Effects of Temperature and Wetness Duration on Infection of Oilseed Rape Leaves by Ascospores of Leptosphaeria maculans (Stem Canker)

In controlled environment experiments, ascospores of Leptosphaeria maculans (stem canker) infected oilseed rape (cv. Nickel) leaves and caused phoma leaf spots at temperatures from 8°C to 24°C and leaf wetness durations from 8 h to 72 h. The conditions that produced the greatest numbers of leaf spot lesions were a leaf wetness duration of 48 h at 20°C; numbers of lesions decreased with decreasing leaf wetness duration and increasing or decreasing temperature. At 20°C with 48 h of leaf wetness, it was estimated that one out of four spores infected leaves to cause a lesion whereas with 8 h of leaf wetness only one out of 300 spores caused a lesion. As temperature increased from 8°C to 20°C, the time from inoculation to the appearance of the first lesions (a measure of the incubation period) decreased from 15 to 5 days but leaf wetness duration affected the length of the incubation period only at sub-optimal temperatures. Analyses suggested that, within the optimal ranges, there was little effect of temperature or wetness duration on incubation period expressed as degree-days; the time until appearance of 50% of the lesions was ca. 145 degree-days. A linear regression of % leaves with lesions (P_l) (square-root transformed) on % plants with lesions (P_p) accounted for 93% of the variance: P_l=1.31+0.061P_p. This relationship was also investigated in winter oilseed rape field experiments in unsprayed plots from October to April in 1995/96 (cv. Envol), 1996/97 (cv. Envol), 1997/98 (cvs Bristol and Capitol) and 1998/99 (cvs Apex, Bristol and Capitol) seasons. The linear regression of % leaves with lesions (square-root transformed) on % plants with lesions accounted for 90% of the variance and had a similar slope to the controlled environment relationship: P_l=0.81+0.051P_p. These results were used to examine relationships between the development of phoma leaf spot on plants in winter oilseed rape crops, the incubation period of L. maculans and the occurrence of infection criteria (temperature, rainfall) in the autumns of 1996, 1997 and 1998.     

Molecular Characterisation of Alternaria linicola and its Detection in Linseed

Nucleotide sequences of the ribosomal DNA (rDNA) internal transcribed spacers (ITS) 1 and 2 and a 1068bp section of the beta-tubulin gene divided seven designated species of Alternaria into five taxa. Stemphylium botryosum formed a sixth closely related taxon. Isolates of A. linicola possessed an identical ITS sequence to one group of A. solani isolates, and two clusters of A. linicola isolates, revealed from beta-tubulin gene data to show minor variation, were as genetically similar to isolates of A. solani as they were to each other. We suggest, therefore, that A. linicola falls within the species A. solani. Similar results suggest that A. lini falls within the species A. alternata. RAPD analysis of the total genomic DNA from the Alternaria spp. concurred with the nucleotide sequence analyses. An oligonucleotide primer (ALP) was selected from the rDNA ITS1 region of A. linicola/A. solani. PCR with primers ALP and ITS4 (from a conserved region of the rDNA) amplified a c. 536bp fragment from isolates of A. linicola and A. solani but not from other Alternaria spp. nor from other fungi which may be associated with linseed. These primers amplified an identical fragment, confirmed by Southern hybridization, from DNA released from infected linseed seed and leaf tissues. These primers have the potential to be used also for the detection of A. solani in host tissues.     

Factors affecting the incubation period of dark leaf and pod spot (Alternaria brassicae) on oilseed rape (Brassica napus)

Experiments to investigate the factors affecting the incubation period of dark leaf and pod spot (Alternaria brassicae) on leaves and pods of oilseed rape (Brassica napus) were done in controlled environment (constant temperatures) and glasshouse conditions (fluctuating temperatures). The length of the incubation period of dark leaf and pod spot decreased as infection and incubation temperatures increased from 6 to 20 °C. The incubation period decreased as wetness period increased from 2 to 12 h, as inoculum concentration increased from 80 to 2 × 10³ spores ml^–1 and as leaf age increased from 4 to 10 days. Asymptotes of leaf age and inoculum concentration, above which the length of the incubation period did not decrease, were 10 days and 2 × 10³ spores ml^–1, respectively. The shortest and longest incubation periods were 1 and 11 days. The mechanism by which the infection conditions influenced the incubation period of dark leaf and pod spot on oilseed rape seemed to be linked to lesion density. Usually, the length of the incubation period decreased greatly with increasing lesion density.     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Biological characteristics and life tables of Neoseiulus umbraticusChant (Acari, Phytoseiidae) at three constant temperatures

The development time, survival and fecundity of the generalist predatory mite, Neoseiulus umbraticusChant, were determined at 20, 25, and 30°C and 65±10% RH. N. umbraticus females completed development in 9.7, 8.0 and 5.9 days, respectively, using a diet of all life stages of Tetranychus cinnabarinus Boisduval. Total developmental times of males were relatively shorter at 25 and 30°C than at 20°C. In general, preoviposition, oviposition, and postoviposition periods of N. umbraticus shortened as temperature increased. The longest survival rate of N. umbraticus of 80.5 days occurred at 20°C, followed by 67.0 and 57.6 days at 25 and 30°C, respectively. Mated females laid an average 0.9, 1.3 and 1.4 eggs per female per day and 33.1, 44.0 and 43.6 eggs over their entire lives at 20, 25 and 30°C, respectively. The sex ratios of this species were 0.57, 0.57 and 0.54 female (female+male) at 20, 25 and 30°C, respectively. The intrinsic rate of increase (r_m) became greater with rising temperatures from 0.123 at 20°C to 0.180 at 30°C. The net reproduction rate (R_o) was highest at 25°C (25.0 females/female) and lowest at 20°C (18.8 females/female), while T_o decreased with increasing temperatures, from 23.8 days at 20°C to 17.5 days at 30°C.     

Effects of light conditions on prodigiosin stability in the biocontrol bacterium Serratia marcescens strain B2

Serratia marcescens strain B2 is a potential biocontrol agent that produces the antibiotic pigment prodigiosin. When this strain was incubated under white or blue light conditions (<100µmol m^–2s^–1), prodigiosin concentration in bacterial cells decreased, but growth did not. However, red and far-red light had no effect on prodigiosin concentration in bacterial cells. Purified prodigiosin was degraded under white or blue light conditions but was not degraded under red and far-red light. Because white and blue light appeared to affect the stability of prodigiosin itself, light conditions may affect the suppressive effects of the biocontrol agent S. marcescens strain B2.     

Germination of Alternaria linicola conidia on linseed: effects of temperature, incubation time, leaf wetness and light regime

Conidia of Alternaria linicola germinated on both water agar and linseed leaves (detached or attached) over a wide range of temperatures (5–25°C) by producing one to several germ tubes. At temperatures between 10°C and 25°C and under continuous wetness in darkness, germination started within 2 h after inoculation and reached a maximum (100%) by 8 to 24 h, depending on temperature. At 5°C, the onset of germination was later and the rate of germ tube elongation was slower than that at 10–25°C. During germination, conidia of A. linicola were sensitive to dry interruptions of wet periods and to light. Short (2 h) or long (12 h) dry interruptions occurring at any time between 2 and 6 h after inoculation stopped conidial germination and germ tube elongation. With continuous wetness, light periods 2 to 12 h long immediately after inoculation inhibited conidial germination, which was resumed only when a dark period followed subsequently. However, germination and germ tube elongation of A. linicola conidia stopped and the viability of the conidia was lost during exposure to dry light periods immediately after inoculation with spore suspensions. Penetration of leaves by A. linicola was evident after 12 h and occurred mainly through epidermal cells (direct) with or without the formation of appressoria.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

The Epidemiology of Purple Leaf Blotch on Leeks in Victoria, Australia

The incidence of purple leaf blotch disease was investigated on seven successive commercial leek crops grown at Cranbourne, Victoria between 1996 and 1997. First symptoms occurred on older leaves, 54–69 days after transplanting. Lesions with typical symptoms were colonised by either Alternaria porri (6%), Stemphylium vesicarium (42%) or mixtures of both pathogens (52%). Purple leaf blotch was caused by a disease complex and was endemic at nobreak Cranbourne due to the continuous cropping of leeks. Disease incidence in all monitored crops increased as plants matured (123–158 days after transplanting) until harvest but never exceeded 11% due to fortnightly applications of mancozeb. Disease levels showed no significant correlation with weekly temperature, precipitation, relative humidity or leaf wetness duration. Disease levels were significantly (P < 0.05) higher on autumn/winter (May/June) 1997 crops when 38 periods of leaf wetness 8 h because of dew and low temperatures (10–13 °C). The weekly rate of increase of disease incidence was significantly (P < 0.01) correlated with days after transplantation. nobreak Concentrations of airborne A. porri and S. vesicarium conidia within leek crops showed a diurnal periodicity and maximum numbers were trapped between 11:00 and 15:00 h. The concentration of airborne S. vesicarium conidia was three to six times the concentration of airborne A. porri conidia. Conidia were more abundant during spring/summer (September–February). Ascospores of Pleospora allii were found during May–September. The greater concentrations of airborne S. vesicarium conidia suggest that it may be the dominant pathogen in the purple leaf blight complex. Fungicide sprays were unnecessary until 8–10 weeks after transplanting, and regular protectant sprays curtailed but did not eradicate purple leaf blight. The results indicated that predictive models, based on temperature and the frequency of leaf wetness periods 8 h, will assist in reducing fungicide inputs as plants mature and, in southern Victoria, fungicide applications on leeks should be timed for autumn/winter when infection periods occur.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Gamma radiation sensitivity of larvae and adults of the red flour beetle, Tribolium castaneum Herbst

In this study, the susceptibility of larvae and adults of Tribolium castaneum (Herbst) to gamma radiation was investigated in the laboratory in Turkey. Gamma radiation was applied at 6 dose levels between 20 and 200Gy to 13–15day-old adults and at 5 dose levels between 20 and 180Gy to 18–20day-old larvae of T. castaneum. All experiments were performed in growth chambers maintained at 27±1°C and 70±5% r.h. The larvae proved to be the more susceptible stage, survival to the adult stage being prevented by exposure of the eggs to 100Gy; the adult stage was less susceptible. LD₅₀ and LD₉₉ values were determined as 19,75 and 42,97Gy for larvae and 33,21 and 64,50Gy for adult stage, respectively. It was concluded that 100Gy is the effective dose for both larval and adult stages.     

Germination and invasion by ascospores and pycnidiospores of <Emphasis Type="Italic">Leptosphaeria maculans</Emphasis> on spring-type <Emphasis Type="Italic">Brassica napus</Emphasis> canola varieties with varying susceptibility to blackleg

The infection processes of ascospores and pycnidiospores of Leptosphaeria maculans were studied on cotyledons of six cultivars of spring-type Brassica napus: one with resistance controlled by a single dominant gene (cv. Surpass 400), three with polygenic resistance (cvs. Dunkeld, Grouse, and Outback), and two susceptible cultivars (Westar and Q2). On all cultivars, ascospore germination, penetration, and development of symptoms on cotyledons were much earlier than that with pycnidiospores. At 2h after inoculation ascospores began to germinate, by 4h about 50% had germinated, and by 6–8h 85%–90% had germinated. In contrast, pycnidiospores began to germinate 1 day after inoculation (dai) and reached only 50% germination by 3 dai. Ascospores began germinating from terminal cells and then later from the interstitial cells. Pycnidiospores germinated predominantly from one end and sometimes from both ends. Germ tubes from ascospores penetrated stomata as early as 4h after inoculation, whereas those from pycnidiospores penetrated at 2 dai. Symptom development with ascospores was 2 days earlier than that with pycnidiospores. Symptoms on Surpass 400 were evident as early as 3–5 dai with ascospores and 5–7 dai with pycnidiospores. However, on other cultivars, symptoms were not evident until 10 dai with ascospores and 12 dai with pycnidiospores. This report is the first on differences in the infection processes by the two spore types. Ascospore and pycnidiospore attachment, germination, and penetration did not differ between resistant and susceptible cultivars, but there were major differences after penetration. Under high humidity, 80%–90% of stomata of susceptible Westar and Q2 had aerial hyphae emerging from stomatal pores. However, fewer stomata (5%–10%) had aerial hyphae on Surpass 400 by 10 dai with ascospores and 12 dai with pycnidiospores, but even these were usually poorly developed. Host differences in spring-type B. napus in relation to production of aerial hyphae have not previously been reported. In Surpass 400, rapid necrosis of guard cells occurred within a few hours of penetration by either type of spore, and subsequently one or a few cells immediately adjacent to the penetration site died. This necrosis then spread to the cells around the penetration site to form a hypersensitive response (in the form of a small, dark lesion) to both ascospores and pycnidiospores. This is the first detailed report on interactions between spring-type B. napus and L. maculans in relation to single dominant gene-based resistance. Neither the cultivars with polygenic resistance nor the susceptible cultivars had such a response.     

Morphological studies on attachment of spores of <Emphasis Type="Italic">Magnaporthe grisea</Emphasis> to the leaf surface of rice

Rice leaves were inoculated with spores of Magnaporthe grisea, and the number of fluorescence-labeled spores that attached to the leaf surface were counted before and after leaves were dipped and then stirred in water. Just 5% of the spores were retained on the leaf surface 1h after inoculation; the percentage retained then increased rapidly between 1.25 and 1.50h, and most had attached by 2h. Scanning electron microscopy revealed that most conidia were lying on a few wart-like protuberances 2–4µm high. Spores became attached when the germ tubes were long enough to reach the leaf surface, at least 3µm, by mucilaginous substances at the tip. Retained spores swayed when water was added under the cover glass from one side, indicating that the attachment was confined to the tips of germ tubes. Spores are attached to the rough leaf surface by mucilaginous substances – not at the tip of spore as reported on smooth artificial substrates but at the tip of the germ tubes.     

Pathogenic variation in poplar rust <Emphasis Type="Italic">Melampsora larici-populina</Emphasis> from England

Using a leaf disc method, 19 isolates of the poplar rust, Melampsora larici-populina , and one isolate of M.populnea from England were inoculated on to 25 poplar clones belonging to Populus nigra and P.trichocarpa, and hybrids between P. deltoides and P. nigra, P. deltoidesand P. trichocarpa, P.tacamahaca and P.trichocarpa, and P. alba and P. tremula. Disease was scored based on the pustule area and inoculum density. In terms of whether sporulating uredinia formed, the 19 isolates showed seven different patterns to the tested poplar clones. The majority of the rust isolates infected P. nigra P3090 and Vereecken, P.nigra×P. deltoides Casale and Tasman, P. tacamahaca×trichocarpa 36 and Balsam Spire, and P.trichocarpa Blom. Populus trichocarpa×P. deltoides 69039/4 was infected by only three isolates collected from southern England. No visible symptoms appeared on P. alba ×P. tremulaTower and P.trichcarpa×P. deltoides×P. deltoides76028/5 in inoculations with M. larici-populina isolates. Populus alba×P.tremula Tower was infected only by M. populnea. When M. larici-populina isolates were tested using AFLP, no differences were found either between isolates from different geographical regions or between those having narrow spectrum of virulence and those showing wide spectrum of virulence on the tested clones. The results suggest that the UK rust populations possess virulences which were found in races E1, E2, E3 and E4 in continental Europe and that rust having virulence patterns similar to race E4 has occurred in UK poplar plantations since 1996.     

Environmental factors influencing sporocarp formation in <Emphasis Type="Italic">Typhula ishikariensis</Emphasis>

Environmental factors influencing sporocarp formation in Typhula ishikariensis were studied under controlled conditions. Sporocarp formation in T. ishikariensis was divided into two stages: stipe elongation from the sclerotium and fertile head development at the tip of the stipe. Factors required for each stage differed. At the stipe elongation stage, low temperature (10°/5°C; day/night) and high humidity were important, but light was not required. In contrast, at the fertile head stage, light and moderate day length (8h/day) were essential. Fertile heads developed at 46µEm^–2s^–1; and high intensity (137µEm^–2s^–1) did not suppress development. Moreover, adding unsterilized soil to the sea sand medium accelerated sporocarp formation. These findings imply that the sclerotium of T. ishikariensis recognizes several physical factors for sporocarp formation. Sporocarps of T. ishikariensis developed within 4 weeks after incubation under optimal conditions. The sporocarp produced basidiospores, and differential mating incompatibility was confirmed among monokaryons derived from basidiospores produced under artificial conditions. This method should be useful for obtaining monokaryons for genetic studies of T. ishikariensis.     

Fusaric Acid Production by Fusarium Oxysporum f.sp. lilii and its Role in the Lily Basal Rot Disease

Fusarium oxysporum f.sp. lilii (Fol), the causal agent of lily basal rot, produced fusaric acid (FA) in aseptic culture. This toxin induced phytotoxicity symptoms on in vitro-grown lily bulblets of two different cultivars: the Fol-susceptible cultivar was more sensitive to the toxin than the Fol-resistant cultivar. When cultured in the presence of FA, the Fol-susceptible cultivar showed a greater tendency to accumulate FA within its tissues than the Fol-resistant cultivar. The polyphenol oxidase activity of the bulblets was inhibited by 1mmolL^–1 FA in both the cultivars, while at lower FA concentrations the enzyme activity increased only in the Fol-susceptible cultivar. Peroxidase showed a steady activity at the 1mmolL^–1 FA concentration in both the cultivars, while at lower FA dosages its activity increased. Within the Fol-infected in vivo tissues of both the lily cultivars, FA was detectable only in traces. The role of this toxin in the lily basal rot disease seems therefore to be of marginal importance.     

The Effect of Soil Moisture and Cabbage Amendment on the Thermoinactivation of Phytophthora nicotianae

The analysis of the effect of soil water matric potential and temperature regimes on the inactivation of chlamydospores of Phytophthora nicotianae in cabbage amended soils was evaluated using three matric potentials (0, -10, and -30kPa), temperature regimes of 1.5h at 44°C, 5h at 41°C and 8h at 35°C, or 3h at 47°C, 5h at 44°C and 8h at 35°C, with a baseline temperature of 25°C during the rest of the day. The results indicated that survival of P. nicotianae was lowest in saturated soil; and as temperature increased, survival of the pathogen decreased at all soil water matric potentials evaluated. Cabbage amendments can enhance the effect of the heat treatment, further decreasing the pathogen population. The soil water matric potentials evaluated represent optimum levels for the study of thermal inactivation. However, under field conditions lower potentials may be found. Extending the range of soil water matric potentials and the treatment time would allow better comparisons with the field data. There is a clear indication that one irrigation period prior to solarization would provide enough moisture to inactivate the primary inoculum of P. nicotianae in the top soil under field conditions; however, other factors may affect the effectiveness of solarization, reducing or enhancing its potential.     

Three distinct groups of isolates of <Emphasis Type="Italic">Tomato yellow leaf curl virus</Emphasis> in Japan and construction of an infectious clone

Complete nucleotide sequences of eight Japanese isolates of Tomato yellow leaf curl virus (TYLCV) were determined and compared with four TYLCV isolates already reported. These isolates separated into three groups – Shizuoka (Sz), Aichi (Ai), Nagasaki (Ng) – and had 99% identities within the groups. Full-length molecules of DNA-A of group Sz consist of 2791nt and those of group Ai contain 2787nt. Both were closely related to TYLCV-Is.M, although those of group Ng had 2793nt and were more closely related to TYLCV-Is. Comparison of common sequences of isolates belonging to groups Sz and Ai had substitutions of 4nt in the intergenic region and nonsynonymous substitutions at open reading frames between the groups. None of the isolates tested had DNA molecules. Agroinfection of four plant species with a DNA-A dimeric infectious clone of TYLCV-SzY, a member of group Sz, resulted in systemic infection. Tomato plants then developed typical yellow leaf curl symptoms.The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB116629, AB116630, AB116631, AB116632, AB116633, AB116634, AB116635, and AB116636     

Environmental conditions influencing Sclerotinia sclerotiorum infection and disease development in lettuce

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.     

Effects of interrupted wet periods and different temperatures on the development of ascochyta blight caused by Mycosphaerella pinodes on pea (Pisum sativum) seedlings

The effect of interrupted wet periods on pycnidiospores of Mycosphaerella pinodes was studied by assessing spore viability, infection and disease development on pea seedlings. Pycnidiospores survived dry periods of up to 21 days after inoculation. Rewetting restored the infection capacity of the pycnidiospore, resulting in high levels of disease. The effects of wet–dry–wet cycles depended on when the dry period occurred during the infection process. No disease symptoms appeared when dry periods occurred during germination. A low level of disease occurred after rewetting in high relative humidity if the interruption of the wet period was long. However, a wet period resulting in leaf wetness after a dry period gave similar levels of infection to those achieved with a continuous wet period. Pycnidiospores formed appressoria but hyphae did not penetrate if a 6–12 h wet period preceded the dry period, and only a few flecks appeared during the dry period. Coalescent necrosis occurred when the dry period followed penetration. The disease was severe in each case when plants were returned to wet conditions after a period of dryness. Lesion development depended on the duration of the initial wet period, and the characteristics (temperature and duration) of both the dry period and the final wet period.