ALBERT

Notes: The growth and development of plant pathogens and their hosts generally respond strongly to the temperature of their environment. However, most studies of plant pathology record pathogen/host measurements against physical time (e.g. hours or days) rather than thermal time (e.g. degree-days or degree-hours). This confounds the comparison of epidemiological measurements across experiments and limits the value of the scientific literature.

Notes: Late blight, caused by Phytophthora infestans, is an ongoing threat to potato and tomato crop production worldwide and considerable fundamental and applied research is conducted with the long-term aim of improved disease control. Understanding the mechanisms, processes and rates of P. infestans evolution is an important factor in predicting the effectiveness and durability of new management practices. A range of phenotypic and genotypic tests has been applied to achieve this goal, but each has limitations and new methods are sought. Recent progress in P. infestans genomics is providing the raw data for such methods and new high-throughput codominant biomolecular markers are currently being developed that have tremendous potential in the study of P. infestans population biology, epidemiology, ecology, genetics and evolution. This paper reviews some key applications, recommends some changes in approach and reports on the status and potential of new and existing methods for probing P. infestans genetic diversity.

Notes: Field trials tested which components of epidemic development of Puccinia striiformis, the cause of yellow rust, were affected by nitrogen (N) fertilizer applied to winter wheat. Both timing and amount of N were varied to affect canopy size and leaf N content, and to provide a supply of mobile N to the pathogen, by causing fresh N uptake after leaf expansion was complete. No N was applied to control plots. A logistic disease-progress function was fitted to disease-severity data, which were assessed in absolute units. Leaf area and specific leaf N (g N per m2 leaf tissue) were quantified. Large and highly significant effects of N on the upper asymptotes, or ‘carrying capacities’ (c) were found. Effects on rates and points of inflection of the epidemics were not significant. Early N resulted in larger shoot numbers and leaf area, but disease was also more severe, so that by grain filling, the remaining green leaf areas were larger without N than with N. Later N treatments did not increase canopy size, but did increase symptom area compared with the control. These effects differ from the concept that N affects disease as a result of its effect on canopy growth, and therefore canopy microclimate, and suggest instead a substrate effect. Linear regression revealed that 51% of the observed differences in c were explained by variation in specific leaf N, suggesting that growth of the rust fungus may depend directly on particular components of total leaf N.

Notes: Fusarium graminearum, one of the causal agents of fusarium head blight (FHB) of wheat in the UK, can be broadly divided into two chemotypes based on the production of the 8-ketotrichothecenes deoxynivalenol (DON) and nivalenol (NIV). DON-producing isolates can be further distinguished on the basis of the predominant acetyl DON derivative that they produce; 3-acetyl DON (3-AcDON) or 15-acetyl DON (15-AcDON). Functional Tri13 and Tri7 are required for the production of NIV and 4-acetyl NIV, respectively, whereas, in isolates that produce only DON and its acetylated derivatives, these genes are nonfunctional or absent. Infections caused by F. graminearum are becoming more frequent in the UK; however, it is unknown whether these represent more than one chemotype. In this study, polymerasae chain reaction (PCR) assays specific for functional and nonfunctional/deleted versions of Tri7 and Tri13 were used to characterize 101 single-spore isolates of Fusarium graminearum as DON or NIV chemotypes. Primer sets developed to Tri3 were used to classify DON chemotypes further by the acetyl derivative produced (3-AcDON or 15-AcDON). Isolates were collected from 65 fields located around England and Wales between 1997 and 2002. All three chemotypes were identified from the F. graminearum population of England and Wales, with 15-AcDON chemotypes predominating overall. All isolates characterized as 3- or 15-AcDON chemotypes had nonfunctional versions of both genes. Where multiple isolates were collected from a field, mixed-chemotype populations were identified. Variation in the number of 11-bp repeats in Tri7 was detected among 3- and 15-AcDON chemotypes. Seventy-two of the 76 DON chemotypes (95%) were classified as 15-AcDON producers and the remaining four isolates (5%) as 3-AcDON producers. In all four isolates with a 3-AcDON chemotype, Tri7 was deleted from the trichothecene gene cluster. There was no evidence of regional variation between 3-AcDON, 15-AcDON or NIV chemotypes within the F. graminearum population.

Notes: Forty-eight chickpea germplasm lines, including 22 differentials used in previous studies, were characterized for disease phenotypes following inoculation with six isolates of Didymella (anamorph Ascochyta) rabiei, representing a wide spectrum of pathogenic variation. Representative isolates were also directly compared with six previously identified races on eight chickpea genotypes. Many of the chickpea differentials reacted similarly to inoculation with each isolate of D. rabiei, and several previously identified races caused similar levels of disease on the differentials. This indicates that the number of differentials can be reduced significantly without sacrificing accuracy in describing pathogenic variation of D. rabiei on chickpea. Pathogenic variation among samples of US isolates allowed classification of the isolates into two pathotypes. The distribution of disease phenotypes of the 48 germplasm lines was bimodal after inoculation with pathotype I isolates, whereas the distribution of disease phenotypes was continuous after inoculation with pathotype II isolates. Such distinct distribution patterns suggest that chickpea plants employ different resistance mechanisms to each pathotype and that the two pathotypes may have different genetic mechanisms controlling pathogenicity. The advantages of using the two-pathotype system in assaying pathogenicity of the pathogen and in studying resistance mechanisms of the host are discussed. Three chickpea accessions, PI 559361, PI 559363 and W6 22589, showed a high level of resistance to both pathotypes, and can be employed as resistance sources in chickpea breeding programmes for resistance to ascochyta blight.

Notes: Data from surveys of winter oilseed rape crops in England and Wales in growing seasons with harvests in 1987–99 were used to construct statistical models to predict, in autumn (October), the incidence of light leaf spot caused by Pyrenopeziza brassicae on winter oilseed rape crops the following spring (March/April), at both regional and individual crop scales. Regions (groups of counties) with similar seasonal patterns of incidence (percentage of plants affected) of light leaf spot were defined by using principal coordinates analysis on the survey data. At the regional scale, explanatory variables for the statistical models were regional weather (mean summer temperature and mean monthly winter rainfall) and survey data for regional light leaf spot incidence (percentage of plants with affected pods) in July of the previous season. At the crop scale, further explanatory variables were crop cultivar (light leaf spot resistance rating), sowing date (number of weeks before/after 1 September), autumn fungicide use and light leaf spot incidence in autumn. Risk of severe light leaf spot (〉 25% plants affected) in a crop in spring was also predicted, and uncertainty in predictions was assessed. The models were validated using data from spring surveys of winter oilseed rape crops in England and Wales from 2000 to 2003, and reasons for uncertainty in predictions for individual crops are discussed.

Notes: The severity of cassava bacterial blight at two sites in the forest-savanna transition (FST) and dry savanna (DS) zones of West Africa were studied by assessing the effects of: (i) shift of planting date; (ii) potassium fertilizer application and mulching; (iii) intercropping cassava with sorghum or cowpea vs. cassava monoculture; and (iv) the combination of these measures. Disease severity of bacterial blight in two genotypes was generally reduced by 20–60% by late planting, without a negative effect on cassava root yield, in monocropping systems in most treatments in the FST zone (reduction in four treatments, and increase in two treatments, out of 19) and the DS zone (two of eight treatments in 1 year). Late planting led to crop failure in the DS zone in the second year. Intercropping cassava with sorghum reduced bacterial blight severity significantly, up to 80% at normal (all treatments) and late planting time (three out of six treatments) in the FST zone, and in some treatments (four out of seven) at normal planting in the DS zone. Intercropping of cassava with cowpea in the DS zone also reduced disease severity. Cassava-sorghum intercropping generally had no effect on root yield compared with cassava monocropping at both planting times in the FST zone and provided an additional harvest of the intercrop, while yield was affected by intercropping in the DS zone at late, and in some treatments (three out of seven) at normal, planting time. Mulching and potassium treatment had no effect on disease severity, but increased or decreased root yield in some treatments in both sites. Analysis of combined data showed that cropping system, year, site, and site combined with planting date were the highest significant determinants of variation in bacterial blight development.

Notes: The effects of temperature–time combinations and other sanitizing factors during composting on 64 plant pathogenic fungi, plasmodiophoromycetes, oomycetes, bacteria, viruses and nematodes were reviewed. In most cases pathogen survival was determined by bioassays of unknown sensitivity and minimum detection limits of 5% infection or more. For 33 out of 38 fungal and oomycete pathogens, all seven bacterial pathogens and nine nematodes, and three out of nine plant viruses, a peak temperature of 64–70°C and duration of 21 days, were sufficient to reduce numbers to below the detection limits of the tests used. Shorter periods and/or lower temperatures than those quoted in these tests may be satisfactory for eradication, but they were not always examined in detail in composting systems. Plasmodiophora brassicae (clubroot of Brassica spp.), Fusarium oxysporum f.sp. lycopersici (tomato wilt) and Macrophomina phaseolina (dry root rot) were more temperature-tolerant, as they survived a peak compost temperature of at least 62°C (maximum 74°C) and a composting duration of 21 days. Synchytrium endobioticum (potato wart disease) survived in water at 60°C for 2 h, but was not examined in compost. For Tobacco mosaic virus (TMV), peak compost temperatures in excess of 68°C and composting for longer than 20 days were needed to reduce numbers below detection limits. However, TMV and Tomato mosaic virus (TomMV) were inactivated over time in compost, even at temperatures below 50°C. Temperatures in excess of 60°C were achieved in different composting systems, with a wide range of organic feedstocks. The potential survival of plant pathogens in cooler zones of compost, particularly in systems where the compost is not turned, has not been quantified. This may be an important risk factor in composting plant wastes.

Notes: This study examined the applicability of rep-PCR genomic fingerprinting to molecular typing of Phaeoacremonium and Phaeomoniella species found on grapevines and associated with the esca syndrome. Fingerprints obtained by rep-PCR were compared with those obtained by microsatellite-primed PCR. The rep-PCR patterns clearly separated the species, and in some cases revealed intraspecific variability. Dendrograms revealed a clear genetic distance between Phaeomoniella chlamydospora and the three Phaeoacremonium species analysed. The results demonstrate that rep-PCR can be a complementary or alternative strategy to other molecular methods as a tool for identification of species and strains of these two genera.

Notes: The effects of powdery mildew (Uncinula necator) on grape yield, juice and wine quality were quantified for cultivar Cabernet Sauvignon (CSa) in 1997 and 1999, and for Sauvignon blanc (Sa) in 1999. Analyses were carried out on batches of healthy berries to which known percentages (0–50%) of diseased berries were added, and on natural clusters that were classified into four visual classes from low (C1) to high (C4) disease severity. CSa diseased berries showed an average weight reduction of 12% (1997) and 20% (1999). The direct consequence of a higher percentage of smaller, diseased berries was a reduction in yield. The accompanying loss of weight in must from C1 to C4 clusters increased from 10 to 45%. Sugar content in diseased berries was not significantly different from disease-free berries in 1997, but was 20–21% (CSa) and 14% (Sa) higher in 1999. Severely infected batches also showed a higher total acidity than healthy ones. The total anthocyanin content of CSa was decreased by 0·91% (1997) and 0·66% (1999) per percentage mildewed berries added by weight. In Sa wines the concentration of 3-mercaptohexanol, a component of varietal aroma, was decreased by powdery mildew. Multidimensional analyses, based on all the variables studied, successfully grouped batches of CSa according to disease severity. Using directional triangular tests wine experts were able to recognize CSa wines produced from berries with ≈25% of powdery mildew; the threshold for nonexperts was 50%. CSa wines obtained from samples with more than 30% of diseased berries by weight were significantly classified as the worst according to preference order criteria, but below this value the preference was not significant. Sa wines with 〈50% mildewed berries could not be differentiated significantly by organoleptic tests performed by nonprofessionals.

Notes: A survey of symptoms of phytophthora root and collar rot of common (Alnus glutinosa) and grey alder (A. incana) in riparian and forest stands in Bavaria was conducted by the Bavarian State Forestry and river authorities. Symptoms were seen in 1041 out of 3247 forest alder stands. The majority of the affected stands (80·9%) were less than 21 years old; 46% of these young stands were growing on nonflooded sites and 92% had been planted. The riparian survey showed that symptoms were widespread along more than 50% of the river systems. Along some rivers the disease incidence exceeded 50%. The ‘alder Phytophthora’ was recovered from 166 of 185 riparian and forest alder stands with symptoms. In 58 of the 60 rivers and streams investigated in detail, the source of inoculum was traced back to infested young alder plantations growing on the river banks or on forest sites that drain into the rivers. Once introduced to a river system, the ‘alder Phytophthora’ infects alders downstream. Baiting tests showed that the ‘alder Phytophthora’ was present in rootstocks of alders from three out of four nurseries which regularly bought in alder plants for re-sale, but not in rootstocks from four nurseries that grew their own alders from seed. In addition, the infected nurseries used water from infested water courses for irrigation. The Bavarian State Ministry for Agriculture and Forestry has developed a code of practice for producing healthy alder plants in forest nurseries. This includes a 3-year fallow period between bare-rooted alder crops because of poor survival of the ‘alder Phytophthora’ in soil.

Notes: Using a previously developed stochastic simulation model for plant disease epidemics, the relationship of the SADIE aggregation statistic Ia with initial epidemic conditions, spore dispersal distance, sampling quadrat size and other spatial statistics was investigated. Most variation in Ia was attributable to the initial spatial pattern of infected plants and sampling quadrat size. The importance of initial spatial pattern on SADIE clustering indices (for patches and gaps) was also demonstrated using a number of selected data sets. Correlation of Ia with clustering indices was close to 1·0. Epidemics arising from the regular and random initial patterns resulted in the smallest and greatest Ia values, respectively, at sampling times after disease spread had occurred. Furthermore, the variability in Ia between simulation runs also varied greatly with initial patterns, being lowest and greatest for the clumped and random initial patterns, respectively. Ia increased initially and then decreased with increasing incidence, especially for the clumped and random initial patterns. Overall, the effect of median spore dispersal distance on Ia was very small, especially for the random initial pattern. The correlation between Ia and intraclass correlation was generally small and varied greatly between initial patterns. However, there was a high positive correlation between Ia and a parameter describing the rate of decline of autocorrelation over spatial lags, indicating that Ia, clustering indices and autocorrelations measure some common properties of patterns.

Notes: A quantitative-competitive PCR (QC-PCR) method with enzyme-linked oligosorbent assay (ELOSA) was developed to monitor Pichia anomala strain K, a biocontrol agent against postharvest diseases on apples. The procedure involved: (i) extraction of strain K DNA; (ii) coamplification of a constant amount of the extracted DNA (containing a strain K DNA marker of 262 base pairs (bp) specifically amplified with SCAR primers K1 and K2) with an internal standard (IS) titration; and (iii) differential hybridization with two specific biotinylated probes either for the target or for the IS sequence on microplate. The IS sequence differed from the target by only a short internal region of 35 bp providing the differential detection between both sequences. Both target and IS sequences proved to be competitive in PCR as well as in sandwich hybridization. Two copies of the target sequence were detected in the strain K genome by means of enzymatic restriction and Southern blot analysis. Varying amounts of strain K cell suspension were quantified in the phosphate buffer used for recovering cells from the apple surface. An accurate estimate of the strain K population was obtained from 103 to 106 yeast cells per apple. The threshold of the method was found to be at 1000 colony-forming units per apple, which was around 100 times more sensitive than the plating method for monitoring strain K.

Notes: A defined medium was developed in which to monitor deoxynivalenol (DON) accumulation, fungal growth and expression of genes involved in trichothecene biosynthesis (designated Tri genes). In liquid culture, DON accumulated shortly after maximum expression of Tri6 and coincident with expression of Tri5. This was generally 96 h after inoculation. The effects of sublethal concentrations of the fungicides azoxystrobin, trifloxystrobin, kresoxim-methyl and tebuconazole on biosynthesis of the trichothecene DON by Fusarium culmorum were studied using this medium. The strobilurin fungicides trifloxystrobin and azoxystrobin significantly reduced the accumulation of DON in culture medium at a range of concentrations. Kresoxim-methyl, also a strobilurin, and tebuconazole, a triazole, did not significantly reduce the accumulation of DON, although levels were lower than those in nonamended cultures. Trifloxystrobin significantly reduced the accumulation of DON when added to cultures before initiation of trichothecene biosynthesis. RT-PCR assays of the expression of Tri6 and Tri5 genes indicated that trifloxystrobin acted by inhibiting the initiation of trichothecene biosynthesis.

Notes: Pseudomonas syringae pv. maculicola causes bacterial leaf spot on cruciferous plants in Australia. This is the first record confirming the identity of seven isolates currently stored as P. syringae pv. maculicola in the herbarium of the Department of Agriculture, Orange, Australia (Herb. DAR). The isolates were identified using pathogenicity testing on cauliflower and fatty acid methyl ester analysis. They clustered together using repetitive sequence-based polymerase chain reaction (rep-PCR) and PCR-restriction fragment-length polymorphism (RFLP) of the 16S−23S internal transcribed spacer region (ITS) using seven restriction enzymes. Identification was confirmed by comparison of these isolates with known overseas isolates of P. syringae pv. maculicola, but there was significant variation in their pathogenicity and genetic structure.

Notes: The spread of Sugarcane yellow leaf virus (SCYLV) in sugarcane plants was studied on Réunion using virus-infected cuttings from four cultivars (R570, R575, R577 and R579). One month after the germination of cuttings in an insect-proof glasshouse, SCYLV was detected by reverse-transcription polymerase chain reaction (RT-PCR) and tissue-blot immunoassay (TBIA) in the leaves, shoots and roots of all cultivars. The distribution of SCYLV in the whole plant did not vary over a 10- to 11-month period of growth. In addition, the spread of SCYLV in sugarcane fields on Réunion was investigated during a survey conducted from 1998 to 2001. Samples were taken in three sugarcane-growing areas, and TBIA was used to detect SCYLV in the three major cultivars (R570, R575 and R579). The percentage of infected stalks varied according to cultivar and growing area, but remained relatively stable for a given cultivar in a given growing area over the 30-month survey period. Cultivar R575 was the most infected cultivar in all three growing areas (mean of 98% infected stalks). The percentage of infected stalks ranged from 16 to 94% in cv. R570 and from 21 to 92% in cv. R579. These results suggested that on Réunion: (i) infected sugarcane stools do not recover from the disease after harvesting; and (ii) the virus is mainly propagated by planting infected cuttings. SCYLV was detected by RT-PCR in the aphid Melanaphis sacchari, a potential vector of this virus. Two months after planting virus-free plants of susceptible cv. R575 in a field surrounded by sugarcane infected with SCYLV, 14% of plants were found infected with the virus. Four months later, 25% of plants were found infected with SCYLV, but no new infections were detected between 6 and 12 months after planting. In the first ratoon crop, 42% of plants were infected with SCYLV after 6 months of growth. Spatial distribution of infected plants suggested that, on Réunion, a small window of time (between 0 and 2 months after planting cuttings) exists during which primary infection can occur. Based on the results obtained in this study, the use of clean planting material for some cultivars and the use of tolerant cultivars should be an efficient means of controlling sugarcane yellow leaf on Réunion.

Notes: A quantitative real-time PCR assay using TaqMan chemistry has been developed to quantify the level of Tilletia spp. contamination in wheat-seed lots. In the UK wheat seed is predominantly contaminated with Tilletia caries (syn. Tilletia tritici), and the probability of detecting other Tilletia spp. is negligible. DNA standards, prepared from T. caries spores, were calibrated using a set of 26 seed samples, with T. caries contamination levels ranging from 0 to 1000 spores per seed. The linear calibration model obtained by the regression of log10 (number of spores per seed + 1) on mean log10 DNA (µg) produced a coefficient of determination (R2) of 0·904. The calibration model was tested using 226 seed samples; of these, 91% fell within the 95% confidence intervals. Of the 21 samples that were outside the limits, 16 were overpredictions and five underpredictions. The five underpredictions were all from seed samples where contamination was less than one spore per seed. The model predicts that samples with 44 pg of DNA will be below one spore per seed with 95% probability. Of the 226 test samples compared with this threshold, 99 contained less than 44 pg DNA, and these were found to have less than one spore per seed by microscopic assay. This real-time assay allows an increase in test throughput and provides the sensitivity required for an advisory threshold of one spore per seed.

Notes: To investigate the ability of black dot symptoms to develop on infected potato tubers during storage, the growth of Colletotrichum coccodes was followed in vitro on malt agar at temperatures ranging from 5–27°C, and in vivo on artificially infected potato tubers kept at 5, 10 and 15°C. In vitro, 13 isolates from different geographical origins grew at all temperatures tested; growth started with a delay of 10 days at 5°C and of 4 days at 10°C, and was fastest at 27°C. All isolates had similar growth patterns and produced conidia and sclerotia at all temperatures. Minitubers were successfully infected at 5, 10 and 15°C by depositing either a mycelial plug or a drop of conidial suspension on the tuber surface. Sclerotia were observed after 7 days at the point of inoculation. Symptoms extended in all cases, although more slowly at 5 and 10 than at 15°C. Latent infections were detected in up to 21% of tubers without black dot symptoms at harvest. These results show that latent infections by C. coccodes are probably quite frequent, and that the pathogen is able to develop at low temperatures in controlled conditions. This suggests that black dot symptoms can increase during storage if stores are not adequately managed.

Notes: Pseudothecial density of the blackleg fungus Leptosphaeria maculans and discharge of ascospores was measured from stubble of a range of Brassica species, including Brassica napus (canola) cultivars, with a range of blackleg resistance. Since ascospores are the primary inoculum, these parameters reflect inoculum potential for blackleg. Stubble from a representative line of each of B. carinata, B. nigra, Sinapis alba and B. napus cv. Surpass 400 (incorporates blackleg resistance from B. rapa ssp. sylvestris) had lower pseudothecial density and discharged fewer ascospores than stubble of other B. napus cultivars (Karoo, Oscar, Emblem, Dunkeld and Columbus). These latter B. napus cultivars and a representative B. juncea line had higher pseudothecial densities and discharged higher numbers of ascospores. If this trait of low blackleg inoculum from stubble could be introgressed into commercial canola cultivars, blackleg disease severity could be substantially reduced, resulting in higher and more stable canola yields. However, the trait of reduced ascospore discharge may not be stable, as demonstrated by the B. rapa ssp. sylvestris-derived resistance already being overcome by the blackleg fungus in Australia.

Notes: Experiments were conducted under growth-chamber conditions to determine if Pratylenchus penetrans systemically alters light use efficiency (LUE) of Russet Burbank potato infected by Verticillium dahliae. Pathogen separation was achieved by inoculating potato roots with the nematode prior to injecting fungal conidia into the stem vasculature. Treatments were P. penetrans alone, V. dahliae alone, nematode and fungus together, and a no-pathogen control. Gas exchange was repeatedly and nondestructively measured on the fifth-youngest leaf with a Li-Cor LI-6200 portable photosynthesis system. By 16 and 20 days after stem injection with the fungus, LUE was synergistically impaired in jointly infected plants. Transpiration in plants infected with both pathogens was significantly reduced. However, the combined effect of nematode and fungus was synergistic in one experiment and additive in the other. Stems were destructively harvested when LUE was synergistically impaired. Coinfected potato plants contained more colony-forming units (CFU) of V. dahliae in stem sap than those infected by the fungus alone in one experiment. Evidence is provided that infection of Russet Burbank roots by P. penetrans systemically affects disease physiology associated with stem vascular infection by V. dahliae. The findings indicate that the role of the nematode in the fungus/host interaction is more than simply to facilitate extravascular and/or vascular entry of the fungus into potato roots.

Notes: The effects of growth and leaf temperature on photosynthesis were evaluated in sweet orange seedlings (Citrus sinensis cv. Pera) infected with Xylella fastidiosa (the bacterium that causes citrus variegated chlorosis, CVC). Measurements of leaf gas exchange and chlorophyll a fluorescence were taken at leaf temperatures of 25, 30, 35 and 40°C in healthy and infected (without visible symptoms) seedlings submitted to two temperature regimes (25/20 or 35/20°C, day/night), not simultaneously. The CO2 assimilation rates (A) and stomatal conductance (gs) were higher in healthy plants in both temperature regimes. Values for A and gs of infected and healthy plants were higher in the 35/20°C regime, decreasing with leaf temperature increase. In addition, differences between healthy and infected plants were higher at 35/20°C, while no differences in chlorophyll a fluorescence parameters were observed except for potential quantum efficiency of photosystem II, which was higher in infected plants. Low A values in infected plants were caused by low gs and probably by biochemical damage to photosynthesis. The high alternative electron sink of infected plants was another effect of reduced A. Both high growth and high leaf temperatures increased differences in A between healthy and infected plants. Therefore this feature may be partially responsible for lower growth and/or productivity of CVC-affected plants in regions with high air temperature.

Notes: Reports of Ceratocystis spp. causing disease of exotic plantation hardwood species have increased in recent years. Ceratocystis fimbriata causes wilt and canker on Eucalyptus spp. in Africa and South America, and C. albofundus results in wilt and death of Acacia mearnsii in Africa. Ceratocystis spp. generally infect wounds on trees, and artificial stem wounding can thus be used to determine the presence of these fungi. The aim of this study was to identify Ceratocystis spp. infecting wounds on Eucalyptus grandis in South Africa. Isolated Ceratocystis spp. were identified using morphological characteristics and comparisons of DNA sequence data for the ITS and 5·8S regions of the rRNA operon. Pathogenicity trials were conducted in the greenhouse to determine the possible role that these Ceratocystis spp. could have in disease development. These trials were also conducted under field conditions. Three Ceratocystis spp. were collected: C. fimbriata, C. moniliformis and C. pirilliformis. This is the first report of C. fimbriata and C. pirilliformis from Eucalyptus spp. in South Africa, and the first report of the latter fungus outside Australia. Both C. fimbriata and C. pirilliformis caused significant lesions on inoculated E. grandis trees. This is the first evidence that C. pirilliformis is a pathogen of Eucalyptus spp. From the results of both greenhouse and field trials, it has the potential to cause serious disease problems in Eucalyptus plantations.

Notes: Batches of two winter wheat cultivars (Riband and Apollo) were inoculated with conidia of Mycosphaerella graminicola at weekly intervals over a 2 year period. Following 72 h incubation, plants were placed in ambient temperatures ranging between −7 and 32°C with mean batch temperatures of 2·9–20·2°C. Latent period until the first visible symptoms ranged between 11 and 42 days. The relationship between development of lesions and accumulated thermal time was described using a shifted cumulative gamma distribution model. The model provided good estimates of lesion development with r2 〉 0·92 for both cultivars. Base temperatures, below which the pathogen did not develop, were estimated from the model as approximately −2·4°C for the two cultivars. Latent period was estimated as being 250 and 301 degree-days above the estimated base temperature, when defined as time from inoculation to first lesion and time to 50% of maximal lesions, respectively, for cv. Riband. The values for cv. Apollo were similar, but with estimates of thermal time periods c. 5% higher. The relationship between mean temperature and inverse latent period, expressed as days either to first lesion or to 50% of maximal lesions, was best described by a linear regression with r2 〉 0·96 for both cultivars. The opportunity for plants to outgrow disease was reduced when prolonged periods of cold temperature occurred, because the base temperature for growth of the pathogen was less than that for the crop.

Notes: There are claims that at least 11 genes confer resistance in barley (Hordeum vulgaris) to one or more components of the soilborne barley mosaic virus complex but, apart from the immunity conferred by the widely used gene rym4, little is known about their mode of action. This study used mechanical (sap) and plasmodiophorid vector-inoculation techniques combined with ELISA, RT-PCR, symptom development and virus transmission to investigate the response of different genotypes to Barley mild mosaic virus (BaMMV). Barley genotypes were grown at 20 and 12°C to test for temperature sensitivity. Plants with the genes rym3 or rym6 were fully susceptible to the virus, whereas those with genes rym1, Rym2, rym5 or rym11 appeared to be immune, as BaMMV was never detected in any tissue type nor was the virus transmitted from them to susceptible genotypes. The remaining genotypes could all be infected to some extent by BaMMV using one or both inoculation methods, and virus could be transmitted from their roots by the plasmodiophorid vector Polymyxa graminis. Plants with the rym7 gene had delayed symptoms compared to susceptible controls at 12°C. Plants with the rym8 gene could be infected by both inoculation methods, but there was no virus in the leaves at 12°C. Plants with the rym9 gene could be infected only by vector inoculation, and virus remained localized in the roots. Plants with the rym10 gene appeared susceptible by mechanical inoculation at both temperatures, but after vector inoculation virus moved to leaves only at 20°C. This suggests the operation of translocation resistance in plants with the rym8, rym9 or rym10 genes, which is temperature-sensitive in rym8 and rym10 and perhaps tissue-specific in rym9. No resistance to P. graminis was observed in any of the genotypes.

Notes: The spread of septoria leaf blotch in wheat caused by Mycosphaerella graminicola is widely reported to depend on the occurrence of splashy rainfall. Previous studies have also implicated an important effect of canopy architecture on the risk of disease spread. This is because architecture affects the proximity of the yield-forming leaves to inoculum present on older diseased leaves within the crop. This study demonstrated that infection of the final three leaves of winter wheat could occur in the absence of splashy rainfall. For cvs Riband and Longbow, the final two leaf layers emerged at a position ≈ 8 cm below older leaves containing sporulating lesions. Under these conditions, infection of new leaves occurred in treatments that simulated dew and nonsplashy rainfall. These treatments resulted in disease incidences of 10–40% above the untreated control on the final two leaf layers. Within a season, the distance between diseased and healthy leaves during the period of stem extension varied substantially across a range of 30 cultivars. While the magnitude of these differences was not the same across seasons, the relative differences between cultivars were generally consistent, suggesting a strong genotype influence on lesion proximity. This study shows how knowledge of the distribution of lesion proximity within a crop can be used to estimate the risk of inoculum transfer for a given maximum splash height. A rapid crop-monitoring method for estimating the distribution of lesion proximity was developed and tested. Lesion proximity was manipulated by plant growth regulator (PGR) treatments. Significant increases in disease, between 14 and 62%, were observed on the upper canopy leaves of plants treated with PGR. The largest differences were observed in treatments where lesion proximity was most affected.

Notes: For individual varieties, tolerance of septoria leaf blotch was quantified by the slope of the relationship between disease and yield. Variation in disease severity and the associated yield responses were provided across two sites and three seasons of field experiments. Slopes were fitted by residual maximum likelihood for two contrasting models: (i) a fixed-effects model, where no prior assumptions were made about the form of the variety slopes; and (ii) a random-effects model, where deviations in individual variety slopes away from the mean variety slope formed a normal random population with unknown variance. The analyses gave broadly similar results, but with some significant differences. The random model was considered more reliable for predicting variety performance. The effects of disease were quantified as symptom area and green canopy duration. Models of the relationship between symptom area and yield were site-specific. When site effects were not taken into account, these models had poor predictive precision. Models based on the canopy green area gave robust predictions of yield and were not site-specific. Differences in disease tolerance were detected in a comparison of 25 commercial winter wheat varieties. Tolerance was not detected directly through symptom measurements, but instead through measurements of canopy green area, which provides a measurement of the effects of disease that accounts for differences in canopy size across sites and seasons. The varieties showing greatest tolerance tended to have lower attainable yield than the intolerant varieties. Presence of the 1BL/1RS chromosome translocation, which has been reported to increase radiation use efficiency, appeared to be associated with intolerance.

Notes: Twenty-four wheat cultivars and breeding lines were screened for isolate-specific resistance to septoria tritici blotch (STB) caused by 12 isolates of Mycosphaerella graminicola. New isolate-specific resistances that could be used in wheat breeding were identified. Major sources of resistance to STB used in world breeding programmes for decades, such as Kavkaz-K4500, Veranopolis, Catbird and TE9111, have several isolate-specific resistances. This suggests that ‘pyramiding’ several resistance genes in one cultivar may be an effective and durable strategy for breeding for resistance to STB in wheat. Several cultivars, including Arina, Milan and Senat, had high levels of partial resistance to most isolates tested as well as isolate-specific resistances. Resistance to isolate IPO323 was common, present in all but one of the major sources of resistance tested. This suggests that resistance to IPO323 may be an indicator of varietal resistance to STB in the field.

Notes: Seed and soil treatment with Pseudomonas fluorescens RGAF 19, P. fluorescens RG 26, Bacillus megaterium RGAF 51 and Paenibacillus macerans RGAF 101 can suppress fusarium wilt of chickpea (Cicer arietinum), but the extent of disease suppression by these rhizobacteria is modulated by soil temperature. In this work, the effect of temperature on plant–rhizobacteria interactions was assessed in relation to biocontrol potential for suppression of fusarium wilt of chickpea. Seed and soil treatment with those rhizobacteria delayed seedling emergence compared with nontreated controls, and either increased or had no deleterious effect on chickpea growth. Pseudomonas fluorescens isolates significantly increased chickpea shoot dry weight at 20°C and root dry weight at 25 and 30°C. All bacterial isolates colonized the chickpea rhizosphere and internal stem tissues at 20, 25 and 30°C, and there was a positive linear trend between bacterial population size in the rhizosphere and temperature increase. The maximum inhibition of mycelial growth and conidial germination of Fusarium oxysporum f. sp. ciceris race 5 in vitro occurred at a temperature range optimal for bacterial growth and production of inhibitory metabolites. These results demonstrate the need to understand the effects of environmental factors on the biological activities of introduced rhizobacteria of significant importance for plant disease suppression.

Notes: The responses of 24 white yam (Dioscorea rotundata) cultivars to mechanical and vector transmission with each of three viruses infecting yams were assessed through enzyme-linked immunosorbent assay (ELISA) and symptom development. The viruses were Dioscorea alata virus (DAV), genus Potyvirus; Dioscorea alata bacilliform virus (DaBV), genus Badnavirus; and Cucumber mosaic virus (CMV), genus Cucumovirus. Only TDr 95-128, a landrace cultivar from Nigeria, developed symptoms of infection with CMV and DaBV following mechanical and vector transmission, respectively. PAS-ELISA showed that nine genotypes remained uninfected by DAV and 11 were uninfected by CMV or DaBV. Genotypes TDr 747 and TDr 1640 both showed resistance to all three viruses.

Notes: Infection of several commercially important potato cultivars with Tobacco rattle virus (TRV) can result in systemic infection with M-type virus, which persists through generations of vegetative propagation, although spraing symptoms in the tubers rarely develop. In field trials of nine such cultivars it was shown that TRV infection can delay plant emergence and retard subsequent growth, reduce tuber yield and mean tuber weight, increase tuber numbers and the proportion of small tubers, increase the incidence of growth cracks and misshapen tubers, diminish dry matter content, and exacerbate after-cooking blackening. Each cultivar was affected differently, showing different combinations and degrees of these effects. Virus concentration in leaf extracts from individual plants was also highly variable. These, and other potato cultivars similarly susceptible to TRV, are grown on large areas of UK and European potato land. The implications of growing such cultivars for the seed, ware and processing sectors of the potato industry are discussed.

Notes: Nonpathogenic strains of Fusarium oxysporum (NPFO) were tested alone, and in combination with rock salt (NaCl), for their ability to suppress fusarium crown and root rot of asparagus in replanted fields. Five NPFO strains were first screened with and without NaCl in the glasshouse using autoclaved asparagus soil mixed with potting mix. Asparagus roots were treated with NPFO suspensions or water, then inoculated 1 week later with pathogenic F. oxysporum; then after 1 week pots received 100 mL 1% NaCl or water. After 12 weeks the NPFO strain CS-20 increased root weight when compared with controls. Rock salt did not affect root growth. Root lesions were reduced relative to the control by all NPFO strains and by NaCl applications, but there were no interactions between NPFO and NaCl. The NPFO strains CS-20, CWB 314 and CWB 318 were compared with no treatment in two 4-year replanted field studies in Hamden, and Windsor, CT, USA. Crowns were treated with NPFO at planting, and NaCl was broadcast at 280 kg ha−1, and at 560 kg ha−1 in later years. CWB 318 reduced disease at both sites and increased marketable yield in Hamden. Treatment with NaCl increased yield by 29% over controls in Windsor, but did not increase yield in Hamden. Recovered root isolates of F. oxysporum categorized by heterokaryon tests revealed that, in the glasshouse, CS-20 colonized roots better than other strains, while CWB 318 was the only strain found on roots from field plots after 4 years. Applications of NaCl did not hinder the suppressive ability of the nonpathogenic strains. The perennial nature of asparagus may require that nonpathogenic strains possess superior survival traits along with biocontrol traits.

Notes: A total of 174 Salix (willow) clones belonging to 57 species and 14 interspecific hybrids were inoculated with seven pathotypes of Melampsora larici-epitea using the leaf disc method. Infection types were scored based on the uredinial pustule area data and the inoculum density. A close correlation (R2 = 0·82) was found between the average pustule area and the average number of spores produced. Most of the willows were also assessed in the field for rust in 1999. Most willow clones belonging to the species native to western Europe were infected by the rust. In inoculation experiments, uredinia developed on 46 S. viminalis clones, out of a total of 47. In the field, all the S. viminalis clones were infected by rust. Within the subgenus Vetrix, eight out of the 17 willow species that originated from North and South America produced rust pustules in inoculation experiments. Of these, S. pellita was most susceptible. Salix irrorata and S. lasiolepsis var. bracelinae produced well developed pustules after inoculation but no rust infections were detected in the field. In both leaf disc tests and field assessments, no rust infections were found on S. candida, S. cordata, S. drummondiana, S. eriocephala, S. hookeriana, S. houghtonii, S. humilis, S. rigida var. mackenziana and S. syrticola. Of 12 species of subgenus Vetrix native to northeast Asia and Japan, only S. kochiana was susceptible both in inoculation tests and in the field. Salix rossica produced no symptoms in leaf disc tests but showed low levels of infection in the field. The maximum infection type scores in leaf disc tests were highly significantly correlated with field disease severity scores (Spearman rank correlation coefficient was 0·76, P 〈 1 × 10−10).

Notes: Mini-Tn5 transposon mutagenesis of a soft-rotting isolate of Pseudomonas fluorescens strain 123 produced six mutant phenotypes with altered pathogenicity and inability to produce a biosurfactant involved in dispersing P. fluorescens cells in a surface aqueous environment. The mutants isolated showed in vivo growth characteristics identical to those of the wild type, but variations in their ability to reduce surface tension of water and to grow in liquid medium containing hexadecane. One of these mutants (EG3) with reduced pathogenicity on broccoli and cauliflower exhibited a restored pathogenic phenotype when complemented with a 7·1-kb segment of P. fluorescens genomic DNA. The loss of aqueous-surface-tension activity in mutants suggests that a biosurfactant may contribute to pathogenicity and enhance pathogen invasion of host tissues.

Notes: A new virus named Nootka lupine vein-clearing virus (NLVCV) was isolated from Lupinus nootkatensis plants that were confined to a relatively small area in the Talkeetna mountains of south-central Alaska. Annual surveys (2000–03) consistently found leaf symptoms of pronounced vein clearing and mosaic on 3- to 4-week-old plants in late June. Spherical particles ≈30 nm in diameter were isolated from these leaves. Virions contained a single-stranded RNA of ≈4·0–4·2 kb and one species of capsid protein estimated to be ≈40 kDa. The double-stranded RNA profile from naturally infected leaves consisted of three major bands ≈4·2, 1·9 and 1·5 kbp. Protein extractions from either sap or virions of diseased plants reacted to polyclonal antiserum made against the virions in Western blot assays. A predicted PCR product ≈500 bp was synthesized from virion RNA using primers specific to the carmovirus RNA-dependent RNA polymerase (RDRP) gene. The nucleotide sequence of the amplified DNA did not match any known virus, but contained short regions of identity to several carmoviruses. Only species belonging to the Fabaceae were susceptible to NLVCV by mechanical inoculation. Based on dsRNA profile, size of virion RNA genome and capsid protein, and similarity of the RDRP gene to that of other carmoviruses, it is suggested that NLVCV is a member of the family Tombusviridae, and tentatively of the genus Carmovirus. As the host range, RDRP gene and dsRNA profile of NLVCV are different from those of known viruses, this is a newly described plant virus.

Notes: A cassava mosaic disease (CMD) pandemic currently affects much of East and Central Africa. To understand the factors driving the pandemic's continued spread, complementary data sets were collected from cassava plots, planted with healthy cuttings, at eight sites along a north–south transect in southern Uganda, through the pandemic's leading edge. Data were collected on virus incidence, symptom severity, populations of the whitefly vector, Bemisia tabaci, their infectivity and ability to transmit different viruses. In 1996, 6 months after planting, CMD incidences were highest at sites 1 and 2, then decreased progressively until site 6, and remained low at sites 7 and 8. The largest B. tabaci populations also occurred at northernmost sites, 1–3. In 1997, CMD incidence increased significantly at sites 5–8 and this was associated with significant increases in the B. tabaci populations. The pandemic's spread was also associated with significant increases in the percentage of dual infections of East African cassava mosaic virus-Uganda and African cassava mosaic virus, which caused the severest symptoms and the greatest reduction in leaf area. Whitefly adults collected from within the pandemic area were infective, whereas those collected ahead of the pandemic were not. The transmission rate of African cassava mosaic virus from plants with dual infections was significantly less than that of East African cassava mosaic virus-Uganda, which may explain the latter's predominance within the pandemic. These results show that the arrival of East African cassava mosaic virus-Uganda into areas affected previously only by African cassava mosaic virus, has resulted in novel virus/vector/host–plant interactions that drive the pandemic's continued spread.

Notes: The effects of crop management patterns on coffee rust epidemics, caused by Hemileia vastatrix, are not well documented despite large amounts of data acquired in the field on epidemics, and much modelling work done on this disease. One main reason for this gap between epidemiological knowledge and understanding for management resides in the lack of links between many studies and actual production situations in the field. Coffee rust epidemics are based on a seemingly simple infection cycle, but develop polycyclic epidemics in a season and polyetic epidemics over successive seasons. These higher-level processes involve a very large number of environmental variables and, as in any system involving a perennial crop, the physiology of the coffee crop and how it affects crop yield. Crop management is therefore expected to have large effects on coffee rust epidemics because of its immediate effect on the infection cycle, but also because of its cumulative effect on ongoing and successive epidemics. Quantitative examples taken from a survey conducted in Honduras illustrate how crop management, different combinations of shade, coffee tree density, fertilization and pruning may strongly influence coffee rust epidemics through effects on microclimate and plant physiology which, in turn, influence the life cycle of the fungus. We suggest there is a need for novel coffee rust management systems which fully integrate crop management patterns in order to manage the disease in a sustainable way.

Notes: The effects of Coniothyrium minitans inoculum quality and an 8-week interval between inoculum application and crop planting on sclerotinia (Sclerotinia sclerotiorum) disease in three successive lettuce crops were investigated in a glasshouse trial. Spore suspensions of three isolates of C. minitans (Conio, IVT1 and Contans) applied at 108 CFU m−2 and a standard Conio maizemeal–perlite application (06 L m−2, 1011 CFU m−2) were assessed for their ability to control S. sclerotiorum. Only the maizemeal–perlite inoculum (isolate Conio) consistently reduced sclerotinia disease. In the third lettuce crop only, isolates IVT1 and Contans formulated by Prophyta and isolate IVT as an oil–water formulation, all applied as spore suspensions, reduced disease at harvest compared with the untreated control. Recovery, viability and C. minitans infection of sclerotia buried during the 8-week period prior to each of the three lettuce crops, and of sclerotia formed on the crop, were tested. Only the maizemeal–perlite inoculum (isolate Conio) reduced the recovery of sclerotia buried in soil for weeks between inoculum application and crop planting, reducing their viability and increasing infection by C. minitans. Eight weeks was sufficient to enable C. minitans to infect sclerotia of S. sclerotiorum, and may account for disease control. After harvest of the second and third crops, maizemeal–perlite treatment (isolate Conio) reduced the number and viability of sclerotia recovered on the soil surface and increased infection by C. minitans compared with spore-suspension treatments. The effect of inoculum concentration and the influence of soil temperature (varying with time of year) on infection of sclerotia by C. minitans are discussed.

Notes: The environmental factors that influence infection of lettuce by ascospores of Sclerotinia sclerotiorum, and subsequent disease development, were investigated in controlled environment and field conditions. When lettuce plants were inoculated with a suspension of ascospores in water or with dry ascospores and exposed to a range of wetness durations or relative humidities at different temperatures, all plants developed disease but there was no relationship between leaf wetness duration or humidity and percentage of diseased plants. Ascospores started to germinate on lettuce leaves after 2–4 h of continuous leaf wetness at optimum temperatures of 15–25°C. The rate of development of sclerotinia disease and the final percentage of plants affected after 50 days were greatest at 16–27°C, with disease symptoms first observed 7–9 days after inoculation, and maximum final disease levels of 96%. At lower temperatures, 8–11°C, disease was first observed 20–26 days after inoculation, with maximum final disease levels of 10%. Disease symptoms were always observed first at the stem base. In field-grown lettuce in Norfolk, 2000 and 2001, inoculated with ascospore suspensions, disease occurred only in lettuce planted in May and June, with a range of 20–49% of plants with disease by 8 weeks after inoculation. In naturally infected field-grown lettuce in Cheshire, 2000, disease occurred mainly in lettuce planted throughout May, with a maximum of 31% lettuce diseased within one planting, but subsequent plantings had little (≤ 4%) or no disease. Lack of disease in the later plantings in both Norfolk and Cheshire could not be attributed to differences in weather factors.

Notes: Gibberella zeae (anamorph Fusarium graminearum) causes head blight of cereals and contaminates grains with mycotoxins such as deoxynivalenol (DON). To determine the correlations among aggressiveness traits, fungal colonization and DON production, 50 progeny from a segregating population of G. zeae were inoculated onto a susceptible winter wheat cultivar in three field environments (year–location combinations). Aggressiveness traits were measured as head-blight rating and plot yield relative to noninoculated plots. Fungal colonization, measured as Fusarium exoantigen (ExAg) content, and DON production were analysed with two ELISA formats. Disease severity was moderate to high based on head-blight rating and relative plot yield. Fusarium ExAg content and DON production ranged from 0·26–1·41 units and from 4·18–43·70 mg kg−1, respectively. Significant (P = 0·01) genotypic variation was found for all traits. Heritability for Fusarium ExAg content was rather low because of high progeny–environment interaction and error. DON/Fusarium ExAg ratio did not vary significantly (P 〉 0·1) among progeny. Correlation between DON production and Fusarium ExAg content across environments was high (r = 0·8, P = 0·01), but no covariation existed between aggressiveness traits and DON/Fusarium ExAg content ratio.

Notes: A total of 360 Pseudomonas savastanoi pv. savastanoi isolates obtained from 11 Italian olive (Olea europaea) cultivars grown in different provinces were assessed with repetitive PCR using short interspersed elements of the bacterial genome as primers (ERIC, BOX and REP primer sets). The population structure of the isolates was determined by using three different hierarchical clustering algorithms: UPGMA, single-link and complete-link methods. REP primers were the most discriminatory. The various fingerprints obtained from the same cultivar and locality persisted over 2 years of knot sampling. Repetitive PCR and UPGMA analysis, using the three data sets combined, revealed 20 patterns with an overall similarity of 81%, with no grouping of the isolates. The resulting dendrogram shows a bush-like topology. Similar results were obtained with the other two clustering methods. In contrast, data obtained from the literature showed that the genetic structure of olive is characterized by bifurcated dendrograms and clear grouping of cultivars. Therefore it appears that the host plant and its pathogen did not cospeciate. The strict adaptation of the bacterium to olive would represent a case of association by colonization.

Notes: In Brazil plants of Pfaffia glomerata with mosaic symptoms were found to be infected with a previously undescribed potyvirus, Pfaffia mosaic virus (PfMV). Virus particles were long and flexuous, c. 10 × 700–800 nm, and cylindrical inclusions typical of potyviruses were present in cells of infected tissue. Partial host-range studies revealed that in addition to P. glomerata, PfMV infected only Chenopodium amaranticolor and Chenopodium quinoa. It was efficiently transmitted by the aphids Aphis gossypii and Myzus persicae. Polyclonal antiserum produced against the PfMV coat protein (CP) reacted with Potato virus Y (PVY), but not with four other potyviruses in PTA-ELISA. The similarity of the nucleotide sequence of the PfMV coat-protein gene (CP) varied from 7 to 76% when compared with other members of the family Potyviridae. Similarity of the 3′ NTR sequence varied from 4 to 23%. In both cases the highest similarity was with PVY. These data indicate that PfMV is a new species in the genus Potyvirus.

Notes: The rust fungus Maravalia cryptostegiae, from south-west Madagascar, was introduced into Australia in 1995 as a classical biological control agent against the highly invasive rubber-vine weed Cryptostegia grandiflora, a woody climber endemic to Madagascar. The rust was released at 69 sites between 1995 and 1997 and is now established throughout the plant's exotic range in Queensland, estimated at over 40 000 km2. Dispersal was low in the first 3–4 months but was virtually linear thereafter, and the rust spread over 100 km within the first year; after 3 years it was recorded 550 km away from the nearest release site. Spraying both dry and aqueous inoculum of uredinioid teliospores from the ground using mist-blowers, as well as from the air by atomizing spore suspensions, resulted in rust-induced defoliation, producing an overall reduction in fecundity and biomass of the weed. In sites with low water tables, weed growth decreased markedly, with a reduction in plant volume from 9 m−3 to 1 m−3 over a 4-year period. Both rust- and drought-induced stress combined to cause up to 75% plant mortality at some sites, and at all monitored sites, seedling recruitment was virtually nil. Improved growth of indigenous grasses amongst rubber-vine thickets has increased fuel loads and created opportunities to use fire as a component of an integrated approach to the management of this economically and ecologically damaging weed.

Notes: A total of 2691 single-lesion isolates of Phytophthora infestans was established from samples of late-blight disease from 354 commercial and garden/allotment sites in Scotland, England and Wales over four growing seasons, 1995–98. The A2 mating type was rare (3·0% of isolates) and was detected at only 34 sites. In vitro tests of sensitivity to the phenylamide fungicide metalaxyl showed that 316 sites yielded isolates with some insensitivity (resistant and/or intermediate); these were more often commercial sites than garden/allotment sites. Over the four seasons, the frequency of isolates with intermediate fungicide sensitivity increased, while the frequency of resistant phenotypes decreased. Resistant isolates were always of A1 mating type. A subset of 1459 isolates from 326 sites was analysed for molecular diversity. Mitochondrial DNA (mtDNA) haplotype Ia predominated (91·0% of isolates); haplotype IIa was present at 54 sites and both haplotypes at 33 sites. The multilocus RFLP probe RG57 detected 30 fingerprints. Four fingerprints were particularly common (RF002, RF006, RF039 and RF040) and 10 were unique to a single site in a single year. The three commonest fingerprints (RF039 〉 RF002 〉 RF006) were of A1 mating type and the fourth (RF040) was A2. RF002 isolates were resistant to the phenylamide metalaxyl and were more common in Scotland than in England and Wales. Small sample sizes limited the usefulness of estimates of diversity. Although approximately half of all sites appeared to be colonized by RF039 genotypes, some sites (both commercial and garden/allotment) showed a higher diversity, having both common and unique genotypes. The genotypic diversity within isolates collected from commercial sites and those from garden/allotment sites were similar. The contributions of sexual reproduction and alternatives to sex to the generation of variation are discussed.

Notes: A time-saving and cost-effective polymerase chain reaction (PCR)-based method was developed for species-specific detection of the scab pathogens (Streptomyces scabies and S. turgidiscabies) prevalent in potato (Solanum tuberosum) in northern Scandinavia. Species specificity of primers was verified using a collection of previously characterized Streptomyces strains isolated from potato scab lesions in Finland and Sweden. A total of 1245 scab lesions was tested from potato cvs Matilda and Sabina grown in the field in two geographic regions of Finland in 2000 and 2001. Freshly harvested or stored potato tubers were incubated at room temperature (18–21°C) under humid conditions for a few days. Bacterial growth was collected from scab lesions for DNA isolation and PCR. The two scab pathogens were detected in the same potato fields, tubers and scab lesions. The relative incidence of S. scabies was high in freshly harvested tubers but was much lower than that of S. turgidiscabies following storage. Both pathogens were seed-transmitted in Matilda and Sabina after 24 weeks of storage at 4°C.