Effects of Weather Variables on Ascospore Discharge from Fusarium graminearum Perithecia

Fusarium graminearum is a predominant component of the Fusarium head blight (FHB) complex of small grain cereals. Ascosporic infection plays a relevant role in the spread of the disease. A 3-year study was conducted on ascospore discharge. To separate the effect of weather on discharge from the effect of weather on the production and maturation of ascospores in perithecia, discharge was quantified with a volumetric spore sampler placed near maize stalk residues bearing perithecia with mature ascospores; the residues therefore served as a continuous source of ascospores. Ascospores were discharged from perithecia on 70% of 154 days. Rain (R) and vapor pressure deficit (VPD) were the variables that most affected ascospore discharge, with 84% of total discharges occurring on days with R≥0.2 mm or VPD≤11 hPa, and with 70% of total ascospore discharge peaks (≥ 30 ascospores/m³ air per day) occurring on days with R≥0.2 mm and VPD≤6.35 hPa. An ROC analysis using these criteria for R and VPD provided True Positive Proportion (TPP) = 0.84 and True Negative Proportion (TNP) = 0.63 for occurrence of ascospore discharge, and TPP = 0.70 and TNP = 0.89 for occurrence of peaks. Globally, 68 ascospores (2.5% of the total ascospores sampled) were trapped on the 17 days when no ascospores were erroneously predicted. When a discharge occurred, the numbers of F. graminearum ascospores sampled were predicted by a multiple regression model with R² = 0.68. This model, which includes average and maximum temperature and VPD as predicting variables, slightly underestimated the real data and especially ascospore peaks. Numbers of ascospores in peaks were best predicted by wetness duration of the previous day, minimum temperature, and VPD, with R² = 0.71. These results will help refine the epidemiological models used as decision aids in FHB management programs.     

Common Leafspot of Alfalfa: Ascospore Discharge and Plant Infection in the Field

Peak daily discharge of Pseudopeziza medicaginis ascospores in the field occurred after sunrise with attendant rises in temperature, wind, speed, and. dissipation of moisture. Protracted discharge over many days rose with initial development of robust, uncrowded apothecia and fell as these became spent and were joined by less productive, small, crowded ones. In the laboratory, detached field-infected leaves discharged ascospores at 5°C to 32.5°C and at relative humidities as low as 90% when fresh, and at 94% or higher when wilted or dried. Pathogen-free alfalfa plants placed overnight in a diseased affalfa plot were inoculated but uninfeeted more frequently in July than they were inoculated and infected. Artificially-inoculated plants placed at the same time in a nearby sugar beet plot became infected every overnight. From early September through early October artificially inoculated plants placed in a plot of sugar beets or a plot of nearly disease-free alfalfa for 24 h periods, became infected in 18 and 19 of 29 such periods. Light rain, fog and dew were frequent during these periods and early morning temperatures fell between - 3 °C and 5 °C- in one-half of the overnights. Dormant ascospores on foliage remained fully infective in the greenhouse for at least 10 days.     

Passage and Survival of Chlamydospores of Phytophthora cinnamomi Rands, the Causal Agent of Forest Dieback Disease, Through the Gastrointestinal Tracts of Termites and Wild Birds

Chlamydospores of Phytophthora cinnamomi Rands have been shown to survive in the intestinal tracts of termites (Nasutitermes exitiosus) and two species of forest birds indigenous to West Australian jarrah forests. Viable chlamydospores were recovered from bird feces within the normal rate of passage time for food through the gut. The above factors would allow these creatures to function as vectors for the spores.     

Ascospore Discharge, Leaf Infestation and Variations in Pathogenicity as Criteria to Predict Impact of Leptosphaeria maculans on Oilseed Rape

The aim of this investigation was to evaluate criteria for the forecast and targeted control of basal stem canker (blackleg) caused by Leptosphaeria maculans on oilseed rape. Ascospore discharge, ratio of aggressive and non-aggressive isolates and leaf and stem infestations were determined during 1991/92–1993/94 at 6–10 sites in Northern Germany. On a 1–9 scale, blackleg intensity varied from 2.3 to 6.3 at BBCH 81 between different sites and years. Ascospore discharge started in September or October, and reached maxima 1 or 2 months later, without an apparent relationship to blackleg or leaf infestation. There was a positive relationship between leaf infestation and blackleg. However, correlation coefficients were too low to be used as a basis for forecasting. On plant residues from the stem base, aggressive isolates were dominant (>80%) on all sites. From higher parts of the stem and from leaves also, non-aggressive isolates were isolated with higher frequencies on some locations, but the proportion of aggressive isolates was not related to the blackleg intensity. Taken all together, the three criteria alone seem to be insufficient for the development of a system of blackleg forecasting and targeted control. Further factors (e.g. climatic factors, seed-and soilborne inoculum, cultural practices) have to be included in models for forecasting the impact of blackleg on oilseed rape.     

The Seed-borne Inoculum of Peronospora manshurica, Causal Agent of Soybean Downy Mildew

The seed-borne inoculum of P. manshurica is described using various microscopic techniques and staining reactions. The inoculum, in addition to the oospores, consists of a thin walled mycelium under the hourglass cell layer of the seed coat and a thick walled resting mycelium in the oospore crust on the seed surface. By a modified Feulgen staining technique it was possible to demonstrate that the thin walled and the thick walled resting mycelium as well as thin and thick walled oogonia contain nuclei which are Feulgen positive suggesting that portions of the mycelium of P. manshurica may survive even on the surface of dry, mature seeds.     

Studies on Pantoea citrea, the Causal Agent of Pink Disease of Pineapple

The causal agent of pink disease of pineapple has been identified as Pantoea citrea, a member of the Enterobacteriaceae. Comparative physiological and biochemical analyses demonstrated that P. citrea isolated from diseased pineapple fruit in the Philippines possesses features identical to those of an American Type Culture Collection type strain of P. citrea and not to those of P. ananas, P. herbicola (formerly Erwinia herbicola), and P. stewartii (formerly Erwinia stewartii). P. citrea induces the production of compounds in pineapple which become pink to reddish-brown upon cooking the fruit, pulp, or juice. This distinct colour is not induced by Escherichia coli, Agrobacterium tumefaciens, Burkholderia gladioli, Pseudomonas fluorescens, Gluconobacter oxydans, Acetobacter aceti, and Acinetobacter calcoaceticus. Like other well characterized bacteria pathogens, such as Pseudomonas syringae pv. phaseolicola, P. citrea elicits the hypersensitive response (HR) in tobacco. By contrast, G. oxydans and A. aceti that have been previously implicated as the causal agents of pink disease, do not elicit HR. Although the nature of the pink colour in pineapple produced by P. citrea has not been elucidated, the locus conferring this activity has been located on its chromosome. The pink colour can be restored in an avirulent, pink colour defective mutant strain, CMC6, by complementation in trans with a specific 3.8 kb genomic DNA fragment of P. citrea. This suggests that P. citrea contains the genetic elements that are required for pink disease.     

Pathogenic Variability in Mycosphaerella brassicicola, the Causal Agent of the Ringspot Disease of Crucifers

Twenty monoascosporic isolates of Mycosphaerella brassicicola from diverse origins were inoculated to 13 different accessions of cauliflower. No hypersensitive response was noticed and the isolates exhibited variable pathogenicity. Three isolates from cauliflower were isolated from the same lesion and had variable pathogenicity. This result suggests that different infection units e.g. ascospores contribute to the development of a single lesion. Furthermore, this finding also indicates that one should work with monoascosporic isolates. Pathogenicity results of this study showed that European isolates from cabbages had reduced pathogenicity compared to French cauliflower ones. Variable levels of pathogenicity were found in French cauliflower isolates. This group had also the most aggressive isolates. Our results suggest that Brittany may be a diversification area for the pathogen. In fact, the climate is mild and humid and crucifers (e.g. cauliflower and broccoli) are continuously grown over the year in this region. Pathotypes found in other growing areas may have moved from Brittany. All these findings supported by other studies suggest that resistance sources available in other countries may not be effective in Brittany. Consequently, screening for resistance sources should be done with cauliflower isolates from Brittany. No accession tested was susceptible or resistant to all isolates tested.     

Biological Control of Protomycopsis phaseoli, the Causal Agent of Leaf Smut of Cowpea

Among the bacteria and fungi associated from the soil where cowpea was grown and tested for antagonism against Protomycopsis phaseoli , Bacillus sp. inhibited the radial growth, Fusarium oxysporum , yeast, Aspergillus fumigatus , Trichoderma harzianum , Trichoderma koningii and Trichoderma sp. reduced radial growth of P. phaseoli . In vitro studies showed that T. harzianum was an active hyperparasite and more effective in reducing the radial growth of P. phaseoli than T. koningii and Trichoderma sp. Spore suspensions of the three Trichoderma spp. prevented the germination of chlamydospores of P. phaseoli . In the field, when applied as spray, Trichoderma sp. was found to be more active in reducing the spread of leaf smut disease than T. harzianum and T. koningii.     

Species-Specific Detection and Identification of Fusarium Species Complex,the Causal Agent of Sugarcane Pokkah Boeng in China

Background

Pokkah boeng disease caused by the Fusarium species complex results in significant yield losses in sugarcane. Thus, the rapid and accurate detection and identification of the pathogen is urgently required to manage and prevent the spreading of sugarcane pokkah boeng.

Methods

A total of 101 isolates were recovered from the pokkah boeng samples collected from five major sugarcane production areas in China throughout 2012 and 2013. The causal pathogen was identified by morphological observation, pathogenicity test, and phylogenetic analysis based on the fungus-conserved rDNA-ITS. Species-specific TaqMan real-time PCR and conventional PCR methods were developed for rapid and accurate detection of the causal agent of sugarcane pokkah boeng. The specificity and sensitivity of PCR assay were also evaluated on a total of 84 isolates of Fusarium from China and several isolates from other fungal pathogens of Sporisorium scitamineum and Phoma sp. and sugarcane endophyte of Acremonium sp.

Result

Two Fusarium species (F. verticillioides and F. proliferatum) that caused sugarcane pokahh boeng were identified by morphological observation, pathogenicity test, and phylogenetic analysis. Species-specific TaqMan PCR and conventional PCR were designed and optimized to target their rDNA-ITS regions. The sensitivity of the TaqMan PCR was approximately 10 pg of fungal DNA input, which was 1,000-fold over conventional PCR, and successfully detected pokkah boeng in the field-grown sugarcane.

Conclusions/Significance

This study was the first to identify two species, F. verticillioides and F. proliferatum, that were causal pathogens of sugarcane pokkah boeng in China. It also described the development of a species-specific PCR assay to detect and confirm these pathogens in sugarcane plants from mainland China. This method will be very useful for a broad range of research endeavors as well as the regulatory response and management of sugarcane pokkah boeng.     

Pathogenicity Process of Pseudomonas fuscovaginae, the Causal Agent of Sheath Brown Rot of Rice

Sheath brown rot of rice caused by Pseudomonas fuscovaginae has been described in areas where low temperatures occur during the rice booting and heading stages. To analyse the relationship between pathogenicity of P. fuscovaginae and low temperatures, pathogenicity process in rice at booting stage was studied in a growth chamber at midrange and low temperatures. Analysis performed at 13°C, 18°C and 23°C in two rice cultivars showed that pathogenicity of P. fuscovaginae was explained by the general model of the independent action. The inoculum dose necessary to obtain 50% of diseased sheaths decreased with increase of temperature Analysis of planta bacterial population dynamics and mean response time pointed out that low temperatures affected pathogen multiplication in host before and after symptoms development. In consideration of our results, it was concluded that low temperatures acted negatively on the pathogenicity process of P. fuscovaginae. Therefore, occurrence of P. fuscovaginae in areas where rice cultivation is restricted by low temperatures can not be explained by a direct effect of temperatures on pathogenicity.     

Nutritional Capability of and Substrate Suitability for Pseudogymnoascus destructans,the Causal Agent of Bat White-Nose Syndrome

Pseudogymnoascus destructans, the causal agent of bat white-nose syndrome, has caused nearly six million deaths in North American bats since its introduction into the United States in 2006. Current research has shown that caves can harbor P. destructans even after the infected bats are removed and bats no longer visit or inhabit previously infected caves. Our research focuses on elucidating reservoir requirements by investigating the nutritional capabilities of and substrate suitability requirements for six different P. destructans isolates from various localities including Illinois, Indiana, New York (Type specimen), and Pennsylvania. Enzyme assays implicate that both urease and b-glucosidase appear to be constitutive, lipase and esterase activity were more rapid than proteinase activity on 6% gelatin, gelatin degradation was accompanied by medium alkalinization, the reduction of thiosulfate generated hydrogen sulfide gas, chitinase and manganese dependent peroxidase activity were not visually demonstrated within eight weeks, and keratinase activity was not evident at pH 8 within eight weeks. We demonstrate that all P. destructans isolates are capable of growth and sporulation on dead fish, insect, and mushroom tissues. Sole nitrogen source assays demonstrated that all P. destructans isolates exhibit Class 2 nitrogen utilization and that growth-dependent interactions occur among different pH and nitrogen sources. Substrate suitability assays demonstrated that all isolates could grow and sporulate on media ranging from pH 5–11 and tolerated media supplemented with 2000 mg/L of calcium and 700 mg/L of three separated sulfur compounds: thiosulfate L-cysteine, and sulfite. All isolates were intolerant to PEG-induced matric potential with delayed germination and growth at −2.5 MPa with no visible germination at −5 MPa. Interestingly, decreasing the surface tension with Tween 80 permitted germination and growth of P. destructans in −5 MPa PEG medium within 14 days suggesting a link between substrate suitability and aqueous surface tension altering substances.     

The Effect of Chitinase on Didymella applanata, the Causal Agent of Raspberry Cane Spur light

In vitro and in vivo studies were conducted to assess the efficacy of the two microbial chitinases Chi I (from Streptomyces sp.) and Chi II (from Serratia marcescens) on Didymella applanata (Niessl.) Sacc., the fungus which causes spur blight of raspberry (Rubus idaeus L.). D. applanata was isolated from canes of diseased raspberries in a plantation in Novosibirsk, Russia. In vitro, the effective concentration of Chi I that reduced the growth of D. applanata was 0.4 U/ml (p = 0.05), but Chi II had no influence on the growth of the fungus in medium. In inoculation experiments on raspberry canes, both chitinases at the rate 0.5 U/ml reduced fungal development. In plantation where canes were inoculated after spraying with chitinase, fruiting bodies of fungus failed to form in all enzyme treatments, whereas a significant number of these fungal fruiting bodies (12.8 per cm²) developed in control treatments lacking chitinases spraying. The chitinases reduced the size of lesions and limited the infection of internal tissues of canes. Field testing of Chi I under natural conditions showed a significant suppression of the independent spur blight. These studies form the basis for further evaluation of ecologically benign control measures for raspberry spur blight.     

Pre-Transplant Depression Is Associated with Length of Hospitalization,Discharge Disposition,and Survival after Liver Transplantation

Depression after liver transplantation has been associated with decreased survival, but the effects of pre-transplant depression on early and late post-transplant outcomes remain incompletely evaluated. We assessed all patients who had undergone single-organ liver transplantation at a single center over the prior 10 years. A diagnosis of pre-transplant depression, covariates, and the outcomes of interest were extracted from the electronic medical record. Potential covariates included demographics, etiology and severity of liver disease, comorbidities, donor age, graft type, immunosuppression, and ischemic times. In multivariable models adjusting for these factors, we evaluated the effect of pre-transplant depression on transplant length of stay (LOS), discharge disposition (home vs. facility) and long-term survival. Among 1115 transplant recipients with a median follow-up time of 5 years, the average age was 56±11 and MELD was 12±9. Nineteen percent of the study population had a history of pre-transplant depression. Pre-transplant depression was associated with longer LOS (median = 19 vs. 14 days, IRR = 1.25, CI = 1.13,1.39), discharge to a facility (36% vs. 25%, OR 1.70,CI = 1.18,2.45), and decreased survival (HR = 1.54,CI = 1.14,2.08) in this cohort, accounting for other potential confounders. In conclusion, pre-transplant depression was significantly associated with longer transplant length of stay, discharge to a facility, and mortality in this cohort.     

Identification of Acetobacter liquefaciens as Causal Agent of Pink-Disease of Pineapple Fruit

Two bacterial strains causing pink-disease of pineapple were identified as Acetobacter liquefaciens and compared with 8 other Acetobacter liquefaciens, 10 Gluconobacter oxydans and 7 Frateuria aurantia strains. The similarieties and differences between these bacteria are discussed.     

Acetaldehyde Is a Causal Agent Responsible for Ethanol-Induced Ripening Inhibition in Tomato Fruit

Inhibition of tomato (Lycopersicon esculentum Mill.) fruit ripening by exogenously applied ethanol was shown to be caused by elevated endogenous levels of acetaldehyde (AA). Exposure of excised pericarp discs of mature-green tomato fruit to ethanol or AA vapors produced elevated levels of both compounds in the tissue, but only the levels of AA were associated with ripening inhibition. Ripening inhibition was dependent on both the applied concentration and the duration of exposure. Discs treated with inhibitory levels of AA had levels of ethanol that were elevated but below that associated with inhibition of ripening. The in vivo activity of alcohol dehydrogenase was inhibited 40 to 60% by 4-methylpyrazole (4-MP), a competitive inhibitor of this enzyme. The inhibitory effect of ethanol on ripening was reduced by the simultaneous application of 4-MP. Tissue treated with 4-MP plus AA vapors had higher endogenous levels of AA and ripening was inhibited longer than in tissue without 4-MP. The tissue AA level resulting from ethanol or AA application appears to be the critical determinant of ripening inhibition.     

Detection and Identification of Brenneria nigrifluens, the Causal Agent of the Shallow Bark Canker of Walnut by, PCR Amplification

A 1 kb DNA band from strains of Brenneria nigrifluens, as shown by amplification of their genomic DNA by polymerase chain reaction (PCR) using minisatellite primer designed on the minisatellite sequence of the M13 phage, was isolated, cloned and sequenced. Specific oligonucleotides (F1–C3) were selected into this 1 kb DNA sequence and used in a PCR assay to detect and identify strains of B. nigrifluens . Several strains of B. nigrifluens were assessed with F1–C3 primers producing a specific band of approximately 250 bp pairs in length. This target was successfully amplified from purified genomic DNA, from bacterial culture and directly from infected walnut bark tissue. No amplification was obtained when the PCR assay was performed on other plant-pathogenic species from the following genera Brenneria, Erwinia, Agrobacterium, Pseudomonas, Ralstonia, Pectobacterium, Xanthomonas and from walnut-associated bacteria, indicating the specificity of these primers. The PCR assay with the primers described here provides a rapid, specific and sensitive diagnostic method for B. nigrifluens and a useful tool for epidemiological studies.     

Plasma Membrane Receptors for Exopolysaccharides of the Ring Rot Causal Agent in Potato Cells

By means of differential centrifugation, microsomal fractions enriched in the plasma membrane were isolated from suspension cell cultures of two cultivars of potato (Solanum tuberosum L.) contrasting in their resistance to the causal agent of ring rot (Clavibacter michiganensis subsp. sepedonicus) (Cms). Electrophoresis of the fractions showed that they comprised a wide range of proteins from 15 to 75 kD. The protein bands were more brightly expressed in the microsomal membranes of the cells of susceptible cultivar. The proteins of 70 and 42 kD were present only in the cellular membranes of the resistant cultivar. In order to visualize the binding of exopolysaccharides (EPS) produced by Cms to the receptors of membrane fractions, a conjugate of EPS with a fluorescent marker was used. The membrane fraction isolated from the cells of the susceptible cultivar was found to be richer in receptors for EPS Cms than the membrane fraction from the resistant cultivar. It is supposed that numerous receptors for EPS present on the plasma membrane may partially account for potato susceptibility to Cms. These receptors may facilitate the binding of bacteria to the plant cells, the formation of colonies, and the development of the disease.