Pathogenicity and Virulence of the Two Dutch VCGs of Verticillium dahliae to Woody Ornamentals

Two experiments were performed in two consecutive years to test whether isolates of different vegetative compatibility groups (VCGs) differ in their ability to cause disease in woody ornamentals, to study the host specificity of the isolates and to get an insight into disease development in woody hosts. A range of woody ornamental plant species, including Acer campestre, Acer platanoides, Acer pseudoplatanus, Catalpa bignonioides, Cotinus coggygria, Robinia pseudoacacia, Rosa canina, Syringa vulgaris and Tilia cordata, were root-dip inoculated with six isolates of Verticillium dahliae, belonging to the two VCGs that occur in the Netherlands (VCG NL-1 and VCG NL-2). Isolates belonging to each VCG caused severe symptoms of verticillium wilt in most plant species tested. Disease progress differed between plant species, but was generally the same for the two VCGs. No overall differences in virulence were observed between the two VCGs for external wilt symptoms, number of dead plants, or shoot length. No significant VCG × plant species interactions were present for these characteristics. However, isolates of VCG NL-1 caused more vascular discolouration than did isolates of VCG NL-2. Isolates within VCGs often differed considerably in their virulence to certain hosts, as shown by highly significant isolate × plant species interactions. Isolates were more virulent on their original host. These findings imply that VCG identification does not contribute to disease prediction for a range of woody hosts.     

Shading effects on spatial growth and biomass partitioning of Cynodon dactylon

Cynodon dactylon is a low-growing C4 weed that is highly sensitive to shading. This species shows severe biomass reductions and highly plastic morphological changes in response to resource availability. The objective of this study was to analyse the effect of shading on the spatial and biomass growth of C. dactylon . Two experiments were carried out exploring different environmental conditions. Six treatments were applied on C. dactylon patches. Daylight and 18%, 41%, 48%, 69% and 85% shading of daylight photosynthetic active radiation (PAR) were imposed in Expt 1 and 0%, 22%, 44%, 50%, 74% and 87% shading of daylight PAR in Expt 2. Patches were slightly elliptical in shape and grew 3.2 and 1.5 cm d⁻¹ with 0% shading, on average, during the whole studied periods in Expts 1 and 2 respectively. Biomass was significantly reduced from 41% shading onwards in Expt 1 and from 50% shading in Expt 2. However, patch extension rate was only significantly reduced from 85% and 74% shading in Expts 1 and 2 respectively. Biomass partitioning was modified by shading in both experiments. A functional analysis of C. dactylon patches showed that the patch extension rate diminished linearly when patch biomass growth rate was lower than 1.66 g d⁻¹; above this value, the extension rate remained constant.     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

Sporulation of Pyrenopeziza brassicae (light leaf spot) on oilseed rape (Brassica napus) leaves inoculated with ascospores or conidia at different temperatures and wetness durations

In controlled environment experiments, sporulation of Pyrenopeziza brassicae was observed on leaves of oilseed rape inoculated with ascospores or conidia at temperatures from 8 to 20°C at all leaf wetness durations from 6 to 72 h, except after 6 h leaf wetness duration at 8°C. The shortest times from inoculation to first observed sporulation ( l ₀), for both ascospore and conidial inoculum, were 11–12 days at 16°C after 48 h wetness duration. For both ascospore and conidial inoculum (48 h wetness duration), the number of conidia produced per cm² leaf area with sporulation was seven to eight times less at 20°C than at 8, 12 or 16°C. Values of Gompertz parameters c (maximum percentage leaf area with sporulation), r (maximum rate of increase in percentage leaf area with sporulation) and l ₃₇ (days from inoculation to 37% of maximum sporulation), estimated by fitting the equation to the observed data, were linearly related to values predicted by inserting temperature and wetness duration treatment values into existing equations. The observed data were fitted better by logistic equations than by Gompertz equations (which overestimated at low temperatures). For both ascospore and conidial inoculum, the latent period derived from the logistic equation (days from inoculation to 50% of maximum sporulation, l ₅₀) of P. brassicae was generally shortest at 16°C, and increased as temperature increased to 20°C or decreased to 8°C. Minimum numbers of spores needed to produce sporulation on leaves were ≈25 ascospores per leaf and ≈700 conidia per leaf, at 16°C after 48 h leaf wetness duration.     

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.     

Genetic analyses of resistance of the wheat differential cultivars Carstens V and Spaldings Prolific to two races of Puccinia striiformis

Genetic analysis of resistance of wheat seedlings to two races of Puccinia striiformis was conducted on F₁, F₂ and F₃ generations from crosses Carstens V (CV) × Lee, Spaldings Prolific (SPA) × Lee and CV × SPA. F₂ generations from crosses of CV and SPA with Strubes Dickkopf (SD) were also studied. The plants were classified into six resistance classes and analysed by factorial correspondence analysis and nonhierarchical classification. The two P. striiformis isolates tested were a French isolate of race 43E138 and a Lebanese isolate of race 2E16, selected for the differences in their virulence spectra for the common differential cultivars Strubes Dickkopf and Nord Desprez. Resistance of CV and SPA was recessive and dominant to races 43E138 and 2E16, respectively. CV possessed three or four resistance genes, one of them being expressed with both races. Two genes of CV had a cumulative effect for resistance to 43E138 and two or three gave dominant resistance to 2E16. SPA had three resistance genes, all of which gave resistance to 2E16 and two of which also gave resistance to 43E138. SPA had one gene in common with CV for resistance to both races. Furthermore, the gene for resistance to race 2E16 in CV and SPA was allelic with a gene in SD, and was probably Yr25 .     

Integrated Pest Management in Cocoa Agroforests in Southern Cameroon: Constraints and Overview

Imports of pesticides have long been subsidised for use in the cocoa agroforests of the humid forest zone of southern Cameroon. With the liberalisation of the cocoa and pesticides sectors and the devaluation of the local currency (CFA franc), farmers are facing fluctuations in the price paid for cocoa and the high cost of farm inputs. Without the support of the extension services, they themselves have developed traditional integrated control methods based on the use of plant extracts mixed with conventional pesticides. From a survey of 300 cocoa farmers, the study assesses the farmers' command of these methods, the pests controlled, the problems encountered and the institutional constraints in the definition and dissemination of integrated control methods. The study concludes with recommendations for research and development towards the better definition and dissemination of integrated control methods. This revised version was published online in July 2006 with corrections to the Cover Date.