Effect of Microsphaeropsis sp. Strain P130A on Germination and Production of Sclerotia of Rhizoctonia solani and Interaction Between the Antagonist and the Pathogen

Microsphaeropsis sp. strain P130A was evaluated for the control of tuber-borne inoculum of Rhizoctonia solani based on the viability of sclerotia produced in vitro and on both the viability and production of tuber-borne sclerotia. The interactions between the antagonist and the pathogen, as well as the effect of the toxins produced by the antagonist on mycelial growth of R. solani were studied using transmission electron microscopy. On sclerotia produced in vitro, for all incubation periods (1 to 42 days), Microsphaeropsis sp. significantly reduced germination. Percent germination of sclerotia treated with Microsphaeropsis sp. decreased with increasing incubation period from an average of 82.0% after 1 day to stabilize at an average of 5.8% after 35 days. Similarly, percent germination of tuber-borne sclerotia was significantly lower when tubers were treated with Microsphaeropsis sp. Both 2% formaldehyde and Microsphaeropsis sp. treatments significantly reduced sclerotia germination to approximately 10% after 42 days of incubation at 4 degrees C. Furthermore, on tubers treated with the antagonist, the number of sclerotia per square centimeter decreased from 1.6 to 0.5 during the 8 months of storage at 4 degrees C, whereas an increase from 1.2 to 7.8 sclerotia per square centimeter was observed on untreated tubers. Microsphaeropsis sp. (strain P130A) colonized hyphae of R. solani within 4 days after contact on culture media. Transmission electron microscopic observations showed that the antagonist induced a rupture of the pathogen plasma membrane and that a chitin-enriched matrix was deposited at sites of potential antagonist penetration. Host penetration was not associated with pathogen cell wall alterations, which occurred at the time of progress of the antagonist in the pathogen cytoplasm. In the presence of a crude extract of Microsphaeropsis sp., cells of R. solani showed cytoplasm disorganization and breakdown of plasma membranes. Antibiosis and mycoparasitism were involved in the antagonism of R. solani by Microsphaeropsis sp., but the sequence by which these events occur, as well as the significance of wall appositions produced by R. solani, is yet to be established.     

Heterogeneity of the aerial concentration and deposition of ascospores of <Emphasis Type="Italic">Venturia inaequalis</Emphasis> within a tree canopy during the rain

Scab is an important disease of apple and its control depends almost exclusively on frequent use of fungicides. Primary scab infection in the spring assumes several steps: ascospore maturation, liberation of ascospores that become airborne, deposition on susceptible tissues, and infection. However, the spatial heterogeneity of ascospores within the tree canopy is unknown. Aerial concentration of ascospore (ACA), ascospore concentration in rain water (ACR) and ascospore deposition (AD) were therefore measured at six heights (20–257 cm from the ground) with rotating-arm air samplers, funnels, and greased glass slides, respectively, during five rain events in 2001 and in 2002. In addition, ACR and AD were measured at eight locations within tree canopy at 196 cm height. Apple scab was assessed at the end of the primary infection period in each sampling location within the apple tree. A similar experimental design was used in 2003 to study the spatial heterogeneity of both AD and primary scab lesions. ACA and AD decreased with increasing height, while ACR increased with increasing height. Based on both variance to mean ratio and the power law relationship in both years, the ACR was heterogeneous, while AD was heterogeneous only during the peaks of ascospore release. The ACR was significantly higher at the centre of the trees and the AD was significantly higher at the centre and at the western edge of the trees. Only the cumulative AD was significantly correlated with apple scab lesions at the same location (r = 0.83). In 2003, a similar pattern of spatial heterogeneity within the tree canopy was observed for AD and primary scab lesion counts and there was a linear relationship (R ² = 0.84) between these two variables. It was concluded that ACR and AD within the tree canopy are not randomly distributed at least during peaks of ascospore release and that AD is a good estimate of primary scab lesion development. This spatial heterogeneity should be considered when estimating ascospore deposition using mathematical models or when quantifying ascosporic inoculum using spore samplers.     

Cytochemical Investigation of the Antagonistic Interaction Between a Microsphaeropsis sp. (Isolate P130A) and Venturia inaequalis

ABSTRACT In an attempt to better understand the mode of action of the antagonistic fungus Microsphaeropsis sp., the interaction between this fungus and Venturia inaequalis was studied, using both light and electron microscopy. Cytological observations indicated that the antagonistic interaction between the two fungi likely involves a sequence of events, including (i) attachment and local penetration of Microsphaeropsis sp. into V. inaequalis hyphae; (ii) induction of host structural response at sites of potential antagonist entry; (iii) alteration of host cytoplasm; and (iv) active multiplication of antagonistic cells in pathogen hyphae, leading to host cell breakdown and release of the antagonist. The interaction was investigated further by gold cytochemistry. The use of gold-complexed beta-1,4-exoglucanase and wheat germ agglutinin/ovomucoid-gold complex to localize cellulosic beta-1,4-glucans and chitin monomers, respectively, resulted in regular labeling of V. inaequalis cell walls. This finding supports other studies refuting the classification of ascomycetes as only a glucan-chitin group. At an advanced state of parasitism, the labeling pattern of cellulose and chitin, which clearly showed that the level of integrity of these compounds was affected, suggested the production of cellulolytic and chitinolytic enzymes by Microsphaeropsis sp. Wall appositions formed in V. inaequalis in response to the antagonist's attack contained both cellulose and chitin. However, penetration of this newly formed material frequently succeeded. This study provides the first detailed picture of the cytological events associated with mycoparasitism in V. inaequalis.     

Spatial heterogeneity,incidence-incidence and incidence-lesion density relationship of apple scab (<Emphasis Type="Italic">Venturia inaequalis</Emphasis>) in managed orchards

The spatial pattern of apple scab was characterized using 10 years of disease incidence and lesion density data collected in managed orchards located in Quebec, Canada. Distributional analyses indicated that scab incidence was better characterized by the beta-binomial than the binomial distribution in 53 and 65% of the data sets at the leaf and shoot scales, respectively. Median values of the beta-binomial parameter θ, a measure of small-scale aggregation, were near 0 (0.003 and 0.028) at both sampling scales, indicating that disease incidence was close to being randomly distributed (low degree of aggregation). For lesion density, the negative binomial distribution fitted the data better than the Poisson distribution in 86% of the data sets at the leaf scale. The median value of the index of dispersion k was 0.068, indicating that aggregation was present. For all apple scab measurements, the power law models provided a good fit to the data. The estimated slope and intercept parameters were significantly greater than 1 and 0, respectively, suggesting that spatial heterogeneity changed systematically with disease incidence. Results of a covariance analysis showed that spatial heterogeneity of scab incidence at both scales and lesion density was not dependent upon shoot type but that spatial heterogeneity of scab incidence and lesion density at the leaf scale was influenced by the sampling period. A hierarchical analysis showed that scab incidence at the tree scale increased as a saturation-type curve with respect to incidence at the leaf or shoot scales. A similar relationship was observed for incidences at the shoot and leaf scales. An effective sample size model based on the binary power law parameters (Madden and Hughes, Phytopathology 89:770–781, 1999) gave the best fit to the leaf and shoot data, respectively. The incidence-lesion density relationship at both scales was well described by a complementary log-log (CLL) and log transformation model ( R_adj² = 0.97 and R_adj² = 0.94 ) \left( {R_{{adj}}^2 = 0.97\,and\,R_{{adj}}^2 = 0.94} \right) , however, the models tended to underestimate lesion density. The information of the spatial relations of apple scab within and between hierarchical scales acquired from this study can be used in developing and evaluating practical disease management strategies and to improve apple scab assessments for fungicide or cultivar susceptibility trials.     

Spatial Distribution of Venturia inaequalis Airborne Ascospores in Orchards

ABSTRACT Apple scab (Venturia inaequalis) causes important economic losses in many apple production areas of the world. The disease is controlled by numerous fungicide applications regardless of the presence of ascospores in the orchard. Airborne ascospore concentration (AAC) can be measured in real time to time fungicide applications. However, the level of heterogeneity of the AAC in commercial orchards was unknown. Consequently, the spatial distribution of V. inaequalis ascospores was studied in a commercial apple orchard of 0.43 ha. The potential ascospore dose (PAD) and AAC were measured in 40 quadrats each of 108 m(2). In each quadrat, the AAC was monitored during the major rain events in spring 1999 and 2000 using spore samplers. The variance-to-mean ratio for the PAD and for most of the AAC sampling dates was >1, indicating an aggregated pattern of distribution. None of the frequency distributions of the most important ascospore ejection events followed the Poisson probability distribution, indicating that the pattern of distribution was not random. For all events, AAC had an aggregated pattern of distribution as suggested by the negative binomial distribution. The PAD followed neither the Poisson nor the negative binomial distribution. Geostatistical analyses confirmed the aggregated pattern of distribution. The cultivars had an effect on the PAD and AAC distribution pattern, but both PAD and AAC were not uniformly distributed within a block of the same cultivar. Therefore, the number, location, and height of samplers required to estimate AAC in orchards need to be investigated before using information on AAC for decision making.     

Risk factors for crop health under global change and agricultural shifts: a framework of analyses using rice in tropical and subtropical Asia as a model

Plant disease epidemiology requires expansion of its current methodological and theoretical underpinnings in order to produce full contributions to global food security and global changes. Here, we outline a framework which we applied to farmers' field survey data set on rice diseases in the tropical and subtropical lowlands of Asia. Crop health risks arise from individual diseases, as well as their combinations in syndromes. Four key drivers of agricultural change were examined: labor, water, fertilizer, and land availability that translate into crop establishment method, water shortage, fertilizer input, and fallow period duration, respectively, as well as their combinations in production situations. Various statistical approaches, within a hierarchical structure, proceeding from higher levels of hierarchy (production situations and disease syndromes) to lower ones (individual components of production situations and individual diseases) were used. These analyses showed that (i) production situations, as wholes, represent very large risk factors (positive or negative) for occurrence of disease syndromes; (ii) production situations are strong risk factors for individual diseases; (iii) drivers of agricultural change represent strong risk factors of disease syndromes; and (iv) drivers of change, taken individually, represent small but significant risk factors for individual diseases. The latter analysis indicates that different diseases are positively or negatively associated with shifts in these drivers. We also report scenario analyses, in which drivers of agricultural change are varied in response to possible climate and global changes, generating predictions of shifts in rice health risks. The overall set of analyses emphasizes the need for large-scale ground data to define research priorities for plant protection in rapidly evolving contexts. They illustrate how a structured theoretical framework can be used to analyze emergent features of agronomic and socioecological systems. We suggest that the concept of "disease syndrome" can be borrowed in botanical epidemiology from public health to emphasize a holistic view of disease in shifting production situations in combination with the conventional, individual disease-centered perspective.     

Age-related susceptibility of strawberry leaves and berries to infection by Podosphaera aphanis

Powdery mildew, caused by Podosphaera aphanis, is a major disease of strawberry worldwide. The importance of the disease varies with production system (June bearing vs day neutral) which could be explained in part by the concurrent presence of susceptible leaves or berries and abundant airborne inoculum. Age-related susceptibility was studied by inoculating strawberry leaves and berries at different growth stages of the June bearing cultivar Jewel and the day neutral cultivar Seascape. On eight occasions in 2007, five plants of each cultivar were inoculated with dry conidia using a settling tower. There was a significant effect of leaf and berry growth stage on the susceptibility, which decreased exponentially as leaves or berries aged to reach almost zero when the leaves were completely expanded or the berries at the pink stage at the time of inoculation. The proportion of maximum mildew severity as a function of leaf or berry growth stage was predicted using non-parametric regression (R² = 0.96–0.97). The predicted values were validated with data collected in fields or tunnels naturally infected by P. aphanis. There was a linear relation between predicted and observed proportion of maximum mildew severity (R² = 0.95–0.99). The results of this study showed that timing fungicide sprays based on periods of high leaf and berry susceptibility should greatly improve management of strawberry powdery mildew.     

Effect of Timing of Application of the Biological Control Agent Microsphaeropsis ochracea on the Production and Ejection Pattern of Ascospores by Venturia inaequalis

ABSTRACT Field and in vitro trials were conducted to establish the influence of the biological control agent Microsphaeropsis ochracea on the ejection pattern of ascospores by Venturia inaequalis and on apple scab development, and to establish the best timing of application. The ejection pattern of ascospores was similar on leaves sprayed with M. ochracea and on untreated leaves. Fall application of M. ochracea combined with a delayed-fungicide program was evaluated in orchards with intermediate and high scab risk. For both orchards, it was possible to delay the first three and two infection periods in 1998 and 1999, respectively, without causing significant increase or unacceptable leaf and fruit scab incidence. To evaluate the best timing of application, sterile leaf disks were inoculated with V. inaequalis and then with M. ochracea 0, 2, 4, 6, 8, 10, 12, 14, and 16 weeks later. After incubation under optimal conditions for pseudothecia development, the number of ascospores was counted. Similarly, M. ochracea was sprayed on scabbed leaves on seven occasions from August to November 1999 and 2000. Leaves were overwintered on the orchard floor and ascospore production was evaluated the following spring. Ascospore production was reduced by 97 to 100% on leaf disks inoculated with M. ochracea less than 6 weeks after inoculation with V. inaequalis, but ascospore production increased with increasing period of time when M. ochracea was applied 8 to 16 weeks after the inoculation with V. inaequalis. In the orchard, the greatest reduction in production of ascospores (94 to 96% in 2000 and 99% in 2001) occurred on leaves sprayed with M. ochracea in August. The production of ascospores was reduced by 61 to 84% in 2000 and 93% in 2001 on leaves sprayed with M. ochracea in September, reduced by 64 to 86% in 2000 and 74 to 89% in 2001 on leaves sprayed in October, and reduced by 54 and 67% in 2000 and 2001, respectively, on leaves sprayed in November. It was concluded that M. ochracea should be applied in August or September and that ascospore maturation models and delayed-fungicide program could be used in orchards treated with this biological control agent.