Effects of wind, relative humidity, leaf movement and colony age on dispersal of conidia of Uncinula necator, causal agent of grape powdery mildew

A wind tunnel was designed to study the effect of wind, relative humidity, leaf movement and colony age on dispersal of conidia of Uncinula necator . Wind speed as low as 2.3 m s⁻¹ instantaneously triggered dispersal of conidia from fixed leaf discs of 18-day-old infections. Conidia were observed on sporulating leaf discs even after exposure to 17 m s⁻¹ wind. The fraction of conidia dispersed at a given wind speed increased with colony age from 12 to 24 days. Conidia of 27-day-old colonies were less easily dispersed. No gradient of maturation of conidia along the conidial chain was observed, suggesting that even newly formed conidia were able to germinate after dispersal. Germination of dispersed conidia decreased slightly with greater colony age. Both wind and simulated rain drops caused dispersal of conidia from infected leaves. Leaf movement at wind speed of 3.5–4 m s⁻¹ increased dispersal, and the first impact of three simulated raindrops caused release of 53% of the total conidia dispersed. Relative humidity had no effect on dispersal of conidia at different wind speeds.     

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.     

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

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

Effect of temperature of production of Botrytis allii conidia on their pathogenicity to harvested white onion bulbs

Botrytis allii colonies incubated at low temperatures have been reported to produce larger conidia that germinate faster and give rise to longer germ-tubes than those grown at room temperature. The present study compared the effect of conidia produced at 20°C and at 0 and –2°C on their pathogenicity to artificially inoculated white onion bulbs, and the effect of conidial concentration (5×10³ and 5×10⁴ conidia/mL) on disease incidence, lesion area, incubation and latent period during storage at 20, 5 and 0°C. At all storage temperatures and periods tested conidia produced at −2°C caused a higher disease incidence and larger areas of rot than those produced at higher temperatures. When the conidial production temperature was raised to 20°C, the duration of incubation on the bulbs inoculated with 5×10⁴ conidia/mL was more than doubled during storage at 0°C, tripled at 5°C, and took 50% longer at 20°C. The incubation period was not significantly affected by conidial concentration at 20°C, and only slightly at 5 and 0°C, but at low temperatures the latent period was longer because of the delay induced in sporulation. These data are consistent with the packers' opinion that cross-infection of spring onions by long-term refrigerated onions in grading lines caused earlier and heavier rotting.     

Effects of temperature and wetness duration on conidial infection, latent period and asexual sporulation of Pyrenopeziza brassicae on leaves of oilseed rape

Experiments in controlled environments were carried out to determine the effects of temperature and leaf wetness duration on infection of oilseed rape leaves by conidia of the light leaf spot pathogen, Pyrenopeziza brassicae . Visible spore pustules developed on leaves of cv. Bristol inoculated with P. brassicae conidia at temperatures from 4 to 20°C, but not at 24°C; spore pustules developed when the leaf wetness duration after inoculation was longer than or equal to approximately 6 h at 12–20°C, 10 h at 8°C, 16 h at 6°C or 24 h at 4°C. On leaves of cvs. Capricorn or Cobra, light leaf spot symptoms developed at 8 and 16°C when the leaf wetness duration after inoculation was greater than 3 or 24 h, respectively. The latent period (the time period from inoculation to first spore pustules) of P. brassicae on cv. Bristol was, on average, approximately 10 days at 16°C when leaf wetness duration was 24 h, and increased to approximately 12 days as temperature increased to 20°C and to 26 days as temperature decreased to 4°C. At 8°C, an increase in leaf wetness duration from 10 to 72 h decreased the latent period from approximately 25 to 16 days; at 6°C, an increase in leaf wetness duration from 16 to 72 h decreased the latent period from approximately 23 to 17 days. The numbers of conidia produced were greatest at 12–16°C, and decreased as temperature decreased to 8°C or increased to 20°C. At temperatures from 8 to 20°C, an increase in leaf wetness duration from 6 to 24 h increased the production of conidia. There were linear relationships between the number of conidia produced on a leaf and the proportion of the leaf area covered by 'lesions' (both log₁₀-transformed) at different temperatures.     

The effect of temperature and water potential on the production of conidia by sclerotia of Botrytis squamosa

The rate of conidiogenic germination of Botrytis squamosa was highest at 16°C and the greatest numbers of conidia per sclerotium (up to 5 × 10⁴) were produced at temperatures of 5–10°C. At temperatures above 20°C, the percentage of sclerotia producing conidia declined rapidly. Decreasing water potential reduced the rate at which conidia were produced and also resulted in fewer conidia produced per sclerotium. However, conidia were produced at water potentials as low as −2 MPa, at which sclerotial germination was at least 60%. A simulation model that included effects of both temperature and water potential was developed from laboratory and field data obtained for conidial production in sclerotia exposed for periods of 1, 2, 3 or 4 weeks during an entire year. There was good agreement between conidiogenic germination predicted by the model and conidial production observed in onion plots artificially inoculated with sclerotia. Temperature and water potential were therefore considered to be the principal microclimatic factors affecting conidial production by B. squamosa. The role of sclerotia in the context of UK onion production is discussed.     

Effects of humidity, leaf wetness, temperature and light on conidial production by Phaeoisariopsis personata on groundnut

Controlled-environment studies of conidial production by Phaeoisariopsis personata on groundnut are described. With constant relative humidity (RH), conidia were only produced above a threshold (94·5% RH) and there was a linear increase between 94·5% RH and 100% RH. Conidial production was less with continuous leaf wetness (resembling heavy dew) than with continuous 98–99% RH, but it was similar with intermittent leaf wetness and intermittent 98–99% RH (8 h at 70% RH each day). With alternate high (≥97% RH) and low humidity, daily conidial production depended both on the duration of high RH and on the low RH value. With 99% RH at night (12 h), night-time conidial production decreased with the previous daytime RH. After conidial production had started, small numbers of conidia were produced even when the RH was well below the threshold (94·5%). Conidia were produced in continuous light when the photon flux density was 2 μmol/m²/s, but production was completely inhibited with 60 μmol/m²/s. With constant RH, more conidia were produced with a 12 h photoperiod than in continuous darkness. However, more than 75% of the conidia were produced in the dark. With continuous darkness, more conidia were produced during the night (18.00–06.00 h) than during the day, but this biological rhythm was overcome with a (light-night)/(dark-day) regime. With constant 98–99% RH there was a linear increase in conidial production with temperature between 10 and 28°C, and virtually no conidia were produced at 33°C. The daily production of conidia increased with time for 2 to 6 days, depending on the treatment.     

Factors influencing infection of eucalypts by Cylindrocladium pteridis

The pattern of Cylindrocladium pteridis adhesion, germination and penetration in eucalypt leaves was assessed using scanning electron microscopy. The effects of inoculum concentration, leaf wetness period, plant age and branch position of cylindrocladium leaf blight and defoliation severity were assessed in greenhouse studies using two Eucalyptus grandis × E. urophylla hybrid clones. Penetration occurred through stomata, and there was no difference in the number of penetrations between young and old leaves. Percentage leaf area with lesions and defoliation increased with the increase in inoculum concentration (1 × 10² to 10⁵ conidia mL⁻¹), duration of leaf wetness period (6 to 48 h) and plant age (60 to 180 days). Branch position in plants also significantly affected the percentage leaf area with lesions and defoliation, the latter variable being significantly higher at the stem base. The highest values of lesion area were also observed on leaves at the stem base in both clones. The Pearson correlation between defoliation and leaf area with lesions was significant in all experiments ( r  > 0·9) indicating a high association between these two variables.     

Effects of humidity and air speed on sporulation of Phytophthora infestans on potato leaves

An apparatus is described for the control of humidity, air speed and temperature around detached potato leaflets and whole plants. Leaflets and plants inoculated with Phytophthora infestans were incubated in air at 15°C. Abundant sporangia were formed in an air speed of 0.3 × 10⁻³ m/s when the ambient humidity was 90–100% RH, but not at 85 or 80% RH. At air speeds of 5.5 × 10⁻³ and 13.7 × 10⁻³ m/s there were numerous sporangia at 100% RH, but not at 95–80% RH. The number of sporangia formed on leaflets kept in air flowing at 5.5 × 10⁻³ m/s with alternating humidities of 80 and 100% RH each for 12 h per day was intermediate between numbers formed on leaflets incubated at each humidity continuously, and higher than numbers formed at a constant 90% RH. Removing leaflets from plants before inoculation did not affect the number of sporangia produced.     

Visual and PCR assessment of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) cultivars

Methods to assess light leaf spot ( Pyrenopeziza brassicae ) on winter oilseed rape cultivars were compared in laboratory, controlled-environment and field experiments. In controlled-environment experiments with seedling leaves inoculated at GS 1,4, the greatest differences in percentage area affected by P. brassicae sporulation were observed with inoculum concentrations of 4 × 10³ or 4 × 10⁴ spores mL⁻¹, rather than 4 × 10² or 4 × 10⁵ spores mL⁻¹, but older leaves had begun to senesce before assessment, particularly where they were severely affected by P. brassicae . In winter oilseed rape field experiments, a severe light leaf spot epidemic developed in 2002/03 (inoculated, September/October rainfall 127·2 mm) but not in 2003/04 (uninoculated, September/October rainfall 40·7 mm). In-plot assessments discriminated between cultivars best in February/March in 2003 and June in 2004, but sometimes failed to detect plots with many infected plants (e.g. March/April 2004). Ranking of cultivar resistance differed between seedling experiments done under controlled-environment conditions and field experiments. The sensitivity of detection of P. brassicae DNA extracted from culture was greater using the PCR primer pair PbITSF/PbITSR than using primers Pb1/Pb2. P. brassicae was detected by PCR (PbITS primers) in leaves from controlled-environment experiments immediately and up to 14 days after inoculation, and in leaves sampled from field experiments 2 months before detection by visual assessment.     

Infection of onion leaves by Alternaria porri and Stemphylium vesicarium and disease development in controlled environments

Infection of onion by Alternaria porri and Stemphylium vesicarium was investigated under a range of controlled temperatures (4–25°C) and leaf wetness periods (0–24 h). Conidia of A. porri and S. vesicarium germinated within 2 h when incubated at 4°C. Terminal and intercalary appressoria were produced at similar frequencies at or above 10°C. The maximum number of appressoria was produced after 24 h at 25°C. Penetration of leaves by both pathogens was via the epidermis and stomata, but the frequency of stomatal penetration exceeded that of epidermal penetration. There was a strong correlation ( R ² > 90%) between appressorium formation and total penetrations at all temperatures. Infection of onion leaves occurred after 16 h of leaf wetness at 15°C and 8 h of leaf wetness at 10–25°C, and infection increased with increasing leaf wetness duration to 24 h at all temperatures. Interruption of a single or double leaf wetness period by a dry period of 4–24 h had little effect on lesion numbers. Conidia of A. porri and S. vesicarium separately or in mixtures caused similar numbers of lesions. Alternaria porri and S. vesicarium are both potentially important pathogens in winter-grown Allium crops and purple leaf blotch symptoms were considered to be a complex caused by both pathogens.     

Differential susceptibility of barnyard grass (Echinochloa crus-galli var. crus-galli) ecotypes to Exserohilum longirostratum

Ten barnyard grass ( Echinochloa crus-galli var. crus-galli ) ecotypes collected from several locations in Malaysian and Indonesian granaries were tested for variation in their susceptibility to the leaf blight pathogen ( Exserohilum longirostratum ). Four phenological growth stages of the ecotypes were sprayed with E. longirostratum at the rate of 1 × 10⁷ conidia mL⁻¹ under glasshouse conditions. The results of the study showed that plants inoculated with 1 × 10⁷ conidia mL⁻¹ and exposed for 24 h dew periods were severely infected and the percentage dry weight reduction of all ecotypes significantly increased. The 1–2 and 2–3-leaf stage plants were completely killed within 6 days after inoculation. However, susceptibility reactions among the ecotypes were observed at the 4–5-leaf stage, while the 6–7-leaf stage was resistant. The ecotypes, K-02, PK-04, KN-02, CJ-01, and L-01, were the most susceptible ecotypes as indicated by a higher area under disease progress curve value. The apparent disease progress rates for these ecotypes were significantly faster compared to the other ecotypes. The dry weight reduction among the four growth stages was variable. These findings show that the 1–2 and 2–3-leaf stages are the most susceptible, while the 6–7-leaf stage was resistant. This study has confirmed that a variable response to the fungal pathogen occurred within an intraspecific barnyard grass collection.     

Biological control of damping-off of sugar beet by Pseudomonas putida applied to seed pellets

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping-off of sugar beet. A density of 6 × 10⁷ 40RNF per pellet reduced Pythium damping-off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 10⁴–6 × 10⁸ per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 10⁶ per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping-off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post-emergence damping-off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).     

Bioassays of glucosinolate-derived isothiocyanates against postharvest pear pathogens

The present study assayed the effect of six isothiocyanates (ITCs), produced by the enzymatic hydrolysis of glucosinolates, on fungal pathogens of pear. Sample pear fruits were artificially inoculated through induced wounds with conidial suspensions of Botrytis cinerea Rhizopus stolonifer Monilinia laxa Mucor piriformis or Penicillium expansum and were then treated with ITCs. Of the six ITCs tested, the ITC from glucoraphenin showed the highest effectiveness after 6 days at 20°C, against M. laxa B. cinerea and M. piriformis . The effectiveness of the ITC from glucoraphenin against M. laxa was assayed in two further trials to test the effect of ITC concentration on different concentrations of inoculum and to determine the duration of the curative effect of this ITC. ITC concentration directly affected fungus control capacity. The highest ITC concentration (3.6 mg mL⁻¹) afforded pathogen control at the highest level of pathogen concentration (10⁶ conidia mL⁻¹) after 6 days at 20°C. Its curative effect was evident up to 40 h after inoculation.     

Development of a spot plow providing complete inversion for effective weed control

Tillage for the "complete inversion" of soil, that is, overturning soil slices 180° was proposed, a "spot plow" was developed and tested to accomplish the task, and a simulation model was evaluated to demonstrate the efficacy of the plow on weed control. A 360 mm wide spot plow was designed to operate at a speed of 1.9 m s⁻¹ for the spot plowing with the least possible lateral displacement of the soil slice by utilizing the inertia of the soil slice and securely rotating it. In field experiments, complete spot inversion required an operating speed of at least 1.6 m s⁻¹; at lower speeds, a portion of the soil block was left half-inverted and further lowering led to considerable lateral displacement. The displacement in the forward and lateral directions was minimal, implying that spot plowing is suitable for potential application to and verification of the weed population dynamics model in the field. A simple linear matrix model of the population dynamics of annual weeds was proposed, whereby four layers of soil were set to describe tillage and other ecological events. The effect of tillage on weed control was evaluated by the equilibrium reproduction rate allowed to sustain a stable population of weeds. The simulation model showed that alternately changing the depth of spot plowing had a significant effect on controlling weeds of low-survival-rate seeds, even when some incomplete inversion of the soil slice was taken into account.     

PCR-based specific and sensitive detection of Pectobacterium carotovorum ssp. carotovorum by primers generated from a URP-PCR fingerprinting-derived polymorphic band

A 24-mer primer pair was generated by sequencing a URP-PCR fingerprinting-derived polymorphic band that is uniquely shared in Pectobacterium carotovorum ssp . carotovorum strains (Pcc). The primer set (EXPCCF/EXPCCR) amplified a single band of expected size (0·55 kb) from genomic DNA obtained from 29 Pcc strains and three Pectobacterium carotovorum ssp. wasabiae (Pcw) strains, but not from other P. carotovorum subspecies atrosepticum , betavasculorum or odoriferum , or from other Erwinia spp. or bacterial genera. The Rsa I digestion profile of the amplified bands divided Pcc strains into five groups with a unique profile from Pcw strains. First-round PCR detected between 5 × 10² and 1 × 10³ colony forming units (CFU) mL⁻¹ and detection sensitivity was increased to as few as 2–4 CFU mL⁻¹ after second-round (nested) PCR. This PCR protocol was used directly to detect Pcc strains in infected plant tissues.     

Factors influencing the production of basidiocarps and the deposition and germination of basidiospores of Crinipellis perniciosa, the causal fungus of witches' broom on cocoa (Theobroma cacao)

The production of basidiocarps by Crinipellis perniciosa on detached, dead witches'brooms from cocoa was assessed in relation to temperature, light, cocoa clone, age of broom and type of tissue, in cabinets with a daily cycle of 8 h wet and 16 h dry. More basidiocarps formed and matured at 20–25°C than at 25–30°C. In the latter regime the pilei were smaller and white, instead of the usual crimson colour, and the stipes were longer. No basidiocarps formed at 30–35°C. At 20–25°C. more basidiocarps formed and matured with light at 100 μE m^-2 s^-1 during the wet period than at 10 μE m^-2 s ^-1. Only one basidiocarp and five primordia developed on 20 brooms kept in the dark. Brooms from 10 cocoa clones at Pichilingue. Ecuador, differed in basidiocarp productivity. most basidiocarps forming on brooms from Seavina and least on ICS clones. The numbers of basidiocarps produced on brooms aged 1.2.3 or 4 months when detached from cocoa trees were similar but time to initiation of the first primordium differed considerably. More basidiocarps formed at nodes than internodes.
The discharge of basidiospores was optimal at 20–25°C and 80% RH: germination was optimal in water agar films. Neither process was dependent on light.     

Antifungal vapour-phase activity of allyl-isothiocyanate against Penicillium expansum on pears

The fungitoxicity of allyl-isothiocyanate (AITC) vapour against Penicillium expansum , the causal agent of blue mould on pears (cvs Conference and Kaiser), was evaluated. The best control of blue mould was obtained by exposing fruits for 24 h in a 5 mg L⁻¹ AITC-enriched atmosphere, the extent of control depending on the inoculum density. Lesion diameter was inversely related to AITC concentration. In treated fruits the percentage of infected wounds increased with conidial concentration, with fewer than 20% affected at 1 × 10³ conidia mL⁻¹ to almost 80% at 1 × 10⁶ conidia mL⁻¹. In comparison, >98% of wounds were infected in untreated fruits irrespective of conidial concentration. AITC treatments were effective up to 24 h after inoculation for Conference and 48 h for Kaiser. AITC treatments also controlled a thiabendazole-resistant strain of P. expansum , reducing the incidence of blue mould by 90% in both cultivars. The use of AITC produced from pure sinigrin or from Brassica juncea defatted meal may be an economically viable alternative to synthetic fungicides against P. expansum .     

An apparatus for collecting total conidia of Blumeria graminis f.sp. hordei from leaf colonies using electrostatic attraction

Conidia from living conidiophores of barley powdery mildew ( Blumeria graminis f.sp. hordei ) on host leaves were collected consecutively using an electrostatic spore collector. The collector consisted of an electrical conductor plate linked to an electrostatic voltage generator and insulator plates placed abreast on a timed conveyer. The conductor plate was negatively charged by the potential supplied from the voltage generator. The negatively charged conductor plate caused dielectric polarization of the insulator plate, and the surface charge on the insulator plate attracted mature conidia abstricted from conidiophores on colonies growing on leaves placed 2 cm from the insulator plate. The surface charge on the insulator plate was proportional to the voltage applied to the conductor plate. Under optimized conditions, abstricted conidia were attracted to the electrostatically activated insulator plates without any detriment to their survival. During a colony's life span of c . 460 h, conidia were released throughout the day and c . 12 × 10⁴ conidia were collected during the lifetime of the colony. This is the first report on the direct quantification of progeny conidia produced by powdery mildew infecting host leaves.     

Differences in cerato-ulmin production between the EAN, NAN and non-aggressive subgroups of Ophiostoma ulmi

A test of comparative in vitro cerato-ulmin wilt toxin production in the aggressive and non-aggressive subgroups of the Dutch elm disease pathogen Ophiostoma ulmi was carried out by turbidity and ELISA methods. Ten non-aggressive, ten EAN aggressive and ten NAN aggressive isolates were tested from a range of geographical sources. In liquid shake cultures the non-aggressive isolates produced the greatest and the NAN aggressives the least mean biomass. Despite considerable variation in cerato-ulmin production by individual isolates in three separate experiments, both the turbidity and ELISA methods showed a clear separation of the non-aggressive and aggressive subgroups. Non-aggressive isolates produced little or no cerato-ulmin (ELISA range of means 0–56.0 ng/ml) and EAN and NAN aggressive isolates moderate to high levels (EAN 1.6–89.0 × 10⁴ ng/ml and NAN 0.2–300 × 10⁴ ng/ml). In the aggressive isolates no correlation was detected between cerato-ulmin production and either biomass or pathogenicity to clonal Commelin elm. The role of cerato-ulmin in the pathogenicity of O. ulmi is discussed.