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Fire blight, which is caused by the bacteria Erwinia amylovora, remains one of the most important diseases limiting the productivity of apple and pear orchards in the United States. In commercial orchards, in-season fire blight management relies exclusively on the use of antibiotic treatments (such as streptomycin and oxytetracycline) and on bacterial biocontrol agents whose efficacy is limited. We hypothesize that the efficacy of the biocontrol agents can be greatly enhanced through targeted delivery to flowers, which serve as initial infection courts, using the Japanese orchard bee, Osmia cornifrons. Many factors, such as the synchrony of life cycle with plant phenology and specificity to pomaceous plants, suggest that O. cornifrons could be an excellent vector of the biocontrol products during bloom in pome tree fruits. However, deployment of this pollinator species to deliver biocontrol agents for fire blight control has not been attempted previously due to the lack of an efficient system to pack the bodies of the bees exiting nesting tubes with the biocontrol products. In this study, we design and test a dispenser system to facilitate the use of O. conifrons as a vector for commercially available biocontrol products for fire blight control. The effectiveness of O. conifrons to deliver biocontrol agents to flowers, and to effect secondary dissemination from treated to untreated flowers is also evaluated in greenhouse experiments. We found that the O. conifrons bees were able to use the nest dispenser designed for the delivery of biological control products, and are effective in vectoring and delivering the Bacillus subtilis-based biological control product (Serenade®) to apple blossoms. We also found that the O. cornifrons were effective in secondary inoculation of this biological control product to newly-opened flowers. These findings suggest the potential use of commercially available O. conifrons and other orchard bees in targeted delivery of biological control products for fire blight, and possibly other diseases, in different fruit crops.

Oxathiapiprolin, a novel oomycete fungicide recently registered by DuPont, was reported to have high intrinsic activity against cucurbit downy mildew (Pseudoperonospora cubensis). The goal of this study was to characterize disease control attributes of oxathiapiprolin-based fungicides critical to effective management of cucurbit downy mildew. In growth chamber and greenhouse studies, oxathiapiprolin-based fungicides were compared with mandipropamid, mefenoxam + mancozeb, fluopicolide + propamocarb, cymoxanil + mancozeb, and ametoctradin + dimethomorph products for pre- and postinfection activity, local systemic movement, and protection of new growth produced after fungicide application. In preventive application, oxathiapiprolin-based fungicides significantly (P < 0.0001) inhibited downy mildew development, with the highest level of disease observed being 0.4% compared with 86.7% observed for mandipropamid. When applied postinfection, oxathiapiprolin-based fungicides significantly (P < 0.0001) suppressed disease development, but disease control was reduced relative to that observed for preventive application. There was a significant effect of formulation on the postinfection activity of oxathiapiprolin, whereby the oil dispersion (OD) formulation was more inhibitory than the water-dispersible granule formulation (0.001 ≤ P ≤ 0.049). Disease severity on the outer half leaf portion protected from spray deposition during fungicide application was lower for oxathiapiprolin-based fungicides (1.6 to 6.6%) than observed for fluopicolide + propamocarb (10.9 to 23.7%), mefenoxam + mancozeb (40.3 to 51.4%), and the nontreated controls (83.3 to 84.9%), which indicates significant acropetal movement within the leaf. Postinfection applications of oxathiapiprolin-based formulations had the greatest effect on lesion growth and sporangia production compared with the other fungicides in the experiment. When applied preventively to rapidly growing plants in a greenhouse, oxathiapiprolin-based fungicides consistently protected new growth that was not present at the time of application, with the OD formulation reducing disease severity by >75% relative to nontreated plants. The practical implications of these observations are discussed.

The impact of cluster compactness and debris retention on harvest bunch rot of Vitis interspecific hybrid ‘Vignoles’ was investigated between 2001 and 2005 near Lake Erie, in Pennsylvania. Cluster compactness was characterized subjectively based on the OIV code 204 standard and objectively by determining the number of berries per centimeter of rachis. In 2001, 2002, and 2004, the median number of berries per centimeter for loose clusters was 6.3, 7.0, and 6.4 compared with 10.2, 12.7, and 12.4 for the compact clusters, respectively. Kolmogorov-Smirnoff and the Mann-Whitney U two-samples tests confirmed that the distribution of the berries per centimeter was significantly (90 ≤ χ2≤ 184.3; P < 0.0001) different between the two subjective compactness categories. Cluster compactness was strongly correlated with bunch rot incidence (χ2= 73.1 and 62.2 for 2001 and 2002, respectively; P < 0.0001), whereby disease incidence was higher in compact than in loose clusters. Logistic regression analysis indicated that every additional berry per centimeter unit of compactness almost doubled the likelihood of a cluster becoming infected with bunch rot (odds ratio = 1.828, 95% confidence interval [CI] = 1.392 to 2.399 in 2001 and odds ratio = 1.705, 95% CI = 1.394 to 2.085 in 2002). In 2004, bunch rot severity in compact clusters was nearly four times that of loose clusters. Linear regression analysis revealed that berries per centimeter accounted for >89% of the variation in bunch rot severity (R2= 0.893, P < 0.0001, n = 30) and >74% in cluster weight (R2= 0.745, P < 0.0001, n = 30). Accumulations of dehiscent floral debris contributed to greater bunch rot severity, and the effect was more pronounced in compact clusters than in loose clusters. Removal of basal leaves at trace bloom reduced berries per centimeter by 13% in 2004 and >25% in 2005, with corresponding reductions in bunch rot severity of 60% in 2004 and 62.5 to 82% in 2005. These results indicate that berries per centimeter is a good indicator of cluster compactness in Vignoles, and that practices that reduce cluster tightness would be effective in an integrated program for control of bunch rot on this cultivar.

Over six seasons from 2007 to 2012, the effects of the timing of cluster zone leaf removal and bloom gibberellin applications on Botrytis bunch rot, cluster morphology, yield, and juice composition were evaluated on Vitis vinifera Chardonnay grapevines. All experimental plots received Botrytis-specific fungicide applications at preclosure and veraison. Leaf removal at veraison, postfruit-set, or trace bloom reduced Botrytis severity by an average of 14, 47, or 71%, respectively, compared to no leaf removal. Leaf removal at trace bloom (LRTB) reduced Botrytis incidence and severity in five and four seasons, respectively, and was as effective as (2007 to 2010), or more effective than (2011 and 2012) two additional fungicide applications (at bloom and at preharvest), suggesting potential to reduce fungicide inputs. Gibberellin was less effective than LRTB, but reduced the incidence of Botrytis in 2008 at 5 mg/L and the severity of Botrytis in 2010 at 25 mg/L. Botrytis development increases with number of berries per cluster and berries per centimeter of cluster (compactness). Berries per cluster was reduced by LRTB in 2007, 2010, and 2011, and by 10 mg/L gibberellin in 2007 and 25 mg/L in 2011. Berries per centimeter was reduced by LRTB and gibberellin in 2007, 2008, and 2011. Leaf removal at postfruit-set produced the most noteworthy effects on juice composition, reducing titratable acidity in 2007, 2011, and 2012. Yields were reduced by LRTB in 2008, but no significant yield effects were observed in subsequent seasons. The efficacy of LRTB for reducing Botrytis bunch rot was highest in years with the most compact clusters, suggesting that the value of this treatment increases with increasing potential for bunch rot development.

Plant stress significantly reduces plant productivity. Automated on-the-go mapping of plant stress would allow for a timely intervention and mitigation of the problem before critical thresholds were exceeded, thereby maximizing productivity. The spectral signature of plant leaves was analyzed by a hyperspectral camera to identify the onset and intensity of plant water stress. Five different levels of water treatment were created in young apple trees (cv. ‘Buckeye Gala’) in a greenhouse. The trees were periodically monitored with a hyperspectral camera along with an active-illuminated spectral vegetation sensor and a digital color camera. Individual spectral images over a 385–1000 nm wavelength range were extracted at a specific wavelength to estimate reflectance and generate spectral profiles for the five different water treatment levels. Various spectral indices were calculated and correlated to stress levels. The highest correlation was found with Red Edge NDVI at 705 and 750 nm in narrowband indices and NDVI at 680 and 800 nm in broadband indices. The experimental results indicated that intelligent optical sensors could deliver decision support for plant stress detection and management.

The aim of this analysis was to estimate the effect sizes and consistency of products evaluated for fire blight control in the eastern United States over the last decade. Because only 3% of the 69 studies published from 2000 to 2008 explicitly presented a measure of within-study variability, a method for estimating the least significant difference (LSD) and, hence the sampling variance, for studies with at least two significant mean separations in the presented mean multiple comparisons was developed. Lin's concordance analysis indicated that the estimated LSD was an accurate predictor of the actual LSD based on 35 studies in a calibration evaluation (ρc = 0.997). Separate multi-treatment random-effects meta-analyses were performed for three control categories: antibiotics, biological control, and plant defense-activating products and mean log response ratios relative to the nontreated controls ([Formula: see text]) were computed for each treatment and then back-transformed to obtain the mean percent disease control. None of the products evaluated performed as well as streptomycin, the standard product for fire blight control, for which the mean disease control was 68.6%. As a group, experimental antibiotics provided the best fire blight control with mean effect sizes ranging from 59.7 to 61.7%. Among the biological controls, the best control was noted for treatments combining the antibiotic streptomycin with a product based on Pantoea agglomerans (55.0% mean disease reduction) or Bacillus subtilis (53.9%). Mean disease control was 31.9, 25.7, and 22.6%, respectively, for products based on B. subtilis, Pantoea agglomerans, and Pseudomonas fluorescens without an antibiotic, suggesting that the higher efficacy of the combination treatments was due to the antibiotic. Among the plant defense-activating products, prohexadione calcium had the highest and most consistent effect size (50.7% control), while other products provided modest mean disease control of between 6.1 and 25.8%. Percent control values were significantly moderated by study location and cultivar used in the study, and were smaller, but more variable, when products were tested under high disease intensity compared with low disease intensity. Results indicate that wide-scale use of biological control and plant defense-activating products in the eastern United States is likely to remain low.

Harvest bunch rot of wine grape, caused primarily by Botrytis cinerea, is a perennial problem limiting the productivity of eastern vineyards, especially on cultivars with compact clusters. The aim of the present study was to evaluate the effectiveness of gibberellic acid (GA) sprays at reducing the compactness of Chardonnay and Vignoles clusters and minimizing bunch rot. Applications of GA reduced the number of berries per centimeter and the incidence and severity of bunch rots in Vignoles and, to a lesser extent, in Chardonnay over three consecutive years; however, the magnitude of GA effects often depended on the timing and rate of application. Bloom GA applications were more effective (P < 0.001) at reducing compactness and bunch rots than prebloom applications. Significantly, negative effects of GA applications on yield were negligible based on data from 4 years of trials on single vines and 2 years of data on 24-vine plots of Vignoles, provided the rates did not exceed 25 ppm. Regression analysis showed that berries per centimeter accounted for between 89 and 94% of variation in the incidence of Botrytis rot on Vignoles. On Chardonnay, compactness accounted for 53% of the variation in incidence, and the estimated compactness level at which no bunch rot would occur was 4.40 ± 1.05 (mean ± standard error) berries per centimeter. The relationship between cluster compactness and spray coverage of berries was also investigated in two separate experiments. Spray coverage of individual berries decreased linearly as cluster compactness increased within the range tested (3 to 18 berries per centimeter). Cluster compactness accounted for two-thirds of the variation in individual berry coverage, and coverage was reduced by 40 to 50% for clusters with about 18 berries per centimeter. These results strongly support the use of GA in integrated management of bunch rot on Vignoles and Chardonnay in eastern U.S. vineyards.

The continuing exponential increase in scientific knowledge, the growing availability of large databases containing raw or partially annotated information, and the increased need to document impacts of large-scale research and funding programs provide a great incentive for integrating and adding value to previously published (or unpublished) research through quantitative synthesis. Meta-analysis has become the standard for quantitative evidence synthesis in many disciplines, offering a broadly accepted and statistically powerful framework for estimating the magnitude, consistency, and homogeneity of the effect of interest across studies. Here, we review previous and current uses of meta-analysis in plant pathology with a focus on applications in epidemiology and disease management. About a dozen formal meta-analyses have been published in the plant pathological literature in the past decade, and several more are currently in progress. Three broad research questions have been addressed, the most common being the comparative efficacy of chemical treatments for managing disease and reducing yield loss across environments. The second most common application has been the quantification of relationships between disease intensity and yield, or between different measures of disease, across studies. Lastly, meta-analysis has been applied to assess factors affecting pathogen–biocontrol agent interactions or the effectiveness of biological control of plant disease or weeds. In recent years, fixed-effects meta-analysis has been largely replaced by random- (or mixed-) effects analysis owing to the statistical benefits associated with the latter and the wider availability of computer software to conduct these analyses. Another recent trend has been the more common use of multivariate meta-analysis or meta-regression to analyze the impacts of study-level independent variables (moderator variables) on the response of interest. The application of meta-analysis to practical problems in epidemiology and disease management is illustrated with case studies from our work on Phakopsora pachyrhizi on soybean and Erwinia amylovora on apple. We show that although meta-analyses are often used to corroborate and validate general conclusions drawn from more traditional, qualitative reviews, they can also reveal new patterns and interpretations not obvious from individual studies.

Bayesian statistical methods are used for meta-analysis in many disciplines, including medicine, molecular biology, and engineering, but have not yet been applied for quantitative synthesis of plant pathology studies. In this paper, we illustrate the key concepts of Bayesian statistics and outline the differences between Bayesian and classical (frequentist) methods in the way parameters describing population attributes are considered. We then describe a Bayesian approach to meta-analysis and present a plant pathological example based on studies evaluating the efficacy of plant protection products that induce systemic acquired resistance for the management of fire blight of apple. In a simple random-effects model assuming a normal distribution of effect sizes and no prior information (i.e., a noninformative prior), the results of the Bayesian meta-analysis are similar to those obtained with classical methods. Implementing the same model with a Student's t distribution and a noninformative prior for the effect sizes, instead of a normal distribution, yields similar results for all but acibenzolar-S-methyl (Actigard) which was evaluated only in seven studies in this example. Whereas both the classical (P = 0.28) and the Bayesian analysis with a noninformative prior (95% credibility interval [CRI] for the log response ratio: –0.63 to 0.08) indicate a nonsignificant effect for Actigard, specifying a t distribution resulted in a significant, albeit variable, effect for this product (CRI: –0.73 to –0.10). These results confirm the sensitivity of the analytical outcome (i.e., the posterior distribution) to the choice of prior in Bayesian meta-analyses involving a limited number of studies. We review some pertinent literature on more advanced topics, including modeling of among-study heterogeneity, publication bias, analyses involving a limited number of studies, and methods for dealing with missing data, and show how these issues can be approached in a Bayesian framework. Bayesian meta-analysis can readily include information not easily incorporated in classical methods, and allow for a full evaluation of competing models. Given the power and flexibility of Bayesian methods, we expect them to become widely adopted for meta-analysis of plant pathology studies.

The risk of spreading fire blight in apples after mechanical thinning with a rotating string blossom thinner was evaluated in field and potted-tree experiments. In the field experiment, using the mechanical thinner on noninoculated trees immediately after operating the equipment on inoculated trees significantly (P < 0.01) increased fire blight incidence resulting in 90 ± 20.01 (mean ± SE) infected shoots compared with 23.5 ± 8.97 diseased shoots in similar trees that were not thinned mechanically. A similar result was obtained in greenhouse experiments whereby healthy apple plants positioned adjacent to diseased plants before the group was subjected to the mechanical thinner developed more than twice the number of infected shoots as that on similar plants that were not thinned. These results indicate that under conditions conducive to infection, the mechanical blossom thinner significantly increases the risk of spreading Erwinia amylovora. The use of the thinner should therefore be limited to orchards with no history of disease in the last 3 years and on days when predicted weather is not suitable for tree infection by E. amylovora; otherwise, a severe fire blight epidemic could develop in the orchard.

The ability to infect host flowers offers important ecological benefits to plant-parasitic fungi; not surprisingly, therefore, numerous fungal species from a wide range of taxonomic groups have adopted a life style that involves flower infection. Although flower-infecting fungi are very diverse, they can be classified readily into three major groups: opportunistic, unspecialized pathogens causing necrotic symptoms such as blossom blights (group 1), and specialist flower pathogens which infect inflorescences either through the gynoecium (group 2) or systemically through the apical meristem (group 3). This three-tier system is supported by life history attributes such as host range, mode of spore transmission, degree of host sterilization as a result of infection, and whether or not the fungus undergoes an obligate sexual cycle, produces resting spores in affected inflorescences, and is r- or K-selected. Across the three groups, the flower as an infection court poses important challenges for disease management. Ecologically and evolutionarily, terms and concepts borrowed from the study of venereal (sexually transmitted) diseases of animals do not adequately capture the range of strategies employed by fungi that infect flowers.

Mimicry is the close resemblance of one living organism (the mimic) to another (the model), leading to misidentification by a third organism (the operator). Similar to other organism groups, certain species of plant-parasitic fungi are known to engage in mimetic relationships, thereby increasing their fitness. In some cases, fungal infection can lead to the formation of flower mimics (pseudo flowers) that attract insect pollinators via visual and/or olfactory cues; these insects then either transmit fungal gametes to accomplish outcrossing (e.g. in some heterothallic rust fungi belonging to the genera Puccinia and Uromyces) or vector infectious spores to healthy plants, thereby spreading disease (e.g. in the anther smut fungus Microbotryum violaceum and the mummy berry pathogen Monilinia vaccinii-corymbosi). In what is termed aggressive mimicry, some specialized plant-parasitic fungi are able to mimic host structures or host molecules to gain access to resources. An example is M. vaccinii-corymbosi, whose conidia and germ tubes, respectively, mimic host pollen grains and pollen tubes anatomically and physiologically, allowing the pathogen to gain entry into the host's ovary via stigma and style. We review these and other examples of mimicry by plant-parasitic fungi and some of the mechanisms, signals, and evolutionary implications.

Application of Serenade, a commercial biofungicide formulation containing the bacterium Bacillus subtilis, to the stigmatic surface of open blueberry Xowers suppresses Xoral infection by the mummy berry fungus Monilinia vaccinii-corymbosi. The deliberate targeting of the stigma with the biocontrol agent in this pathosystem prompted us to evaluate potential negative impacts on pollination and pollination-related fruit characteristics. Application of Serenade to the stigmatic surface of detached blueberry Xowers in the laboratory had no eVect (P > 0.05) on the number of pollen tubes entering the style or their growth rates within the stylar canal. There was also no reciprocal eVect, i.e., population dynamics of B. subtilis were unaltered by the presence of pollen. Application of the biocontrol product to open Xowers, regardless of whether it was done 1 day before or immediately prior to pollination, did not impact fruit set or the number of seeds per berry, but marginally (P D 0.048) aVected fruit weight in one of two experimental runs in the greenhouse; fruit weights in the two Serenade timing treatments were signiWcantly diVerent from each other but neither was diVerent from that of the control that received pollen only. In a Weld experiment in which honey bees were utilized to vector the biocontrol product to open Xowers, application of Serenade did not aVect fruit weight but signiWcantly reduced fruit set from 49.1 to 38.1% (P D 0.0382) and seed number to about half of that of the untreated control (P D 0.0109). However, fruit weights and seed numbers in the experiment were low even in treatments receiving no Serenade, indicative of poor pollination overall. Taken together, these results indicate that application of Serenade has no inherently adverse eVects on pollination and associated fruit characteristics, but caution should be exercised in applying this product in conditions otherwise unfavorable for adequate pollination.

Infection of blueberry flowers by Monilinia vaccinii-corymbosi , which causes mummy berry disease, occurs via the gynoecium and involves conidial germination on the stigmatic surface followed by hyphal ingress into the stylar canal and subsequent colonization of the ovary. The extent to which these events mimic pollen–pistil interactions during pollination was investigated. Similar to blueberry pollen tubes, conidial germ tubes of M. vaccinii-corymbosi adhered selectively to imprints of stylar transmitting tract tissue on nitrocellulose membrane, with adhesion in both cases occurring at the tips of the tubes. By contrast, hyphae of the related Monilinia fructicola , which is nonpathogenic on blueberry and does not cause gynoecial infection, adhered indiscriminately to the entire membrane. Using monoclonal antibodies, the presence of epitopes of esterified and unesterified pectins and of arabinogalactan proteins (AGPs), which have been implicated in adhesion and pollen tube guidance in other plant species, was documented on blueberry pollen tubes in vitro. Epitopes of certain AGPs, but not of pectins, were also localized on conidia and hyphae of M. vaccinii-corymbosi . However, such epitopes were also detected on M. fructicola , suggesting that they are unlikely to be a discriminatory factor between fungi capable or not capable of gynoecial infection. Microscopic observation of inoculated pistils showed that similar to pollen tubes, hyphae of M. vaccinii-corymbosi tracked the lobes of the stylar lumen, grew directionally (i.e. with very limited branching) in close proximity to cells of the stylar adaxial epidermis and to one another, and were surrounded by extracellular matrix. By contrast, hyphae of M. fructicola , while being able to ingress into the style, branched profusely within the stylar canal, showing no directional growth or affinity to specific regions of the lumen. We propose these results as evidence of specialized opportunism by M. vaccinii-corymbosi , whereby fungal hyphae appear to mimic host pollen tubes and take advantage of an infrastructure intended to support host reproduction in order to facilitate infection of the ovary.

Monilinia vaccinii-corymbosi , the fungus that causes mummy berry disease, infects open blueberry flowers via the stigma–style pathway, followed by internal colonization and subsequent mummification of the developing fruit. The potential for use of the commercial biocontrol products Serenade ( Bacillus subtilis QRD137) and BlightBan ( Pseudomonas fluorescens A506) to suppress floral infection in this pathosystem was investigated. B. subtilis exhibited marked antibiosis against the pathogen in vitro, reducing radial growth rates of M. vaccinii-corymbosi to less than one-half of those of the untreated control after 1 week of co-culture on potato-dextrose agar ( P =0.0029). In contrast, P. fluorescens did not reduce fungal growth compared with the untreated control ( P =0.3182). When Serenade was applied to stigmatic surfaces of detached blueberry flowers in the laboratory, population densities of B. subtilis , determined by dilution-plating, were variable 1 and 2 days after application, followed by a consistent drop in bacterial numbers after 3 and 4 days. In contrast, population densities of P. fluorescens increased exponentially 1 and 2 days after application of BlightBan and remained constant thereafter. The greatest mean population densities observed were 1.6 × 10 5 and 1.5 × 10 5 colony-forming units per stigma for B. subtilis and P. fluorescens , respectively. In a separate experiment, conidia of M. vaccinii-corymbosi were applied to detached flowers 24 h before or after application of the biocontrol products to flower stigmas. Serenade significantly reduced the number and growth rates of hyphae penetrating the stylar canal to less than one-fifth of those measured in the untreated control, whereas fungal suppression following application of BlightBan was less pronounced. Reductions in fungal growth rate in flowers treated with B. subtilis isolated from Serenade and reformulated by freeze-drying in skim milk were intermediate between the untreated control and the Serenade treatment, suggesting that part of the observed activity of Serenade was due to the formulation of that product. The results of these experiments identify Serenade as a promising candidate for further testing against mummy berry disease.

Pseudosclerotia (infected, mummified fruit) of Monilinia vacciniicorymbosi overwinter on the orchard floor and germinate to produce apothecia in early spring, providing the only source of primary inoculum for mummy berry disease of blueberry. Three experiments were carried out to develop a model for the relative efficacy of mechanical cultivation in reducing the risk associated with primary inoculum. In the first experiment, apothecial emergence from pseudosclerotia buried 0, 1.5, 3, 6, and 10 cm below the soil surface was monitored to determine the critical depth necessary to inhibit emergence. No apothecia emerged from pseudosclerotia buried at depths of ≥3 cm, and the critical depth of burial was determined at 2.6 cm by regression analysis. In the second experiment, pseudosclerotia or plastic beads (used as surrogates for pseudosclerotia) were placed on the soil surface of experimental plots before cultivation with an in-row rotary cultivator, a disc harrow, or a rotary cultivator with each implement operated in a single pass. Vertical distribution profiles of pseudosclerotia or beads in the topsoil were characterized after excavation with a custom-built sampling device. The proportion of pseudosclerotia placed below the critical depth of 2.6 cm was 20.9, 52.6, and 78.6% for the in-row rotary cultivator, the disc harrow, and the rotary cultivator, respectively. For all three implements, vertical distribution profiles of pseudosclerotia and plastic beads were very similar, allowing the latter to be used in subsequent experiments in commercial fields. In the third experiment, two blueberry plantings were surveyed to determine the horizontal distribution of pseudosclerotia on the orchard floor with distance from the crowns of the plants. The greatest frequency of pseudosclerotia occurred between 30 and 40 cm from the plants. Based on measurements of the distance from plants within which different implements can operate, the proportion of pseudosclerotia accessible by cultivation ranged from 58.7% for the disc harrow to 87.2% for the in-row rotary cultivator. Taken together, results from the three experiments indicated that cultivation with a single implement can reduce risk of apothecial emergence by about 50%. More effective risk reductions may be obtained by combining implements that result in deep burial of pseudosclerotia with those that have access to pseudosclerotia near the plants. This was demonstrated by a commercial cultivation method that utilized three passes of different implements and resulted in extensive reshaping of plant beds, placing 88.2% of beads below the critical depth of 2.6 cm.

Four isolates of Bacillus spp. were tested in a 2-year field study for biological control of pre- and postharvest diseases of apple. For the preharvest test, bacteria were applied to ‘Golden Delicious’ and ‘Rome Beauty’ trees in May or May and June. Foliar apple scab severity was assessed weekly. After harvest, fruit were wounded and then either left untreated or given a postharvest application of the bacteria. Wounded fruit were then inoculated with the bitter rot pathogen and lesion size was measured over 8 days. Bacillus megaterium isolate A3-6, B. mycoides isolate A1-1, and B. cereus isolate FLS-5 applied in May or May and June significantly reduced fruit and foliar apple scab severity in both years. A postharvest application of the bacteria (alone or in combination with a preharvest application) resulted in the greatest suppression of bitter rot on both cultivars (P < 0.04). The May + June + postharvest application of isolate A3-6 resulted in the greatest suppression of bitter rot, with an average of 45 and 95% reduction in lesion size compared with nontreated controls on ‘Golden Delicious’ and ‘Rome Beauty’ fruit, respectively. Results from this study indicate that preharvest applications of the bacteria were able to reduce foliar and fruit scab and an additional postharvest application was effective in reducing bitter rot severity.

Background Desirable apple varieties are clonally propagated by grafting vegetative scions onto rootstocks. Rootstocks influence many phenotypic traits of the scion, including resistance to pathogens such as Erwinia amylovora, which causes fire blight, the most serious bacterial disease of apple. The purpose of the present study was to quantify rootstock-mediated differences in scion fire blight susceptibility and to identify transcripts in the scion whose expression levels correlated with this response. Results Rootstock influence on scion fire blight resistance was quantified by inoculating three-year old, orchard-grown apple trees, consisting of 'Gala' scions grafted to a range of rootstocks, with E. amylovora. Disease severity was measured by the extent of shoot necrosis over time. 'Gala' scions grafted to G.30 or MM.111 rootstocks showed the lowest rates of necrosis, while 'Gala' on M.27 and B.9 showed the highest rates of necrosis. 'Gala' scions on M.7, S.4 or M.9F56 had intermediate necrosis rates. Using an apple DNA microarray representing 55,230 unique transcripts, gene expression patterns were compared in healthy, un-inoculated, greenhouse-grown 'Gala' scions on the same series of rootstocks. We identified 690 transcripts whose steady-state expression levels correlated with the degree of fire blight susceptibility of the scion/rootstock combinations. Transcripts known to be differentially expressed during E. amylovora infection were disproportionately represented among these transcripts. A second-generation apple microarray representing 26,000 transcripts was developed and was used to test these correlations in an orchard-grown population of trees segregating for fire blight resistance. Of the 690 transcripts originally identified using the first-generation array, 39 had expression levels that correlated with fire blight resistance in the breeding population. Conclusions Rootstocks had significant effects on the fire blight susceptibility of 'Gala' scions, and rootstock-regulated gene expression patterns could be correlated with differences in susceptibility. The results suggest a relationship between rootstock-regulated fire blight susceptibility and sorbitol dehydrogenase, phenylpropanoid metabolism, protein processing in the endoplasmic reticulum, and endocytosis, among others. This study illustrates the utility of our rootstock-regulated gene expression data sets for candidate trait-associated gene data mining.

Orchard management practices, such as destroying of overwintered inoculum and limiting the number of fungicide applications, are often recommended as tactics for slowing the development of resistance to sterol demethylation-inhibitor (DMI) fungicides in populations of Venturia inaequalis. However, there is little quantitative evidence relating the use of such practices to levels of resistance in orchards. The aim of this study was to evaluate the sensitivity of V. inaequalis isolates from Pennsylvania to DMI fungicides, and to identify orchard management factors related to the incidence of resistant isolates. In total, 644 single-spore V. inaequalis cultures obtained from 20 apple orchards in 2008 or 2009 were tested for sensitivity to myclobutanil, fenbuconazole, or difenoconazole. Growers provided management history of the sampled plots. Widespread shifts toward resistance to the three fungicides were noted, with mean effective concentration for 50% inhibition (EC50) values of 2.136, 0.786, and 0.187 μg/ml for myclobutanil, fenbuconazole, and difenoconazole, respectively. Cross resistance to the three fungicides was documented in high correlation (Spearman's r > 0.6) between mean EC50 values for 14 orchards. Based on a 0.5-μg/ml threshold, 66 and 26% of isolates were resistant to myclobutanil and fenbuconazole, respectively, and 22% were cross resistant to the two fungicides. A significant between-year shift toward increased resistance was noted in two of three orchards surveyed in both years. Failure to use dormant copper sprays, older trees, larger orchards, orchards with ≤10 cultivars, and application of >4 DMI sprays were positively correlated (0.0001 < P < 0.05) with the incidence of resistant isolates. Isolates from orchards with >4 DMI sprays were four times as likely to be resistant to fenbuconazole (odds ratio = 4.57; P = 0.015). Isolates from orchards without dormant copper sprays were twice as likely to be cross-shifted toward resistance to all three fungicides (odds ratio = 1.76; P = 0.048). Results identify management practices that can reduce the risk of V. inaequalis developing resistance to DMI fungicides.

The fibrillins are a large family of chloroplast proteins that have been linked with stress tolerance and disease resistance. FIBRILLIN4 (FIB4) is found associated with the photosystem II light-harvesting complex, thylakoids, and plastoglobules, which are chloroplast compartments rich in lipophilic antioxidants. For this study, FIB4 expression was knocked down in apple (Malus 3 domestica) using RNA interference. Plastoglobule osmiophilicity was decreased in fib4 knockdown (fib4 KD) tree chloroplasts compared with the wild type, while total plastoglobule number was unchanged. Compared with the wild type, net photosynthetic CO2 fixation in fib4 KD trees was decreased at high light intensity but was increased at low light intensity. Furthermore, fib4 KD trees produced more anthocyanins than the wild type when transferred from low to high light intensity, indicating greater sensitivity to high light stress. Relative to the wild type, fib4 KD apples were more sensitive to methyl viologen and had higher superoxide levels during methyl viologen treatment. Arabidopsis (Arabidopsis thaliana) fib4 mutants and fib4 KD apples were more susceptible than their wild-type counterparts to the bacterial pathogens Pseudomonas syringae pathovar tomato and Erwinia amylovora, respectively, and were more sensitive to ozone-induced tissue damage. Following ozone stress, plastoglobule osmiophilicity decreased in wild-type apple and remained low in fib4 KD trees; total plastoglobule number increased in fib4 KD apples but not in the wild type. These results indicate that FIB4 is required for plastoglobule development and resistance to multiple stresses. This study suggests that FIB4 is involved in regulating plastoglobule content and that defective regulation of plastoglobule content leads to broad stress sensitivity and altered photosynthetic activity.

Plant stress significantly reduces plant productivity. Automated on-the-go mapping of plant stress allows for timely intervention and mitigation of the problem before critical thresholds are exceeded, thereby maximizing productivity. A hyperspectral camera analyzed the spectral signature of plant leaves to identify the plant water stress. Five different levels of water treatment were created on young apple trees (Buckeye Gala) in a greenhouse and continuously monitored with a hyperspectral camera along with an active-illuminated spectral vegetation sensor and a digital color camera. Individual spectral images over a 400 – 1000 nm wavelength range were extracted at a specific wavelength to estimate reflectance and generate spectral profiles for five groups of apple trees at different water treatment levels. Various spectral indices were investigated and correlated to stress levels. The highest correlation was found with Red Edge NDVI at 705 nm and 750 nm in narrowband indices and NDVI at 680 nm and 800 nm in broadband indices. The experimental results indicate that intelligent optical sensors could deliver decision support for plant stress detection and management.

Plant stress has been estimated by spectral signature using both passive and active sensors. As optical sensors measure reflected light from a target, changes in illumination characteristics critically affect sensor response. Active sensors minimize the illumination effects by producing their own illumination that is reflected from the target and measured by the detector. Although active sensors use modulated radiation that can be differentiated from ambient illumination, in order to validate data and increase the accuracy, sensor performance characteristics must be well understood and examined in different target conditions of plant leaves. The performance of an active NDVI sensor was evaluated to study the effect of: 1) partial canopy coverage, 2) target off-center, 3) standoff distance, 4) target surface tilting, 5) solar bidirectional effect, 6) temperature, and 7) diurnal radiation change. These evaluations provide a valid range of sensor measurements and a motivation to improve the measurement accuracy by using selective data that can be validated by supplemental sensors.

We review trends and advances in three specific areas of theoretical plant epidemiology: models of temporal and spatial dynamics of disease, the synergism of epidemiology and population genetics, and progress in statistical epidemiology. Recent analytical modelling of disease dynamics has focused on SIR (susceptible–infected–removed) models modified to include spatial structure, stochasticity, and multiple management-related parameters. Such models are now applied routinely to derive threshold criteria for pathogen invasion or persistence based on pathogen demographics (e.g., Allee effect or fitness of fungicide-resistant strains) and/or host spatial structure (e.g., host density or patch size and arrangement). Traditionally focused on the field level, the scale of analytical models has broadened to range from individual plants to landscapes and continents; however, epidemiological models for interactions at the cellular level, e.g., during the process of virus infection, are still rare. There is considerable interest in the concept of scaling, i.e., to what degree and how data and models from one scale can be transferred to another (smaller or larger) scale. Despite assertions to the contrary, the linkages between epidemiology and population genetics are alive and well as exemplified by recent efforts to integrate epidemiological parameters into population genetics models (and vice versa) and by numerous integrated studies with an applied focus (e.g., to quantify sources and types of primary and secondary inoculum). Statistical plant epidemiology continues to rely heavily on the medical and ecological fields for inspiration and conceptual advances, as illustrated by the recent surge in papers utilizing ROC (receiver operating characteristic), Bayesian, or survival analysis. Among these, Bayesian analysis should prove especially fruitful given the reliance on uncertain and subjective information for practical disease management. However, apart from merely adopting statistical tools from other disciplines, plant epidemiologists should be more proactive in exploring potential applications of their concepts and procedures in rapidly expanding disciplines such as statistical genetics or bioinformatics. Although providing the scientific basis for disease management will always be the raison d’etre for plant epidemiology, a broader perspective will help the discipline to remain relevant as more resources are being devoted to genomic and ecosystem-level science.