Survey of fungi associated with subterranean clover leaves and petioles in Western Australia

A wide range of fungi was recorded from leaves and petioles of subterranean clover ( Trifolium subterraneum L.), with and without symptoms of disease, in Western Australia. Leptosphaerulina trifolii (Rostrup) Petrak and Phoma medicaginis Malb. & Roum. were the two most frequently isolated fungi. Ascochyta caulicola Lamb., Colletotrichum truncatum (Schw.) Andus & Moore, Curvularia trifolii (Kauffm.) Boedijn., Fusarium avenaceum (FR.) Sacc, Myrothecium verrucaria (Fr.) Sacc, Phoma medicaginis , and Stemphylium globuliferum (Vestergr.) Simmons were new records on subterranean clover foliage in Western Australia. Phoma medicaginis was demonstrated to be pathogenic and this is the first record as a pathogen on subterranean clover foliage in Western Australia, M. verrucaria and F. avenaceum caused leaf spotting or more extensive lesions as a consequence of undetermined phytotoxic products produced on agar plate cultures.     

Early development of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape (Brassica napus) in relation to temperature and leaf wetness.

In controlled environment experiments to study early development of light leaf spot, lesions developed with leaf wetness durations of 16 to 48 h after inoculation of oilseed rape with conidial suspensions of Pyrenopeziza brassicae at 12 or 18°C, but not with leaf wetness durations of 0 to 13h. The incubation period was 21 to 22 days at 12°C and 14 to 18 days at 18°C for leaf wetness durations of 16 to 48 h. The latent period was 21 to 23 days at 12°C and 18 to 19 days at 18°C, and the total number of lesions increased with increasing leaf wetness duration at both temperatures. In field experiments, light leaf spot always developed on oilseed rape with a leaf wetness duration of 48 h after inoculation in both 1990/1991 and 1991/1992, but the percentage leaf area affected was less on plants placed in an oilseed rape crop than on those placed in a glasshouse. Plants moved to an oilseed rape crop immediately after inoculation nearly always developed light leaf spot symptoms when they were inoculated between 19 October 1990 and 1 March 1991 or between 27 September 1991 and 14 February 1992, but plants inoculated between 31 August and 16 October 1990 or on 20 September 1991, when estimated leaf wetness duration was less than 16 h for several days after they were placed in crops, did not develop symptoms. The latent period of light leaf spot on plants transferred to the oilseed rape crop was 15 to 40 days, and there was an approximately linear relationship between 1 (latent period) and mean temperature during this period. The accumulated temperature during the latent period ranged from c. 150 to 250 day-degrees. The severity of lesions on these plants increased with increasing temperature from 5 to 15°C.     

Effects of temperature on germination and hyphal growth from conidia of Ramularia rhei and Ascochyta rhei, causing spot diseases of rhubarb (Rheum rhaponticum)

Rhubarb leaf and petiole spot disease, caused by Ramularia rhei and Ascochyta rhei , has gradually become more noticeable in the UK field crop. Conidial germination and subsequent colony growth of R. rhei and A. rhei were investigated under in vitro conditions on potato dextrose agar and in vivo on leaf discs. Results indicated that the two fungi responded differently to temperature. Ramularia rhei was better adapted to temperatures ≤ 25°C, with an optimum around 20°C, whereas A. rhei was more adapted to temperatures ≥ 15°C, with an optimum > 25°C. Overall, conidia of R. rhei germinated and subsequent colonies grew at greater rates than those of A. rhei on leaf discs at temperatures ≤ 25°C. These results indicated that it is important to identify the causal agent of leaf and petiole spot diseases in rhubarb field crops in order to estimate disease risks accurately.     

Leaf blotch severity on spring barley infected by isolates of Rhynchosporium secalis under different temperature and humidity regimes

The infection efficiency and severity of leaf blotch on spring barley inoculated with three pathotypes of Rhynchosporium secalis from central Norway were studied under different temperature and humidity regimes. Seedlings of the cultivar Arve were subjected to two constant temperatures, 13° or 18°C. Dry periods of 8 h or longer before or after a wet period of 4 h, carried out in the first 48 h postinoculation, reduced disease severity assessed 16 days after inoculation. The effect of dry periods of up to 24 h was nullified when plants were subjected to high humidity for 48 h after the dry treatment. The disease developed most rapidly when the wet period was 48 h and the temperature 18°C. At or near the optimum temperature for R. secalis (18°C), leaf wetness duration as short as 2 h resulted in considerable disease. Isolates reacted differently to temperature. The most aggressive isolate caused severe disease irrespective of temperature (56–70% of the leaf area infected); however, disease severity caused by the least aggressive isolate was significantly higher at the optimum temperature compared with a lower temperature (13°C). This information can facilitate evaluation of weather data in relation to predicting leaf blotch for advisory purposes.     

Relationships between temperature and latent periods of rust and leaf-spot diseases of groundnut

The effect of temperature on the latent periods of rust, late leaf spot and early leaf spot diseases of groundnut caused by Puccinia arachidis, Phaeoisariopsis personata and Cercospora arachidicola , respectively, was studied. The latent periods (LP) of rust, late leaf spot and early leaf spot ranged from 12–49 days, 13–38 days and 13–39 days, respectively, between 12°C and 33°C An equation relating the rate of pathogen development (1/LP) to temperature was fitted using daily mean temperatures to provide three cardinal temperatures: the minimum (T_mln), optimum (T_opl), and maximum (T_max), T_mln was about 12°C for rust and about 10°C for the two leaf-spot diseases. T_opt, for all three diseases was close to 25°C. T_max was 31°C for early leaf spot, and extrapolated values for late leaf spot and rust were about 35 and 40°C, respectively.
For P. personata , a temperature response curve was fitted using data only from controlled environment experiments. This curve was used to simulate latent periods from both mean daily and mean hourly temperatures in the field. There was substantially better agreement between observed and simulated latent period with hourly temperatures, provided the developmental rate of the pathogen was determined at a constant temperature.     

Effects of root- and stolon-infecting fungi on root-colonizing nematodes of white clover

The effects on white clover ( Trifolium repens ) of different combinations of the nematodes Meloidogyne trifoliophila , Helicotylenchus dihystera and Heterodera trifolii and nine stolon-infecting and three root-infecting fungi were studied in a glasshouse experiment. The presence of the fungus Phytophthora megasperma alone increased ( P  < 0·001) root-rot severity and reduced ( P  < 0·001) plant growth. Other species combinations, such as Phoma nebulosa and Alternaria alternata , interacted and increased root-rot severity. Combinations of P. megasperma with Pythium irregulare , and P. nebulosa with Phoma medicaginis or A. alternata , increased M. trifoliophila populations. Several other fungi ( P. irregulare , P. nebulosa , Colletotrichum coccodes , Macrophomina phaseolina , P. medicaginis and Phoma sp.) interacted with the nematode M. trifoliophila causing severe root-knot symptoms. The results indicated that fungi and nematodes interacted to cause root and stolon rot and reduced yields, and that poor persistence of white clover in pastures is likely to be a problem with a complex etiology.     

The effect of humidity on potato gangrene development in naturally contaminated tubers

During the winters of 1977/78 and 1978/79 tubers of cv. Arran Banner naturally contaminated with Phoma exigua var, foveata were kept at either high or low humidity and at 5, 10, 15 or 20 C. During the 1978/79 winter cv. Majestic tubers were stored under similar conditions at 5 and 10°C only. At monthly intervals tubers from each lot were damaged to encourage gangrene development and returned to their storage condition except the Majestic tubers which were switched from high to low humidity or vice versa and returned to the appropriate temperature. No gangrene rots developed in tubers kept at 1 5 or 20°C at either humidity. At 5 and 10°C fewer rots developed at high humidity compared with low humidity including Majestic tubers kept Initially at high humidity. Inoculum levels in soil on tubers were not markedly affected by storage conditions.     

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.     

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.     

Phenotypic variability of Leptosphaeria lindquistii (anamorph: Phoma macdonaldii), a fungal pathogen of sunflower

Growth of 17 isolates of Phoma macdonaldii , the causal agent of sunflower black stem, was investigated for response to pH and temperature, and for morphology and asexual morphogenesis (pycnidiogenesis and pycnidium size). For all isolates, the optimum pH for growth was between 4 and 5, and the optimum temperature varied between 20 and 30°C and radial growth was slowest at 5 and 35°C. Significant differences in the number and size of pycnidia were observed between isolates. Pycniospore germination was investigated under various conditions in five isolates chosen for their geographical origins, pigmentation, optimum growth temperature and pycnidiogenesis. Increasing the concentration from 10⁶ to 10⁷ pycniospores per mL decreased the germination rate. The optimum temperature for pycniospore germination varied between 15 and 30°C, depending on the isolate, and the optimum and maximum pH values were 5 and 7, respectively. The optimum and minimum relative humidities allowing pycniospore germination were 100 and 95%, respectively. Pycniospore germination was photo-independent. An artificial inoculation method was developed and the aggressiveness of the pathogen was assessed on a susceptible sunflower cultivar, using a 1–9 scale that integrated the percentage of necrotic area on the cotyledon petiole at the stage when the first pair of leaves was fully developed. Significant differences in aggressiveness were observed among the 17 isolates. The parameters investigated clearly suggest the occurrence of a wide phenotypic variability in Phoma macdonaldii .     

Identification of Isolates that Cause a Leaf Spot Disease of Brassicas as Xanthomonas campestris pv. raphani and Pathogenic and Genetic Comparison with Related Pathovars

ABSTRACT Twenty-five Xanthomonas isolates, including some isolates received as either X. campestris pv. armoraciae or pv. raphani, caused discrete leaf spot symptoms when spray-inoculated onto at least one Brassica oleracea cultivar. Twelve of these isolates and four other Xanthomonas isolates were spray- and pin-inoculated onto 21 different plant species/cultivars including horseradish (Armoracia rusticana), radish (Raphanus sativus), and tomato (Lycopersicon esculentum). The remaining 13 leaf spot isolates were spray-inoculated onto a subset of 10 plant species/cultivars. The leaf spot isolates were very aggressive on several Brassica spp., radish, and tomato causing leaf spots and dark sunken lesions on the middle vein, petiole, and stem. Based on the differential reactions of several Brassica spp. and radish cultivars, the leaf spot isolates were divided into three races, with races 1 and 3 predominating. A differential series was established to determine the race-type of isolates and a gene-for-gene model based on the interaction of two avirulence genes in the pathogen races and two matching resistance genes in the differential hosts is proposed. Repetitive-DNA polymerase chain reaction-based fingerprinting was used to assess the genetic diversity of the leaf spot isolates and isolates of closely related Xanthomonas pathovars. Although there was variability within each race, the leaf spot isolates were clustered separately from the X. campestris pv. campestris isolates. We propose that X. campestris isolates that cause a nonvascular leaf spot disease on Brassica spp. should be identified as pv. raphani and not pv. armoraciae. Race-type strains and a neopathotype strain for X. campestris pv. raphani are proposed.     

山楂花腐病的研究Ⅱ.山楂花腐病的症状

 山桂花腐病菌为害叶片、新梢和幼果。叶片上,展叶后4~5天即可出现褐色小班,经6~7天后病斑可扩展到病叶的1/2~2/3。天气潮湿时,在病斑上生出灰白色霉层,即病菌的分生孢子。在新梢上,初生褐色至红褐色病斑,当病斑环绕新梢一周时,新梢上部萎蔫枯死。在幼果上,落花后10天左右幼果果面出现褐色病斑,逐渐扩大,直至整个果实变成暗褐色脱落。     

Effect of temperature and leaf wetness duration on infection of sweet oranges by Asiatic citrus canker

Asiatic citrus canker, caused by Xanthomonas smithii ssp. citri , formerly X. axonopodis pv. citri , is one of the most serious phytosanitary problems in Brazilian citrus crops. Experiments were conducted under controlled conditions to assess the influence of temperature and leaf wetness duration on infection and subsequent symptom development of citrus canker in sweet orange cvs Hamlin, Natal, Pera and Valencia. The quantified variables were incubation period, disease incidence, disease severity, mean lesion density and mean lesion size at temperatures of 12, 15, 20, 25, 30, 35, 40 and 42°C, and leaf wetness durations of 0, 4, 8, 12, 16, 20 and 24 h. Symptoms did not develop at 42°C. A generalized beta function showed a good fit to the temperature data, severity being highest in the range 30–35°C. The relationship between citrus canker severity and leaf wetness duration was explained by a monomolecular model, with the greatest severity occurring at 24 h of leaf wetness, with 4 h of wetness being the minimum duration sufficient to cause 100% incidence at optimal temperatures of 25–35°C. Mean lesion density behaved similarly to disease severity in relation to temperature variation and leaf wetness duration. A combined monomolecular-beta generalized model fitted disease severity, mean lesion density or lesion size as a function of both temperature and duration of leaf wetness. The estimated minimum and maximum temperatures for the occurrence of disease were 12°C and 40°C, respectively.     

Evaluation of susceptibility of an olive progeny (Picholine x Meski) to olive leaf spot disease caused by Fusicladium oleagineum

Forty five descendants from a crossing between olive cultivar Picholine de Languedoc and the local cultivar Meski were evaluated for their susceptibility to olive leaf spot (Fusicladium oleagineum) in field and laboratory studies. Disease incidence varied greatly among the descendants and was correlated with the severity of leaf symptoms. Based on field observations and laboratory tests, the hybrids were classified into five groups: 1) very susceptible to the disease: Meski and 17H, 2) susceptible to the disease : 20J, 8I, 11I, 21I, 14H, 16H, 23H, 6J, 8J and 23J, 3) moderately resistant to the disease : 15I, 16I, 22I, 23I, 8K, 13H, 18H, 22H, 9J, 10J, 11J, 12J, 14J, 16J, 18J, 4) resistant to the disease: 21J, 12I, 13I, 18I, 19I, 9K, 10K, 12H, 15H, 19H, 15J and 22J, and completely resistant to the disease : cultivar Picholine and hybrids 6I and 19I. The analysis of anatomical parameters showed that the polyphenol content and composition were not correlated with disease incidence. However, the cuticle thickness and the trichome density are very important physical parameters that can determine the degree of susceptibility of hybrids to the disease. The assessment method may be useful to screen olive cultivars and hybrids for Fusicladium resistance.     

Effects of temperature on the development of light leaf spot (Pyrenopeziza brassicae) on oilseed rape (Brassica napus)

The effects of temperature on the development of light leaf spot (Pyrenopeziza brassicae) on winter oilseed rape were investigated in controlled-environment experiments. The proportion of conidia which germinated on leaves, the growth rate of germ tubes, the severity of light leaf spot and the production of conidia increased with increasing temperature from 5 to 15 C. The time to 50% germination of conidia and the incubation and latent periods of light leaf spot lesions decreased when temperature increased from 5 to 15°C. At 20°C, however, light leaf spot severity and production of conidia were less and the incubation and latent periods were longer than at 15 C. There were differences between P brassicae isolates and oilseed rape cultivars in the severity of light leaf spot, the production of conidia and the length of the incubation period but not in the length of the latent period. The responses to temperature for lesion severity and incubation and latent periods appeared to be approximately linear over the temperature range 5-15°C and could be quantified using linear regression analysis.     

Interactions between Cercospora arachidicola and Phoma arachidicola, and their effects on defoliation and kernel yield of groundnut

Cercospora arachidicola and Phoma arachidicola spread independently on groundnuts when leaf area was not limiting, but where C. arachidicola colonized leaves early, it usually spread at the expense of P. arachidicola. Juice extracted from leaflets infected with C. arachidicola and separated by thin-layer chromatography contained unidentified compounds which were not present in juice from uninfected leaflets, and prevented the growth of P. arachidicola. Cercospora leaf spot reduced photosynthesis and was the major contributor to premature leaf drop, which had an adverse effect on yield. Well blotch ( P. arachidicola ) only caused defoliation and depressed yield where there was very little Cercospora leaf spot present.     

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.     

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).     

Phoma black stem severity and phytoestrogen production in annual Medicago spp. is primarily determined by interaction of cultivar and pathogen isolate

Phoma black stem and leaf spot disease (caused by Phoma medicaginis) not only diminishes forage and seed yield but stimulates production of detrimental phytoestrogens in annual Medicago spp. This study aimed to evaluate relationships between disease development from five isolates of P. medicaginis on 16 cultivars with production of coumestrol and 4′-O-methylcoumestrol. In the presence of P. medicaginis, Sava had the highest coumestrol and 4′-O-methylcoumestrol (640 and 85 mg/kg, respectively) followed by Caliph (253 and 15 mg/kg, respectively). In the absence of P. medicaginis, Jemalong and Paragosa showed highest and lowest coumestrol (137 and 0 mg/kg, respectively). 4′-O-methylcoumestrol was not produced in disease-free plants, but coumestrol was. Disease incidence and severity on leaves and on petiole/stems, and consequent leaf yellowing severity ranged from 5%–98.7%, 0%–100%, 4.4%–98.7%, 1.7%–100%, and 0%–85%. Sava, Paraponto, Harbinger, and Serena were most susceptible, while Tornafield and Caliph were least susceptible. There was significant overall positive correlation of disease incidence/severity factors across cultivars (p < 0.01) with both coumestrol and 4′-O-methylcoumestrol. Jemalong was least responsive and Paragosa and Sava most responsive to coumestrol production following P. medicaginis inoculation. Coumestrol in inoculated Paragosa increased to 373 mg/kg in comparison with 0 mg/kg in controls. These findings are of critical importance towards developing less disease-susceptible annual Medicago spp. producing less detrimental phytoestrogens. Least susceptible cultivars like Tornafield and Caliph can be used to manage yield loss, whilst least responsive cultivars to phytoestrogen production like Caliph also can help to reduce phytoestrogen production.     

Effects of bean line pattern mosaic virus on the monocyclic components of rust and angular leaf spot of Phaseolus bean at different temperatures

Monocyclic components (development rate during the incubation period or latent period, lesion density, lesion size and disease severity) of rust ( Uromyces appendiculatus ) and of angular leaf spot ( Phaeoisariopsis griseola ) in two bean ( Phaseolus vulgaris ) cultivars (Rosinha G-2 and Carioca), pre-infected or not with bean line pattern mosaic virus (BLPMV), were determined. Trials were conducted at temperatures in the range from 9 to 27°C for rust and from 12 to 30°C for angular leaf spot. Regardless of viral pre-infection, the effect of temperature on the four monocyclic components followed an optimum curve and could be described by a generalized beta function. Generally, angular leaf spot was favoured by higher temperatures with an optimum for disease severity between 24.2 and 28.3°C compared with 15.9–18.5°C for rust. Pre-infection with BLPMV did not change the shape of the optimum curves for all components, but significantly reduced lesion density and disease severity on both cultivars. The development rates during incubation and latent periods for both fungal diseases were not affected by BLPMV. Pre-infection with virus did not alter the ranking of cultivars with respect to resistance to both fungal diseases.