Prepenetration stages in infection of clematis by Phoma clematidina

A detailed study of conidial germination, germ-tube growth and the formation of infection structures in Phoma clematidina , the causal agent of clematis wilt, is described for two clematis varieties differing in disease resistance. On both the resistant and susceptible varieties, the fungus entered leaves and stems by direct penetration of the cuticle, often, but not always, following the formation of infection structures. More germ tubes per conidium were formed on the susceptible host, but these germ tubes were on average shorter than on the resistant host. Although germ tubes regularly entered the plant via trichomes, stomata were not found to be sites of entry. Following penetration of the cuticle of resistant plants, germ-tube growth was sometimes restricted to the subcuticular region, and halo formation occurred at the sites where penetration was attempted. Subcuticular growth and halo formation were not observed on susceptible plants. These observations may partly explain the resistance of small-flowered clematis varieties to P. clematidina .     

Identification and characterization of Pepino mosaic potexvirus in tomato

At the beginning of 1999, a new virus disease occurred in protected tomato crops in The Netherlands. Initial diagnostic tests revealed the presence of a potexvirus but serological tests ruled out the presence of Potato X potexvirus (PVX). Tests for other potexviruses reported from solanaceous crops provisionally identified the virus as Pepino mosaic potexvirus (PepMV). The virus was purified, and an antiserum was produced, which showed strong reactions with both the type isolate of PepMV from pepino and two other isolates from tomato. Host range and symptomatology of the pepino and tomato isolates of PepMV revealed clear differences from PVX. However, differences were also observed between the pepino and tomato isolates of PepMV. Sequence alignment of DNA fragments of 584 bp derived from the RNA polymerase cistron showed almost 95% identity with the pepino isolate, whereas the identity with PVX appeared to be < 60%. Together, these results identified PepMV as the causal agent of the new virus disease in tomato. Based on the differences from the type isolate from pepino ( Solanum muricatum ), the isolates from tomato should be considered as a distinct strain of PepMV for which the name tomato strain is proposed.     

Functional Analysis of ABC Transporter Genes From Botrytis cinerea Identifies BcatrB as a Transporter of Eugenol

The role of multiple ATP-binding cassette (ABC) and major facilitator superfamily (MFS) transporter genes from the plant pathogenic fungus Botrytis cinerea in protection against natural fungitoxic compounds was studied by expression analysis and phenotyping of gene-replacement mutants. The expression of 11 ABC (BcatrA–BcatrK) and three MFS genes (Bcmfs1, Bcmfs2 and Bcmfs4) was studied. All genes showed a low basal level of expression, but were differentially induced by treatment with cycloheximide and the plant defence compounds camptothecin, eugenol, psoralen, resveratrol and rishitin. The latter compounds induced expression of BcatrB at a high level. Eugenol was more toxic to BcatrB gene-replacement mutants than to the control isolates. Eugenol also caused an instantaneous increase in mycelial accumulation of the fungicide fludioxonil, a known substrate of BcatrB. However, there was no difference in virulence between the wild-type and BcatrB gene-replacement mutants on Ocimum basilicum, a plant known to contain eugenol. The results indicate that BcatrB is a transporter of lipophilic compounds, such as eugenol, but its role in virulence remains uncertain.     

Further Contributions to the Development of a Differential Set of Pea Cultivars (Pisum sativum) to Investigate the Virulence of Isolates of Aphanomyces euteiches

Common root rot (Aphanomyces euteiches Drechs.) has become a very destructive disease in the French pea crops since 1993. For an accurate investigation of the virulence variability among French A. euteiches populations and between French and foreign populations, a new set of differential pea genotypes was developed. Thirty-three American and European pea lines, displaying different levels of resistance, were screened in a growth chamber against two French isolates. Symptoms (disease severity from 0 to 5, evaluating symptom surface on roots and epicotyl) and percentage of top fresh weight (inoculated/uninoculated top fresh weight ratio) were measured. From this screening 12 relatively resistant lines, from various genetic backgrounds, were identified along with a highly susceptible control. This set of 13 genotypes was inoculated under controlled conditions with 14 isolates from France, Sweden, USA, Canada and New Zealand, to investigate genotype–isolate interactions. Root symptoms were rated (disease severity), and a susceptibility/resistance threshold was established at disease severity = 1. Significant quantitative interactions were observed, and five 'resistance patterns' were identified, leading to a set of six pea genotypes: Baccara (susceptible), Capella, MN313, 902131, 552 and PI180693. Fields trials of this set in 1999 and 2000 gave the same resistance rankings than in growth chamber conditions. This set will allow more accurate assessments of the variability in virulence/aggressiveness of A. euteiches isolates from France and foreign countries, and further investigations of the epidemiological and genetic basis of pea–A. euteiches interactions.     

Modelling Plant Disease Epidemics

An epidemic is the progress of disease in time and space. Each epidemic has a structure whose temporal dynamics and spatial patterns are jointly determined by the pathosystem characteristics and environmental conditions. One of the important objectives in epidemiology is to understand such spatio-temporal dynamics via mathematical and statistical modelling. In this paper, we outline common methodologies that are used to quantify and model spatio-temporal dynamics of plant diseases, with emphasis on developing temporal forecast models and on quantifying spatial patterns. Several examples of epidemiological models in cereal crops are described, including one for Fusarium head blight.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).     

Comparison of an in vitro and a damping-off assay to test soils for suppressiveness to Pythium aphanidermatum

Testing of soil samples in greenhouse assays for suppressiveness to soilborne plant pathogens requires a considerable investment in time and effort as well as large numbers of soil samples. To make it possible to process large numbers of samples efficiently, we compared an in vitro growth assay with a damping-off assay using Pythium aphanidermatum as the test organism on tomato seedlings. The in vitro test compares the radial growth or relative growth of the fungus in soil to that in autoclaved soil and reflects suppressiveness of soils to the pathogen. We used soils from a field experiment that had been farmed either organically or conventionally and into which a cover crop (oats and vetch in mixture) had been incorporated 0, 10, 21, and 35 days previously. We obtained a significant, positive correlation between damping-off severities of tomato seedlings in damping-off assays and both relative and radial growth in vitro. In addition, radial and relative growth of P. aphanidermatum in the in vitro assay were positively correlated with several carbon and nitrogen variables measured for soil and incorporated debris. We did not find differences between the two farming systems for either growth measures of P. aphanidermatum or disease severities on tomato at different stages of cover crop decomposition. The in vitro assay shows potential for use with any fungus that exhibits rapid saprophytic growth, and is most suitable for routine application in suppressiveness testing.     

Quantifying patch distribution at multiple spatial scales: Applications to wildlife-habitat models

Multiscale analyses are widely employed for wildlife-habitat studies. In most cases, however, each scale is considered discrete and little emphasis is placed on incorporating or measuring the responses of wildlife to resources across multiple scales. We modeled the responses of three Arctic wildlife species to vegetative resources distributed at two spatial scales: patches and collections of patches aggregated across a regional area. We defined a patch as a single or homogeneous collection of pixels representing 1 of 10 unique vegetation types. We employed a spatial pattern technique, three-term local quadrat variance, to quantify the distribution of patches at a larger regional scale. We used the distance at which the variance for each of 10 vegetation types peaked to define a moving window for calculating the density of patches. When measures of vegetation patch and density were applied to resource selection functions, the most parsimonious models for wolves and grizzly bears included covariates recorded at both scales. Seasonal resource selection by caribou was best described using a model consisting of only regional scale covariates. Our results suggest that for some species and environments simple patch-scale models may not capture the full range of spatial variation in resources to which wildlife may respond. For mobile animals that range across heterogeneous areas we recommend selection models that integrate resources occurring at a number of spatial scales. Patch density is a simple technique for representing such higher-order spatial patterns.     

Using traffic flow theory to model traffic mortality in mammals

Traffic has a considerable effect on population and community dynamics through the disruption and fragmentation of habitat and traffic mortality. This paper deals with a systematic way to acquire knowledge about the probabilities of successful road crossing by mammals and what characteristics affect this traversability. We derive a model from traffic flow theory to estimate traffic mortality in mammals related to relevant road, traffic and species characteristics. The probability of successful road crossing is determined by the pavement width of the road, traffic volume, traversing speed of the mammals and their body length. We include the traversability model in a simple two-patch population model to explore the effects of these road, traffic and species characteristics on population dynamics. Analysis of the models show that, for our parameter ranges, traffic volume and traversing speed have the largest effect on traffic mortality. The population size is especially negatively affected when roads have to be crossed during the daily movements. These predictions could be useful to determine the expected effectiveness of mitigating measures relative to the current situation. Mitigating measures might alter the road and traffic characteristics. The effects of these changes on traffic mortality and population dynamics could be analysed by calculating the number of traffic victims before and after the mitigating measures. This revised version was published online in July 2006 with corrections to the Cover Date.     

Diagnosis of Congenital Cardiac Right-to-left Shunts with 99mTc-macroaggregated Albumin

Diagnosing right-to-left congenital cardiac shunts can be difficult. Cardiac catheterization and angiocardiography represent the traditional gold standard for diagnosis, but they are invasive. Nuclear scintigraphy using 99mTc-macroaggregated albumin (MAA) has been employed in humans as an alternate method of diagnosis. This study reviews eight dogs presented for evaluation of a suspect right-to-left cardiac shunt that were examined using 99mTc-MAA. In all, 2-4 mCi (74-148 MBq) of reduced particle 99mTc-MAA were injected IV in a cephalic vein and static images of the whole body, including right and left lateral, dorsal, and ventral views, were acquired for 60 s and stored into a 256 x 256 x 16 matrix. Shunt fractions were calculated. One dog with radiopharmaceutical distribution limited to the lungs did not have a shunt. Seven dogs had distribution of the radiopharmaceutical outside the pulmonary capillary bed, indicating bypassing of the pulmonary capillary circulation due to a right-to-left shunt. Four dogs had 99mTc-MAA within the brain. Three dogs that did not have brain uptake, but instead had a sharp cutoff of radioactivity at the level of the front limbs and neck, were diagnosed with reverse patent ductus arteriosus (PDA). The asymmetric distribution of the radiopharmaceutical is due to the location of the shunt, distal to the brachiocephalic trunk and left subclavian artery. Shunt fractions of dogs with extrapulmonary radioactivity ranged from 40% to 59%. Nuclear scintigraphy with 99mTc-MAA is a quick alternative method of diagnosing right-to-left cardiac shunts that permits quantification of shunt fraction. Distinguishing between reverse PDA and other right-to-left shunts may be possible based on the radiopharmaceutical distribution.