Modelling spatio-temporal dynamics of herbicide resistance

A mathematical model has been developed for the risk assessment of the spread of genes conferring herbicide resistance in plant populations. The model combines an age-and-stage-structured population dynamic model, a population genetic model and a model of spatial spread. This is achieved by embedding a local matrix population model into a cellular automaton model with raster cells as spatial units. The dynamics of each cell is determined by both its local dynamics and the interaction with neighbouring cells. The model is applied to the evaluation of management strategies to delay or even to prevent long-term evolution of resistance in an annual grass weed. The results show that the appearance and spread of resistant genes is a highly non-linear process exhibiting threshold phenomena, which occur for a wide range of parameters. The properties of the seed survival curve constitute the `genetic memory' of the system and thus determine its long-term dynamics. It is possible to delay the evolution of resistance by suspension of treatment, reduction in herbicide application rate and introducing fallow periods. Spatial spread from an infested plot is inhibited by leaving untreated strips between adjacent fields.     

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.     

Identification of ALS inhibitor‐resistant Amaranthus biotypes using polymerase chain reaction amplification of specific alleles

Summary Polymerase chain reaction (PCR) amplification of specific alleles (PASA) was adapted as a molecular marker‐based method for the rapid detection of point mutations in Amaranthus retroflexus and Amaranthus rudis leading to ALS inhibitor resistance. Two pairs of primers were designed for the specific amplification of alleles of the ALS gene of susceptible and resistant biotypes. The allele‐specific primer matched the desired allele, but mismatched the different allele at its 3′ end. Differentiation was carried out by comparison of the amplified DNA fragments in gel electrophoresis after PASA‐PCR. In A. rudis, differentiation was possible with one PCR and genomic DNA as probe. A ‘nested’ PCR was necessary for the differentiation of sensitive and resistant A. retroflexus. PASA is useful for the identification of resistant weed biotypes and also as a monitoring tool to map resistance occurrence and distribution. Advantages include the fast and clear separation of those plants with and without mutations at an early stage of development, its easy and consistent performance and quick results compared with existing resistance detection tests. These advantages, when combined with management strategies, enable further activities to reduce herbicide resistance.     

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.     

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.     

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.     

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.     

Comparison of two sampling methods for leafhoppers (Homoptera, Cicadellidae) associated with sesame in the east Mediterranean region of Turkey

Yellow sticky-traps and a modified Johnson-Taylor suction-trap were used to index the relative population dynamic of leafhoppers associated with sesame in the east Mediterranean region of Turkey. Comparison of the obtained results signified important differences between both traps. 32 leafhoppers species were caught in the suction-trap while only 18 species were determined on yellow sticky-traps. For most of the leafhopper species, relatively more males were determined on the yellow-traps than in the suction-trap, indicating a higher activity of male leafhoppers during daytime hours. A close relationship between the seasonal flight pattern as indexed by sticky-traps and by suction-trap was only observed forAsymmetrasca decedens (Paoli) andEmpoasca decipiens Paoli. For most other leafhopper species no or only a very poor correlation for the relative population dynamic was determined between both traps. Important leafhopper vector species, e.g.,Circulifer haematoceps (Mulsant et Rey) andOrosius orientalis (Matsumura) were better represented in yellow sticky-trap catches than they were in the suction trap.

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Zusammenfassung  Die relative Populationsdynamik der mit Sesam assozüerten Zwergzikaden wurde mit beleimten Gelbtafeln und einer stationären Johnson-Taylor-Saugfalle an der südöstlichen Mittelmeerküste der Türkei untersucht. Beim Vergleich beider Methoden zeigten sich deutliche Unterschiede zwischen den Fallentypen. In der Saugfalle konnten 32 Zwergzikadenarten erfaßt werden, während dies auf den Gelbtafeln nur 18 Arten waren. Die meisten Zwergzikadenarten zeigten auf den Gelbtafeln ein stark in Richtung der Männchen verschobenes Geschlechterverhältnis, was eine erhöhte Flugaktivität der Männchen während des Tageslichts vermuten läßt. Auch der Verlauf der saisonalen Flugaktivität war für die meisten Zwergzikadenarten zwischen den beiden Fallentypen sehr unterschiedlich, und es konnten zwischen Saug- und Gelbfalle keine deutlichen Korrelationen hergestellt werden. Nur für die beiden zusammen erfaßten ArtenAsymmetrasca decedens (Paoli) undEmpoasca decipiens Paoli wurde ein enger Zusammenhang zwischen der mit der Saugfalle und den gelben Leimtafeln erfaßten relativen Populationsdynamik festgestellt. Wichtige Vektorenarten unter den Zwergzikaden, z. B.Circulifer haematoceps (Mulsant et Rey) undOrosius orientalis (Matsumura) waren auf den Gelbtafeln besser repräsentiert als in der Saugfalle.

     

Auxins increase the occurrence of leaf-colour variants in Caladium regenerated from leaf explants

Leaf explants of Caladium ‘Pink Cloud’ were cultured in vitro on MS medium containing various auxins (NAA, IBA, IAA, 2,4,5-T and 2,4-D) in combination with cytokinin (BA). NAA gave the most vigorous in vitro propagation of this plant, and only 15% of the plants were leaf-colour variants on the medium containing 0.5 μmol NAA. Leaf colour variation was observed in all plants regenerated on the medium containing 2,4-D at 0.5–4.5 μmol. In hormone-free medium, only a few leaf-colour variants (6%) occurred, but the rate of plant regeneration was very low. Application of 0.5 μmol NAA together with 4.5 μmol BA seemed to be the most appropriate for in vitro propagation of Caladium ‘Pink Cloud’ with only a few leaf-colour variants.