Pathogenicity of <Emphasis Type="Italic">Mycochaetophora gentianae</Emphasis>, causal fungus of gentian brown leaf spot,as affected by host species,inoculum density,temperature, leaf wetness duration,and leaf position

When the influence of host species, inoculum density, temperature, leaf wetness duration, and leaf position on the incidence of gentian brown leaf spot caused by Mycochaetophora gentianae, was examined, the fungus severely infected all seven Gentiana triflora cultivars, but failed to infect two cultivars of G. scabra and an interspecific hybrid cultivar. Inoculum density correlated closely with disease incidence, and a minimum of 10² conidia/mL was enough to cause infection. In an analysis of variance, temperature and leaf wetness duration had a significant effect upon disease incidence, which increased with higher temperature (15–25°C) and longer duration of leaf wetness (36–72 h). No disease developed at temperatures lower than 10°C or when leaf wetness lasted <24 h. At 48-h leaf wetness, disease incidence was 0, 28, 77, and 85% at 10, 15, 20, and 25°C, respectively. Middle and lower leaves on the plant were more susceptible than upper leaves. In microscopic observations of inoculated leaves, >50% of conidia germinated at temperatures >15°C after 24-h leaf wetness. More appressoria formed at higher temperatures (15–25°C) with extended duration of leaf wetness (24–72 h). At 48-h leaf wetness, appressorium formation was 0, 8, 26, and 73% at 10, 15, 20, and 25°C, respectively. These results suggest that temperature and leaf wetness duration were important factors for infection of gentian leaves.     

Effects of inoculum density, leaf age, moisture, temperature, and wetness duration on black streak of edible burdock

Inoculum density, temperature, leaf age, and wetness duration were evaluated for their effects on the development of black streak (Itersonilia perplexans) on edible burdock (Arctium lappa L.) in a controlled environment. The effect of relative humidity (RH) on ballistospores production by I. perplexans was also evaluated. Symptoms of black streak on leaves increased in a linear fashion as the inoculum density of I. perplexans increased from 10² to 10⁶ ballistospores/ml. Rugose symptoms on young leaves were observed at densities of ≥10⁴ ballistospores/ml. Disease severity of I. perplexans in relation to leaf age followed a degradation curve when the leaves were inoculated with ballistospores. Disease severity was high in newly emerged leaves up to 5 days old, declined as leaf age increased to 29 days, and was zero when leaf age increased from 30 to 33 days. Disease development of edible burdock plants exposed to ballistospores of I. perplexans was evaluated at various combinations of temperature (10°, 15°, 20°, 25°C) and duration of leaf wetness (12, 24, 36, 48, and 72 h). Disease was most severe when plants were in contact with the ballistospore sources at 15° or 20°C. The least amount of disease occurred at 25°C regardless of wetness duration. Ballistospores required 24–36 h of continuous leaf wetness to cause visible symptoms by infection on edible burdock. Ballistospores production in infected lesions required at least 95.5% RH.     

Effects of temperature,inoculum concentration,leaf age,and continuous and interrupted wetness on infection of olive plants by Spilocaea oleagina

Experiments were conducted on olive plants in controlled environments to determine the effect of conidial concentration, leaf age, temperature, continuous and interrupted leaf wetness periods, and relative humidity (RH) during the drier periods that interrupted wet periods, on olive leaf spot (OLS) severity. As inoculum concentration increased from 1·0 × 10² to 2·5 × 10⁵ conidia mL^?1, the severity of OLS increased at all five temperatures (5, 10, 15, 20 and 25°C). A simple polynomial model satisfactorily described the relationship between the inoculum concentration at the upper asymptote (maximum number of lesions) and temperature. The results showed that for the three leaf age groups tested (2–4, 6–8 and 10–12 weeks old) OLS severity decreased significantly (P < 0·001) with increasing leaf age at the time of inoculation. Overall, temperature also affected (P < 0·001) OLS severity, with the lesion numbers increasing gradually from 5°C to a maximum at 15°C, and then declining to a minimum at 25°C. When nine leaf wetness periods (0, 6, 12, 18, 24, 36, 48, 72 and 96 h) were tested at the same temperatures, the numbers of lesions increased with increasing leaf wetness period at all temperatures tested. The minimum leaf wetness periods for infection at 5, 10, 15, 20 and 25°C were 18, 12, 12, 12 and 24 h, respectively. The wet periods during early infection processes were interrupted with drying periods (0, 3, 6, 12, 18 and 24 h) at two levels of RH (70 and 100%). The length of drying period had a significant (P < 0·001) effect on disease severity, the effect depending on the RH during the interruption. High RH (100%) resulted in greater disease severity than low RH (70%). A polynomial equation with linear and quadratic terms of temperature, wetness and leaf age was developed to describe the effects of temperature, wetness and leaf age on OLS infection, which could be incorporated as a forecasting component of an integrated system for the control of OLS.     

Biological control of bacterial spot on peach by nonpathogenic Xanthomonas campestris strains AZ98101 and AZ98106

A suspension of nonpathogenic Xanthomonas campestris (nXc) strain AZ98101 or AZ98106 was sprayed on peach trees in biological control tests against bacterial spot caused by Xanthomonas arboricola pv. pruni. Treatment with each strain reduced the number of leaves with spots. Meta-analysis of the results from seven field trials showed an integrated risk ratio (the ratio of leaf spot incidence on inoculated leaves to incidence on control leaves) of 0.46 by treatment with AZ98101 and 0.54 by AZ98106, indicating that leaf spot incidence was significantly reduced by each strain. Each also significantly reduced the incidence of fruit spot. The risk ratio with AZ98101 was significantly lower than that with AZ98106. The population of AZ98101R-1, a rifampicin-resistant mutant of AZ98101, was 6.3 × 10³ CFU/g (fresh mass) on leaves and 8.0 × 10³ CFU/g on fruit at 12 days after inoculation, but was undetectable (<10² CFU/g) by 24 days. This is the first report that nXc strains AZ98101 and AZ98106 effectively control bacterial spot on peach.     

Modelling the effects of temperature and leaf wetness on monocyclic infection in a tropical fungal pathosystem

Modelling the epidemiology of water yam anthracnose (Dioscorea alata) caused by the fungus Colletotrichum gloeosporioides is an important research goal, as it will allow the investigation of a wide range of scenarios of new practices to reduce the disease impact before experimentation in the field. Developing such a model requires a prior knowledge of the fungus’s response to the environmental conditions, which will be affected by pest management. In this work, we first measured the response of the fungus to the main physical environmental factors controlling its development, namely temperature (ranging from 18 °C to 36 °C) and wetness duration (from 2 h to 72 h). As response variables, we measured the percentage of formed appressoria (relative to the total number of spores), the length of the latent period (time lag between inoculation and first symptoms observed), and the rate of necrotic lesion extension (percentage of diseased leaf surface at different time steps). These variables allow us to estimate the effects of temperature and wetness duration on the success of infection (appressoria formation) and the subsequent rate of disease development (latent period length and lesion extension rate). The data were fitted to non-linear models chosen for their ability to describe the observed patterns. From our data and model analyses, we were able to estimate parameters such as the optimal and maximal temperatures (25–28 °C and 36 °C, respectively), the required wetness duration to reach 20 % of infection success and the time to reach 5 % disease severity as a function of temperature.     

Effects of inoculum density,leaf wetness duration and nitrate concentration on the occurrence of lettuce leaf spot

In Ehime Prefecture, Japan, lettuce leaf spot (Septoria lactucae) caused huge losses in marketable lettuce yields. To explore potential measures to control disease outbreaks, the effects of inoculum density, leaf wetness duration and nitrate concentration on the development of leaf spot on lettuce (Lactuca sativa) were evaluated. Conidia were collected from diseased plants in an infested field by single-spore isolation and were used to inoculate potted lettuce plants with different conidial concentrations. Lesions developed on inoculated lettuce plants at inoculum concentrations from 10⁰ to 10⁶ conidia/ml. The disease was more severe when the inoculum exceeded 10² conidia/ml, and severity increased with increasing concentrations. Assessment of the relationship between disease development and the duration of postinoculation leaf wetness revealed that symptoms appeared when the inoculated plants remained wet for 12 h or longer. The number of lesions and total nitrogen content in the lettuce leaves both increased when nitrate was applied.     

不同湿润持续时间及叶片温度对温室黄瓜霜霉病发生的影响

为探明不同湿润持续时间及叶片温度与黄瓜霜霉病发生的关系,通过观察不同湿润条件下黄瓜霜霉病的初显症时间,计算逐日显症率及累积显症率,并利用热红外成像仪对显症后叶片温度进行连续监测。结果显示,不同湿润持续时间对黄瓜霜霉病的初显症时间、逐日显症率产生影响。叶片湿润持续4 h,黄瓜霜霉病在接种后7.00 d显症;叶片湿润持续12 h,黄瓜霜霉病初显症时间最早,仅为3.25 d。叶片湿润持续时间不同,黄瓜霜霉病初显症时的叶片温度存在显著差异。回归分析表明,初显症时间与最大温差呈显著正相关,与平均温度呈显著负相关。叶片湿润持续4、6 h的病斑出现高峰在显症后第2、3天,逐日显症率分别是37.50%和41.18%,比叶片湿润持续10、12 h的早。显症后期,湿润持续时间4、6、8、10、12 h的病斑累积显症率分别是87.94%、93.71%、90.25%、84.24%和88.36%,差异不显著。表明接种黄瓜霜霉菌Pseudoperonospora cubensis后叶片湿润持续时间越长,潜育期越短,叶片最大温差越小,叶片平均温度越大。     

Effect of interrupted leaf wetness periods on suppression of sporulation ofBotrytis allii andB. cinerea by antagonists on dead onion leaves

Saprophytic antagonists were evaluated for suppression of sporulation ofBotrytis allii andB. cinerea on artificially killed segments of onion leaves that were pre-inoculated with the pathogens. During incubation of the antagonisttreated leaf segments in moist chambers, periods of leaf wetness and leaf dryness were alternated to simulate conditions in the field. Interruption of humid conditions with dry periods had a differential effect on antagonists.Alternaria alternata, Chaetomium globosum, Ulocladium atrum andU. chartarum suppressed sporulation ofB. allii almost completely under continuously wet conditions, and when the leaf wetness periods were interrupted with drying periods of 9h imposed 16, 40, and 64 h after the antagonists were applied. When leaf wetness was interrupted 16 h after antagonist application, the number of conidia ofB. allii produced cm^–2 leaf surface after eight days was under the detection limit of 5.2 × 10³ conidia on leaves treated with these antagonists compared to 3.7 × 10⁵ conidia on leaves that were not treated. On the other hand,Gliocladium roseum, G. catenulatum andSesquicillium candelabrum, all highly efficient under continuously wet conditions, were of low to moderate efficiency when leaf wetness periods had been interrupted 16 h after application of the antagonists. The antagonists showed the same differentiation and sensitivity to interrupted wetness periods when tested withB. cinerea.     

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.     

Effect of temperature,relative humidity,leaf wetness and leaf age on <Emphasis Type="Italic">Spilocaea oleagina</Emphasis> conidium germination on olive leaves

The effects of temperature, relative humidity (RH), leaf wetness and leaf age on conidium germination were investigated for Spilocaea oleagina, the causal organism of olive leaf spot. Detached leaves of five ages (2, 4, 6, 8 and 10 weeks after emergence), six different temperatures (5, 10, 15, 20, 25 and 30°C), eight wetness periods (0, 6, 9, 12, 18, 24, 36 and 48 h), and three RH levels (60, 80 and 100%) were tested. Results showed that percentage germination decreased linearly in proportion to leaf age (P < 0.001), being 58% at 2 weeks and 35% at 10 weeks. A polynomial equation with linear term of leaf age was developed to describe the effect of leaf age on conidium germination. Temperature significantly (P < 0.001) affected frequencies of conidium germination on wet leaves held at 100% RH, with the effective range being 5 to 25°C. The percent germination was 16.1, 23.9, 38.8, 47.8 and 35.5% germination at 5, 10, 15, 20 and 25°C, respectively, after 24 h. Polynomial models adequately described the frequencies of conidium germination at these conditions over the wetness periods. The rate of germ tube elongation followed a similar trend, except that the optimum was 15°C, with final mean lengths of 175, 228, 248, 215 and 135 μm at 5, 10, 15, 20 and 25°C, respectively after 168 h. Polynomial models satisfactorily described the relationships between temperature and germ tube elongation. Formation of appressoria, when found, occurred 6 h after the first signs of germination. The percentage of germlings with appressoria increased with increasing temperature to a maximum of 43% at 15°C, with no appressoria formed at 25°C after 48 h of incubation. Increasing wetness duration caused increasing numbers of conidia to germinate at all temperatures tested (5–25°C). The minimum leaf wetness periods required for germination at 5, 10, 15, 20 and 25°C were 24, 12, 9, 9 and 12 h, respectively. At 20°C, a shorter wetness period (6 h) was sufficient if germinating conidia were then placed in 100% RH, but not at 80 or 60%. However, no conidia germinated without free water even after 48 h of incubation at 20°C and 100% RH. The models developed in this study should be validated under field conditions. They could be developed into a forecasting component of an integrated system for the control of olive leaf spot.     

Effect of pea canopy architecture on microclimate and consequences on ascochyta blight infection under field conditions

In order to investigate the impact of pea canopy architecture and development on microclimate and infection by Mycosphaerella pinodes, two field experiments were conducted in 2009 and 2010 at Le Rheu (France) to obtain canopies contrasted in height, closure dynamic, leaf area index (LAI) and leaf area density (LAD). Three pea cultivars (Athos, Antares, Gregor) were sown at two (80 and 40 seeds/m² in 2009) and three densities (80, 40 and 30 seeds/m² in 2010) and microclimatic sensors were located inside the canopy (at the bottom and in the middle) and outside. Two main sources of wetness were identified: rainfall and dew. During rainfall periods, average daily leaf wetness duration (LWD) was about 15 h, and 3 to 10 h longer inside than outside the canopies. LWD was positively correlated with LAI until canopy closure during these periods. During dry periods when dew was the only source of leaf wetness, average daily LWD was short, decreasing as the canopy developed. Shorter LWDs were observed at the base than at the mid-level of the canopies and longer LWDs were observed outside the canopy and inside the less dense canopies irrespective of the cultivar. LWD was negatively correlated with canopy height and LAI during these periods. Slow wind speeds were recorded inside the canopies (less than 0.5 km/h) and no significant canopy effect was observed on air temperature. An infection model was developed and showed that only rainfall periods which induced long LWDs inside the canopy, were favourable to M. pinodes infection under our climatic conditions and suggested a more favourable microclimate inside dense canopies.     

Early detection of sugar beet pathogen Ramularia beticola in leaf and air samples using qPCR

A quantitative PCR method (qPCR) was developed for the detection and quantification of Ramularia beticola causing Ramularia leaf spot in sugar beet. R. beticola specific primers were designed based on the internal transcribed spacer region 2 (ITS2). The assay was applied on DNA extracted from spores trapped on tape from Burkard spore traps placed in an artificially inoculated sugar beet field trial and in two sugar beet fields with natural infections. R. beticola DNA was detected at variable amounts in the air samples 14 to 16 days prior to first visible symptoms. R. beticola DNA was detected in air samples from fields with natural infection at significant and increasing levels from development of the first symptoms, indicating that spore production within the crop plays a major role in the epidemic development of the disease. Sugar beet leaves sampled from the inoculated field trial were also tested with the qPCR assay. It was possible to detect the presence of R. beticola in the leaves pre-symptomatic at least 10 days before the occurrence of the visible symptoms of Ramularia leaf spot. This is the first report of a molecular assay, which allows screening for the presence of R. beticola in plant material and in air samples prior to the appearance of visible symptoms. An early detection has potential as a tool, which can be part of a warning system predicting the onset of the disease in the sugar beet crop and helping to optimise fungicide application.     

Effects of host resistance, temperature, leaf wetness, and leaf age on infection and lesion development of pecan scab

ABSTRACT The effects of partial host resistance, temperature, leaf wetness duration, and leaf age on infection and lesion development of pecan scab were evaluated. Trees of cultivars Wichita (susceptible) and Sumner (resistant) were inoculated with conidia of Cladosporium caryigenum and placed in mist chambers set at 15, 25, or 35 degrees C. The trees were removed from the chambers after 3, 6, 12, 24, 36, or 48 h of leaf wetness and placed in a greenhouse to allow disease development. After 8 to 16 days, disease began to develop on both 'Wichita' and 'Sumner'. Logistic regression analysis showed that the probability of a leaf becoming infected was greatest for 'Wichita' it decreased with increasing leaf age and temperature and increased with increasing leaf wetness. Leaves on 'Wichita ' were susceptible to infection between 2 and 23 days after budbreak, while leaves on 'Sumner' were susceptible to infection from 2 to 18 days after budbreak. Infection frequency, lesion size, and conidia production decreased proportionately with increasing leaf age. The magnitude of this effect was greatest on 'Sumner'. Conidia production was positively correlated with lesion size, and both were positively correlated with infection frequency on both cultivars.     

Effects of leaf wetness and temperature on late leaf-spot infection of groundnut

Experiments are described to quantify the effects of temperature and leaf wetness duration on infection of groundnut by Phaeoisariopsis personata. Temperature response curves for conidial germination and infection were similar, with optima close to 20°C and minimum and maximum temperatures of about 8°C and 34 C, respectively. The effect of temperature on infection between 15°C and 26°C was slight. Lesions developed only if the leaf wetness period exceeded about 20 h, and the total wetness period necessary for maximum infection exceeded 160 h. The number of lesions resulting from a fixed amount of inoculum was several times greater if leaves were exposed to alternate wet and dry periods (intermittent wetness), compared with continuous wetness. With intermittent wetness the length of the dry period had little effect on the number of lesions, providing it exceeded 2 h. The response curve relating total wetness periods to lesion density was an exponential asymptote.     

Silicon suppresses tan spot development on wheat infected by <Emphasis Type="BoldItalic">Pyrenophora tritici-repentis</Emphasis>

Tan spot caused by Pyrenophora tritici-repentis is the main foliar diseases of wheat in Brazil. The effect of silicon (Si) on the components of resistance of a susceptible (Fundacep Horizonte) and a moderately resistant (Quartzo) wheat cultivar was studied in a controlled environment. Silicon was supplied as calcium silicate in the soil 30 days before sowing. At the booting stage, a conidial suspension of the fungus was sprayed onto the flag leaves of potted plants, which were incubated under moist conditions for 48 h. Afterwards, inoculated leaves were assessed for: incubation period (IP), infection efficiency (IE), area under lesion size curve (AULSC), lesion size (LS), severity (SEV) and area under severity curve (AUSC). Foliar Si concentrations were quantified at the end of the evaluations. Si supply to plants increased leaf Si concentration in 233% for Fundacep Horizonte (from 4.8 to 16.0 g kg^?1 of dry matter) and 211% for Quartzo (from 5.3 to 16.5 g kg^?1 of dry matter). In the Si + treatments, IP was longer by 24 and 17 h, IE declined by 53.5 and 65.5%, LS (at 264 h after inoculation) by 4.6 mm (from 9.5 to 4.9 mm) and 5.9 mm (from 8.2 to 2.3 mm), and SEV by 53% (from 54.4 to 18.8%) and 88% (from 47.7 to 5.5%) respectively, for the Fundacep Horizonte and Quartzo cultivars. The Si x cultivar interaction was not significant for AULSC and AUSC, and these variables were reduced by 55.8 and 80.8%, respectively, in plants supplied with Si. In conclusion, Si enhanced the resistance of wheat plants to tan spot development by affecting several resistance components, regardless of the resistance level of the cultivar. However, the greatest reduction in tan spot development by Si supply was observed when using a moderately resistant cultivar.     

Response of leaf acetolactate synthase from different leaf positions and seedling ages to sulfonylurea herbicide

Responses of acetolactate synthase (ALS) from grass and broadleaf weed to sulfonylurea (SU) herbicide were compared in relation to the leaf position in a seedling and seedling age. The responses of Echinochloa crus-galli (L.) P. Beauv. and Eclipta prostrata L., dominant grass and broadleaf weed in paddy fields in Korea, respectively, to azimsulfuron were examined. In this study, in vivo ALS assay was used to verify the responsibility of selected weed species at different leaf stages to SU-herbicides. The data from in vivo ALS assay could be used for discriminating the degree of tolerance between weeds showed different susceptibility. In E. crus-galli and E. prostrata there was no apparent relationship between the chlorophyll concentrations and herbicide concentrations treated on leaves. Both in E. crus-galli and E. prostrata, the free amino acid concentrations, however, were increased as herbicide concentration increased in the younger leaves. The free amino acid concentrations were generally higher in older leaves than young leaves and were significantly increased concomitantly with increasing herbicide concentration. The ALS activity was decreased rapidly with higher azimsulfuron rates in old but not senescent leaves compared to juvenile leaves. Generally, ALS activity was less sensitive at the early leaf stage than late leaf stage. The activity of ALS in E. prostrata was highly responsive to application time and more susceptible to the herbicide as compared to E. crus-galli. The highest levels of acetoin were observed in the uppermost and youngest leaf in all species tested.     

Conidial production and viability of Calonectria pseudonaviculata on infected boxwood leaves as affected by temperature,wetness, and dryness periods

Calonectria pseudonaviculata causes lesions on boxwood leaves and twigs. Controlled-environment experiments were conducted to determine the effects of temperature and leaf wetness period on C. pseudonaviculata sporulation on diseased (cv. Suffruticosa) leaves and of dryness periods and high temperature on conidial survival. Infected leaves were incubated in moist chambers and subjected to six temperatures (9, 13, 17, 21, 25, and 29°C) and six leaf wetness periods (0, 12, 24, 40, 48, and 72 h). Spore production was influenced significantly by wetness period, temperature, and their interaction. Increasing duration of leaf wetness and increasing temperature generally increased sporulation, with no sporulation occurring at 29°C or 9 and 13°C, except at 72 h of wetness exposure, while it was optimal at 21°C. Detached leaves with profuse conidia were subjected to a range of drying (relative humidity at 65%) times (0, 2, 4, 6, and 8 h) at two temperatures of 21 and 29°C. Conidia were then harvested and plated on water agar. Germinating conidia were counted to measure the spore viability. Spore mortality increased with increasing dryness duration at both temperatures but occurred more quickly and severely at 29 than 21°C. Overall, this study extended biological knowledge of conditions required for crucial stages of the C. pseudonaviculata disease cycle and the obtained results will be vital for developing boxwood blight forecasting and management tools.     

Infection of Linseed by Alternaria linicola; Effects of Inoculum Density, Temperature, Leaf Wetness and Light Regime

Controlled environment studies were conducted to determine the effects of inoculum density, temperature, leaf wetness and light regime on the infection of linseed by Alternaria linicola. The % cotyledons and leaves with symptoms, and the disease severity (% leaf area with symptoms) increased linearly when the inoculum density increased from 1×10³ to 1×10⁵ conidiaml^–1. The first symptoms appeared on cotyledons and leaves 4 and 6 days after inoculation, respectively. Eight hours of leaf wetness were sufficient to initiate the disease at 25°C but not at 15°C, when 10-h periods of leaf wetness were required. % leaf area with symptoms was lower at 15°C than that at 25°C irrespective of the leaf wetness periods tested. Interruption of a continuous leaf wetness period by a 12-h dry period, occurring at any time between 1 and 18h after inoculation, decreased the % cotyledons with symptoms and the disease severity, with the greatest reductions (60% and 100%, respectively) being observed when the dry period began 6h after inoculation. A. linicola conidia were able to exploit successive 12-h periods of leaf wetness cumulatively to infect linseed plants. Disease incidence and severity were positively correlated with the dark period following inoculation, but they were negatively related to the length of the initial light period. Our findings suggest that infection of linseed by A. linicola and further development of symptoms can occur under unfavourable environmental conditions.