological Society, 1988. (2) J. B. Jones et al. Plant Dis. 70:151, 1986.
Epidemiology and Chemical Control of Take-All on Seminal and Adventitious Roots of Wheat
Authors: Bailey D. J. , Paveley N. , Pillinger C. , Foulkes J. , Spink J. , Gilligan C. A.
Published: May 2007

Epidemiological modeling is used to examine the effect of silthiofam seed treatment on field epidemics of take-all in winter wheat. A simple compartmental model, including terms for primary infection, secondary infection, root production, and decay of inoculum, was fitted to data describing change in the number of diseased and susceptible roots per plant over thermal time obtained from replicated field trials. This produced a composite curve describing change in the proportion of diseased roots over time that increased monotonically to an initial plateau and then increased exponentially thereafter. The shape of this curve was consistent with consecutive phases of primary and secondary infection. The seed treatment reduced the proportion of diseased roots throughout both phases of the epidemic. However, analysis with the model detected a significant reduction in the rate of primary, but not secondary, infection. The potential for silthiofam to affect secondary infection from diseased seminal or adventitious roots was examined in further detail by extending the compartmental model and fitting to change in the number of diseased and susceptible seminal or adventitious roots. Rates of secondary infection from either source of infected roots were not affected. Seed treatment controlled primary infection of seminal roots from particulate inoculum but not secondary infection from either seminal or adventitious roots. The reduction in disease for silthiofam-treated plants observed following the secondary infection phase of the epidemic was not due to long-term activity of the chemical but to the manifestation of disease control early in the epidemic.
Effects of Latent Infection, Temperature, Precipitation, and Irrigation on Panicle and Shoot Blight of Pistachio in California
Authors: Mila A. L. , Driever G. F. , Morgan D. P. , Michailides T. J.
Published: May 2007

Panicle and shoot blight, caused by a Fusicoccum sp., is an economically important disease of pistachio in California. Between 1999 and 2001, the disease severity was monitored throughout the growing season in 10 pistachio orchards, irrigated with drip, microsprinklers, low-angled (12°) sprinklers, or flood. The effect of temperature, precipitation pattern, irrigation system, and incidence of Fusicoccum sp. latent infection on panicle and shoot blight severity was quantified with a generalized linear model for repeated measures. The number of continuous rainy days in April and May and the cumulative daily mean temperatures from June to early September had a significant positive effect on panicle and shoot blight of pistachio leaves and fruit. Drip irrigation significantly decreased disease risk. Other factors, such as the number of discontinuous rainy days in April and May, the cumulative deviation from the 30-year average temperature during the dry days of April and May, the incidence of latent infection (only on leaves), and irrigation with microsprinklers or lowangled (12°) sprinklers were weak explanatory variables of panicle and shoot blight severity. Knowledge of panicle and shoot blight risk may contribute significantly to decisions regarding the appropriate application of fungicides, especially in years or fields of low risk.
Variation in Occurrence of Dematiaceous Hyphomycetes on Forage Bermudagrass over Years, Sampling Times, and Locations
Authors: Pratt R. G.
Published: May 2007

Leaf samples of forage bermudagrass with symptoms of infection by species of Bipolaris, Curvularia, and Exserohilum (dematiaceous hyphomycetes) were collected from three swine waste application sites in Mississippi at eight sampling times during each of 3 years. Samples were assayed for pathogens by observing sporulation on plated leaf tissue. Among 3,600 leaves assayed, eight species of the three genera were observed. Features and criteria for the practical identification of species on plated leaf samples are described. Sporulation by dematiaceous hyphomycetes was observed on 97% of leaves; a single pathogen was observed on 20% and two to five pathogens were observed on 77% of leaves. Distributions of leaves among classes with one to five pathogens per leaf, for sites within years, always differed significantly (P = 0.01) from a Poisson distribution and usually included fewer leaves than expected with four or five pathogens. Significant (P = 0.05) variation in frequencies of occurrence of pathogens among 72 samples of 50 leaves each was attributed to pathogen species, sampling times, and species-time interactions. Exserohilum rostratum, Curvularia lunata, and Bipolaris cynodontis were the most frequent pathogens across years and sites; B. spicifera and C. geniculata were intermediate; and B. hawaiiensis, B. sorokiniana, and B. stenospila were least frequent. For the five most common pathogens, significant differences in frequency among locations were commonplace. Six pathogens exhibited significant (P = 0.05) positive and negative correlations with others in overall frequencies of occurrence across years, sampling times, and sites. However, χ2tests of dual occurrence indicated that interactions between specific pairs of pathogens in or on leaves are not likely to be major causes for overall frequency correlations. Results indicate that dematiaceous hyphomycetes typically infect forage bermudagrass on swine waste application sites in complexes rather than as individual species; that E. rostratum, C. lunata, and B. cynodontis are the predominant pathogens; and that frequencies of pathogens often differ significantly between locations.
Identifying Risk Factors for European Stone Fruit Yellows from a Survey
Authors: Thébaud Gaël , Sauvion Nicolas , Chadœuf Joël , Dufils Arnaud , Labonne Gérard
Published: May 2007

European stone fruit yellows (ESFY) is becoming a major economic problem for Prunus growers in Europe. The causal agent (“Candidatus Phytoplasma prunorum”) and its vector (Cacopsylla pruni) have been identified, but the present knowledge of the risk factors for this disease relies, at best, on specific experiments. To assess the relative significance of several factors correlated with ESFY incidence in the field, an exhaustive survey was performed on apricot and Japanese plum orchards in the Crau plain (France). After a preliminary multivariate exploration of the data, we used a logistic regression model to analyze and predict the cumulative number of diseased trees on the basis of a set of quantitative (age, planting density, and area of the orchard) and categorical variables (species, cultivar, and rootstock). Because of the nature of the data, we used an overdispersed binomial model and we developed a parametric bootstrap procedure based on the beta-binomial distribution to obtain confidence intervals. Our results indicated that the age, species, and cultivar of the scion were the major factors explaining the observed number of diseased trees. The planting density and the rootstocks used in the zone under study were less significant, and the area of the orchard had no effect. The residuals of the model showed that some explanatory variables had not been taken into account, because part of the remaining variability could be explained by a grower effect. The spatial distribution of the residuals suggested that one of the reasons for this grower effect was the correlation between orchards closer than 100 m, possibly caused by the flight behavior of infectious vectors.
Individual-Based Approach to Modeling Hyphal Growth of a Biocontrol Fungus in Soil
Authors: Knudsen G. R. , Stack J. P. , Schuhmann S. O. , Orr K. , LaPaglia C.
Published: May 2007

Fungi in soil perform beneficial roles that include biological control of soilborne plant pathogens. However, relatively little predictive information is available about the growth and activity of fungal hyphae in soil habitats. A stochastic computer simulation model (“Fungmod”) was developed to predict hyphal growth of the biocontrol fungus Trichoderma harzianum ThzID1 in soil. The model simulates a fungal colony as a population of spatially referenced hyphal segments, and is individual-based, in that records of spatial location and branching hierarchy are maintained for individual hyphal nodes. In this way, the entire spatial structure of the fungal colony (hyphal network) can be explicitly reconstructed at any point in time. Also, the soil habitat is modeled as a population of spatially referenced 1-mm3soil cells, allowing for the simulation of a spatially heterogeneous environment. Initial hyphal growth parameters were derived from previously published results, and the model was tested against new data derived from image analysis of hyphal biomass accumulation in soil. The ability to predict fungal growth in natural habitats will help to improve the predictability of successful myco-parasitic events in biological control systems.
Etiology and Real-Time Polymerase Chain Reaction-Based Detection ofGremmeniella- andPhomopsis-Associated Disease in Norway Spruce Seedlings
Authors: Børja Isabella , Solheim Halvor , Hietala Ari M. , Fossdal Carl Gunnar
Published: May 2007

In spring 2002, an unusual disease outburst was recorded on Norway spruce seedlings in southeast Norway. Extensive damage was recorded on 1- and 2-year-old Norway spruce seedlings that either had wintered in nursery cold storage or had been planted out in autumn 2001. The damage was characterized by leader shoot dieback and stem necroses on the upper or lower part of the shoot from 2001. Gremmeniella abietina and a Phomopsis sp. frequently were isolated from the diseased seedlings. Internal transcribed spacer (ITS) ribosomal (r)DNA sequence analysis and random amplified microsatellites profiling indicated that the G. abietina strains associated with diseased nursery seedlings belonged to the large-tree type (LTT) ecotype of the European race of G. abietina var. abietina, and inoculation tests confirmed their pathogenicity on Norway spruce. Based on ITS rDNA sequence analysis, the Phomopsis strains associated with diseased seedlings did not represent any characterized Phomopsis spp. associated with conifers. The Phomopsis sp. was not pathogenic in inoculation tests, indicating that it may be a secondary colonizer. ITS-based real-time polymerase chain reaction assays were developed in order to detect and quantify G. abietina and Phomopsis in the nursery stock. We describe here the G. abietina-associated shoot dieback symptoms on Norway spruce seedlings and conclude that the unusual disease outburst likely was related to the G. abietina var. abietina epidemic caused by the LTT on large Scots pines in 2001.
Ascospore Release and Infection of Apple Leaves by Conidia and Ascospores of Venturia inaequalis at Low Temperatures
Authors: Stensvand Arne , Gadoury David M. , Amundsen Terje , Semb Lars , Seem Robert C.
Published: May 2007

Mills' infection period table describes the number of hours of continuous leaf wetness required at temperatures from 6 to 25°C for infection of apple leaves by ascospores of Venturia inaequalis and reports that conidia require approximately two-thirds the duration of leaf wetness required by ascospores at any given temperature. Mills' table also provides a general guideline that more than 2 days of wetting is required for leaf infection by ascospores below 6°C. Although the table is widely used, infection times shorter than those in the table have been reported in lab and field studies. In 1989 a published revision of the table eliminated a potential source of error, the delay of ascospore release until dawn when rain begins at night, and shortened the times reported by Mills for ascospore infection by 3 h at all temperatures. Data to support the infection times below 6°C were lacking, however. Our objective was to quantify the effects of low temperatures on ascospore discharge, ascospore infection, and infection by conidia. In two of three experiments at 1°C, the initial release of ascospores occurred after 131 and 153 min. In the third experiment at 1°C, no ascospores were detected during the first 6 h. The mean time required to exceed a cumulative catch of 1% was 143 min at 2°C, 67 min at 4°C, 56 min at 6°C, and 40 min at 8°C. At 4, 6, and 8°C, the mean times required to exceed a cumulative catch of 5% were 103, 84, and 53 min, respectively. Infection of potted apple trees by ascospores at 2, 4, 6, and 8°C required 35, 28, 18, and 13 h, respectively; substantially shorter times than previously were reported. In parallel inoculations of potted apple trees, conidia required approximately the same periods of leaf wetness as ascospores at temperatures from 2 to 8°C, rather than the shorter times reported by Mills or the longer times reported in the revision of the Mills table. We propose the following revisions to infection period tables: (i) shorter minimum infection times for ascospores and conidia at or below 8°C, and (ii) because both ascospores and conidia are often present simultaneously during the season of ascospore production and the required minimum infection times appear to be similar for both spore types, the adoption of a uniform set of criteria for ascosporic and conidial infection based on times required for infection by ascospores to be applied during the period prior to the exhaustion of the ascospore supply. Further revisions of infection times for ascospores may be warranted in view of the delay of ascospore discharge and the reduction of airborne ascospore doses at temperatures at or below 2°C.
Improved Medium for Selecting Nitrate-Nonutilizing (nit) Mutants of Verticillium dahliae
Authors: Korolev Nadia , Katan Talma
Published: May 2007

Nitrate-nonutilizing (nit) mutants are commonly used to determine vegetative compatibility between isolates of Verticillium dahliae by complementation (heterokaryon) testing. These mutants emerge spontaneously as chlorate-resistant sectors growing out of partially restricted, wild-type colonies on chlorate-amended media. The commonly used chlorate media are based on minimal medium (MMC) or cornmeal agar (CMC), amended with potassium chlorate. nit mutants recovered on these media constituted 10 to 36%(on MMC) and 25 to 45%(on CMC) of the apparently resistant sectors. An improved water agar chlorate medium (WAC) is described that is more effective for selecting chlorate-resistant nit mutants. WAC medium consists of agar (2%), glucose (0.02%), and potassium chlorate (2 to 5%). On WAC, growth of most V. dahliae isolates was strongly inhibited, and 66 to 100%(average >80%) of the chlorate-resistant sectors formed were nit mutants. Most mutants were characterized as nit1, and about 6% as NitM.
Detection and Quantification of Resistance of Venturia inaequalis Populations to Sterol Demethylation Inhibitors
Authors: Köller Wolfram , Wilcox W. F. , Barnard J. , Jones A. L. , Braun P. G.
Published: May 2007

Monoconidial isolates of Venturia inaequalis were collected in 1990 and 1991 from orchards in New York, Michigan, and Nova Scotia that had never or only sporadically been treated with fungicides acting as sterol demethylation inhibitors (DMIs). Sensitivities of isolates to two representative DMIs (fenarimol and myclobutanil) were determined by a sensitivity test based on the relative growth (RG) of mycelial colonies at one discriminatory dose. Mean isolate sensitivities were not significantly different (P > 0.2) for the majority of the populations tested, and all sensitivity data obtained from these sites were combined to provide a baseline distribution of isolate sensitivities for both fenarimol and myclobutanil. The baseline distributions were compared with isolate sensitivities determined for an experimental orchard in Nova Scotia with a documented case of DMI resistance and for a commercial orchard in Michigan with a long history of DMI use and first evidence of practical DMI resistance. For both DMIs tested and in both treated orchards, frequencies of isolates with RG values <80 had decreased or only slightly increased compared to the baseline population. In contrast, frequencies of isolates with RG values >80 had increased more than 20-fold over baseline levels. Thus, isolates with RG values >80 were rated DMI resistant. The validity of a qualitative isolate classification was tested in controlled infection studies. At doses of fenarimol and myclobutanil recommended for commercial control of apple scab, reproduction of a typical sensitive isolate on treated apple leaves was suppressed completely. Lesions caused by a resistant isolate continued to expand and produced abundant conidia. Statistical analysis of orchard sensitivities revealed that the analysis of isolate counts grouped into the categories DMI sensitive or resistant was most indicative in comparisons of orchard sensitivities aimed at detection of practical DMI resistance. A high degree of cross-resistance between fenarimol and myclobutanil indicated that sensitivity tests with one of the DMIs employed as the diagnostic tool in this study can serve as a test for other DMIs.
Phenology of Apothecium Production in Populations ofMonilinia vaccinii-corymbosifrom Early- and Late-Maturing Blueberry Cultivars
Authors: Lehman Jeffrey S. , Oudemans Peter V.
Published: May 2007

Pseudosclerotia were evaluated for differences in timing of apothecium development in four controlled experiments conducted over a 2-year period. In a separate experiment, conidia from 10 randomly selected isolates from both of the fungal populations were used to inoculate open flowers. Germination of pseudosclerotia produced from these artificial inoculations also was evaluated. The timing and rate of shoot elongation for cvs. Weymouth and Jersey were assessed in one greenhouse and two field experiments. Average development times for the fungal population from cv. Weymouth were 8 to 15 days earlier or 33 to 42% less than those for the population from cv. Jersey. The fungal population from Weymouth also exhibited less variation in development times for each developmental stage measured. Similarly, germination of pseudosclerotia produced in artificial inoculations differed between populations. On average, pseudosclerotia derived from the Weymouth population produced apothecia 16 days earlier. During spring 1995 and 1996, vegetative and truss buds on cv. Weymouth developed 4 to 16 days earlier than those on cv. Jersey. These results demonstrate that M. vaccinii-corymbosi exhibits variation in timing of pseudosclerotia germination and apothecium development within and between populations. We hypothesize that differences observed in the timing of apothecium development are related to the fitness of the populations on their original host cultivars and were selected by host phenology.
Genetic Analysis and Identification of Amplified Fragment Length Polymorphism Markers Linked to the alm1 Avirulence Gene of Leptosphaeria maculans
Authors: Pongam Patchara , Osborn Thomas C. , Williams Paul H.
Published: May 2007

A gene-for-gene interaction was previously suggested by mapping of a single major locus (LEM 1) controlling cotyledon resistance to Leptosphaeria maculans isolate PHW1245 in Brassica napus cv. Major. In this study, we obtained further evidence of a gene-for-gene interaction by studying the inheritance of the corresponding avirulence gene in L. maculans isolate PHW1245. The analysis of segregating F1 progenies and 14 test crosses suggested that a single major gene is involved in the interaction. This putative avirulence gene was designated alm1 after the resistance locus identified in B. napus. Amplified fragment length polymorphism (AFLP) markers were used to generate a rudimentary genetic linkage map of the L. maculans genome and to locate markers linked to the putative avirulence locus. Two flanking AFLP markers, AC/TCC-1 and AC/CAG-5, were linked to alm1 at 3.1 and 8.1 cM, respectively. Identification of markers linked to the avirulence gene indicated that the differential interaction is controlled by a single gene difference between parental isolates and provides further support for the gene-for-gene relationship in the Leptosphaeria-Brassica system.
Rapid Population Analysis of Magnaporthe grisea by Using rep-PCR and Endogenous Repetitive DNA Sequences
Authors: George M. L. C. , Nelson R. J. , Zeigler R. S. , Leung H.
Published: May 2007

DNA samples from Magnaporthe grisea isolates were fingerprinted by using repetitive element-based polymerase chain reaction (rep-PCR) with two outwardly directed primer sequences from Pot2, an element found in approximately 100 copies in the fungal genome. Variable length fragments, defining the sequences lying between these elements, were generated, and fingerprint patterns specific for individual strains were established. “Long PCR” conditions, including higher pH (9.2) and increased extension time (10 min) were used to amplify DNA fragments ranging from 400 bp to longer than 23 kb. Polymorphisms specific to M. grisea strains were generated, allowing inference of their genetic relationships. Segregation analysis was used to confirm single-locus inheritance for the fragments amplified by rep-PCR. Cluster analysis revealed robust groupings that corresponded to previously determined MGR586 restriction fragment length polymorphism lineages of the rice-infecting strains of the pathogen. We have also demonstrated the utility of rep-PCR to differentiate isolates that infect rice from those that infect nonrice hosts. DNA fingerprinting by Pot2 rep-PCR provides an efficient means to monitor the population dynamics of the blast pathogen. Because of the method's low cost and ease in application, it is now feasible to conduct large-scale population studies to understand the impact of host genotypes on pathogen evolution.
Specificity of Prehaustorial Resistance to Puccinia hordei and to Two Inappropriate Rust Fungi in Barley
Authors: Hoogkamp T. J. H. , Chen W.-Q. , Niks R. E.
Published: May 2007

To elucidate the specificity of prehaustorial resistance to inappropriate rust fungi, we studied two populations of recombinant inbred lines of barley that segregated for partial resistance (PR) to Puccinia hordei and for the resistance to the inappropriate rust species P. recondita f. sp. tritici and P. hordei-murini. PR to P. hordei is prehaustorial and nonhypersensitive, and its level can be assessed accurately by measuring the latent period of the fungus. The resistance to the inappropriate rust species is a combination of prehaustorial (nonhypersensitive) and posthaustorial (hypersensitive) mechanisms. The amount of nonhypersensitive, early abortion of P. recondita f. sp. tritici and P. hordei-murini sporelings reflects the degree of prehaustorial defense to the two inappropriate rust species. All lines showing a long latent period of P. hordei also had a relatively high level of early abortion of the growth of P. recondita f. sp. tritici and P. hordei-murini. This indicates that genes for PR to P. hordei are also effective against these two inappropriate rust species. The reverse was not necessarily true; some lines showing a high level of early abortion of P. recondita f. sp. tritici and P. hordei-murini had a low level of PR to P. hordei. Moreover, lines with a similar level of prehaustorial resistance to P. recondita f. sp. tritici could differ considerably in their prehaustorial resistance to P. hordei-murini. This indicates that genes for prehaustorial resistance may exhibit rust species specificity.
Quantification of Within-Field Spread of Soybean Mosaic Virus in Soybean Using Strain-Specific Monoclonal Antibodies
Authors: Nutter Forrest W. , Schultz Patricia M. , Hill John H.
Published: May 2007

Strain-specific monoclonal antibodies were used to follow the temporal increase and spatial spread of soybean mosaic virus (SMV) strain G-5 released from a point source. The use of strain-specific monoclonal antibodies allowed discrimination of within-field temporal and spatial spread of SMV strain G-5 from non-G-5 SMV isolates that originated from exogenous field sources. SMV isolates originating from exogenous sources have potential to alter the temporal and spatial pattern of within-field virus spread, which could potentially affect the choice of models used to quantify within-field pathogen spread. Analysis of SMV epidemics in field-plot experiments indicated that the logistic model was the most appropriate model to describe and compare the temporal spread of SMV among years. On the basis of ordinary runs analyses, within-field spread of SMV strain G-5 was random in 1991 and 1994, but was mostly aggregated in 1992 and 1993. Non-G-5 SMV isolates arising from exogenous sources displayed a random spatial pattern over time. This is the first study in which pathogen incidence (detection of SMV using strain-specific monoclonal antibodies) as opposed to disease incidence (based on symptoms) was employed to monitor and model SMV spread in time and space.
Use of Reciprocal Grafting to Study Brown Stem Rot Resistance in Soybean
Authors: Bachman M. S. , Nickell C. D.
Published: May 2007

Resistance to brown stem rot of soybean has been identified and utilized in cultivar development and germ plasm enhancement. However, little is known about the nature of resistance to this disease. In an attempt to better understand the resistance mechanism by locating the tissue or tissues responsible for resistance, reciprocal grafts were made between brown stem rot-resistant and -susceptible genotypes including near-isogenic lines for resistance gene Rbs2. Results of brown stem rot evaluation of grafted plants indicate that brown stem rot resistance is conditioned by the root in all resistance sources evaluated.
Near Immunity to Rice Tungro Spherical Virus Achieved in Rice by a Replicase-Mediated Resistance Strategy
Authors: Huet H. , Mahendra S. , Wang J. , Sivamani E. , Ong C. A. , Chen L. , de Kochko A. , Beachy R. N. , Fauquet C.
Published: May 2007

Rice tungro disease is caused by rice tungro bacilliform virus (RTBV), which is responsible for the symptoms, and rice tungro spherical virus (RTSV), which assists transmission of both viruses by leafhoppers. Transgenic japonica rice plants (Oryza sativa) were produced containing the RTSV replicase (Rep) gene in the sense or antisense orientation. Over 70% of the plants contained one to five copies of the Rep gene, with integration occurring at a single locus in most cases. Plants producing antisense sequences exhibited significant but moderate resistance to RTSV (60%); accumulation of antisense RNA was substantial, indicating that the protection was not of the homology-dependent type. Plants expressing the full-length Rep gene, as well as a truncated Rep gene, in the (+)-sense orientation were 100% resistant to RTSV even when challenged with a high level of inoculum. Accumulation of viral RNA was low, leading us to conclude that RTSV Rep-mediated resistance is not protein-mediated but is of the cosuppression type. Resistance was effective against geographically distinct RTSV isolates. In addition, RTSV-resistant transgenic rice plants were unable to assist transmission of RTBV. Such transgenic plants could be used in an epidemiological approach to combat the spread of the tungro disease.
Microbial Antagonism at the Root Level Is Involved in the Suppression of Fusarium Wilt by the Combination of Nonpathogenic Fusarium oxysporum Fo47 and Pseudomonas putida WCS358
Authors: Duijff Ben J. , Recorbet Ghislaine , Bakker Peter A. H. M. , Loper Joyce E. , Lemanceau Philippe
Published: May 2007

Two biological control agents, nonpathogenic Fusarium oxysporum Fo47 and Pseudomonas putida WCS358, were evaluated for suppression of Fusarium wilt of flax grown in nutrient solution and for suppression of the population density and metabolic activity of the causal organism F. oxysporum f. sp. lini strain Foln3GUS on root surfaces. Due to the presence of an introduced gusA reporter gene construct in Foln3GUS, the pathogen expressed β-glucuronidase activity that was related to its carbon metabolism. At a Fo47 to Foln3GUS inoculum ratio of 100:1, both the population density of the pathogen and the β-glucuronidase activity on and in flax roots were reduced by the nonpathogenic strain, and Fusarium wilt was suppressed. At a Fo47 to Foln3GUS inoculum ratio of 10:1, Fo47 decreased the severity of Fusarium wilt to a smaller extent and it also reduced β-glucuronidase activity without reducing the density of Foln3GUS on flax roots. At a nonpathogenic to pathogenic Fusarium strains ratio of 10:1, the addition of P. putida WCS358 further suppressed Fusarium wilt and the density of the pathogen at the root level, whereas a mutant of WCS358 deficient in pseudobactin production had no significant effect. Iron availability to WCS358 on flax roots, assessed by ice-nucleation activity conferred from a transcriptional fusion (pvd-inaZ) of an ice-nucleation reporter gene to an iron-regulated promoter, was sufficiently low to allow pseudobactin production. P. putida WCS358 did not reduce the severity of Fusarium wilt of flax when inoculated without Fo47, and it did not improve disease suppression achieved by high inoculum doses of Fo47 (a Fo47 to Foln3GUS ratio of 100:1). Together, these data provide evidence that (i) suppression of Fusarium wilt of flax by Fo47 is related to reductions in the population density and metabolic activity of the pathogen on the root surface; (ii) WCS358 can enhance the biological control activity of Fo47, but this enhancement depends on the population of Fo47 relative to the pathogen; and (iii) pseudobactin contributes to suppression of Fusarium wilt by the combination of Fo47 and WCS358 on roots in which conditions are conducive to pseudobactin production by the bacterium.
Transmission and the Role of Honeybees in Field Spread of Blueberry Shock Ilarvirus, a Pollen-Borne Virus of Highbush Blueberry
Authors: Bristow P. R. , Martin R. R.
Published: May 2007

The rapid and random spread of blueberry shock ilarvirus (BlShV) in commercial plantings suggested that insects played a role in transmission. Transmission from infected field plants to trap plants only occurred during bloom, indicating that flowers were the avenue for infection and flower-visiting insects including pollinators were involved. Trap plants readily became infected during bloom when a honeybee hive was included in cages with diseased field plants and trap plants. There was limited transmission in cages without a hive and no transmission when healthy field bushes were caged with trap plants either with or without honeybees. Infective pollen survived within the hive for 1 but not 2 weeks. Western flower thrips did not transmit BlShV when they were allowed to feed on caged flowering trap plants in the presence of a source of infected pollen. Results of pollen washes, cell disruption, and electron microscopy show that particles of BlShV were on, in, and between the cells of the pollen tetrad. Pollen from infected plants germinated as readily as pollen from healthy plants. BlShV was seedborne, but at a low level. All 42 cultivars tested were susceptible to BlShV when inoculated by grafting. The primary mechanism of transmission appears to be the transfer of BlShV-contaminated pollen by honeybees from flowers on infected plants to flowers on healthy plants.
Pathogenicity of Leptosphaeria maculans Isolates on a Brassica napus-B. juncea Recombinant Line
Authors: Somda Irénée , Delourme Régine , Renard Michel , Brun Hortense
Published: May 2007

The Brassica napus-B. juncea recombinant line (MX), resistant to Leptosphaeria maculans, was produced by interspecific crosses and bears one gene (Jlm1) from the B. juncea B genome. We investigated whether this new resistance was race specific by characterizing protection against a large sample of L. maculans isolates. The pathogenicity of 119 isolates of L. maculans comprising 105 A-group isolates and 14 B-group isolates was studied at the cotyledon stage under controlled conditions using the MX line, the susceptible B. napus cultivar Westar, and the resistant B. juncea cultivar Picra. All but one of the isolates were pathogenic on ‘Westar’. Only 3 of the 105 A-group isolates caused very mild symptoms on ‘Picra’. Two of these strains were isolated from the MX line and the other from Sinapis arvensis. The other 102 strains caused hypersensitive-type responses. Most B-group isolates were pathogenic on ‘Picra’. There were differences in pathogenicity among A-group isolates tested on the MX line, whereas all B-group isolates were pathogenic on this line. A-group isolates obtained from the MX line were more frequently pathogenic on the MX line than those obtained from B. napus cultivars. One isolate from S. arvensis infected the MX line. These results suggest that the resistance of the MX line is unlikely to be durable. Thus, the new resistance gene Jlm1 should probably be used in association with other sources of resistance, in plant breeding schemes, to prevent the breakdown of this resistance.
Distribution of Cranberry Fruit-Rotting Fungi in New Jersey and Evidence for Nonspecific Host Resistance
Authors: Stiles C. M. , Oudemans P. V.
Published: May 2007

A survey was conducted over a 3-year period to determine the frequencies and distributions of fruit-rotting fungi in New Jersey cranberry beds. In the first 2 years of the study, Physalospora vaccinii and Glomerella cingulata were the most prevalent and widespread field-rotting fungi. In the third year, the frequency of G. cingulata declined markedly. Other species such as Coleophoma empetri, Phyllosticta vaccinii, and Phomopsis vaccinii were isolated at high frequencies from a limited number of locations. Storage-rotting fungi including Allantophomopsis cytisporea and A. lycopodina were isolated at low frequencies, but were widely distributed within the growing region. On sound fruit, a somewhat different profile emerged. Fungi such as Phyllosticta elongata, Alternaria spp., and Physalospora vaccinii were commonly isolated. In comparisons among different cranberry cultivars, no differences in the fungal profiles were seen. This was interpreted to indicate that if differences in fruit-rot resistance exist, they are likely to be general forms of resistance rather than fungal species-specific mechanisms.
Two Paths of Sequence Divergence in the Citrus Tristeza Virus Complex
Authors: Hilf Mark E. , Karasev Alexander V. , Albiach-Marti Maria R. , Dawson William O. , Garnsey Stephen M.
Published: May 2007

Comparison of a sampling of complementary DNA (cDNA) sequences from the Florida citrus tristeza virus (CTV) isolates T3 and T30 to the sequence of the genome of the Israeli isolate VT showed a relatively consistent or symmetrical distribution of nucleotide sequence identity in both the 5′ and 3′ regions of the 19.2-kb genome. In contrast, comparison of these sequences to the sequence of isolate T36 showed a dramatic decrease in sequence identity in the 5′ proximal 11 kb of the genome. A cDNA probe derived from this region of the T36 genome hybridized to double-stranded RNA (dsRNA) of only 3 of 10 different Florida CTV isolates. In contrast, analogous probes from T3 and T30 hybridized differentially to the seven isolates not selected by the T36 probe. Primers designed from cDNA sequence for polymerase chain reaction (PCR) selectively amplified these 10 isolates, allowing them to be classified as similar to T3, T30, or T36. In contrast, individual cDNA probes derived from the 3′ terminal open reading frames of the T3, T30, and T36 genomes all hybridized to dsRNA from all Florida CTV isolates tested, and PCR primers designed from the T36 capsid protein gene sequence amplified successfully from all isolates. Based on these data, we propose the creation of two groups of CTV, exemplified by the VT and T36 isolates, respectively. Isolates in the VT group, which include isolates VT, T3, and T30, have genomic sequence divergence that is relatively constant in proportion and distribution throughout the genome, and candidate isolates for that group could be considered strains of the same virus. The T36 group is differentiated from the VT group by the highly divergent 5′ genomic sequence. This 5′ region of the CTV genome, thus, can serve as a measure of the extent of sequence divergence and can be used to define new groups and group members in the CTV complex.
Homology-Dependent Virus Resistance in Transgenic Plants with the Coat Protein Gene of Sweet Potato Feathery Mottle Potyvirus: Target Specificity and Transgene Methylation
Authors: Sonoda S. , Mori M. , Nishiguchi M.
Published: May 2007

Nicotiana benthamiana plants were transformed with the coat protein (CP) coding sequence and the 3′ nontranslated region (NTR) of the severe strain of sweet potato feathery mottle potyvirus (SPFMV-S). Regenerated lines were screened for virus resistance using recombinant potato virus X (PVX) engineered to contain the sequence homologous to the transgene. Out of 19 transgenic lines, 7 showed virus resistance after inoculation by the recombinant PVX. In most of the resistant lines, relatively low steady-state accumulation of the CP gene mRNA and little or no protein products were observed, suggesting that the resistance was manifested by a post-transcriptional gene-silencing mechanism. The resistant lines could be divided into two groups according to the target specificity of the silencing mechanism; one group recognizing the 3′ part of the transgene mRNA and the other not only the 3′ part, but also the 5′ and the central part of the transgene mRNA. Particular regions of the transgene corresponding to the RNA target in the resistant lines were differentially methylated compared with the transgene sequence in a susceptible line.
Aerial Transmission of Thielaviopsis basicola, a Pathogen of Corn-Salad, by Adult Shore Flies
Authors: Stanghellini M. E. , Rasmussen S. L. , Kim D. H.
Published: May 2007

Chlamydospores of Thielaviopsis basicola were consistently observed in frass excreted by adults and larvae of shore flies that were collected in the immediate vicinity of naturally infected corn-salad plants obtained from a commercial greenhouse production facility. Approximately 95% of the adult flies and 85% of the larvae were internally infested with the pathogen. Pathogen-free adult shore flies were subsequently shown to acquire the pathogen by ingestion after feeding on naturally infected plants. Viable propagules of the pathogen were excreted by these internally infested adults and were capable of transmitting the pathogen to healthy seedlings, which subsequently became infected.
Tomato Spotted Wilt Tospovirus Adapts to the TSWV N Gene-Derived Resistance by Genome Reassortment
Authors: Qiu Wenping , Moyer James W.
Published: May 2007

Pathogen- and host-derived resistance have been shown to suppress infection by many plant viruses. Tomato spotted wilt tospovirus (TSWV) is among these systems; however, it has easily overcome nearly all host resistance genes and has recently been shown to overcome resistance mediated by the TSWV N gene. To better understand the resistance-breaking mechanisms, we have chosen TSWV N gene-derived resistance (TNDR) as a model to study how plant viruses defeat resistance genes. A defined viral population of isolates TSWV-D and TSWV-10, both suppressed by TNDR, was subjected to TNDR selection by serial passage in an N-gene transgenic plant. The genotype analysis demonstrated that the mixed viral population was driven to form a specific reassortant, L10M10SD, in the presence of TNDR selection, but remained as a heterogeneous mixture in the absence of the selection. A genotype assay of 120 local lesion isolates from the first, fourth, and seventh transfers confirmed the shift of genomic composition. Further analysis demonstrated that the individual L10, M10, and SD RNA segments were each selected independently in response to TNDR selection rather than to a mutation or recombination event. Following the seventh transfer on the N-gene transgenic plants, TSWV S RNA remained essentially identical to the S RNA from TSWV-D, indicating that no intermolecular recombination occurred between the two S RNAs from TSWV-10 and TSWV-D nor with the transferred N gene. These results support the hypothesis that TSWV utilizes genome reassortment to adapt to new host genotypes rapidly and that elements from two or more segments of the genome are involved in suppression of the resistance reaction.
Cytology of Infection of Maize Seedlings by Fusarium moniliforme and Immunolocalization of the Pathogenesis-Related PRms Protein
Authors: Murillo I. , Cavallarin L. , Segundo B. San
Published: May 2007

We have investigated the histology of infection of maize seedlings by Fusarium moniliforme in association with a biochemical host defense response, the accumulation of the PRms (pathogenesis-related maize seed) protein. Light microscopy of trypan blue-stained sections and scanning electron microscopy revealed direct penetration by F. moniliforme hyphae through the epidermal cells of the seedling and colonization of the host tissue by inter- and intracellular modes of growth. Pathogen ingress into the infected tissue was associated with the induction of defense-related ultrastructural modifications, as exemplified by the formation of appositions on the outer host cell wall surface, the occlusion of intercellular spaces, and the formation of papillae. Cellular and subcellular immunolocalization studies revealed that PRms accumulated at very high levels in those cells types that represent the first barrier for fungal penetration such as the aleurone layer of germinating seeds and the scutellar epithelial cells of isolated germinating embryos. A highly localized accumulation of PRms within papillae of the inner scutellar parenchyma cells also occurred, suggesting that signaling mechanisms that lead to the accumulation of PRms in papillae of cell types that are distant from the invading pathogen must operate in the infected maize tissues. Our study also revealed the presence of a large number of fungal cells with an abnormal shape that showed PRms-specific labeling. PRms was found to accumulate in clusters over the fungal cell wall. Taken together, the occurrence of PRms in cell types that first establish contact with the pathogen, as well as in papillae, and in association with fungal cell walls suggests that PRms may have a function in the plant defense response.