Molecular detection and quantification of Colletotrichum abscissum in sweet orange propagative material

Postbloom fruit drop disease (PFD) has caused serious impacts on citrus production in the Americas, occurring sporadically and suddenly during rain in the flowering period. In São Paulo State, Brazil, Colletotrichum abscissum is the causal agent responsible for over 80% of the disease incidence. Pathogen dispersal over long distances and the origin of primary inoculum are still unclear for PFD epidemics. We tested the hypothesis that citrus propagation material can harbour C. abscissum DNA by quantifying it in leaves of budwood increase block trees (BIB) and young citrus plants (YP) using multiplex quantitative PCR (qPCR). C. abscissum DNA was detected in all citrus nurseries, regardless of the type of propagative material, the sweet orange variety, or the nursery location. Overall, 73.4% of all samples from citrus nurseries have DNA from the pathogen, with a detection limit of 10 conidia. The average of 155 conidia found in YP was higher than the conidia observed on leaf samples from BIB (p = 0.03), although leaf samples from cultivars Valencia and Pera did not differ significantly, with means around 127 and 118 conidia, respectively (p = 0.75). This is the first molecular detection of C. abscissum in citrus propagative material. The multiplex qPCR assay may be used as a protocol for an accurate diagnosis of C. abscissum in citrus propagative material, may assist a better understanding of the pathogen dispersal over long distances, and may be used for further studies involving the quantification of C. abscissum in citrus orchards.     

The relative importance of conidia and ascospores as primary inoculum of Venturia inaequalis in a southeast England orchard

Apple scab, caused by Venturia inaequalis, can lead to large losses of marketable fruit if left uncontrolled. The disease appears in orchards during spring as lesions on leaves. These primary lesions are caused by spores released at bud burst from overwintering sources; these spores can be sexually produced ascospores from the leaf litter or asexual conidia from mycelium in wood scab or within buds. The relative importance of conidia and ascospores as primary inoculum were investigated in an orchard in southeast England, UK. Potted trees not previously exposed to apple scab were placed next to (c. 1 m) orchard trees to trap air‐dispersed ascospores. Number and position of scab lesions were assessed on the leaves of shoots from both the potted trees (infection by airborne ascospores) and neighbouring orchard trees (infection by both ascospores and splash‐dispersed, overwintered conidia). The distribution and population similarity of scab lesions were compared in the two tree types by molecular analysis and through modelling of scab incidence and count data. Molecular analysis was inconclusive. Statistical modelling of results suggested that conidia may have contributed approximately 20–50% of the primary inoculum in early spring within this orchard: incidence was estimated to be reduced by 20% on potted trees, and lesion number by 50%. These results indicate that, although conidia are still a minority contributor to primary inoculum, their contribution in this orchard is sufficient to require current management to be reviewed. This might also be true of other orchards with a similar climate.     

Genetic variation and demographic history analysis of Pestalotiopsis,Pseudopestalotiopsis, and Neopestalotiopsis fungi associated with tea (Camellia sinensis) inferred from the internal transcribed spacer region of the nuclear ribosomal DNA

To elucidate the demographic history within and among the genera Pseudopestalotiopsis, Neopestalotiopsis, and Pestalotiopsis and their genetic diversity, we studied internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA) with no saturation effects in 172 Pestalotiopsis-like fungal accessions isolated from Camellia sinensis planted in nine provinces in China (64 from Yunnan, 4 from Guangxi, 2 from Guizhou, 42 from Jiangxi, 3 from Sichuan, 3 from Zhejiang, 1 from Henan, 18 from Fujian, 12 from Chongqing, and 18 from other provinces) and in Japan (1), France (1), Turkey (1), and Thailand (2). Among these, 44 Pestalotiopsis-like isolates from Yunnan in this study formed a clade in phylogenetic trees with Pestalotiopsis accessions in GenBank, despite their cultural characteristics on potato dextrose agar displaying no genus specificity. Based on estimated sequence polymorphism parameters of average pairwise differences (K), haplotype diversity (H_d), nucleotide diversity (Pi), θ (per site) from η, and recombination event rate (R_m), we identified moderate genetic variation in these three genera. Molecular evolution analysis with neutrality tests and mismatch distribution demonstrated that Pseudopestalotiopsis and Pestalotiopsis ribosomal ITS sequences evolved under a neutral model. However, the recent demographic expansion of the three genera promoted geographical isolation but showed no clear geographical structure. Pairwise genetic divergences between haplotypes and a median-joining network revealed Yunnan or Jiangxi as a possible centre of origin, and that Pseudopestalotiopsis and Neopestalotiopsis may derive from Pestalotiopsis.     

Characterization of monilia disease caused by Monilinia linhartiana on quince in southern Spain

This study elucidates the aetiology and epidemiology of monilia disease of quince caused by the fungus Monilinia linhartiana in Spain. Disease incidence and the dynamics of apothecial development and ascospore discharge were quantified and the pathogen was characterized using morphological and molecular methods. The pathogen did not produce conidia or apothecia on agar media but produced conidia on leaves showing symptoms and apothecia on mummified young quince fruit. Monilinia linhartiana was not pathogenic on ripe quince fruit but was readily isolated from developing, mummified fruit (pseudosclerotia). Phylogenetic analysis based on 5·8S‐ITS region sequences placed M. linhartiana in the Disjuntoriae section of Monilinia species infecting rosaceous hosts. Studies during 2004–2008 in four commercial orchards in southern Spain determined two major infection periods for the disease. The first coincided with the unfolding of the first leaves and resulted in leaf blotch and shoot blight. The second coincided with flowering and led to mummification of developing young fruit. Foliar infection was apparently initiated by airborne ascospores produced on pseudosclerotia that overwintered on the soil surface, while flower infection was probably initiated by conidia produced on leaf lesions. Incidence of diseased shoots ranged from 1 to 91% and was correlated with calculated inoculum potential, based on the density and maturity of apothecia formed on pseudosclerotia. This epidemiological study has made it possible to characterize the life cycle of monilia disease on quince in southern Spain, which will help the development of new control strategies.     

Development of a grower‐conducted inoculum detection assay for management of grape powdery mildew

Management of grape powdery mildew (Erysiphe necator) and other polycyclic diseases often relies on calendar‐based pesticide application schedules that assume the presence of inoculum. An inexpensive, loop‐mediated isothermal amplification (LAMP) assay was designed to quickly detect airborne inoculum of E. necator to determine when to initiate a fungicide application programme. Field efficacy was tested in 2010 and 2011 in several commercial and research vineyards in the Willamette Valley of Oregon from pre‐bud break to véraison. In each vineyard, three impaction spore traps were placed adjacent to the trunk. One trap was maintained and used by the grower to conduct the LAMP assay (G‐LAMP) on‐site and the other two traps were used for laboratory‐conducted LAMP (L‐LAMP) and quantitative PCR assay (qPCR). Using the qPCR as a gold standard, L‐LAMP was comparable with qPCR in both years, and G‐LAMP was comparable to qPCR in 2011. Latent class analysis indicated that qPCR had a true positive proportion of 98% in 2010 and 89% in 2011 and true negative proportion of 96% in 2010 and 64% in 2011. An average of 3·3 fewer fungicide applications were used when they were initiated based on spore detection relative to the grower standard practice. There were no significant differences in berry or leaf incidence between plots with fungicides initiated at detection or grower standard practice plots, suggesting that growers using LAMP to initiate fungicide applications can use fewer fungicide applications to manage powdery mildew compared to standard practices.     

The widespread occurrence of overwintered conidial inoculum of <Emphasis Type="Italic">Venturia inaequalis</Emphasis> on shoots and buds in organic and integrated apple orchards across the Netherlands

A 2-year study was conducted to determine the widespread occurrence of overwintered conidial inoculum of Venturia inaequalis and its impact on the apple scab control in 18 apple orchards (organic and integrated) with various levels of scab in the Netherlands. Autumn assessments of scab lesions showed that the integrated orchards had a significantly lower scab incidence (<20%) compared to that of the organic orchards (>60%). At the bud-break phenological stage, the mean numbers of nonviable and viable conidia on 1 cm pieces of shoots ranged from 1 to about 90 and from 6 to more than 1000 in the integrated and the organic orchards, respectively, for both years. However, viable conidia on shoots were found only in 2 integrated and 6 organic orchards out of the 18 and the viability of conidia was below 2%. The mean numbers of viable and nonviable conidia per 100 buds ranged from 24 to more than 1000 and from 230 to almost 5000 in the integrated and the organic orchards, respectively, for both years. In both years, some 60–85% of the conidia was found on the outer bud scales. The percentage viability associated with the outer bud tissues was below 2% for all the orchards. However, the percentage of viable conidia within the inner bud tissues ranged from 0% to 6% in the integrated and from 2% to 11% in the organic orchards for both years. Differences between the organic and the integrated orchards were clearly demonstrated for overwintered conidia associated with both shoot and bud samples. The relationship between autumn scab incidence and numbers of overwintered conidia associated with shoots or buds was exponential. If the autumn scab incidence was above 40%, then the number of overwintered conidia markedly increased. We conclude that specific treatments for overwintering conidia of Venturia inaequalis may not be necessary in integrated orchards with a low scab incidence in the previous autumn. However, the risk of early scab epidemics initiated by overwintered conidia potentially is high in organic orchards. Preventative measures in early spring and also in the previous year must be established in these orchards.     

Phomopsis husk rot of macadamia in Australia and South Africa caused by novel Diaporthe species

Phomopsis husk rot (PHR) in macadamia is a disease of economic importance in major commercial production areas in Australia and South Africa. Effective control of PHR is hindered by limited knowledge about its aetiology and epidemiology. The diversity and pathogenicity of more than 50 isolates of Diaporthe associated with PHR in macadamia orchards in Australia and South Africa was assessed. Multilocus phylogenetic analyses of DNA sequences of the ITS, tef1α, and tub2 gene loci revealed four novel clades that are described as Diaporthe australiana sp. nov., D. drenthii sp. nov., D. macadamiae sp. nov., and D. searlei sp. nov. Pathogenicity tests with representative isolates found that all four species caused PHR of varying severity between and within species, as well as between the two macadamia cultivars HAES 344 and HAES 816. The Australian species, D. australiana, was the most aggressive species compared with the three South African species. This study improves our understanding of the aetiology of PHR in macadamia and paves the way for more effective disease management.     

Development of a qPCR test to detect and quantify Elsinoë piri in unsprayed and organic apple orchards and assessment of apple cultivar susceptibility to the disease

Recently, Elsinoë piri has become an important pathogen in unsprayed and organic apple orchards. The increasing demand for organic apples as well as the limited use of fungicides imposed by European legislation has turned this pathogen, which has been known for over a century in Europe, into a resurgent phytosanitary threat. It also represents a new trait to be taken into account in apple breeding programmes. As E. piri is extremely difficult to isolate and causes symptoms that can be confused with those caused by other pathogens, a quantitative PCR (qPCR) test has been developed based on the internal transcribed spacer (ITS) region of the rDNA gene. This test, which displays a low limit of detection, allows the early detection of the fungus in apple leaves. As a quantitative method, it is a promising tool for breeding purposes as it can be used on leaves to assess the level of quantitative resistance of apple cultivars to the disease. The molecular test is also useful to gain knowledge on the epidemiology of this re-emerging fungal disease. Spore trapping conducted in 2015, 2016 and 2020 using Rotorod devices and the qPCR test showed that airborne inoculum is released at the end of the growing season. Tests were also carried out to demonstrate that the pathogen was detected in buds during winter. A comparison of 18 E. piri isolates based on the sequencing of five genome regions highlighted a high genotypic diversity. All these isolates were detected with the qPCR test developed in this study.     

Is the onset of septoria tritici blotch epidemics related to the local pool of ascospores?

To elucidate the early epidemic stages of septoria tritici blotch, especially the relationship between the onset of epidemics, the local availability of primary inoculum, and the presence of wheat debris, the early disease dynamics and airborne concentration in Zymoseptoria tritici ascospores were concomitantly assessed at a small spatiotemporal scale and over two years, using spore traps coupled with a qPCR assay. One plot, with the crop debris left, provided a local source of primary inoculum, while the other plot, without debris, lacked any. According to the assay's limits of detection, daily spore trap samples were classified as not detectable or not quantifiable, detectable, and quantifiable. The proportions of samples assigned to the different classes and numbers of spores in samples classified as quantifiable were significantly different between years, time periods (from November to March) and spore trap location (field with or without debris). The effect of year on the airborne ascospore concentration was high: 22 daily peaks with more than 230 ascospores m^?3 day^?1 were identified in the autumn of 2012/13, but none in the autumn of 2011/12. The local presence of wheat debris had no obvious effect on the amount of airborne ascospores or on the earliness of the two epidemics, especially in the year with high inoculum pressure (2012/13). These results suggest that the amount of primary airborne inoculum available in a wheat crop is not a limiting factor for the onset of an epidemic.     

A multiplex real-time PCR assay enables simultaneous rapid detection and quantification of bacteria associated with acute oak decline

Acute oak decline (AOD) is a syndrome affecting mature oak trees and is characterized by stem bleeds from vertical fissures on trunks, and inner bark necrosis caused by a polybacterial consortium, in which Gibbsiella quercinecans and Brenneria goodwinii, and to a lesser extent Rahnella victoriana and Lonsdalea britannica, play key roles. Here we report a novel multiplex real-time PCR assay that enables simultaneous and rapid detection and quantification of these four bacterial species from stem bleed swabs. Experiments with axenic cultures were performed to determine specificity and sensitivity of the multiplex quantitative PCR (qPCR). Whilst the primer/probe set for B. goodwinii was species-specific, primer/probe sets for the other three species were able to identify other members of their respective genera. There was no cross detection of genera within the multiplex qPCR, and non-target bacteria were not detected. The multiplex AOD assay had differential sensitivity for each bacterial species. The assay was evaluated on swab samples collected from stem bleeds of declining oak trees at a site in south-east England and was able to detect all four bacterial species. Absolute quantification of the bacteria from swab samples was possible through the inclusion of a standard curve prepared from dilutions of gene copy standards. This diagnostic tool will facilitate rapid detection of AOD-associated bacteria from samples that can easily be taken by non-specialists without specific training, and will also find application in other experimental work such as pathogenicity and control trials.     

Monitoring conidial density of <Emphasis Type="Italic">Monilinia fructigena</Emphasis> in the air in relation to brown rot development in integrated and organic apple orchards

In a three-year Hungarian study, conidial density of Monilinia fructigena in the air determined from mid-May until harvest was related to brown rot disease progress in integrated and organic apple orchards. Conidia of M. fructigena were first trapped in late May in both orchards in all years. Number of conidial density greatly increased after the appearance of first infected fruit, from early July in the organic and from early August in the integrated orchard. Conidial number continuously increased until harvest in both orchards. Final brown rot incidence reached 4.3–6.6% and 19.8–24.5% in the integrated and organic orchards, respectively. Disease incidence showed a significant relationship with corresponding cumulative numbers of trapped conidia both in integrated and organic orchards, and was described by separate three-parameter Gompertz functions for the two orchards. Time series analyses, using autoregressive integrated moving average (ARIMA) models, revealed that the temporal patterns of the number of airborne conidia was similar in all years in both integrated and organic orchards. Conidia caught over a 24-h period showed distinct diurnal periodicity, with peak spore density occurring in the afternoon between 13.00 and 18.00. Percent viability of M. fructigena conidia ranged from 48.8 to 70.1% with lower viability in dry compared to wet days in both orchards and all years. Temperature and relative humidity correlated best with mean hourly conidial catches in both integrated and organic apple orchards in each year. Correlations between aerial spore density and wind speed were significant only in the organic orchard over the 3-year period. Mean hourly rainfall was negatively but poorly correlated with mean hourly conidial catches. Results were compared and discussed with previous observations.     

Prevalence of Phytophthora species in macadamia orchards in Australia and their ability to cause stem canker

In Australia, Phytophthora cinnamomi is the only species reported as the causal agent of stem canker and root rot in macadamia. In other countries, five Phytophthora species have been reported to cause diseases in macadamia, which led us to question if more than one Phytophthora species is responsible for poor tree health in macadamia orchards in Australia. To investigate this, samples were collected from the rhizosphere, stem, and root tissues of trees with and without symptoms, nurseries, and water sources from 70 commercial macadamia orchards in Australia. Phytophthora isolates were identified based on morphological characteristics and DNA sequencing. P. cinnamomi was the most predominant and widely distributed species, and was obtained from the different types of samples including symptomless root tissues. In addition to P. cinnamomi, only P. multivora was isolated from diseased tissue (stem canker) samples. Six other Phytophthora species were obtained from the rhizosphere samples: P. pseudocryptogea, P. citrophthora, P. nicotianae, P. gondwanense, P. sojae, and a new Phytophthora taxon. Only P. cinnamomi was obtained from macadamia nursery samples, while five Phytophthora species were obtained from water sources. Of the heterothallic Phytophthora species, mating type A2 isolates were dominant in P. cinnamomi isolates, whereas only mating type A1 isolates were obtained for P. nicotianae, P. pseudocryptogea, and P. citrophthora. Pathogenicity assays revealed that P. cinnamomi and P. multivora caused significantly larger stem and leaf lesions than P. citrophthora, P. nicotianae, and P. pseudocryptogea. Phytophthora sp. and P. sojae were nonpathogenic towards leaves and stems.     

Characterization of fungi (Fusarium and Rhizoctonia) and oomycetes (Phytophthora and Pythium) associated with apple orchards in South Africa

Several species of fungi and oomycetes including Fusarium, Rhizoctonia, Phytophthora and Pythium have been reported as root pathogens of apple where they contribute to a phenomenon known as apple replant disease. In South Africa, little is known about specific species in these genera and their pathogenicity toward apple. Therefore, these aspects were investigated along with the development and optimization of qPCR tests for detection and quantification of the most virulent oomycete species. In eight investigated orchards, the oomycete Phythophthora cactorum was widely distributed, while nine Pythium species were differentially distributed among the orchards. Pythium irregulare was the most widely distributed and the most virulent species along with P. sylvaticum, P. vexans and Ph. cactorum. Seven binucleate Rhizoctonia anastomosis groups (AGs) were also differentially distributed among the orchards, with the majority appearing to be non-pathogenic while certain AG-I and AG-F isolates exhibited low virulence on apple. In the genus Fusarium, F. oxysporum was widely distributed, but isolates were non-pathogenic. Fusarium solani and F. avenaceum were less frequently encountered, with only some isolates having low virulence. qPCR data obtained from seedling roots inoculated with the most virulent Pythium species (P. irregulare, P. sylvaticum and P. vexans) and the genus Phytophthora were not always reproducible between trials, or isolates of the same species. In general, seedling growth inhibition was associated with the presence of a low amount of pathogen DNA (±40 fg μl⁻¹ to 2 pg μl⁻¹) in roots. Pythium irregulare, although having the lowest DNA concentrations in roots, was the only species for which a significant negative correlation was found between seedling weight and pathogen DNA concentration.     

Multiplex qPCR assay for simultaneous quantification of CYP51-S524T and SdhC-H152R substitutions in European populations of Zymoseptoria tritici

Demethylation inhibitor (DMI) and succinate dehydrogenase inhibitor (SDHI) fungicides are currently relied upon for the control of septoria tritici blotch (STB) in European wheat fields. However, multiple mutations have occurred over time in the genes encoding the targeted proteins that have led to a practical loss of fungicide efficacies. Among the different amino acid substitutions in Zymoseptoria tritici associated with resistance to these fungicides, S524T in CYP51 (DMI target) and H152R in SdhC (SDHI target) are regarded as conferring the highest resistance factors to DMI and SDHI, respectively. To facilitate further studies on the monitoring and selection of these substitutions in Z. tritici populations, a multiplex allele-specific quantitative PCR (qPCR) assay allowing for estimation of both allele frequencies in bulk DNA matrices was developed. The assay was then used on complex DNA samples originating from a spore trap network set up in Belgium, Denmark, Sweden, and Ireland in 2017 and 2018, as well as on leaf samples with symptoms. The S524T allele was present in all field samples and its proportion was significantly higher in Ireland than in Belgium, whereas the proportion of H152R was only sporadically present in both countries. The frequency of S524T varied greatly in the airborne inoculum of all four countries; however, the H152R allele was never detected in the airborne inoculum. The method developed in this study can be readily adopted by other laboratories and used for multiple applications including resistance monitoring in field populations of Z. tritici.     

Evaluation of the temporal distribution of <Emphasis Type="Italic">Fusarium graminearum</Emphasis> airborne inoculum above the wheat canopy and its relationship with Fusarium head blight and DON concentration

With the aim of unravelling the role of airborne Fusarium graminearum inoculum in the epidemic of Fusarium head blight (FHB) caused by this species in wheat spikes, a network of Burkard air samplers was set up in five wheat fields distributed in Belgium from 2011 to 2013. Each year from April to July, the daily amounts of F. graminearum inoculum above the wheat canopy were quantified using a newly developed TaqMan qPCR assay. The pattern of spore trapping observed was drastically different per year and per location with a frequency of detection between 9 and 66% and a mean daily concentration between 0.8 and 10.2 conidia-equivalent/m³. In one location, air was sampled for a whole year. Inoculum was frequently detected from the wheat stem elongation stage until the end of the harvesting period, but high inoculum levels were also observed during the fall. Using a window-pane analysis, different periods of time around wheat flowering (varying in length and starting date) were investigated for their importance in the relation between airborne inoculum and FHB parameters (FHB severity, frequency of F. graminearum infection and DON). For almost all the combinations of variables, strong and significant correlations were found for multiple window lengths and starting times. Inoculum quantities trapped around flowering were highly correlated with F. graminearum infection (up to R?=?0.84) and DON (up to R?=?0.9). Frequencies of detection were also well correlated with both of these parameters. DON concentrations at harvest could even be significantly associated with the F. graminearum inoculum trapped during periods finishing before the beginning of the anthesis (R?=?0.77). Overall, these results highlight the key role of the airborne inoculum in F. graminearum epidemics and underline the importance of monitoring it for the development of disease forecasting tools.     

Detection and quantification of Fusarium commune in host tissue and infested soil using real‐time PCR

Fusarium wilt caused by Fusarium commune is a major limiting factor for Chinese water chestnut (Eleocharis dulcis) production in China. A SYBR Green I real‐time quantitative polymerase chain reaction (qPCR) assay was developed based on the mitochondrial small subunit rDNA of F. commune. Assay specificity of the FO1/FO2 primer set was tested on 41 fungal isolates, and only a single PCR band of c. 178 bp from F. commune was amplified. The detection limits of the assay were 1 fg μL^?1 pure F. commune genomic DNA, 1 pg μL^?1 F. commune genomic DNA mixed with host plant genomic DNA (0·5 ng μL^?1), and 1000 conidia/g soil (artificially inoculated). The amount of F. commune DNA in stem tissues detected by qPCR was significantly correlated with the disease severity (DS) ratings; however, the qPCR assay showed no significant positive correlation between spore densities in soil of different fusarium wilt DS groupings and the DS ratings. The qPCR assay was further applied to 76 soil samples collected from commercial fields of E. dulcis during the 2011 and 2012 growing seasons. The spore density of F. commune detected was positively correlated with disease index in the 2012 growing season but not in 2011. The qPCR method can be used for rapid and specific detection of F. commune in plant and soil samples, which will facilitate monitoring of the pathogen and improvement of disease management.     

Real‐time quantitative PCR assays for the rapid detection and quantification of Fusarium oxysporum f. sp. phaseoli in Phaseolus vulgaris (common bean) seeds

Fusarium oxysporum f. sp. phaseoli (Fop) is a devastating pathogen that can cause significant economic losses and can be introduced into fields through infested Phaseolus vulgaris (common bean) seeds. Efficient seed health testing methods can aid in preventing long‐distance dissemination of this pathogen by contaminated seeds. In order to improve detection of Fop in seed, a rapid, accurate and sensitive real‐time PCR assay (qPCR) protocol was developed for detection of Fop in common bean seeds. Seed lots of seven cultivars with infection incidence ranging from 0·25 to 20% were prepared by mixing known amounts of Fop‐infected seeds with Fop‐free seeds. Direct comparisons between SYBR Green and TaqMan qPCR methods were performed using primers based on the Fop virulence factor ftf1. The primers developed in this study produced a 63 bp product for highly virulent strains of Fop but did not produce an amplicon for nonpathogenic or weakly pathogenic isolates of F. oxysporum from P. vulgaris or other hosts. Under optimized conditions, both qPCR assays detected Fop infection at low levels (0·25%); however, the results suggest the TaqMan assay was more reliable at quantification than the SYBR Green assay. Linear regression models were fitted to the relationships between results of qPCR assays and infection incidence, but the models differed among cultivars. Fungal biomass per seed differed among cultivars and was related to seed size. The results indicate that the TaqMan assay developed in this study is a useful tool for the detection and quantification of Fop in bean seeds.     

Conidial density of <Emphasis Type="Italic">Monilinia</Emphasis> spp. on peach fruit surfaces in relation to the incidences of latent infections and brown rot

To evaluate the effect of conidial density of Monilinia spp. on the fruit surface on the incidence of latent infection and brown rot in peaches, eleven field surveys were performed in commercial orchards located in Cataluña, Spain over four growing seasons from 2002 to 2005, and nine surveys were conducted to determine the sources of overwintered Monilinia spp. inoculum. There was a significant positive relationship (r?=?0.69) between the numbers of conidia of Monilinia spp. on the fruit surface and the incidence of latent infections, but not with brown rot at harvest. Although mummified fruit, twigs and pits have been identified as being able to carry the pathogen from year to year in peaches grown in Spanish orchards, no relationships between any of these sources and the numbers of conidia on the fruit surface, or incidence of latent infection or brown rot were found. The effect of temperature (T), solar radiation (SR), rainfall (R) and wind speed (WS) on the area under the number of conidia of Monilinia spp. curve (AUncC) on peach surfaces was analysed. Regression analysis revealed that T, SR, R, and WS could account for 99% of the total variation in the area of the AUncC on peach surfaces. Thus, in order to reduce the incidence of latent infection and brown rot it is essential not only to remove the sources of primary inoculum but also to reduce the number of Monilinia spp. conidia on the fruit surface. Furthermore, the sources of airborne conidia of Monilinia spp. should be taken into consideration in disease management programmes in Spain.     

Secondary inoculum dynamics of Monilinia spp. and relationship to the incidence of postharvest brown rot in peaches and the weather conditions during the growing season

Eight field surveys over three growing seasons (April to October in 2006, 2007, and 2008) were done in three commercial peach and nectarine orchards in order to determine the secondary inoculum dynamics of Monilinia spp. and relationship to the weather conditions and incidence of postharvest brown rot in the Ebro Valley, which is the main peach fruit growing region in Spain. After regression analysis of the epidemiological data of postharvest brown rot and the climatic variables, a disease outbreak can be predicted from (i) the times of the first appearance of airborne conidia, the first appearance of conidia on the surface of flowers and fruits, and the first latent infection, all of which occur 2?months before harvest (ii) the number of conidia on the fruit surface, 2?weeks and 1?week before harvest, (iii) the preharvest incidence of brown rot, and (iv) the mean environmental temperature from popcorn to harvest. From these results, we confirmed the importance of the secondary inoculum dynamics of Monilinia spp. and the utility of these dynamics to predict an outbreak of brown rot in peaches on the day of their harvest and after their harvesting.     

The role of seeds and airborne inoculum in the initiation of leaf blotch (Rhynchosporium secalis) epidemics in winter barley

Both airborne spores of Rhynchosporium secalis and seed infection have been implied as major sources of primary inoculum for barley leaf blotch (scald) epidemics in fields without previous history of barley cropping. However, little is known about their relative importance in the onset of disease. Results from both quantitative real‐time PCR and visual assessments indicated that seed infection was the main source of inoculum in the field trial conducted in this study. Glasshouse studies established that the pathogen can be transmitted from infected seeds into roots, shoots and leaves without causing symptoms. Plants in the field trial remained symptomless for approximately four months before symptoms were observed in the crop. Covering the crop during part of the growing season was shown to prevent pathogen growth, despite the use of infected seed, indicating that changes in the physiological condition of the plant and/or environmental conditions may trigger disease development. However, once the disease appeared in the field it quickly became uniform throughout the cropping area. Only small amounts of R. secalis DNA were measured in 24 h spore‐trap tape samples using PCR. Inoculum levels equivalent to spore concentrations between 30 and 60 spores per m³ of air were only detected on three occasions during the growing season. The temporal pattern and level of detection of R. secalis DNA in spore tape samples indicated that airborne inoculum was limited and most likely represented rain‐splashed conidia rather than putative ascospores.