Effects of nitrogen, phosphorus, potassium and calcium nutrition on strawberry anthracnose

The effects of a range of concentrations of four nutrients – nitrogen, phosphorus, potassium and calcium – in fertilizer solutions on the severity of anthracnose on strawberry cv. Nyoho cultivated under a noncirculation hydroponics system were determined after inoculation with Colletotrichum gloeosporioides . Crop growth and tissue nitrogen, phosphorus, potassium and calcium contents of the entire above-ground parts of the plant were also investigated. Elevated nitrogen and potassium concentrations in the fertilizer solution increased disease severity in contrast to phosphorus and calcium. Treatment with either NH₄ or NO₃ nitrogen was not significantly different. The dry weight of the strawberry plants increased significantly with elevated concentrations of nitrogen ( R ² = 0·9078) and phosphorus ( R ² = 0·8842), but was not influenced by the elevated amounts of potassium ( R ² = 0·8587) and calcium ( R ² = 0·6526) concentrations.     

Heterothallism in Plasmopara viticola

Sexuality in the oomycete Plasmopara viticola , the causal agent of grapevine downy mildew, was studied using isolates from five populations from North America and Europe. Leaf discs of Vitis vinifera cv. 'Chardonnay' were inoculated with either individual single-sporangiophore isolates, or in all possible pairwise combinations of 25 isolates from New York State, USA. The occurrence of oospores in leaf discs indicated that the pathogen was heterothallic with two mating types, P1 and P2 in a ratio of 11 : 14 for this population. Heterothallism was confirmed when three representative isolates of each mating type from New York were coinoculated with each of 40 isolates from populations of P. viticola from Michigan, Missouri (USA), Germany and Italy. For each isolate tested, oospores formed with either test isolates of P1 or test isolates of P2 mating types, indicating that the isolates were exclusively P1 or P2 only. For these same isolates, no oospores formed as a result of self-crosses. The ratio of P1 : P2 mating types for all isolates in the study was 27 : 38, statistically equivalent to a 1 : 1 ratio according to χ² analysis ( P  = 0·68).     

Effect of temperature on latent period of septoria leaf blotch on winter wheat under outdoor conditions

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 r ² > 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 r ² > 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.     

Effect of water on germination of Plasmopara viticola oospores

The effect of moisture in grape leaf litter holding overwintering Plasmopara viticola oospores was investigated. Oospores were incubated under different regimes of water activity ( a_W 0·991 to 0·123) for 2 to 15 days and their ability to germinate and cause infection was determined using a sensitive leaf disk assay. Reduction of a_W caused a significant shift in the infection dynamics, with maximum effect when a_W  ≤ 0·56. Dynamics of a_W in the leaf litter under natural conditions were estimated from moisture data using a Chen-Clayton equation. Daily patterns of leaf litter moisture (M in % weight) were determined in non rainy periods between mid February and mid June, while the Chen-Clayton equation was calculated using data of a_W and M measured in both sorption and desorption conditions, at different temperatures. Water activity was highest at 08·00 hours, decreased progressively until 14·00 hours, and then increased. Water activity was favourable for oospores to develop in about 25% of the measurements, all made between 18·00 hours and 08·00 hours. A close relationship was found between vapour pressure deficit (VPD in hPa) and a_W of the leaf litter, so that when VPD is lower than 2·13 hPa there is sufficient water for oospores to develop. Results showed that leaf litter moisture due to water from the atmosphere makes oospore development possible during non rainy periods.     

Correlative analysis of Mycosphaerella graminicola pathogenicity and cell wall-degrading enzymes produced in vitro: the importance of xylanase and polygalacturonase

Eight Mycosphaerella graminicola isolates were investigated for correlations between pathogenicity and the in vitro production of cell wall-degrading enzymes. Isolate pathogenicity was evaluated in terms of lesion and production of pycnidia in wheat leaves. Additionally, the isolates were compared over time for their ability to produce in vitro significant levels of xylanase (EC 3·2·1·8), β-xylosidase (EC 3·2·1·37), β-1,3-glucanase (EC 3·2·1·6), cellulose (EC 3·2·1·4) and polygalacturonase (EC 3·2·1·15) activities when grown in a liquid medium. Correlation tests and principal component analysis revealed a significant correlation between the in vitro production of xylanase and pectinase and pathogenicity components. Xylanase was correlated to necrosis frequency ( r  = 0·795), β-xylosidase was correlated to the mean of the lesion length ( r  = −0·787), whereas polygalacturonase was correlated to the time when 50% of the leaves contained a lesion ( r  = 0·776), the lesion frequency ( r  = 0·646) and the time when 50% of the leaves showed pycnidia ( r  = −0·711). The results suggest that these two groups of cell wall-degrading enzymes are therefore likely to be key determinants of pathogenicity in M. graminicola .     

Factors affecting the control of Pythium ultimum damping-off of sugar beet by Pythium oligandrum

Application of Pythium oligandrum to a soil-based compost as a mycelial suspension (5 × 10² CFU g⁻¹ of dry compost) and oospore alginate pellets (10⁵ oospores/g of dry compost) controlled pre- and postemergence damping-off of sugar beet caused by Pythium ultimum to a level similar to metalaxyl seed treatment. Oospore seed treatments and aqueous suspensions of oospores applied to compost failed to control disease. Problems in the use of P. oligandrum oospore inocula for the control of damping-off were highlighted. It was shown that treatment of oospores with cellulase (20 g L⁻¹) increased germination approximately three-fold in comparison to untreated spores. Untreated and cellulase pretreated oospores were subsequently evaluated as seed treatments for their ability to control damping-off of sugar beet. The highest rate of pretreated oospores (10⁴ oospores/seed) gave levels of emergence and establishment in infested compost that were not significantly different from the uninfested controls, whereas seed treatment with untreated oospores gave no significant reduction in disease. In a trial carried out in a controlled environment to assess the effect of pH (4.5–8.0), P. oligandrum (10⁴ cellulase pretreated oospores/seed) was shown to control pre- and postemergence damping-off of sugar beet at pH 7.0 and 7.5 only.     

Assessing mango anthracnose using a new three-dimensional image-analysis technique to quantify lesions on fruit

An accurate image-analysis method was developed to assess quantitatively the spot-like lesions on fruits resulting from pathogen attack. The technique was applied to evaluation of the development and severity of anthracnose of mango fruit, caused by the fungus Colletotrichum gloeosporioides . In this method, a stepper motor rotates the mango fruit along its longitudinal axis while acquiring a sequence of 360 images of its total surface (one image for each degree). This set of images is used to create a pseudocylindrical 'equal-area' projection of the fruit in a two-dimensional map containing complete morphometrical and photometrical information of its surface. The lesion area can easily be evaluated from this map with image-analysis procedures. Quantitative data (percentage of area affected) can be used to establish an assessment scale for the disease based on lesion spots measured, as well as for detailed laboratory studies of mango anthracnose development. The average error of the method is −0·1%, standard deviation 0·44 ( r ² = 0·99), and it may be adapted for use with most commercial image analysers and for other diseases with spot-like symptoms.     

Pathogenicity of the root-knot nematode Meloidogyne javanica on potato

Host–parasite relationships and pathogenicity of Meloidogyne javanica on potatoes (newly recorded from Malta) were studied under glasshouse and natural conditions. Potato cvs Cara and Spunta showed a typical susceptible reaction to M. javanica under natural and artificial infections, respectively. In potato tubers, M. javanica induced feeding sites that consisted of three to four hypertrophied giant cells per adult female. Infection of feeder roots by the nematode resulted in mature large galls which usually contained at least one mature female and egg mass. In both tubers and roots, feeding sites were characterized by giant cells containing granular cytoplasm and many hypertrophied nuclei. Cytoplasm in giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P ) [0–64 eggs + second-stage juveniles (J2s) per cm³ soil] and growth of cv. Spunta potato seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^{( P − T )}] was fitted to fresh shoot weight and shoot height data of nematode-inoculated and control plants. Tolerance limits ( T ) for fresh shoot weight and shoot height of cv. Spunta plants infected with M. javanica were 0·50 and 0·64 eggs + J2s per cm³ soil, respectively. The m parameter in that model (i.e. the minimum possible y -values) for fresh shoot weight and shoot height were 0·60 and 0·20, respectively, at P  = 64 eggs + J2s per cm³ soil. Root galling was proportional to the initial nematode population density. Maximum nematode reproduction rate was 51·2 at a moderate initial population density ( P  = 4 eggs + J2s per cm³ soil).     

Sensitivity of Sclerotinia homoeocarpa to demethylation-inhibiting fungicides in Ontario, Canada, after a decade of use

In late 2003, nine populations of Sclerotinia homoeocarpa in Ontario Canada (seven of which had been previously sampled in early 1994, prior to the registration of sterol demethylation-inhibiting (DMI) fungicides for turf disease control in Canada) were sampled and tested for sensitivity to propiconazole. Four of the nine populations had not been treated with DMI fungicides during the intervening years, and isolates from these locations were sensitive to propiconazole (geometric mean EC₅₀ values of 0·005–0·012 µ g mL⁻¹, compared with 0·005–0·008 µ g mL⁻¹ for the original 1994 populations). Among the five populations from 2003 that had been exposed to DMI fungicides, mean EC₅₀ values were significantly greater, ranging from 0·020 to 0·048 µ g mL⁻¹. A significant correlation of determination was found between estimated number of fungicide applications and log EC₅₀ ( R ² = 0·832, P  = 0·0001), and the equation predicted that 42·3 applications of propiconazole would be needed to bring a sensitive population (EC₅₀ < 0·01  µ g mL⁻¹) to a resistant level (EC₅₀ > 0·10  µ g mL⁻¹). Fungicide sensitivity vs. duration of fungicide efficacy was also tested, and it was found that isolates with decreased sensitivity were able to more quickly overcome the inhibitory effects of fungicide application, reducing the duration of control from 3 weeks to 2 weeks.     

Effect of temperature on head blight of wheat caused by Fusarium culmorum and F. graminearum

The effect of small temperature differentials (16 vs. 20°C) on the pathogenicity of deoxynivalenol producing single isolates of Fusarium culmorum and F. graminearum and on the fusarium head blight (FHB) response of eight wheat cultivars was examined. Fusarium culmorum inoculation caused greater visual disease symptoms at 20°C than at 16°C, both overall and on an individual cultivar basis (overall AUDPC = 13·5 and 9·6, respectively) ( P  < 0·05). In contrast, F. graminearum inoculation caused greater overall visual disease symptoms at 16°C than at 20°C, both overall and at the individual cultivar level (overall AUDPC = 12·8 and 10·9, respectively) ( P  < 0·05). Results showed both F. culmorum and F. graminearum inoculations caused a greater loss in yield at 20°C (54·3 and 46·9% relative 1000-grain weight, respectively) compared with 16°C (73·3 and 66·9% relative 1000-grain weight, respectively) ( P  < 0·05). Fusarium culmorum -inoculated heads contained similar amounts of fungal DNA at both 16 and 20°C (1·9 and 1·7 ng mg⁻¹ of plant material, respectively) (not significant), while for F. graminearum inoculation, plants contained higher amounts of fungal DNA at 20°C (2·0 and 1·0 ng mg⁻¹ of plant material, respectively) ( P  < 0·05). Overall, there was a significant negative correlation between AUDPC and percentage relative 1000-grain weight at both 16 and 20°C ( r  =−0·693 and −0·794, respectively, P  < 0·01).     

Effect of water potential on mycelial growth and perithecial production of Monosporascus cannonballus in vitro

The effects of osmotic water potential (Ψ_s) on mycelial growth and perithecial production of Monosporascus cannonballus , the cause of root rot and vine decline of melons, were examined at 25°C on potato dextrose agar (PDA) amended with KCl, NaCl or sucrose. Patterns of the growth responses of four isolates to decreasing Ψ_s were similar for each of the osmotica. Compared with growth on nonamended PDA (−0·3 MPa), growth of all isolates increased as Ψ_s was reduced to −0·8 MPa. Maximum growth occurred at Ψ_s values of −0·6 to −0·8 MPa. Growth was not reduced below that on nonamended PDA until Ψ_s was reduced to −1·8 MPa, and a 50% reduction in growth did not occur until Ψ_s was reduced to < −2·5 MPa. Reproduction was much more sensitive to reduced Ψ_s than was mycelial growth, and perithecia were produced only at Ψ_s ≥ −0·7 or −0·8 MPa on PDA amended with KCl or NaCl, respectively. Three isolates produced perithecia on PDA amended with sucrose only at Ψ_s ≥ −0·6 MPa, but the fourth isolate produced perithecia at ≥ −1·9 MPa. Colonization of the xylem early in disease development may provide an essential source of water for subsequent reproduction in the root cortex during plant senescence. Postharvest cultivation to expose and desiccate roots may prevent reproduction even when temperatures lethal to hyphae are not attained.     

Effect of wind on the dispersal of oospores of Peronosclerospora sorghi from sorghum

The effect of wind on the dispersal of oospores of Peronosclerospora sorghi , cause of sorghum downy mildew (SDM) is described. The oospores are produced within the leaves of aging, systemically infected sorghum plants. These leaves typically undergo shredding, releasing oospores into the air. Oospores are produced in large numbers (6.1 × 10³ cm⁻² of systemically infected leaf) and an estimate of the settling velocity of single oospores (0.0437 m s⁻¹) of P. sorghi indicated their suitability for wind dispersal. In wind tunnel studies wind speeds as low as 2 m s⁻¹ dispersed up to 665 oospores per m³ air from a group of leaves previously exposed to wind and displaying symptoms of leaf shredding. The number of oospores dispersed increased exponentially with increasing wind speed. At 6 m s⁻¹, up to 12 890 oospores per m³ air were dispersed. Gusts increased oospore dispersal. A constant wind speed of 3 m s⁻¹ dispersed a mean of 416 oospores per m³. When gusts were applied the mean was 15 592 oospores per m³. In field experiments in Zimbabwe, oospores were sampled downwind from infected plants in the field and at a height of 3.8 m above ground level immediately downwind of an infected crop. These data indicate that wind could play a major role in the dispersal of oospores from infected plants in areas where SDM infects sorghum, perhaps dispersing oospores over long distances.     

Aggressiveness and host specificity of Brazilian isolates of Phytophthora infestans

The population of Phytophthora infestans in Brazil consists of two clonal lineages, US-1 associated with tomatoes and BR-1 associated with potatoes. To assess whether host specificity in these lineages resulted from differences in aggressiveness to potato and tomato, six aggressiveness-related epidemiological components – infection frequency (IF), incubation period (IP), latent period (LP), lesion area (LA), lesion expansion rate (LER) and sporulation at several lesion ages (SSLA) – were measured on detached leaflets of late blight-susceptible potato and tomato plants. Infection frequency of US-1 was similar on potato and tomato leaflets, but IF of BR-1 was somewhat reduced on tomato. Incubation period was longer on both hosts with US-1, although this apparent lineage affect was not significant. Overall there was no host effect on IP. On potato, BR-1 had a shorter LP (110·3 h) and a larger LA (6·5 cm²) than US-1 (LP = 162·0 h; LA = 2·8 cm²). The highest LER resulted when isolates of BR-1 (0·121 cm² h⁻¹) and US-1 (0·053 cm² h⁻¹) were inoculated on potato and tomato leaflets, respectively. The highest values of the area under the sporulation capacity curve (AUSC) were obtained for isolates of US-1 inoculated on tomato leaflets (6146) and for isolates of BR-1 on potato leaflets (3775). In general, higher values of LA, LER, SSLA and AUSC, and shorter values of LP were measured when isolates of a clonal lineage were inoculated on their original host than with the opposite combinations. There is evidence that there are quantitative differences in aggressiveness components between isolates of US-1 and BR-1 clonal lineages that probably contribute to host specificity of P. infestans populations in Brazil.     

Host–parasite relationships of Meloidogyne incognita on spinach

Host–parasite relationships in root-knot disease of spinach caused by Meloidogyne incognita race 1 were studied under glasshouse conditions. Nematode-induced mature galls were large and usually contained one or more females and egg masses with eggs. Feeding sites were characterized by the development of giant cells containing granular cytoplasm and many hypertrophied nuclei. The cytoplasm in these giant cells was aggregated alongside the thickened cell walls. Stelar tissues within galls appeared disorganized. The relationship between initial nematode population density ( P _i) in a series from 0–128 eggs and second-stage juveniles per cm³ soil and growth of spinach cv. Symphony F₁ seedlings was tested under glasshouse conditions. A Seinhorst model [ y = m  + (1 −  m ) z ^P–T ] was fitted to fresh top- and total plant-weight data for inoculated and control plants. Tolerance limits ( T ) of spinach cv. Symphony F₁ to M. incognita race 1 for fresh top and total plant weights were 0·25 and 0·5 eggs and second-stage juveniles per cm³ soil, respectively. The minimum relative values for fresh top and total plant weights were zero in both cases at P _i ≥ 32 eggs and second-stage juveniles per cm³ soil. Root galling was least at low initial population densities and greatest at 16 eggs and second-stage juveniles per cm³ soil. Maximum nematode reproduction rate was 33·1-fold at the lowest P _i.     

Susceptibility of grapevine buds to infection by powdery mildew Erysiphe necator

Bud colonization and perennation of powdery mildew ( Erysiphe necator ) was studied by inoculating shoots of grapevine ( Vitis vinifera cv. Carignane) at different phenological stages. Disease incidence and severity assessments indicated that buds were most susceptible at the three- to six-unfolded-leaf stage. Incidence of powdery mildew colonies on the surface of buds collected from these shoots 7 weeks postinoculation was highest at these stages (68 and 62%, respectively), which indicates that colonization of the bud interior via the infected bud surface is likely to occur within this period. Histological analyses of buds revealed hyphae with haustoria, conidiophores and conidia on all parts of the bud interior except for the meristems. In particular, trichomes were frequently parasitized by haustoria. In total, 13·2% of all buds analysed, and 32·3% of all buds originating from shoots inoculated at the three-unfolded-leaf stage, were infected by E. necator . In the spring of the following year, buds from inoculated shoots yielded 18 flag shoots (1·6% of all emerging shoots). These primary infections caused an epidemic 28 days after the appearance of the first flag shoot. A linear regression analysis on the frequency of infections of the bud exterior, bud interior and flag shoots revealed that incidence of external bud infection in the first season is strongly correlated with flag shoot incidence in the following season ( R ² = 0·94). Hence predictions of flag shoot incidence may be reliably based on the incidence of infection on the outer bud scales in the preceding season.     

Effect of metalaxyl on formation and germination of oospores of Phytophthora infestans

Oospores of Phytophthora infestans were produced in potato leaf discs floating on metalaxyl solution (100 μg mL⁻¹ a.i.) and inoculated with all combinations of two metalaxyl-sensitive and two -resistant parental isolates. Numbers of oospores produced varied between different matings, depending on parents, in the absence of the fungicide and when metalaxyl was added 0, 7, 14 and 21 days after inoculation. Oospores were not produced when metalaxyl was added at the time of inoculation (0 days) when either one or both parents were sensitive to metalaxyl. In two of three such matings further oospore formation was arrested when metalaxyl was added either 7 or 14 days after inoculation. Oospores extracted from leaf discs 14, 21 and 28 days after inoculation were assessed for germination on water agar after 21 days. Germination of oospores from water control treatments varied between 6 and 30% depending on the cross. Germination was significantly reduced in oospores of metalaxyl-sensitive parents extracted 28 days after inoculation of leaf discs treated with metalaxyl 0, 7 and 14 days after inoculation compared with the 21-day treatment. Minimal differences in germination were observed for oospores from the mating of resistant parents irrespective of metalaxyl treatment, although germination was generally low, not exceeding 8.5%.     

Effect of soil physical and chemical properties on the sorption-desorption hysteresis of the diketonitrile metabolite of isoxaflutole

Isoxaflutole, [4-(2-methanesulfonyl-4-trifluoromethylbenzoyl)-5-cyclopropyl isoxazole] is a relatively new pre-emergence herbicide which undergoes rapid conversion to a diketonitrile metabolite in soil. The half-life of isoxaflutole is very short but the half-life of diketonitrile is much longer and hence, diketonitrile remains for a extended period of time in soil. Sorption-desorption studies were conducted with five soils varying in physical and chemical properties. The batch equilibration technique was used for the sorption experiments, while completely mixed batch reactor systems with the decant and refill method was used for the desorption experiments. Four subsequent desorptions were examined after the sorption process in each soil with an equilibration period of seven days. An apparent sorption-desorption hysteresis was observed in all five soils. Organic matter content and the clay content of the soils were the two determining factors for hysteresis. In soils with high organic matter content, the sorption-desorption hysteresis was mainly governed by organic matter content, but in soils with low organic matter clay content played an important role. With the exception of the Chelsea soil, which had a very high organic matter content (57.4%), all other soils exhibited a high correlation between the clay content and hysteresis index (HI) values calculated at 0.75 ( r ² = 0.960), 25 ( r ² = 0.934) and 150 mg L⁻¹ ( r ² = 0.928). In conclusion, the potential for leaching through soil and crop injury due to isoxaflutole and its metabolite would decrease as soil organic matter and clay content increases.     

Estimating relative crop yield loss resulting from herbicide damage using crop ground cover or rated stunting, with maize and sethoxydim as a case study

The research goal was to determine whether crop damage from herbicides measured early in the growing season soon after treatment could be used to estimate relative crop yield loss. Percentage stunting was rated visually and percentage crop ground cover (i.e. percentage of the ground surface covered by vegetation) was determined from video photographs taken 2–4 weeks after sethoxydim-susceptible maize ( Zea mays L.) was sprayed with sethoxydim at various rates plus crop oil concentrate. Averaged over 3 years, relative percentage maize yield was a negative sigmoidal function of relative sethoxydim rates from 0.065× to 0.5×, where the 1× rate was 420 g a.i. ha⁻¹ ( r ² = 0.80). Relative maize yield was positively linearly related to percentage crop ground cover and negatively linearly related to rated percentage stunting averaged over 3 years. Linear regression models of relative maize yield vs. percentage maize ground cover explained only slightly more data variability ( r ² = 0.86) than did rated stunting ( r ² = 0.82) over 3 years. The advantages and disadvantages of rated stunting and crop ground cover as scientific measurements are discussed.     

Effect of inoculum type and timing of application of Coniothyrium minitans on Sclerotinia sclerotiorum: influence on apothecial production

The effects of different inocula of the mycoparasite Coniothyrium minitans on carpogenic germination of sclerotia of Sclerotinia sclerotiorum at different times of year were assessed. A series of three glasshouse box bioassays was used to compare the effect of five spore-suspension inocula of C. minitans , including three different isolates (Conio, IVT1 and Contans), with a standard maizemeal–perlite inoculum. Apothecial production, as well as viability and C. minitans infection of S. sclerotiorum sclerotia buried in treated soil, were assessed. Maizemeal–perlite inoculum at 10⁷ CFU per cm³ soil reduced sclerotial germination and apothecial production in all three box bioassays, decreasing sclerotial recovery and viability in the second bioassay and increasing C. minitans infection of sclerotia in the first bioassay. Spore-suspension inocula applied at a lower concentration (10⁴ CFU per cm³ soil) were inconsistent in their effects on sclerotial germination in the three box bioassays. Temperature was an important factor influencing apothecial production. Sclerotial germination was delayed or inhibited when bioassays were made in the summer. High temperatures also inhibited infection of sclerotia by C. minitans . Coniothyrium minitans survived these high temperatures, however, and infected the sclerotia once the temperature decreased to a lower level. Inoculum level of C. minitans was an important factor in reducing apothecial production by sclerotia. The effects of temperature on both carpogenic germination of sclerotia and parasitism of sclerotia by C. minitans are discussed.     

Real-time PCR identification and detection of Fuscoporia torulosa in Quercus ilex

Fuscoporia torulosa is the causal agent of white alveolar wood decay on several species including a large number of forest trees. Early detection of the fungus is essential to identify diseased trees before spread occurs to healthy plants. However, current detection methods based on isolation from infected tissues on semi-selective media are laborious, time consuming and require expertise in identifying the pathogen after isolation. In the present study, a rapid and reliable Scorpion-PCR based molecular method to identify and detect F. torulosa in planta was developed in a highly polymorphic portion of the internal transcribed spacer (ITS) regions. Specificity of primers and probe was assessed by means of both BLAST analyses and by using genomic DNA from 131 F. torulosa isolates and 43 other fungi and oomycetes from different hosts and geographic areas. In Scorpion-PCR the limit of detection was 1 pg of total DNA and a high correlation ( r ² = 0·996) was achieved between target DNA quantity and cycle threshold (Ct). Real-time PCR combined with effective procedures for DNA extraction enabled the detection of F. torulosa from naturally infected tissues of oaks with and without fruit bodies in approximately 6 h. Comparative testing showed that detection of F. torulosa in wood samples is more sensitive and reliable with real-time PCR than with conventional isolation.