Soil organic carbon fractions and humic substances are affected by land uses of Caatinga forest in Brazil

Caatinga is a Brazilian dry ecosystem that occupies around 1 million km² and is one of the largest tropical dry forests of the world. About 46% of the area that was originally covered has been deforested. Land use can cause pronounced reduction in soil carbon stocks that play a major role in the global carbon cycle. The objective of this study was to improve our understanding of the effect of land use on oxidizable carbon fractions, total carbon stocks and humic substances in different layers of soil in a Brazilian semi-arid region. We analyzed soils from tropical dry forest (TDF), forest succession with Anadenanthera falcata (ANA), with Tabebuia alba (TAB), secondary scrubby regeneration (SCR), and non-irrigated maize (MS). Forests showed larger fractions of more labile carbon, except for TDF. The most recalcitrant fraction of carbon stock, humin fraction stock, with the different land use decreased by 38–53% compared to TDF. Oxidizable carbon fractions, carbon stocks, and humic fraction stocks were able to differentiate the successional land uses and agricultural cover from TDF, mainly in the 0–5?cm layer. Our results show that changes in land use, especially with ANA forest succession, showed a larger labile carbon fraction, indicating easy decomposition and loss. Our results provide an alternative tool for the management of deforested areas in tropical dry caatinga ecosystems. This would contribute to the conservation of dry forest systems and could serve as guideline for sustainable management of agriculturally impacted caatinga areas.     

Relationship between disease severity and escape of Pseudoperonospora cubensis sporangia from a cucumber canopy during downy mildew epidemics

Fundamental to the development of models to predict the spread of cucurbit downy mildew is the ability to determine the escape of Pseudoperonospora cubensis sporangia from infected fields. Aerial concentrations of sporangia, C (sporangia m^?3), were monitored using Rotorod samplers deployed at 0·5 to 3·0 m above a naturally infected cucumber canopy in two sites in central and eastern North Carolina in 2011, where disease severity ranged from 1 to 40%. Standing crop of sporangia was assessed each morning at 07·00 h EDT and ranged from 320 to 16 170 sporangia m^?2. Disease severity and height above the canopy significantly (P < 0·0001) affected C with mean concentration (C_m) being high at moderate disease. Values of C_m decreased rapidly with canopy height and at a height of 2·0 m, C_m was only 7% of values measured at 0·5 m when disease was moderate. Daily total flux (F_D) was dependent on disease severity and ranged from 5·9 to 2242·3 sporangia m^?2. The fraction of available sporangia that escaped the canopy increased from 0·028 to 0·171 as average wind speed above the canopy for periods of high C increased from 1·7 to 3·6 m s^?1. Variations of C_m and F_D with increasing disease were well described (P < 0·0001) by a log‐normal model with 15% as the threshold above which C_m and F_D decreased as disease severity increased. These results indicate that disease severity should be used to adjust sporangia escape in spore transport simulation models that are used to predict the risk of spread of cucurbit downy mildew.     

Application of fuzzy logic for the simulation of Plasmopara viticola using agrometeorological variables*

A mechanistic model called PLASMO was developed earlier to simulate grapevine downy mildew (Plasmopara viticola) and has been applied in several viticultural areas of Italy since 1988 by the collaboration of several research institutions of Firenze. In this study, a new simulation model based on fuzzy logic has been developed for the same structure (biological cycle of P. viticola). This approach allows classical quantitative information to be used together with qualitative information. Vague concepts can also be handled. Agrometeorological data is used, with an hourly time step, starting from budbreak to the end of the growing season. Air temperature, relative humidity, rainfall and leaf wetness are required. The simulated processes are the growth of grapevine leaf area and the main phases of the biological cycle of the pathogen: incubation, sporulation, germination, spore survival and inoculation. The main epidemiological outputs are timing of infection events and disease intensity. The performance of the model is evaluated and the mechanistic and fuzzy logic approaches are compared.     

Quantification of airborne spores of <Emphasis Type="Italic">Monilinia fructicola</Emphasis> in stone fruit orchards of California using real-time PCR

The fungal pathogen Monilinia fructicola causes blossom blight and fruit brown rot of stone fruits in California. In this study, spore densities in the air were monitored in six orchard/year combinations with Burkard spore traps. A real-time PCR assay was developed to efficiently quantify the dynamics of spore density in these orchards during the growing season. Different patterns of dynamics of spore density were observed in these orchards. A linear relationship between numbers of spores counted with a compound microscope and those determined with the real-time PCR assay was obtained, using the same samples of spore traps. Spore density in five of six orchard/year combinations ranged from 0.0 to 0.05 spores l⁻¹, except for that in orchard 4, which showed much higher values of spore density in the air, as well as higher values and wider range of incidences of blossom infection and fruit rot than those in the other orchards. The results demonstrated a potential method to quantitatively determine spore inoculum potential in orchards by using a real-time PCR assay.     

Type and quantity of organic amendments determine the amount of carbon stabilized in particle-size fractions of a semiarid degraded soil

A 9-month lab experiment was carried out with three different amendments (vine pruning wastes, PW; composted vine pruning wastes, cPW; and sewage sludge, SS) added at three different rates (90, 180, and 240 t ha^?1, dry weight) in order to test whether the type or the quantity of the amendments applied to a semiarid, degraded soil determined the C_org accumulation in its particle-size fractions (coarse sand, 200–2,000 µm; fine sand, 63–200 µm; silt, 2–63 µm; and clay, 0.1–2 µm). All amendments, independently of their C/N ratios, resulted in similar C_org content and accumulation in coarse sand and silt-sized fractions after 9 months. In the clay-sized fraction, enrichment in C_org produced the incorporation of particles from this particle-size fraction into the silt-sized fraction. Likewise, increasing the application rates of the amendments led to larger C_org contents into the particle-size fractions of all amended soils except for the clay-sized fraction. The application of SS resulted in lower basal respiration-to-C_org ratios in the clay-sized fraction than the application of PW and cPW, suggesting a higher protection of the C_org in the SS treatment. These results indicate that organic amendments from woody plants with C/N ratios higher than 30, such as PW, favor C_org accumulation in the fine sand-sized fractions. Furthermore, our findings suggest that the application rate of such amendments, rather than the C/N ratios and amendment origin (from sludge or woody plants), is the key factor for promoting C_org accumulation in the silt-sized fractions of semiarid degraded soils.     

Spore germination and disease development after application of pycnidiospores ofAscochyta caulina toChenopodium album plants

Ascochyta caulina is considered a potential mycoherbicide againstChenopodium album. Disease development ofC. album plants and plants of 14 other species after application of pycnidiospores ofA. caulina was studied in climate chamber experiments. The experiments were carried out to analyse disease development with time, and to recognize factors that may limit disease development. Two time courses of necrosis ofC. album plants were observed, (1) an increase of necrosis followed by a decline, and (2) an increase of necrosis up to completion with subsequent plant death. Courses of necrosis with time could be described by a non-monotonic, critically damped model when plants survived infection and by a monomolecular model when plants died from infection. Disease development was influenced by interactions between wetness period, density of the spore suspension applied, plant development stage at the time of spore application, and temperature. Disease was favoured by a long wetness period, a high number of spores applied, an early plant development stage at the time of spore application, and temperatures of 18 C and higher. Disease development was limited to plant species of the generaChenopodium, Atriplex andSpinacia with differences between the species. Pathogenicity differed significantly between threeA. caulina isolates tested. Variation in resistance between four source populations ofC. album was small. Prospects forA. caulina as a mycoherbicide are discussed.     

Combining permanent aerobiological networks and molecular analyses for large-scale surveillance of forest fungal pathogens: A proof-of-concept

Forest disease management relies principally on a preventive approach in which epidemiological surveillance plays a crucial role. However, efficient and cost-effective surveillance methods are not currently available for large spatial scales. Nevertheless, aerobiological networks have been set up for several decades in many countries to monitor pollen dispersal and provide real-time assessments of allergenic risk. Here, we suggest that the same approach could be used for the surveillance of forest pathogens. Using molecular methods, we analysed samples from 12 sites of the French aerobiological network, at different dates. Both metabarcoding by high-throughput sequencing (using two markers and two different bioinformatics approaches) and real-time PCR targeting eight important forest pathogens were conducted. To validate the approach, temporal and spatial trends of spore detection were compared with field disease data. The metabarcoding approach demonstrated that many fungal plant pathogens could be found in aerobiological samples. Moreover, five of the eight targeted forest pathogens were detected by real-time PCR, with temporal and spatial trends of spore capture consistent with field data. In particular, Hymenoscyphus fraxineus was detected at high frequency in aerobiological samples in the areas where ash dieback has been present for the longest period of time, and at lower frequency in areas with more recent invasion. Spore detection of seasonal pathogens showed a temporal pattern similar to that of disease reports. Overall, our study provides a proof of concept that permanent aerobiological networks combined with molecular methods may provide a useful tool for large-scale surveillance of forest pathogens.     

一株对桃蚜有高致病性球孢白僵菌的分离、筛选与鉴定

为筛选出对桃蚜Myzus persicae具有高致病性的生防真菌,以从陕西省秦岭原始森林采集到的鳞翅目僵虫虫体中分离获得的5株真菌为研究对象,测定其对桃蚜的致病性并筛选出高致病性菌株表现最佳杀蚜活性时的孢子悬浮液浓度,同时结合形态学及18S rDNA和ITS-rDNA序列分析对致病性最高的病原菌进行鉴定。结果显示,从5株菌株中初步筛选出1株高致病性菌株BQ-63,处理7 d后桃蚜的死亡率为80.33%,校正死亡率为81.58%,僵虫率也达到最高,为80.78%;当菌株BQ-63的孢子浓度为108个/mL时,对桃蚜的致病性达到最高,处理7 d后死亡率为89.53%,校正死亡率为90.10%,僵虫率为89.84%;通过形态学和分子生物学鉴定,确定菌株BQ-63为球孢白僵菌Beauveria bassiana。表明菌株BQ-63对桃蚜具有高致病性,可作为生防真菌进行进一步的研究。     

Temporal and spatial dispersal of Thekopsora areolata basidiospores,aeciospores, and urediniospores

Cherry spruce rust causes huge yield losses in Norway spruce seed production in Fennoscandia. The causal agent, Thekopsora areolata, has three types of spores that disperse during spring: basidiospores are produced on basidia that grow out from teliospores in overwintered bird cherry leaf litter to infect new pistillate spruce cones, aeciospores are released from old diseased spruce cones to infect bird cherry leaves, and urediniospores are produced from new bird cherry leaves for reinfection. No study has examined the dispersal of T. areolata spores, including the basidiospores that cause primary infection in spruce cones. In this study, teliospores of T. areolata were germinated in the laboratory and the morphology of basidiospores was described. T. areolata spores were sampled in Ultuna, Sweden and Joutsa, Finland with 21 spore traps at each site. Peaks in aeciospores were observed from 11 to 25 May and from 2 to 8 June at the Finnish site, and from 4 to 18 May at the Swedish site. Urediniospores were first observed 2–3 weeks after the peaks in aeciospores and they were mainly distributed within 10 m from the bird cherry trees. Peaks of 1–2 weeks in basidiospore detection coincided with multiple rain events. The basidiospore peak overlapped with the spruce pollen peak in Finland but not in Sweden. The quantities of basidiospores from different spore traps within 100 m from the spore source had no gradient. Information on spatial and temporal spore release is important for making decisions on disease management strategies.     

Dynamics of cercospora leaf spot disease determined by aerial spore dispersal in artificially inoculated sugar beet fields

Cercospora beticola is one of the most important fungal pathogens of sugar beet, causing cercospora leaf spot (CLS) disease. Due to the decreasing efficacy of various fungicides caused by resistance traits, the development of a sustainable disease management strategy has become more important. Therefore, detailed knowledge about the epidemiology of the pathogen is crucial. Until now, little was known about the spatiotemporal dispersal of C. beticola spores from the primary inoculum source. Rapid detection of C. beticola spores could facilitate a more precise and targeted disease control. Therefore, a TaqMan real-time PCR assay for detection and quantification of C. beticola spores caught with Rotorod spore traps was established. In 2016 and 2017, field trials were conducted to monitor C. beticola aerial spore dispersal and disease development within an inoculated field and in the adjacent noninoculated area. With the established detection method, C. beticola spores were successfully quantified and used as a measure for aerial spore dispersal intensity. The analysis of the spatiotemporal spread of C. beticola spores revealed a delay and decrease of aerial spore dispersal with increasing distance from the inoculated area. Consequently, disease incidence and severity were reduced in a similar manner. These results imply that spore dispersal occurs mainly on a small scale within a field, although long distances can be overcome by C. beticola spores. Moreover, secondary aerial spore dispersal from sporulating leaf spots seems to be the main driver for CLS disease development. These results provide an important basis for further improvement of CLS control strategies.     

Suppression of potato cyst nematode root penetration by the endoparasitic nematophagous fungusHirsutella rhossiliensis

The endoparasitic nematophagous fungusHirsutella rhossiliensis was tested for its ability to suppress root penetration and cyst formation by the potato cyst nematode speciesGlobodera pallida. Isolates ofH. rhossiliensis were obtained from infected potato cyst nematode juveniles from different starch potato fields in The Netherlands. The isolates showed no difference in spore adhesion to juveniles on agar plates (adhesion rate: ±90%). The most rapid growing isolate, CBS 108.94, was used for experiments. Vegetative mycelial colonies ofH. rhossiliensis CBS 108.94, grown in potato dextrose broth, were used as soil inoculum. During submerged cultivation the mycelial colonies produced phialides (spore-bearing cells) but no spores. Exposed to the air, however, spores were rapidly formed. The effect of different soil inoculum densities of mycelial colonies on root penetration byGlobodera pallida was examined in an experiment in 250-ml pots. Up to a mycelial colony concentration representing a potential spore density of 10⁴ g^–1 soil no suppression occurred. At approximated densities of 2.5×10⁴ and 10⁵ spores g^–1 soil the numbers of juveniles which penetrated roots were reduced by 30% and 34%, respectively. The distribution of the inoculum could be improved by fragmentation of the mycelial colonies before soil inoculation. Using mycelial fragments, again no suppression of root penetration was observed up to a potential spore density of 10⁴ g^–1 soil, but at densities of 10⁵ and 10⁶ g^–1 a suppression of 54% and 88%, respectively, was measured. In a greenhouse experiment, soil inoculation with mycelial colonies with a potential spore production of 2.5×10⁵ g^–1 soil resulted in a suppression of root penetration of 37% and 51% after 5 and 6 weeks, respectively, but the number of newly formed cysts after 18 weeks in soil was not different for control and inoculated pots. It is concluded thatH. rhossiliensis may be useful for the reduction of root damage caused by juveniles of potato cyst nematodes, but the usefulness for population control is doubtful.     

Detection and quantification of airborne inoculum of Hymenoscyphus pseudoalbidus using real‐time PCR assays

A method based on real‐time polymerase chain reaction (PCR) and the use of rotating‐arm spore traps was developed for quantifying airborne Hymenoscyphus pseudoalbidus ascospores. The method was sensitive and reproducible, and the collection efficiency was 10% of the spores present in the air. The temporal ascospore dispersal pattern was studied over 3 years by collecting spores every 15 days for a 24 h air‐sampling period during the ash‐growing season. The highest production was detected from the end of June to the beginning of September. The overall ascospore production did not differ significantly among stands within a specific year but there were differences from year to year. There was a positive correlation between air temperature and the number of ascospores trapped, with most of the positive samples being observed at temperatures above 12°C. The vertical profile of ascospore dispersal showed a strong decrease in ascospore density within a height of 3 m, regardless of date of collection. An analysis of the spore traps installed at increasing distances from an infected stand showed that most of the ascospores were deposited downwind within 50 m of the stand. These data are discussed in context of the epidemiology of the disease.     

Multiplex real-time PCR assays for the detection and identification of Heterobasidion species attacking conifers in Europe

Four species of the destructive forest pathogen Heterobasidion annosum sensu lato (s.l.) are present in Europe: H. annosum sensu stricto (s.s.), H. abietinum and H. parviporum are native species, while H. irregulare is a non-native invasive species currently reported only in Italy, yet recommended for regulation throughout Europe. In this study, real-time PCR detection tests were developed for each of the four species, which can be used simultaneously or individually thanks to probes labelled with species-specific fluorescent dyes. The different performance criteria of each assay were evaluated, and it was determined that they were theoretically capable of detecting amounts of DNA corresponding to 311, 29 and 29 cell nuclei in H. annosum s.s., H. irregulare and H. parviporum, respectively. The specificity of each assay was assessed with a wide set of strains. Real-time PCR tests successfully detected Heterobasidion species from 36 fruiting bodies taken from the forest, as well as from artificially inoculated or naturally infected wood samples. The multiplex real-time PCR assays developed in this study could have practical applications both in forest management and in phytosanitary monitoring.     

Comparison of techniques for estimation of resting spores of Plasmodiophora brassicae in soil

Clubroot (Plasmodiophora brassicae) is an important disease of canola (Brassica napus) and other brassica crops. Accurate estimation of inoculum load in soil is important for evaluating producer risk in planting a susceptible crop, but also for evaluation of management practices such as crop rotation. This study compared five molecular techniques for estimating P. brassicae resting spores in soil: quantitative polymerase chain reaction (qPCR), competitive positive internal control PCR (CPIC-PCR), propidium monoazide PCR (PMA-PCR), droplet digital PCR (ddPCR) and loop-mediated isothermal DNA amplification (LAMP). For ddPCR and LAMP, calibrations were developed using spiked soil samples. The comparison was carried out using soil samples collected from a long-term rotation study at Normandin, Québec, with replicated plots representing 0-, 1-, 2-, 3-, 5- and 6-year breaks following susceptible canola infested with clubroot. CPIC-PCR and ddPCR provided repeatable estimates of resting spore numbers in soil compared with estimates from qPCR or LAMP alone. CPIC-PCR provided the most robust measurement of spore concentration, especially in the 2 years following a crop of susceptible canola, because it corrected for effects of PCR inhibitors. PMA-PCR demonstrated that a large proportion of the DNA of P. brassicae detected in soil after the susceptible canola crop was derived from spores that were immature or otherwise not viable. Each assay provided a similar pattern of spore concentration in soil, which supported the conclusion of a previous study at this site that resting spore numbers declined rapidly in the first 2 years after a susceptible crop, but much more slowly subsequently.     

Detection of viridiofungin A and other antifungal metabolites excreted by <Emphasis Type="Italic">Trichoderma harzianum</Emphasis> active against different plant pathogens

Antibiosis is assumed to be an essential mechanism exerted by potential biocontrol agents (BCAs) of Trichoderma spp. Therefore, in the present study, we report for the first time on the elucidation and production of viridiofungin A (VFA) from T. harzianum isolate T23 cultures and investigate the antifungal potential of VFA and some other secondary metabolites purified from T. harzianum cultures against Fusarium moniliforme. The bioautography assay revealed that T. harzianum isolates T16 and T23 excreted several secondary metabolites with antifungal activity. Following isolation and purification of the antifungal zones, three fractions (F223, F323 and F423) from extracts of isolate T23 and two fractions (F416 and F516) from extracts of isolate T16 exhibited pronounced fungitoxic activity in the bioautography and antibiotic disk assays against Cladosporium spp. and F. moniliforme, respectively. The structure of the antifungal metabolite in fraction F323 was identified as viridiofungin A (VFA), the first report of production of VFA by isolate T23 of T. harzianum. Following cultivation of isolate T23 in PDB medium for 9 days, 94.6 mg l⁻¹ of VFA were determined. VFA and fraction F516 retarded the mycelial growth of F. moniliforme in the non-volatile phase assay by >90% for each 250 μg ml⁻¹ 7 days post-inoculation (dpi). While VFA and fraction F416 showed both volatile and non-volatile effects, fraction F516 seemed to exhibit mainly non-volatile activity. Microscopic examination revealed that hyphae of F. moniliforme grown on VFA-amended medium were less branched and appeared thicker than untreated hyphae. Furthermore, in the presence of VFA, formation of chlamydospores by F. moniliforme was increased. Finally, the antifungal spectrum of VFA towards various important plant pathogens was evaluated. Germination of propagules of a variety of fungal pathogens in vitro was differentially inhibited by VFA. While in the presence of 100 μg ml⁻¹ VFA conidial germination of V. dahliae was completely inhibited, a slightly higher concentration (150 μg ml⁻¹) of the inhibitor was required to suppress germination of Phytophthora infestans sporangia or sclerotia of Sclerotinia sclerotiorum. Contrary to several reports in the literature, VFA proved to be fungistatic rather than fungicidal. However, neither VFA nor the other Trichoderma metabolites, such as 6PAP, F416 and F516, exhibited any antibacterial activity against Gram-positive and Gram-negative bacteria.     

Image analysis for the identification of the quarantine pest Tilletia indica*

Diagnosis of the teliospores of Tilletia indica (Karnal bunt) detected in wash tests is complicated by the fact that wheat grain can be contaminated by ‘look‐alike’ species, namely Tilletia walkeri (ryegrass bunt) and Tilletia horrida (rice smut). Although morphological diagnosis is possible when there are relatively large numbers of teliospores present, it is difficult with only a few spores as there are significant overlaps in the characteristics of each species. Molecular methods can confirm presumptive identifications, but these take 2–3 weeks. Image analysis offers the potential for more rapid confirmation. An image analysis system was developed for use on bleached spores. Bleaching spores reveals additional morphological characters (spore profile and spore wall layers) that may be used in an image analysis system to discriminate species. The image‐processing software automatically locates spores on a given image and calculates perimeter, surface area, number of spines and spine size, maximum and minimum ray radius, aspect ratio and roundness. Principal components analysis (PCA) is performed on the parameters to obtain a linear separation of spore species. Accuracy of 97% in separating T. indica and T. walkeri has been achieved in preliminary tests using PCA, but further evaluation is required.     

Local dispersal of Puccinia striiformis f. sp. tritici from isolated source lesions

Understanding how disease foci arise from single source lesions has not been well studied. Here, single wheat leaves were inoculated with Puccinia striiformis f. sp. tritici urediniospores, and all wheat leaves within two intersecting 0.3 × 3.0 m transects were sampled in eight replicates over 3 years. The lesions observed on each of the top three leaves on plants within 1.5 m from the source lesion were three‐dimensionally mapped. The total number of lesions within a 1.5 m radius was estimated by dividing the number of lesions observed within each 0.025 m‐wide annulus by the fraction of the annulus sampled. The estimated total number of lesions produced within 1.5 m of a single source lesion ranged from 27 to 776, with a mean of 288 lesions. Eighty percent of the lesions were recorded within 0.69 m of the source infection. The proportion of total lesions observed at a given distance from the source was fitted well by the Lomax and Weibull distributions, reflecting the large proportion of lesions arising close to the source, and when fitted by an inverse power distribution had a slope (b) of 2.5. There were more lesions produced on leaves higher in the canopy than on lower leaves, with more lesions being detected above than below the point of inoculation. Simultaneous measurement of lesion gradients and spore dispersal in the final year of the study suggests that this pattern is due to greater susceptibility of upper leaves, rather than increased dispersal to upper leaves.     

Variation in rates of spore deposition of Fusarium circinatum, the causal agent of pine pitch canker, over a 12-month-period at two locations in Northern California

Patterns of spore deposition by Fusarium circinatum, the causal agent of pine pitch canker (PPC) of Monterey pine (Pinus radiata) and other conifers, were studied between May 2003 and April 2004 at two sites in Northern California using a novel spore trapping method combined with a real-time polymerase chain reaction (PCR) approach. At each study site, two plots were sampled by placing spore traps at 100 m intervals along transects 600 m in length. The air was sampled continuously by exchanging the spore traps every 2 weeks. The spore deposition rate (DR), ranged from 0 to 1.3 x 10(5) spores m(2). Spores were detected throughout the year, with higher trapping frequencies (TF) during the rainy season (November to April), than during the dry season (May to October). The detection of spores on traps at distances larger than 200 m from any Monterey pine, suggests at least midrange aerial dispersal. Finally, different inoculum loads were associated with trees displaying different levels of disease symptoms, suggesting infectiousness of the pathogen varies as the disease progresses. This study represents one of the first documenting continuous inoculum pressure values over an entire year for a forest pathogen, and provides important epidemiological information that will be invaluable in the development of disease progression models.     

Life-table analysis of faba bean rust

In controlled near-optimum conditions (18 °C), monocyclic sporulation capacity and spore infection efficiency were assessed for faba bean rust on the first and second leaves of field bean. After a latency period of 8–10 days, lesions sporulated duringc. 50 days. Spore production on the second leaf,c. 9×10⁴ spores per lesion, was two times as high as spore production on the first leaf. Infection efficiency was similar for both leaf layers, with a mean value of 0.11 lesion per inoculated spore. Infection efficiency decreased strongly when spores originated from mother lesions older than 20 days. Three life-table statistics (the net reproduction numberR _o , the mean generation timeT _g , and the maximum relative growth rater _max ) were calculated.R _o was larger andT _g was longer for the second than for the first leaf, butr _max was nearly the same for both leaf layers (0.31–0.33 day^–1).r _max was compared with the exponential growth rater measured in a field experiment. From the difference between the two rates, the fraction of inoculum lost in field conditions was estimated at 0.54–0.94. The life-table statistics were also compared to those of other legume rusts, and implications of life-table analysis for comparative epidemiology were discussed.     

Detection of plant pathogens using real‐time PCR: how reliable are late Ct values?

Effective detection of pathogens from complex substrates is a challenging task. Molecular approaches such as real‐time PCR can detect pathogens present even in low quantities. However, weak real‐time PCR signals, as represented by high cycle threshold (C_t) values, may be questionable. Therefore, setting a reliable C_t threshold to declare a positive reaction is important for specific detection. In this study, five methods were assessed for their performance in determining a C_t cut‐off value. These methods were based on the widely used probability of detection (POD) or receiver‐operating characteristic (ROC) approaches. Two important forest pathogens, Hymenoscyphus fraxineus and Fusarium circinatum, were used to set up three experimental frameworks that combined two types of substrates (seed lots and spore traps) and different PCR machines. The ROC‐based method emerged as the most complete and flexible method under various experimental conditions. It was demonstrated that the ROC method leads to a cut‐off value below which late C_t results can reliably be considered indicative of positive test results. This cut‐off value must be determined for each experimental approach used. The method based on the distribution of a previously determined set of C_t values corresponding to false‐positives appeared to be better adapted to detecting false‐negative results, and thus useful for testing potentially invasive pathogens.