bluuebunny/crossref_metadata_2025 huggingface dataset:
Spatiotemporal Analysis of Epiphytotics of Downy Mildew of Oilseed Poppy in Tasmania, Australia
Authors: Scott J. B. , Hay F. S. , Wilson C. R. , Cotterill P. J. , Fist A. J.
Published: May 2007

Downy mildew, caused by Peronospora arborescens, has become the major disease affecting oilseed poppy (Papaver somniferum) since its first record in Tasmania in 1996. Two field trials conducted in 2000 and 2001 studied the progression and spatial distribution of downy mildew epiphytotics. The logistic and exponential models best described the progression of disease incidence and severity, respectively. Incidence and severity increased rapidly following canopy closure. In 2001, incidence increased from 0.16%, prior to canopy closure, to 100% at late flowering (40 days). Spatial analyses of epiphytotics were conducted by fitting the beta-binomial and binomial distributions, median runs analysis, and the spatial analysis by distance indices. All analyses demonstrated that the distribution of incidence and severity was strongly spatially aggregated from canopy closure until at least late flowering. These results suggest that secondary spread from a few primary infections is the major factor in epiphytotics.
Expression of Oryzacystatin I and II in Alfalfa Increases Resistance to the Root-Lesion Nematode
Authors: Samac Deborah A. , Smigocki Ann C.
Published: May 2007

Digestive cysteine proteinases have been isolated from plant-parasitic nematodes as well as coleopteran and hemipteran insects. Phytocystatins, inhibitors of cysteine proteinases, are found in a number of plants where they may play a role in defense against pathogens and pests. The cDNAs of the phytocystatins from rice, oryzacystatin I (OC-I) and oryzacystatin II (OC-II), were expressed in alfalfa (Medicago sativa) plants under the control of the potato protease inhibitor II (PinII) promoter and the plants were evaluated for resistance to the root-lesion nematode (Pratylenchus penetrans). A PinII-β-glucuronidase (GUS) gene was introduced into alfalfa to determine the pattern of gene expression from this promoter. Constitutive GUS expression was observed in leaf and root vascular tissue, and in some plants, expression was observed in leaf mesophyll cells. Mechanical wounding of leaves increased GUS expression approximately twofold over 24 h. Inoculation with root-lesion nematodes resulted in localized GUS expression. Populations of root-lesion nematodes in alfalfa roots from one line containing the PinII::OC-I transgene and one line containing the PinII::OC-II transgene were reduced 29 and 32%, respectively, compared with a transgenic control line. These results suggest that oryzacystatins have the potential to confer increased resistance to the root-lesion nematode in alfalfa.
Ethylene-Insensitive Tobacco Shows Differentially Altered Susceptibility to Different Pathogens
Authors: Geraats Bart P. J. , Bakker Peter A. H. M. , Lawrence Christopher B. , Achuo Enow A. , Höfte Monica , van Loon L. C.
Published: May 2007

Transgenic tobacco plants (Tetr) expressing the mutant etr1-1 gene from Arabidopsis thaliana are insensitive to ethylene and develop symptoms of wilting and stem rot when grown in nonautoclaved soil. Several isolates of Fusarium, Thielaviopsis, and Pythium were recovered from stems of diseased Tetr plants. Inoculation with each of these isolates of 6-week-old plants growing in autoclaved soil caused disease in Tetr plants but not in nontransformed plants. Also, when 2-week-old seedlings were used, nontransformed tobacco appeared nonsusceptible to the Fusarium isolates, whereas Tetr seedlings did develop disease. Tetr seedlings were not susceptible to several nonhost Fusarium isolates. In contrast to results with Fusarium isolates, inoculation of 2-week-old seedlings with a Thielaviopsis isolate resulted in equal symptom development of nontransformed and Tetr tobacco. In order to explore the potential range of pathogens to which Tetr tobacco plants display enhanced susceptibility, the pathogenicity of several root and leaf pathogens was tested. Tetr plants were more susceptible to the necrotrophic fungi Botrytis cinerea and Cercospora nicotianae and the bacterium Erwinia carotovora, but only marginally more to the bacterium Ralstonia solanacearum. In contrast, the biotrophic fungus Oidium neolycopersici, the oomycete Peronospora tabacina, and Tobacco mosaic virus caused similar or less severe symptoms on Tetr plants than on nontransformed plants. Total peroxidase activity of Tetr plants was lower than that of nontransformed plants, suggesting a role for peroxidases in resistance against necrotrophic microorganisms. A comparable range of pathogens was examined on Arabidopsis and its ethylene-insensitive mutants etr1-1 and ein2-1. With the exception of one Fusarium isolate, ethylene insensitivity increased susceptibility of Arabidopsis plants to a similar spectrum of necrotizing pathogens as in tobacco. Thus, both ethylene-insensitive tobacco and Arabidopsis plants appear to be impaired in their resistance to necrotrophic pathogens.
Composition and Distribution of Pythium Communities in Wheat Fields in Eastern Washington State
Authors: Paulitz T. C. , Adams K.
Published: May 2007

Pythium spp. were isolated from a mixture of soil and roots collected from 80 wheat fields in eastern Washington in the summer of 2000 from an area encompassing approximately 27,000 km2. These sites covered a range of soil textures (coarse to fine, silty loess), average annual precipitation (200 to 600 mm), and average annual temperatures (7 to 11°C). Soil type and annual precipitation run in an east-west gradient, while temperature has a north-south gradient. Species were identified using classical methods and by sequencing the internal transcribed spacer (ITS)-1 region of the rDNA and comparing these sequences to a database from a worldwide collection of Pythium spp. The species with the highest frequency of occurrence among all the sites were P. abappressorium sp. nov. (A) (50%), P. rostratum (R) (40%), P. debaryanum (D) (37.5%), P. heterothallicum (H) (33.7%), P.oligandrum (O) (31.2%), an unidentified P. sp. (aff. echinulatum) (E) (25%), and P. ultimum (U) (18%). P. intermedium, P. irregulare, P. paroecandrum, P. sylvaticum, P. dissimile, and P. dissoticum were isolated at a low frequency. From one to six species were isolated at each site, and there were 46 different species combinations detected. The species presence/absence data from all sites were analyzed with Jaccard's similarity coefficient hierarchical cluster analysis. Six communities were identified (species within each community designation in order of frequency among the sites within the community)-AD, AOU, AR, DEH, HE, and RU. In general, P. abappressorium was evenly distributed over all zones. AOU was more prevalent in zones with lower precipitation and coarser soil, while DEH and HE were associated with zones with higher precipitation and finer-textured soils on the basis of comparison of frequency distributions with the expected distribution over all the sites. The RU community was more prevalent in higher temperature zones. Canonical correspondence analysis was performed to examine the relationship between species and environmental variables. Soil type and precipitation were highly correlated with each other and with axis 1, which separated P. ultimum and P. abappressorium (lower variable values) from P. heterothallicum (higher variable values). Axis 2 and 3 were most correlated with temperature, and these axes separated P. oligandrum (higher value) from P. debaryanum (lower value) and P. ultimum-P. rostratum from the other species. The results suggest that Pythium species composition, distributions, and associations on a given crop may be influenced by environmental factors at a mesoscale level (100 to 1,000,000 ha).
Diversity of Epidemic Populations of Gibberella zeae from Small Quadrats in Kansas and North Dakota
Authors: Zeller Kurt A. , Bowden Robert L. , Leslie John F.
Published: May 2007

Gibberella zeae (anamorph Fusarium graminearum) causes Fusarium head blight (FHB) of wheat and barley and has been responsible for several billion dollars of losses in the United States since the early 1990s. We isolated G. zeae from the top, middle, and bottom positions of wheat spikes collected from 0.25-m2 quadrats during severe FHB epidemics in a single Kansas (KS) field (1993) and in a single North Dakota (ND) field (1994). Three amplified fragment length polymorphism (AFLP) primer pairs were used to resolve 94 polymorphic loci from 253 isolates. Members of a subset of 26 isolates also were tested for vegetative compatibility groups (VCGs). Both methods indicated high levels of genotypic variability and identified the same sets of isolates as probable clones. The mean number of AFLP multilocus haplotypes per head was approximately 1.8 in each population, but this value probably underestimates the true mean due to the small number of samples taken from each head. Isolates with the same AFLP haplotype often were recovered from different positions in a single head, but only rarely were such apparently clonal isolates recovered from more than one head within a quadrat, a pattern that is consistent with a genetically diverse initial inoculum and limited secondary spread. The KS and ND samples had no common AFLP haplotypes. All G. zeae isolates had high AFLP fingerprint similarity (>70%, unweighted pair group method with arithmetic means similarity) to reference isolates of G. zeae lineage 7. The genetic identity between the KS and ND populations was >99% and the estimated effective migration rate was high (Nm ≈70). Tests for linkage disequilibrium provide little evidence for nonrandom associations between loci. Our results suggest that these populations are parts of a single, panmictic population that experiences frequent recombination. Our results also suggest that a variety of population sampling designs may be satisfactory for assessing diversity in this fungus.
Evolution of Concepts in Forest Pathology
Authors: Manion Paul D.
Published: May 2007

Foundation concepts in forest pathology are based on experiences evolving over time. Three examples will be addressed. (i) The primary concept behind education and research in forest pathology is the widely accepted attitude that disease-causing agents limit full utilization of forest resources. Therefore, we study diseases to find a weak link and then utilize this information to enhance our portion of the shared resource. The sustainable environmental issues of today have changed this concept, in my mind, to one of addressing what is the appropriate “healthy amount of disease” in a sustainable forest ecosystem. (ii) The initial concept that weakened understory trees and poorly managed forests deteriorate and decline over time because of numerous insults from biotic and abiotic agents has evolved into a decline disease stabilizing selection concept whereby healthy dominant trees in the forest (the survivors) are selectively killed by a combination of specifically ordered factors. (iii) The concept that heart-rot decay is initiated by infection through wounds that expose heartwood has evolved into the concept of infection in the sapwood that is compartmentalized over time in the center of the tree.
Testing a Rapid Diagnostic Medium for Erwinia amylovora and Development of a Procedure for Sampling Blossoms in Pear Orchards
Authors: Kritzman G. , Shwartz H. , Marcus R. , Manulis S. , Klietman F. , Oppenheim D. , Zilberstaine M. , Shtienberg D.
Published: May 2007

The coliform agar produced by Merck was tested for rapid diagnosis of Erwinia amylovora (the causal agent of fire blight) in pear blossoms. The medium enabled the diagnosis to be completed within 36 h. Diagnoses performed with the medium were confirmed by the BIOLOG and the fatty-acid profile methods. The diagnostic medium was used to determine the spatial distribution of colonized blossoms in the orchards and it was found that E. amylovora may be distributed both in clusters and at random. These findings were used in the development of a statistical model for sampling blossoms in the orchard. The model determines the number of trees to be sampled in the orchard and the number of blossoms be taken from each tree, which would enable the true colonization incidence of blossoms in the orchard to be estimated at desired levels of accuracy and confidence. Parameters included in the model are: the total number of trees in the orchard (T), the number of trees to be sampled in the orchard (t), the number of blossoms to be sampled from each tree (n), the true colonization incidence of blossoms (π), a coefficient of aggregation (ρ), the required level of confidence (1 - α), and the required level of accuracy (L). Sensitivity analyses revealed that the parameter governing sample size is the required level of accuracy. Sampling of 20 blossoms from each of several hundred trees is required to achieve an accuracy of ±1%, but only a few single trees are needed for an accuracy level of ±10%. A sampling procedure then was developed, validated with an independent data set, and found to be accurate. It was concluded that sampling of pear blossoms and estimation of the incidence of blossom colonization by E. amylovora could improve fire blight management, but not in all cases.
Presence of Xylella fastidiosa in Sweet Orange Fruit and Seeds and Its Transmission to Seedlings
Authors: Li W.-B. , Pria W. D. , Lacava P. M. , Qin X. , Hartung J. S.
Published: May 2007

Xylella fastidiosa, a xylem-limited bacterium, causes several economically important diseases in North, Central, and South America. These diseases are transmitted by sharpshooter insects, contaminated budwood, and natural root-grafts. X. fastidiosa extensively colonizes the xylem vessels of susceptible plants. Citrus fruit have a well-developed vascular system, which is continuous with the vascular system of the plant. Citrus seeds develop very prominent vascular bundles, which are attached through ovular and seed bundles to the xylem system of the fruit. Sweet orange (Citrus sinensis) fruit of cvs. Pera, Natal, and Valencia with characteristic symptoms of citrus variegated chlorosis disease were collected for analysis. X. fastidiosa was detected by polymerase chain reaction (PCR) in all main fruit vascular bundles, as well as in the seed and in dissected seed parts. No visual abnormalities were observed in seeds infected with the bacterium. However, the embryos of the infected seeds weighed 25% less than those of healthy seeds, and their germination rate was lower than uninfected seeds. There were about 2,500 cells of X. fastidiosa per infected seed of sweet orange, as quantified using real-time PCR techniques. The identification of X. fastidiosa in the infected seeds was confirmed by cloning and sequencing the specific amplification product, obtained by standard PCR with specific primers. X. fastidiosa was also detected in and recovered from seedlings by isolation in vitro. Our results show that X. fastidiosa can infect and colonize fruit tissues including the seed. We also have shown that X. fastidiosa can be transmitted from seeds to seedlings of sweet orange. To our knowledge, this is the first report of the presence of X. fastidiosa in seeds and its transmission to seedlings.
Effect of 2,4-Diacetylphloroglucinol on Pythium: Cellular Responses and Variation in Sensitivity Among Propagules and Species
Authors: de Souza Jorge T. , Arnould Christine , Deulvot Chrystel , Lemanceau Philippe , Gianinazzi-Pearson Vivienne , Raaijmakers Jos M.
Published: May 2007

The antibiotic 2,4-diacetylphloroglucinol (2,4-DAPG) plays an important role in the suppression of plant pathogens by several strains of Pseudomonas spp. Based on the results of this study, there is variation within and among Pythium spp. to 2,4-DAPG. Also, various propagules of Pythium ultimum var. sporangiiferum, that are part of the asexual stage of the life cycle, differ considerably in their sensitivity to 2,4-DAPG. Mycelium was the most resistant structure, followed by zoosporangia, zoospore cysts, and zoospores. Additionally, we report for the first time that pH has a significant effect on the activity of 2,4-DAPG, with a higher activity at low pH. Furthermore, the level of acetylation of phloroglucinols is also a major determinant of their activity. Transmission electron microscopy studies revealed that 2,4-DAPG causes different stages of disorganization in hyphal tips of Pythium ultimum var. sporangiiferum, including alteration (proliferation, retraction, and disruption) of the plasma membrane, vacuolization, and cell content disintegration. The implications of these results for the efficacy and consistency of biological control of plant-pathogenic Pythium spp. by 2,4-DAPG-producing Pseudomonas spp. are discussed.
Interactions Between Strains of 2,4-Diacetylphloroglucinol-Producing Pseudomonas fluorescens in the Rhizosphere of Wheat
Authors: Landa Blanca B. , Mavrodi Dmitri M. , Thomashow Linda S. , Weller David M.
Published: May 2007

Strains of fluorescent Pseudomonas spp. that produce the antibiotic 2,4-diacetylphoroglucinol (2,4-DAPG) are among the most effective rhizobacteria controlling diseases caused by soilborne pathogens. The genotypic diversity that exists among 2,4-DAPG producers can be exploited to improve rhizosphere competence and biocontrol activity. Knowing that D-genotype 2,4-DAPG-producing strains are enriched in some take-all decline soils and that P. fluorescens Q8r1-96, a representative D-genotype strain, as defined by whole-cell repetitive sequence-based polymerase chain reaction (rep-PCR) with the BOXA1R primer, is a superior colonizer of wheat roots, we analyzed whether the exceptional rhizosphere competence of strain Q8r1-96 on wheat is characteristic of other D-genotype isolates. The rhizosphere population densities of four D-genotype strains and a K-genotype strain introduced individually into the soil were significantly greater than the densities of four strains belonging to other genotypes (A, B, and L) and remained above log 6.8 CFU/g of root over a 30-week cycling experiment in which wheat was grown for 10 successive cycles of 3 weeks each. We also explored the competitive interactions between strains of different genotypes inhabiting the same soil or rhizosphere when coinoculated into the soil. Strain Q8r1-96 became dominant in the rhizosphere and in nonrhizosphere soil during a 15-week cycling experiment when mixed in a 1:1 ratio with either strain Pf-5 (A genotype), Q2-87 (B genotype), or 1M1-96 (L genotype). Furthermore, the use of the de Wit replacement series demonstrated a competitive disadvantage for strain Q2-87 or strong antagonism by strain Q8r1-96 against Q2-87 in the wheat rhizosphere. Amplified rDNA restriction analysis and sequence analysis of 16S rDNA showed that species of Arthrobacter, Chryseobacterium, Flavobacterium, Massilia, Microbacterium, and Ralstonia also were enriched in culturable populations from the rhizosphere of wheat at the end of a 30-week cycling experiment in the presence of 2,4-DAPG producers. Identifying the interactions among 2,4-DAPG producers and with other indigenous bacteria in the wheat rhizosphere will help to elucidate the variability in biocontrol efficacy of introduced 2,4-DAPG producers and fluctuations in the robustness of take-all suppressive soils.
Factors that Affect Spread of Pseudomonas syringae in the Phyllosphere
Authors: Upper Christen D. , Hirano Susan S. , Dodd Kimberly K. , Clayton Murray K.
Published: May 2007

Successful spread of an organism to a new habitat requires both immigration to and growth on that habitat. Field experiments were conducted to determine the relative roles of dispersal (i.e., immigration) and bacterial multiplication in spread of Pseudomonas syringae pv. syringae in the phyllosphere. To study spread, individual plots consisted of three nested concentric squares with the inner 6 m2 planted to snap beans serving as the sink. Each sink, in turn, was surrounded by a barrier zone, usually 6 m wide, which was surrounded by a 6-m-wide source area. The source areas were planted with snap bean seeds inoculated with doubly marked strains derived from wild-type P. syringae pv. syringae B728a. The treatments were designed to test the effects of the nature and width of the barrier zone and suitability of the habitat in the sinks on spread of P. syringae pv. syringae. The marked strains introduced into the source areas at the time of planting were consistently detected in sink areas within a day or two after emergence of bean seedlings in the sources as assessed by leaf imprinting and dilution plating. The amounts of spread (population sizes of the marked strain in sinks) across barrier zones planted to snap bean (a suitable habitat for growth of P. syringae pv. syringae), soybean (not a favorable habitat for P. syringae pv. syringae), and bare ground were not significantly different. Thus, the nature of the barrier had no measurable effect on spread. Similarly, spread across bare-ground barriers 20 m wide was not significantly different from that across barriers 6 m wide, indicating that distance on this scale was not a major factor in determining the amount of spread. The suitability of the sink for colonization by P. syringae pv. syringae had a measurable effect on spread. Spread to sinks planted to clean seed was greater than that to sinks planted with bean seeds inoculated with a slurry of pulverized brown spot diseased bean leaves, sinks planted 3 weeks before sources, and sinks planted to a snap bean cultivar that does not support large numbers of P. syringae pv. syringae. Based of these results, we conclude that the small amount of dispersal that occurred on the scale studied was sufficient to support extensive spread, and suitability of the habitat for multiplication of P. syringae pv. syringae strongly influenced the amount of spread.
Identification and Molecular Mapping of a Gene in Wheat Conferring Resistance to Mycosphaerella graminicola
Authors: Adhikari Tika B. , Anderson Joseph M. , Goodwin Stephen B.
Published: May 2007

Septoria tritici leaf blotch (STB), caused by the ascomycete Mycosphaerella graminicola (anamorph Septoria tritici), is an economically important disease of wheat. Breeding for resistance to STB is the most effective means to control this disease and can be facilitated through the use of molecular markers. However, molecular markers linked to most genes for resistance to STB are not yet available. This study was conducted to test for resistance in the parents of a standard wheat mapping population and to map any resistance genes identified. The population consisted of 130 F10 recombinant-inbred lines (RILs) from a cross between the synthetic hexaploid wheat W7984 and cv. Opata 85. Genetic analysis indicated that a single major gene controls resistance to M. graminicola in this population. This putative resistance gene is now designated Stb8 and was mapped with respect to amplified fragment length polymorphism (AFLP) and microsatellite markers. An AFLP marker, EcoRI-ACG/MseI-CAG5, was linked in repulsion with the resistance gene at a distance of approximately 5.3 centimorgans (cM). Two flanking microsatellite markers, Xgwm146 and Xgwm577, were linked to the Stb8 gene on the long arm of wheat chromosome 7B at distances of 3.5 and 5.3 cM, respectively. The microsatellite markers identified in this study have potential for use in marker-assisted selection in breeding programs and for pyramiding of Stb8 with other genes for resistance to M. graminicola in wheat.
Quantitative Models for Describing Temperature and Moisture Effects on Sporulation of Phomopsis amygdali on Peach
Authors: Lalancette N. , Foster K. A. , Robison D. M.
Published: May 2007

Twigs with constriction cankers were pruned from a Prunus persica ‘Jerseyglo’ orchard and placed in incubators under high humidity (>95%) at constant temperatures of −6, 1, 10, 17, 24, 31, 38, and 45°C. Cankers were removed for observation after 2, 6, 24, 48, 72, and 96 h incubation. Sporulation was expressed as the percentage of pycnidia producing cirri and the number of conidia per pycnidium or canker. The experiment was first performed during 1998 to 1999 and then repeated in 1999 to 2000 and 2000 to 2001. Sporulation was modeled by fitting the Richards function to each dependent variable with duration of high relative humidity as the independent variable. The asymptote and rate parameters of the Richards model were expressed as Gaussian and quadratic functions of temperature, respectively. Models created from the first two experimental repetitions were validated by statistical comparison to those created independently from the third repetition. When models were fit to the pooled data, temperature and moisture described 69 to 80% of the variation in sporulation. The models specified a temperature ranging from 0 to 37°C (optimum temperatures for cirri formation and conidia production range from 19 to 20 and 22 to 23°C, respectively), and the majority of sporulation occurred between 16 and 48 h from initiation of the high humidity period. These results show that the environmental criteria for sporulation coincide with those that prolong the susceptibility of infection courts during fall and spring.
Logistic Regression Modeling of Prevalence of Soybean Sclerotinia Stem Rot in the North-Central Region of the United States
Authors: Mila A. L. , Carriquiry A. L. , Yang X. B.
Published: May 2007

Regional prevalence of soybean Sclerotinia stem rot (SSR), caused by Sclerotinia sclerotiorum, was modeled using tillage practices, soil texture, and weather variables (monthly air temperature and monthly precipitation from April to August) as inputs. Logistic regression was used to estimate the probability of stem rot prevalence with historical disease data from four states of the north-central region of the United States. Potential differences in disease prevalence between states in the region were addressed using regional indicator variables. Two models were developed: model I used spring (April) weather conditions and model II used summer (July and August) weather conditions as input variables. Both models had high explanatory power (78.5 and 77.8% for models I and II, respectively). To investigate the explanatory power of the models, each of the four states was divided into small geographic areas, and disease prevalence in each area was estimated using both models. The R2 value of the regression analysis between observed and estimated SSR prevalence was 0.65 and 0.71 for models I and II, respectively. The same input variables were tested for their significance to explain the within-field SSR incidence by using Poisson regression analysis. Although all input variables were significant, only a small amount of variation of SSR incidence was explained, because R2 of the regression analysis between observed and estimated SSR incidence was 0.065. Incorporation of available site-specific information (i.e., fungicide seed treatment, weed cultivation, and manure and fertilizer applications in a field) improved slightly the explained amount of SSR incidence (R2 = 0.076). Predicted values of field incidence generally were overestimated in both models compared with the observed incidence. Our results suggest that preseason prediction of regional prevalence would be feasible. However, prediction of field incidence would not, and a different site-specific approach should be followed.
Novel Cytopathological Structures Induced by Mixed Infection of Unrelated Plant Viruses
Authors: Martin E. M. , Cho J. D. , Kim J. S. , Goeke S. C. , Kim K. S. , Gergerich R. C.
Published: May 2007

When two unrelated plant viruses infect a plant simultaneously, synergistic viral interactions often occur resulting in devastating diseases. This study was initiated to examine ultrastructural virus-virus interactions of mixed viral infections. Mixed infections were induced using potyviruses and viruses from other plant virus families. Novel ultrastructural paracrystalline arrays composed of co-infecting viruses, referred to as mixed virus particle aggregates (MVPAs), were noted in the majority of the mixed infections studied. When the flexuous rod-shaped potyvirus particles involved in MVPAs were sectioned transversely, specific geometrical patterns were noted within some doubly infected cells. Although similar geometrical patterns were associated with MVPAs of various virus combinations, unique characteristics within patterns were consistent in each mixed infection virus pair. Centrally located virus particles within some MVPAs appeared swollen (Southern bean mosaic virus mixed with Blackeye cowpea mosaic virus, Cucumber mosaic virus mixed with Blackeye cowpea mosaic virus, and Sunn hemp mosaic virus mixed with Soybean mosaic virus). This ultrastructural study complements molecular studies of mixed infections of plant viruses by adding the additional dimension of visualizing the interactions between the coinfecting viruses.
Improved Sampling Methods for Real-Time Polymerase Chain Reaction Diagnosis of Citrus Canker from Field Samples
Authors: Mavrodieva Vessela , Levy Laurene , Gabriel Dean W.
Published: May 2007

Citrus bacterial canker disease has been introduced at least three times into Florida in the last 15 years and, despite federal and state quarantine and eradication efforts, continues to spread in Florida. Accurate, fast, and reliable detection of the causal agent is of great importance. However, citrus bacterial canker is caused by at least two groups of phylogenetically distinct Xanthomonas citri strains, and there is host range variation within both groups. We developed a fast, sensitive and reliable real-time polymerase chain reaction (PCR) assay using a portable, field-hardened RAPID machine and primers designed to detect all canker-causing strains. Single-lesion sampling methods were developed that required minimal handling and allowed complete real-time PCR diagnosis in a total time of 4 h and with an apparent sensitivity of less than 10 CFU of target cells from diseased lesions. This sensitivity allowed molecular detection for the first time of X. citri in a herbarium sample from a 1912 canker outbreak. Sensitivity was improved significantly by the use of CaCO3 and Silwet L-77, and by either minimizing the amount of citrus lesion tissue sampled or by soaking or swiping but not grinding the lesions. Primer design also was of significant importance in both specificity and sensitivity.
Identification of Pathotypes of Xanthomonas axonopodis pv. manihotis in Africa and Detection of Quantitative Trait Loci and Markers for Resistance to Bacterial Blight of Cassava
Authors: Wydra K. , Zinsou V. , Jorge V. , Verdier V.
Published: May 2007

Cassava suffers from bacterial blight attack in all growing regions. Control by resistance is unstable due to high genotype-environment interactions. Identifying genes for resistance to African strains of Xanthomonas axonopodis pv. manihotis can support breeding efforts. Five F1 cassava genotypes deriving from the male parent ‘CM2177-2’ and the female parent ‘TMS30572’ were used to produce 111 individuals by backcrossing to the female parent. In all, 16 genotypes among the mapping population were resistant to stem inoculation by four strains of X. axonopodis pv. manihotis from different locations in Africa, and 19 groups with differential reactions to the four strains were identified, suggesting that the strains represent different pathotypes. Four genotypes were resistant to leaf inoculation, and three were resistant to both stem and leaf inoculations. Genotypes with susceptible, moderately resistant, and resistant reactions after leaf and stem inoculation partly differed in their reactions on leaves and stems. Based on the genetic map of cassava, single-markeranalysis of disease severity after stem-puncture inoculation was performed. Eleven markers were identified, explaining between 16 and 33.3% of phenotypic variance of area under disease progress curve. Five markers on three and one linkage groups from the female- and male-derived framework of family CM8820, respectively, seem to be weakly associated with resistance to four strains of X. axonopodis pv. manihotis. Based on the segregation of alleles from the female of family CM8873, one marker was significantly associated with resistance to two X. axonopodis pv. manihotis strains, GSPB2506 and GSPB2511, whereas five markers were not linked to any linkage group. The quantitative trait loci (QTL) mapping results also suggest that the four African strains belong to four different pathotypes. The identified pathotypes should be useful for screening for resistance, and the QTL and markers will support breeding for resistance.
Evolution of an Osmosensing Histidine Kinase in Field Strains of Botryotinia fuckeliana (Botrytis cinerea) in Response to Dicarboximide Fungicide Usage
Authors: Cui Wei , Beever Ross E. , Parkes Stephanie L. , Templeton Matthew D.
Published: May 2007

DNA sequence polymorphisms in the putative two-component histidine protein kinase encoded by the Daf1 gene have been identified within a sample of 5 sensitive and 27 dicarboximide-resistant field strains of Botryotinia fuckeliana (anamorph Botrytis cinerea). The gene of 3948 bp is predicted to encode a 1315-amino acid protein comprising an N-terminal region, an amino acid repeat region, which has been hypothesized to be the binding site for dicarboximide fungicide, and a C-terminal region encompassing kinase and response regulator domains. Two amino acid variants were distinguished among the sensitive strains characterized by alanine (group 1), or threonine (group 2), at position 1259 in the C-terminal region. All resistant strains could be classified into either group 1 or group 2 but, in addition, all showed changes in the second amino acid repeat region. On the basis of the differences in this repeat region, four classes of resistant strains were recognized; class 1 characterized by an isoleucine to serine mutation, class 2 by an isoleucine to asparagine mutation, class 3 by an isoleucine to arginine mutation (all at position 365), and class 4 by an isoleucine to serine mutation (position 365) as well as a glutamine to proline mutation (position 369). All classes showed similar low levels of resistance to iprodione and to vinclozolin, except for class 3 and class 4 strains, which show low resistance to iprodione but moderate (class 3) or high (class 4) resistance to vinclozolin. The classes as a group did not differ from sensitive strains in osmotic sensitivity measured as mycelial growth response, but some class 1 strains showed an abnormal morphology on osmotically amended medium. The evolution of the amino acid differences is discussed in relation to field observations. It is proposed that class 1 and class 2 strains arose by single mutations within the sensitive population, whereas classes 3 and 4 arose by single mutations within a resistant population.
Barley Traits Associated with Resistance to Fusarium Head Blight and Deoxynivalenol Accumulation
Authors: Choo Thin Meiw , Vigier Bernard , Shen Qiu Quan , Martin Richard A. , Ho Keh Ming , Savard Marc
Published: May 2007

Fusarium head blight (FHB) or scab is a destructive disease of barley in many countries. A better understanding of the interrelationships between plant traits and FHB resistance should help in the development of effective and efficient breeding strategies for FHB-resistant cultivars. Recent mapping studies indicate that many of the quantitative trait loci (QTL) for FHB resistance coincide with the QTL for plant height, heading date, and spike characteristics. Therefore, a study was conducted to investigate the relationship of morphological and physiological traits to FHB infection and deoxynivalenol (DON) accumulation in a barley doubled-haploid (DH) population derived from a Léger × CI9831 cross. Approximately 190 DH lines were grown at Ottawa (Ontario) for 2 years, Charlottetown (Prince Edward Island) for 1 year, and Hangzhou (Zhejiang) for 2 years. The field plots were inoculated with Fusarium graminearum at each location. FHB incidence was positively correlated with DON content. Resistance to FHB was associated with two-row spike, purple lemma, long glume awn, tall stature, and resistance to lodging, but it was not associated with long rachilla hairs, rough lemma awn, or heading date. Two-row spike was associated with tall stature and resistance to lodging. These associations as well as its spike characteristics helped reduce FHB infection and DON accumulation in two-row lines compared with six-row lines. The association between long glume awn and FHB resistance could be due to genetic linkages. Therefore, trait associations should be taken into consideration when breeding for FHB resistance and interpreting data from FHB experiments.
An Extracellular Protease of Pseudomonas fluorescens Inactivates Antibiotics of Pantoea agglomerans
Authors: Anderson L. Meadow , Stockwell Virginia O. , Loper Joyce E.
Published: May 2007

Pseudomonas fluorescens A506 and Pantoea agglomerans strains Eh252 and C9-1 are biological control agents that suppress fire blight, an important disease of pear and apple caused by the bacterium Erwinia amylovora. Pseudomonas fluorescens strain A506 suppresses disease largely through competitive exclusion of E. amylovora on surfaces of blossoms, the primary infection court, whereas Pantoea agglomerans strains Eh252 and C9-1 produce antibiotics that are toxic to E. amylovora. In this study, an extracellular protease produced by A506 is characterized and evaluated for its capacity to inactivate the antibiotics produced by the strains of Pantoea agglomerans. Activity of the extracellular protease was optimal at pH 9 and inhibited by zinc- or calcium-chelators, indicating that the protease is an alkaline metalloprotease. In an agar plate bioassay, partially purified extracellular protease inactivated the antibiotics mccEh252 and herbicolin O, which are produced by Pantoea agglomerans strains Eh252 and C9-1, respectively. Derivatives of A506 deficient in extracellular protease production were obtained by transposon mutagenesis, and the aprX gene encoding the protease was cloned and sequenced. Strain A506 inactivated mccEh252 and herbicolin O in agar plate bioassays, whereas the aprX mutant did not inactivate the antibiotics. Both A506 and the aprX mutant were insensitive to antibiosis by C9-1 and Eh252; thus, the protease was not required to protect A506 from antibiosis. These data highlight a previously unknown role of the extracellular protease produced by Pseudomonas fluorescens A506 in interactions among plant-associated microbes.
Formulation of Bacillus spp. for Biological Control of Plant Diseases
Authors: Schisler D. A. , Slininger P. J. , Behle R. W. , Jackson M. A.
Published: May 2007

Maximizing the potential for successfully developing and deploying a biocontrol product begins with a carefully crafted microbial screening procedure, proceeds with developing mass production protocols that optimize product quantity and quality, and ends with devising a product formulation that preserves shelf-life, aids product delivery, and enhances bioactivity. Microbial selection procedures that require prospective bio-control agents to possess both efficacy and amenability to production in liquid culture increase the likelihood of selecting agents with enhanced commercial development potential. Scale-up of biomass production procedures must optimize product quantity without compromise of product efficacy or amenability to stabilization and formulation. Formulation of Bacillus spp. for use against plant pathogens is an enormous topic in general terms but limited in published specifics regarding formulations used in commercially available products. Types of formulations include dry products such as wettable powders, dusts, and granules, and liquid products including cell suspensions in water, oils, and emulsions. Cells can also be microencapsulated. Considerations critical to designing successful formulations of microbial biomass are many fold and include preserving biomass viability during stabilization, drying, and rehydration; aiding biomass delivery, target coverage, and target adhesion; and enhancing biomass survival and efficacy after delivery to the target. Solutions to these formulation considerations will not necessarily be compatible. Data from several biocontrol systems including the use of B. subtilis OH 131.1 (NRRL B-30212) to reduce Fusarium head blight of wheat are used to illustrate many of these issues. Using our recently described assay for efficiently evaluating biomass production and formulation protocols, we demonstrate the effectiveness, in vitro, of UV protectant compounds lignin (PC 1307) and Blankophor BBH in reducing OH 131.1 morbidity when cells were exposed to UV light from artificial sunlight.
The Role of Bacillus-Based Biological Control Agents in Integrated Pest Management Systems: Plant Diseases
Authors: Jacobsen B. J. , Zidack N. K. , Larson B. J.
Published: May 2007

Bacillus-based biological control agents (BCAs) have great potential in integrated pest management (IPM) systems; however, relatively little work has been published on integration with other IPM management tools. Unfortunately, most research has focused on BCAs as alternatives to synthetic chemical fungicides or bactericides and not as part of an integrated management system. IPM has had many definitions and this review will use the national coalition for IPM definition: “A sustainable approach to managing pests by combining biological, cultural, physical and chemical tools in a way that minimizes economic, health and environmental risks.” This review will examine the integrated use of Bacillus-based BCAs with disease management tools, including resistant cultivars, fungicides or bactericides, or other BCAs. This integration is important because the consistency and degree of disease control by Bacillus-based BCAs is rarely equal to the control afforded by the best fungicides or bactericides. In theory, integration of several tools brings stability to disease management programs. Integration of BCAs with other disease management tools often provides broader crop adaptation and both more efficacious and consistent levels of disease control. This review will also discuss the use of Bacillus-based BCAs in fungicide resistance management. Work with Bacillus thuringiensis and insect pest management is the exception to the relative paucity of reports but will not be the focus of this review.
Postharvest Biological Control ofBotrytis cinereaon Kiwifruit by Volatiles of [Isabella] Grapes
Authors: Kulakiotu Eleni K. , Thanassoulopoulos Constantine C. , Sfakiotakis Evangelos M.
Published: May 2007

The potential of volatile substances emitted by ‘Isabella’ grapes (Vitis labrusca) to control gray mold (Botrytis cinerea) on ‘Hayward’ kiwifruit (Actinidia deliciosa) was studied. The closed Mariotte system was used as a bioassay method to analyze quantitatively the biological action of these volatiles on B. cinerea growth. In vivo experiments compared the effects of volatiles from ‘Isabella’ grapes versus volatiles from ‘Roditis’ grapes (V. vinifera) and a B. cinerea control on the growth and disease development of B. cinerea on kiwifruit. The effect of the volatiles on the growth of B. cinerea was tested at various temperatures and times of inoculation after the wounding of kiwifruit, as well as using various weights and developmental stages of the grapes. The ‘Isabella’ volatiles limited the incidence of infection by reducing both the inoculum density and the activity of the pathogen. The weight and developmental stage of the grapes were important in the degree of inhibitory action of the ‘Isabella’ volatiles. The inhibitory action was more pronounced at 21°C irrespective of the inoculation time after wounding. The study shows the potential for successful biological control of B. cinerea on kiwifruit by volatiles from ‘Isabella’ grapes.
Rain Splash Dispersal of Gibberella zeae Within Wheat Canopies in Ohio
Authors: Paul P. A. , El-Allaf S. M. , Lipps P. E. , Madden L. V.
Published: May 2007

Rain splash dispersal of Gibberella zeae, causal agent of Fusarium head blight of wheat, was investigated in field studies in Ohio between 2001 and 2003. Samplers placed at 0, 30, and 100 cm above the soil surface were used to collect rain splash in wheat fields with maize residue on the surface and fields with G. zeae-infested maize kernels. Rain splash was collected during separate rain episodes throughout the wheat-growing seasons. Aliquots of splashed rain were transferred to petri dishes containing Komada's selective medium, and G. zeae was identified based on colony and spore morphology. Dispersed spores were measured in CFU/ml. Intensity of splashed rain was highest at 100 cm and ranged from 0.2 to 10.2 mm h-1, depending on incident rain intensity and sampler height. Spores were recovered from splash samples at all heights in both locations for all sampled rain events. Both macroconidia and ascospores were found based on microscopic examination of random samples of splashed rain. Spore density and spore flux density per rain episode ranged from 0.4 to 40.9 CFU cm-2 and 0.4 to 84.8 CFU cm-2 h-1, respectively. Spore flux density was higher in fields with G. zeae-infested maize kernels than in fields with maize debris, and generally was higher at 0 and 30 cm than at 100 cm at both locations. However, on average, spore flux density was only 30% lower at 100 cm (height of wheat spikes) than at the other heights. The log of spore flux density was linearly related to the log of splashed rain intensity and the log of incident rain intensity. The regression slopes were not significantly affected by year, location, height, and their interactions, but the intercepts were significantly affected by both sampler height and location. Thus, our results show that spores of G. zeae were consistently splash dispersed to spike heights within wheat canopies, and splashed rain intensity and spore flux density could be predicted based on incident rain intensity in order to estimate inoculum dispersal within the wheat canopy.
Spatial and Temporal Analyses of Bacterial Blight of Onion Caused by Xanthomonas axonopodis pv. allii
Authors: Roumagnac P. , Pruvost O. , Chiroleu F. , Hughes G.
Published: May 2007

Bacterial blight of onion is a severe disease, which emerged over the past decade in several onion-producing areas. This disease currently is observed in both the Old and New Worlds. Although the causative agent, Xanthomonas axonopodis pv. allii, is potentially seedborne, seed transmission and significance of seedborne initial inoculum for the development of bacterial blight of onion previously has not been assessed. This article describes experimental work designed to evaluate the biological importance of seedborne X. axonopodis pv. allii as an initial inoculum source, and examine the temporal and spatial dynamics of the disease. Over 3 years, outbreaks of bacterial blight of onion always were induced in experimental plots sown with naturally contaminated seed lots, with a contamination rate determined as 0.04%. Analyses of disease patterns indicated a likely seedborne origin for the inoculum associated with the early stages of epidemics. Spatial analyses performed with several statistical methods indicated aggregated patterns of disease incidence data. Primary foci enlarged over time, and a few distinct secondary foci sometimes were established after occurrence of wind-driven rains (with gusts up to 15 m s-1). Distances between primary and secondary foci ranged from less than 1 m (satellite foci) to 25 m. It remains possible that longdistance dispersal of inoculum was at least partly involved in the later stages of epidemics.