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Curvularia americana and C. tropicalis are described as causes of leaf and crown necroses on Bermuda grass (Cynodon dactylon x Cynodon transvaalensis) in Veneto, Northern Italy. These pathogens were characterized using morphological characters, and a multilocus molecular phylogenetic analysis based on the nuclear ribosomal internal transcribed spacer (ITS) region, the partial glyceraldehyde-3-phosphate dehydrogenase (gapdh) and translation elongation factor-1α (tef1-α) genes. Pathogenicity tests and the fulfilment of Koch’s postulates confirmed C. americana and C. tropicalis as foliar pathogens of Bermuda grass. This is the first report of C. americana and C. tropicalis as pathogens of Bermuda grass, and the first record these two fungi in Europe.

Colletotrichum ocimi causes black spot of basil (Ocimum basilicum) and is a serious threat to basil cultivation as it compromises leaf production. The pathogen also infects seeds, which could become primary sources of inoculum for spread of black spot. A SYBR Green real-time PCR assay was developed to detect Colletotrichum ocimi in basil leaves and seeds, based on the partial β-tubulin (tub2) gene sequence. Two primer sets were designed and tested. The selected primer pairs produced amplicons of 130 bp. The real-time PCR assay was validated for analytical specificity, sensitivity, selectivity, repeatability and reproducibility. The assay was specific for C. ocimi with respect to ten Colletotrichum spp. and to another 12 pathogens of basil plants. Sensitivity was 1 pg µL-1 of genomic fungal DNA and amplification analyses were not influenced by basil genomic DNA. The assay detected and quantified C. ocimi in artificially inoculated basil leaves. This is the first specific primer set for C. ocimi, which allows rapid detection and quantification of the pathogen is a useful tool for diagnostics in plants. Detection in seeds would also be possible, but will require an optimized extraction method. The qPCR detection of C. ocimi in planta can contribute to adoption of effective preventive disease management strategies.

Strawberry (Fragaria × ananassa Duch.) is widely cultivated in Italy. During May-June 2022, mild symptoms of an unknown leaf spot disease appeared on 5-10% of June-bearing strawberry (cv. Elodì) plants transplanted in July 2021 in a commercial farm located in the province of Cuneo, North Italy. During September-November 2022, the symptoms appeared also on 10-15% of the plants transplanted in July 2022. The disease was scattered throughout the field, large 600 m2, both on new and senescent leaves. Fungicides (sulphur, Tiovit Jet; penconazole, Topas 10 EC) were applied to the plants according to integrated pest management during the growing period. The disease symptoms were purplish to brown necrotic leaf spots up to 1-3 mm in diameter and chlorotic leaf margins. Black lesions were occasionally observed on the petioles, appearing as small necrotic or larger elongated lesions causing leaf death. Peritechia were observed in planta after about 4 months from sampling and measured (144 to 239 μm and 200 to 291 μm, n = 10). Diseased leaves and petioles from about 10 plants were surface disinfested for 1 min in 1% NaClO, rinsed with sterile water and plated on potato dextrose agar (PDA) amended with 25 mg streptomycin sulphate/liter. A fungus with white cottony colonies was repeatedly recovered and maintained in pure culture on PDA. Biguttulate conidia with rounded ends were measured (4.3 to 8.0 μm and 1.2 to 2.9 μm, average 6.1× 2.3 μm, n = 50) from 21-day old colonies grown in PDA at 22°C and 12 h photoperiod. According to colony and conidia morphology, the isolate was identified as Gnomoniopsis sp. (Walker et al., 2010). The fungal DNA was extracted from a pure culture of one isolate selected as a representative (code FR2-22), by using the E.Z.N.A. Fungal DNA Mini Kit (Omega Bio-Tek, Darmstadt, Germany). The identification was carried out by amplifying and sequencing the internal transcribed spacer (ITS) region and the partial translation elongation factor 1-α (TEF) gene using the primers ITS1/ITS4 and EF-728F/EF2 (Udayanga et al., 2021), respectively. The purified PCR products were sequenced at the BMR Genomics Centre (Padova, Italy) obtaining 551bp (ITS) and 652bp (TEF) sequences deposited in GenBank (Accession nos. OQ179950 and OQ190173, respectively). A BLASTn search of both sequences revealed to be 100% identical to the ITS and TEF loci of Gnomoniopsis fructicola sequences of the isolates VPRI_15547 and CBS 275.51 deposited in GenBank with accession Nos. MT378345 and MT383092. The pathogenicity of the isolate FR2-22 was assessed in two trials by biological tests (3 replicates with 1 plant per replicate/pot) in two greenhouse compartments, kept at temperature 20-24°C and at humidity 80-90%. Healthy leaves of forty-day-old strawberry plants (cv. Elodì) were sprayed with 1-5 x106 conidia/ml obtained from the FR2-22 isolate grown on PDA at 25°C for 20 days. The control (water-sprayed plants) was kept in the same conditions. Small leaf spots similar to the symptoms previously observed in the farm were observed 15 days post inoculation. Furthermore, 30 to 40% of leaves developed symptoms similar to those observed in the field after 25-40 days, while the control remained health. The same fungal isolate was repeatedly reisolated from the affected leaves and petioles and identified based on TEF sequencing. Gnomoniopsis fragariae comb. nov., designed as new name for Gnomoniopsis fructicola (Udayanga et al., 2021), has previously been reported on Fragaria × ananassa plants in Australia and in the USA (Farr and Rossman, 2023). To the best of our knowledge, this is the first report of G. fragariae on strawberry in Italy. The impact of the disease caused by this pathogen could be of high importance in the future of strawberry production in Italy. Healthy propagation material and strict disease management practices in nurseries is a requirement to avoid disease epidemics.

Red raspberry (Rubus idaeus L.) represents a relevant crop which production has largely increased worldwide during the last decade. Cane blight is one of the most common diseases of red raspberry and it can produce considerable losses. During 2019–2021, surveys were conducted in red raspberry orchards in Cuneo province, Northern Italy, to investigate the etiology and pathogen diversity in association with cane blight. Eleven isolates were collected from symptomatic plants of the cultivars ‘Diamond Jubilee’ and ‘Grandeur’, and were identified through multi-locus phylogenies and morphological features. Three fungal species were identified: Paraconiothyrium fuckelii, Diaporthe eres and Neocosmospora parceramosa. Four different genomic regions were included in the molecular analyses: ITS, tef1, tub2 and rpb2. The species pathogenicity was confirmed and P. fuckelii was revealed as the most aggressive. This study provides an insight into raspberry cane blight in Italy and the genomic loci sequencing lay the basis to develop specific diagnostic protocols to monitor the presence of pathogens and to prevent disease spread, adopting effective integrated control strategies.

Women first began to take a stance in science over 50 years ago, and since then, they have produced a great number of publications. However, the first step towards science was a matter of accrediting women as rightful members of the scientific community. During the late nineteenth and early twentieth centuries, women started to enter into graduate programmes, and such a step helped them to make a difference. In many cases and many countries, botanists and mycologists were, in some way, pioneering in the plant pathology discipline. This manuscript considers some of the women who led the way in plant pathology, with a special focus on those who also dealt with the early aspects of plant disease management. Women who were active in different geographical areas are featured, and an attempt has also been made to provide some less well-known stories. Moreover, the importance of women behind the scenes, as is the case for many lab and field technicians, whose lives have not been chronicled, is acknowledged. Finally, some suggestions are given on how to improve the present situation and increase the number of women in science as well as in technical positions, with special emphasis on the positive effect of female mentors and role models.

Climate change already challenges people?s livelihood globally and it also affects plant health. Rising temperatures facilitate the introduction and establishment of unwanted organisms, including arthropods, pathogens, and weeds (hereafter collectively called pests). For example, a single, unusually warm winter under temperate climatic conditions may be sufficient to assist the establishment of invasive plant pests, which otherwise would not be able to establish. In addition, the increased market globalization and related transport of recent years, coupled with increased temperatures, has led to favorable conditions for pest movement, invasion, and establishment worldwide. Most published studies indicate that, in general, pest risk will increase in agricultural ecosystems under climate-change scenarios, especially in today?s cooler arctic, boreal, temperate, and subtropical regions. This is also mostly true for forestry. Some pests have already expanded their host range or distribution, at least in part due to changes in climate. Examples of these pests, selected according to their relevance in different geographical areas, are summarized here. The main pathways used by them, directly and/or indirectly, are also discussed. Understanding these path-ways can support decisions about mitigation and adaptation measures. The review concludes that preventive mitigation and adaptation measures, including biosecurity, are key to reducing the projected increases in pest risk in agriculture, horticulture, and forestry. Therefore, the sustainable management of pests is urgently needed. It requires holistic solutions, including effective phytosanitary regulations, globally coordinated diagnostic and surveillance systems, pest risk modeling and analysis, and preparedness for pro-active management. Made available in DSpace on 2022-09-29T17:05:20Z (GMT). No. of bitstreams: 1 Climate-change-and-pathways-used-by-pests-as-challenges-to-plant-health-in-agriculture-and-forestry-2022.pdf: 461955 bytes, checksum: 6cdb6958897691a0c6b9962fc1a6f01f (MD5) Previous issue date: 2022

Seed health is one of the quality characteristics that must be assessed during the seed certification process. In this study, the freezing blotter method was used to detect fungi present in a total of 102 and 75 maize seed lots produced in 2019 and 2020 respectively, by eight companies in different geographic areas. The analysis of the seed samples produced in 2019 showed significant differences among companies and countries in the percentage of infection with Aspergillus spp. and Fusarium spp. The results obtained with the samples produced in 2020 were not statistically different. PCR-based tests were carried out on 90 isolates to identify the fungal species and differentiate the mycotoxigenic isolates. The results showed that Fusarium verticillioides and Aspergillus flavus were the most common species infecting seed and that 46.5% of F. verticillioides and 57.1% of A. flavus isolates were toxigenic. The most effective seed treatments against Fusarium spp. and Aspergillus spp. contained azoxystrobin and prothioconazole.

Aspergillus flavus may colonize hazelnuts and produce aflatoxins in the field and during storage. The main purpose of this study was to investigate the influence of drying temperature and exposure times on the viability of A. flavus and its ability to produce aflatoxins during the drying process and storage. Hazelnuts were inoculated with A. flavus and dried at different temperatures to reach 6% moisture content and a water activity (aw) of 0.71, a commercial requirement to avoid fungal development and aflatoxin contamination. Hazelnuts were dried at 30, 35, 40, 45, and 50°C and subsequently stored at 25°C for 14 days. After drying at 30, 35, and 40°C, increased amounts of A. flavus were evident, with the highest concentration occurring after drying at 35°C ([6.1 ± 2.4] × 106A. flavus CFU/g). At these temperatures, aflatoxins were detected only at 30 and 35°C. Aflatoxins, however, were present at higher levels after drying at 30°C, with concentrations of 1.93 ± 0.77 μg/g for aflatoxin B1 (AFB1) and 0.11 ± 0.04 μg/g for aflatoxin B2 (AFB2). After 14 days of storage, the highest A. flavus concentration and the highest levels of mycotoxins were detected in samples treated at 35°C ([8.2 ± 2.1] × 107A. flavus CFU/g and 9.30 ± 1.58 μg/g and 0.89 ± 0.08 μg/g for AFB1 and AFB2, respectively). In hazelnuts dried at 45 or 50°C, no aflatoxins were found either after drying or storage, and a reduction of A. flavus viable conidia was observed, suggesting that a shorter and warmer drying is essential to guarantee nut safety. The lowest temperature that guarantees the lack of aflatoxins should be selected to maintain the organoleptic quality of hazelnuts. Therefore, 45°C should be the recommended drying temperature to limit A. flavus growth and aflatoxin contamination on hazelnuts. HIGHLIGHTS

The impact of combined environmental factors, such as temperature and CO₂, on the control of the powdery mildew of zucchini, caused by Podosphaera xanthii, and of different control measures has been studied on plants grown in phytotrons. Five experimental trials were conducted, and the powdery mildew severity of both treated and untreated zucchini plants was found to be significantly affected by the interaction between temperature (three different regimes: 16–18; 18–22; 22–26°C), CO₂ (two concentrations: 400–450 and 800–850 ppm) and the treatments. However, at the end of the trials, the efficacy of all the products was not affected by the different, tested environmental conditions. Sulphur consistently provided the highest disease control (75%–85% efficacy). Among the resistant inducers that were tested, calcium oxide was the most effective, in terms of powdery mildew control under all the conditions tested in phytotrons, reducing disease severity from 46% to 61%. Foliar applications of phosphite (14%–28% efficacy), Ampelomyces quisqualis (12%–23% efficacy) and potassium silicate (13%–24% efficacy) only slightly reduced the disease severity for all the tested temperature regimes and CO₂ concentrations, compared to the untreated control. The results obtained under our experimental conditions show that a possible increase in CO₂ concentration and temperature, which is expected for the next few years, should not influence the efficacy of the tested resistance inducers or of sulphur against powdery mildew on zucchini. Moreover, the suppressive effect of calcium oxide is in light of its possible use in greenhouses for zucchini powdery mildew control under 400–450 ppm of CO₂ and under enriched condition of 800–850 ppm of CO₂.

Among the many responsibilities of the worldwide scientific community are advancing the knowledge base that underpins each scientific discipline, addressing the pressing scientific issues of the day (e.g., emerging infectious diseases, food security, and climate change), and perhaps most importantly, educating and training subsequent generations of scientists. Yet, around the globe, advances in scientific and communications technology, proliferation and mining of data, and increasing financial constraints of university systems have led to fundamental changes in our institutions of higher learning. Increasing emphasis on interdisciplinary and multidisciplinary approaches to problem solving in agriculture add to the complexity of providing robust preparation for the plant pathologists of the future. Thus, as the U.N. recognizes the year 2020 as the International Year of Plant Health, it is fair to ask if current approaches to graduate education in plant pathology are adequate to meet current and anticipated challenges and if the outcomes can be improved.

Plants are frequently exposed to prolonged and intense drought events. To survive, species must implement strategies to overcome progressive drought while maintaining sufficient resources to sustain the recovery of functions. Our objective was to understand how stress rate development modulates energy reserves and affects the recovery process. Grenache Vitis vinifera cultivar was exposed to either fast-developing drought (within few days; FDD), typical of pot experiments, or slow-developing drought (few weeks, SDD), more typical for natural conditions. FDD was characterized by fast (2-3 days) stomatal closure in response to increased stress level, high abscisic acid (ABA) accumulation in xylem sap (>400 μg L-1 ) without the substantial changes associated with stem priming for recovery (no accumulation of sugar or drop in xylem sap pH). In contrast, SDD was characterized by gradual stomatal closure, low ABA accumulation (

Background Bio-hydrogen production via dark fermentation of low-value waste is a potent and simple mean of recovering energy, maximising the harvesting of reducing equivalents to produce the cleanest fuel amongst renewables. Following several position papers from companies and public bodies, the hydrogen economy is regaining interest, especially in combination with circular economy and the environmental benefits of short local supply chains, aiming at zero net emission of greenhouse gases (GHG). The biomasses attracting the largest interest are agricultural and urban green wastes (pruning of trees, collected leaves, grass clippings from public parks and boulevards), which are usually employed in compost production, with some concerns over the GHG emission during the process. Here, an alternative application of green wastes, low-value compost and intermediate products (partially composted but unsuitable for completing the process) is studied, pointing at the autochthonous microbial consortium as an already selected source of implementation for biomass degradation and hydrogen production. The biocatalysts investigated as mainly relevant for hydrogen production were the [FeFe]-hydrogenases expressed in Clostridia, given their very high turnover rates. Results Bio-hydrogen accumulation was related to the modulation of gene expression of multiple [FeFe]-hydrogenases from two strains (Clostridium beijerinckii AM2 and Clostridium tyrobutyricum AM6) isolated from the same waste. Reverse Transcriptase quantitative PCR (RT-qPCR) was applied over a period of 288 h and the RT-qPCR results showed that C. beijerinckii AM2 prevailed over C. tyrobutyricum AM6 and a high expression modulation of the 6 different [FeFe]-hydrogenase genes of C. beijerinckii in the first 23 h was observed, sustaining cumulative hydrogen production of 0.6 to 1.2 ml H2/g VS (volatile solids). These results are promising in terms of hydrogen yields, given that no pre-treatment was applied, and suggested a complex cellular regulation, linking the performance of dark fermentation with key functional genes involved in bio-H2 production in presence of the autochthonous consortium, with different roles, time, and mode of expression of the involved hydrogenases. Conclusions An applicative outcome of the hydrogenases genes quantitative expression analysis can be foreseen in optimising (on the basis of the acquired functional data) hydrogen production from a nutrient-poor green waste and/or low added value compost, in a perspective of circular bioeconomy.

Highbush blueberry is an increasingly important crop due to its economic value and demonstrated health benefits of blueberries. Leaf spots are considered as minor diseases of blueberry plants, but they adversely affect blueberry productivity, causing reduced photosynthetic activity, flower bud formation and berry production. Surveys of blueberry crops were conducted in Piedmont, Northern Italy, during 2019-2020. Fungi isolated from leaf spots of the blueberry cultivar ‘Blue Ribbon’ were identified as Colletotrichum helleniense through a robust multi-locus phylogeny. Eight genomic loci were considered: tub, gapdh, act, cal, his3, chs-1, ApMat and gs. Morphological characters of a representative strain were assessed. Pathogenicity was confirmed on four blueberry cultivars, although with different levels of aggressiveness to the cultivars. This study shows the importance of a polyphasic approach to investigate species of Colletotrichum, and the relevance of molecular tools for the species-level characterization within the ‘Kahawae’ clade. This is the first report of Colletotrichum helleniense causing leaf anthracnose on Vaccinium corymbosum.

A monitoring was conducted in 35 commercial vineyards located in Piedmont (North-West Italy) in September 2018 to test the sensitivity of Botrytis cinerea populations to the fungicides that were commonly used in the past and at present: benzimidazoles, dicarboximides, anilinopyrimidines, phenylpyrroles, hydroxyanilides and SDHI fungicides. Sensitivity was tested by evaluating the spore germination on a medium amended with a discriminatory concentration of the different tested botryticides. Conidial suspensions were prepared from infected, randomly sampled bunches and used for the tests. In comparison with the years 2008–2014, the following trend was recorded: benzimidazole resistance was quite stable and was found in about 50% of the vineyards involved in the monitoring; dicarboximide resistance had disappeared in the tested samples, while the frequency of resistance to anilinopyrimidines had increased; the resistance to fenhexamid had decreased; in the case of boscalid, the presence of resistance was quite stable; no resistance to phenylpyrroles was found, although the high discriminatory dose used for fludioxonil cannot exclude the presence in the tested vineyards of low levels of resistance to this fungicide, typical of multidrug-resistant strains.

Monilinia laxa and M. fructicola are two causal agents of brown rot, one of the most important diseases in stone fruit. Two species cause blight on blossoms and twigs and brown rot on fruit in pre- and postharvest. Both species are distributed worldwide in North and South America, Australia, and Japan. In Europe, M. laxa is endemic, while M. fructicola was introduced in 2001 and it is now widespread in several countries. Currently, both species coexist in European stone fruit orchards. Monilinia spp. overwinter in cankers and mummified fruit. Mummy monitoring during winter permits growers to understand which species of Monilinia will be prevalent in an orchard during the following season, permitting planning of an appropriate crop protection. Traditionally, the identification has been carried out using morphological features and even with polymerase chain reaction (PCR)-based assays that requires time and well-equipped laboratories. In this study, two isothermal-based methods were designed to identify these pathogens in a faster way than using traditional methods. The loop-mediated amplification (LAMP) assays were validated on some isolates of Monilinia spp. coming from the mummy monitoring according to the international European and Mediterranean Plant Protection Organization standard (PM7/98), taking into account specificity, sensitivity, repeatability, and reproducibility. The sensitivity of both assays was checked by monitoring (at different time points) two nectarine varieties artificially inoculated and stored at two different temperatures. The reliability of both LAMP assays against the quantification of the inoculum was compared with previously published quantitative PCR assays. Both LAMP methods were able to detect a low number of cells. These LAMP methods could be a useful tool for monitoring brown rot causal agents in the field and during postharvest.

Background Aspergillus fumigatus, the causal agent of aspergillosis in humans, is commonly present as a saprophyte in various organic substrates, such as spoiled silages. Aspergillosis is generally combated with demethylation inhibitor (DMI) fungicides, but the recent appearance of resistant medical and environmental strains made current treatment strategies less reliable. The goal of this study was to determine the evolution of A. fumigatus populations during the ensiling process of whole-crop corn, high moisture corn and wet grain corn, and to monitor the sensitivity of isolates from treated and untreated fields to one medical and one agricultural DMI fungicide. Results A. fumigatus was isolated from fresh forage at harvest at rather low concentrations (102 cfu g-1 ). The low frequency lingered during the silage process (at 60 and 160 days), whereas it significantly increased during air exposure (at 7 and 14 days of air exposure). Field treatment of corn with a mixture of prothioconazole and tebuconazole did not affect the sensitivity of A. fumigatus isolates. One of 29 isolates from the untreated plot was resistant to voriconazole. A unique amino acid substitution (E427K) was detected in the cyp51A gene of 10 of 12 sequenced isolates, but was not associated with DMI resistance. Conclusion A. fumigatus significantly increased during aerobic deterioration of ensilaged corn after silo opening, compared with the low presence in fresh corn and during ensiling. Field treatment of corn with DMI fungicides did not affect the sensitivity of A. fumigatus isolates collected from fresh and ensiled corn. © 2019 Society of Chemical Industry.

The genera Paramyrothecium and Albifimbria have been established from the former genus Myrothecium and they generally comprise common soil-inhabiting and saprophytic fungi. Within these genera, only two fungi have been recognized as phytopathogenic thus far: P. roridum and A. verrucaria, both of which cause necrotic leaf spots and plant collapse. Severe leaf necrosis and plant decay have been observed in Northern and Southern Italy on leafy vegetable crops. Thirty-six strains of Paramyrothecium- and Albifimbria-like fungi were isolated from affected plants belonging to eight different species. Based on morphological characteristics, 19 strains were assigned to A. verrucaria, whereas the remaining strains, which mostly resembled Paramyrothecium-like fungi, could not be identified precisely. Molecular characterization of six loci (internal transcribed spacer [ITS], β-tubulin [tub2], calmodulin [cmdA], translation elongation factor 1-alpha [tef1], large subunit ribosomal RNA [LSU], and mitochondrial ATP 6synthase 6 [ATP6]) of the 36 new isolates and three previously ITS-characterized isolates assigned all strains to four species: A. verrucaria, P. roridum, P. foliicola, and P. nigrum. Single and concatenated phylogenetic analyses were conducted, and they clearly distinguished the isolated fungi into four different groups. A. verrucaria, P. roridum, P. foliicola, and P. nigrum were able to induce leaf necrosis singly, and they were confirmed to be the causal agents of the leaf spot disease through pathogenicity assays. The involvement of fungi previously considered saprophytic (i.e., P. foliicola and P. nigrum) in the development of plant disease for the first time deserves particular attention because of the possibility of their transmission by seeds and the limited knowledge of their management with chemicals.

Some plant species are capable of significant reduction of xylem embolism during recovery from drought despite stem water potential remains negative. However, the functional biology underlying this process is elusive. We subjected poplar trees to drought stress followed by a period of recovery. Water potential, hydraulic conductivity, gas exchange, xylem sap pH, and carbohydrate content in sap and woody stems were monitored in combination with an analysis of carbohydrate metabolism, enzyme activity, and expression of genes involved in sugar metabolic and transport pathways. Drought resulted in an alteration of differential partitioning between starch and soluble sugars. Upon stress, an increase in the starch degradation rate and the overexpression of sugar symporter genes promoted the efflux of disaccharides (mostly maltose and sucrose) to the apoplast. In turn, the efflux activity of the sugar-proton cotransporters caused a drop in xylem pH. The newly acidic environment induced the activity of apoplastic invertases leading to the accumulation of monosaccharides in the apoplast, thus providing the main osmoticum necessary for recovery. During drought and recovery, a complex network of coordinated molecular and biochemical signals was activated at the interface between xylem and parenchyma cells that appeared to prime the xylem for hydraulic recovery.

Maize (Zea mays L.) is a cereal crop of great economic importance in Italy; production is currently of 62,587,469 t, with an area that covers 628,801 ha, concentrated in northern Italy (ISTAT 2020). Fusarium species are associated with root and crown rot causing failures in crop establishment under high soil moisture. In 2019 maize seedlings collected in a farm located in San Zenone degli Ezzelini (VI, Italy) showed root and crown rot symptoms with browning of the stem tissues, wilting of the seedling, and collapsing due to the rotting tissues at the base of the stem. The incidence of diseased plants was approximately 15%. Seedlings were cleaned thoroughly from soil residues under tap water. Portions (about 3-5 mm) of tissue from roots and crowns of the diseased plants were cut and surface disinfected with a water solution of NaClO at 0.5% for 2 minutes and rinsed in sterile H20. The tissue fragments were plated on Potato Dextrose Agar (PDA) amended with 50 mg/l of streptomycin sulfate and incubated for 48-72 hours at 25oC. Over the 80 tissue fragments plated, 5% were identified as Fusarium verticillioides, 60% as Fusarium spp., 35% developed saprophytes. Fusarium spp. isolates that showed morphological characteristics not belonging to known pathogenic species on maize were selected and used for further investigation while species belonging to F. oxysporum were discarded. Single conidia of the Fusarium spp. colonies were cultured on PDA and Carnation Leaf Agar (CLA) for pathogenicity tests, morphological and molecular identification. The colonies showed white to pink, abundant, densely floccose to fluffy aerial mycelium. Colony reverse showed light violet pigmentation, in rings on PDA. On CLA the isolates produced slightly curved macronidia with 3 septa 28.1 - 65.5 µm long and 2.8-6.3 µm wide (n=50). Microconidia were cylindrical, aseptate, 4.5 -14.0 µm long and 1.5-3.9 µm wide (n=50). Spherical clamydospores were 8.8 ± 2.5 µm size (n=30), produced singly or in pairs on the mycelium, according to the description by Skovgaard et al. (2003) for F. commune. The identity of two single-conidia strains was confirmed by sequence comparison of the translation elongation factor-1α (tef-1α), and RNA polymerase II subunit (rpb2) gene fragments (O'Donnell et al. 2010). BLASTn searches of GenBank, and Fusarium-ID database, using the partial tef-1α (MW419921, MW419922) and rpb2 (MW419923, MW419924) sequences of representative isolate DB19lug07 and DB19lug20, revealed 99% identity for tef-1α and 100% identity to F. commune NRRL 28387(AF246832, AF250560). Pathogenicity tests were carried out by suspending conidia from a 10-days old culture on PDA in sterile H2O to 5×104 CFU/ml. Fifty seeds were immersed in 50 ml of the conidial suspension of each isolate for 24 hours and in sterile water (Koch et al. 2020). The seeds were drained, dried at room temperature, and sown in trays filled with a steamed mix of white peat and perlite, 80:20 v/v, and maintained at 25°C and RH of 80-85% for 14 days with 12 hours photoperiod. Seedlings were extracted from the substrate, washed under tap water, and observed for the presence of root and crown rots like the symptoms observed on the seedlings collected in the field. Control seedlings were healthy and F. commune was reisolated from the symptomatic ones and identified by resequencing of tef-1α gene. F. commune has been already reported on maize (Xi et al. 2019) and other plant species, like soybean (Ellis et al. 2013), sugarcane (Wang et al. 2018), potato (Osawa et al. 2020), indicating that some attention must be paid in crop rotation and residue management strategies. To our knowledge this is the first report of F. commune as a pathogen of maize in Italy. References Ellis M L et al. 2013. Plant Disease, 97, doi: 10.1094/PDIS-07-12-0644-PDN. ISTAT. 2020. http://dati.istat.it/Index.aspx?QueryId=33702. Accessed December 28, 2020. Koch, E. et al. 2020. Journal of Plant Diseases and Protection. 127, 883-893 doi: 10.1007/s41348-020-00350-w O'Donnell K et al. 2010. J. Clin. Microbiol. 48:3708. https://doi.org/10.1128/JCM.00989-10 Osawa H et al. 2020. Journal of General Plant Pathology, doi.org/10.1007/s10327-020-00969-5. Skovgaard K 2003. Mycologia, 95:4, 630-636, DOI: 10.1080/15572536.2004.11833067. Wang J et al. 2018. Plant Disease, 102, doi/10.1094/PDIS-07-17-1011-PDN Xi K et al. 2019. Plant Disease, 103, doi/10.1094/PDIS-09-18-1674-PDN.

Brown rot is a common apple disease in Italy, caused by Monilinia fructicola, M. laxa and M. fructigena (Martini et al. 2013). In September 2020, in a 'Jeromine' apple orchard under integrated pest management located in Scarnafigi (44°39'N, 7°33'E, north-western of Italy), fruits (8.6%) showing brown to blackish firm lesions (6.0 to 8.0 cm diameter) were observed. In some fruits, rots were covered by yellowish stromata. Two isolates (MPI1; MPI2) were obtained from two symptomatic apples and cultured on potato dextrose agar (PDA) for 7 days at 25°C in 12-h light/12-h dark regime. A white-to-greyish mycelium with slightly undulate margins and irregular, black stromata developed on PDA after 12 days incubation. Conidia, observed in branched monilioid chains, (Suppl. Fig. 1) were one-celled, globose, limoniform, hyaline, 38 to 58 μm (mean: 48) × 20 to 44 μm (mean: 33). Based on morphology, the isolates were tentatively identified as Monilinia polystroma (G.C.M. Leeuwen) Kohn. A polymerase chain reaction with primers ITS1 and ITS4 was performed on internal transcribed spacer (ITS) region 1 and 2 and 5.8S gene. The sequenced amplicons (435 bp - 445 bp; GenBank Accession No. MW600854; MW600855) showed 100% identity to the reference isolate of M. polystroma (HQ846944) and to other isolates from apples (AM937114; JX315717) and plum (GU067539). The ITS region of M. polystroma had five nucleotides to distinguish it from the closest species M. fructigena (Zhu et al. 2016; MH862738) (Suppl. Fig. 2). The pathogenicity of both isolates was tested on mature 'Jeromine' apples (10.1% total soluble solids). Three replicates of six apples per isolate were surface disinfected with 1% NaClO. A mycelial plug (5 mm) from colony grown on PDA was inserted using a cork borer into a hole (6 mm) in each fruit (Vasić et al. 2016). Apples inoculated with sterile PDA plugs were used as control. Fruits were placed at 22 ± 1 °C, 85% relative humidity and 12 h light/12 h dark regime. Lesion size was measured after 3, 6 and 9 days of incubation. All inoculated fruits developed typical brown rot symptoms 6 days after inoculation and yellowish stromata appeared on the surface; control fruit remained healthy (Suppl. Fig. 3). The virulence of both isolates was statistically similar (Suppl. Table 1). M. polystroma was reisolated from all inoculated fruits and confirmed by molecular methods. This is the first report of M. polystroma on apple in Italy. M. polystroma was previously reported on apple in Hungary (Petróczy et al. 2009), on apricot in Switzerland (Hilber-Bodmer et al. 2012), on peach and pear in Italy (Martini et al. 2014; 2015), on plum in China (Zhu et al. 2016), and on apple in Serbia (Vasić et al. 2018). The emergence of this pathogen for pome and stone fruit production in Europe stimulates to study its biology and epidemiology, and its fitness and management, as compared to the other endemic Monilinia species.

This study represents the first survey studying the occurrence, genetic diversity, and pathogenicity of Botryosphaeriaceae species associated with symptomatic citrus species in citrus-production areas in five European countries. Based on morphological features and phylogenetic analyses of internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA), translation elongation factor 1-alpha (TEF1) and β-tubulin (TUB2) genes, nine species were identified as belonging to the genera Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum. Isolates of Neofusicoccum parvum and Diplodia pseudoseriata were the most frequently detected, while Dothiorella viticola had the widest distribution, occurring in four of the five countries sampled. Representative isolates of the nine Botryosphaeriaceae species used in the pathogenicity tests caused similar symptoms to those observed in nature. Isolates assayed were all re-isolated, thereby fulfilling Koch’s postulates. Isolates of Diplodia pseudoseriata and Diplodia olivarum are recorded for the first time on citrus and all species found in our study, except N. parvum, are reported for the first time on citrus in Europe.

This study was aimed at identifying Alternaria species associated with heart rot disease of pomegranate fruit in southern Italy and characterizing their mycotoxigenic profile. A total of 42 Alternaria isolates were characterized. They were obtained from pomegranate fruits with symptoms of heart rot sampled in Apulia and Sicily and grouped into six distinct morphotypes based on macro- and microscopic features. According to multigene phylogenetic analysis, including internal transcribed spacer (ITS), translation elongation factor 1-α (EF-1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and a SCAR marker (OPA10-2), 38 isolates of morphotypes 1 to 5 were identified as Alternaria alternata, while isolates of morphotype 6, all from Sicily, clustered within the Alternaria arborescens species complex. In particular, isolates of morphotype 1, the most numerous, clustered with the ex-type isolate of A. alternata, proving to belong to A. alternata. No difference in pathogenicity on pomegranate fruits was found between isolates of A. alternata and A. arborescens and among A. alternata isolates of different morphotypes. The toxigenic profile of isolates varied greatly: in vitro, all 42 isolates produced tenuazonic acid and most of them other mycotoxins, including alternariol, alternariol monomethyl ether, altenuene and tentoxin.

In 1845, when the potato late blight reached Ireland, potato was a big staple food in the country. Two varieties of this species, selected over time for their high productivity and popularity among consumers, were cultivated on countless acres. In fact, in 1800s, the Irish had developed a complete dependence on this crop, which came from South America, mostly from the region of Lake Titicaca, between Peru and Bolivia.

Urban and industrial waste management is an ongoing major issue for environment and economy, involving not only public authorities but also enterprises.

In 1979 I had been working for a few months at the Institute of Plant Pathology and already Angelo Garibaldi and Alberto Matta were proposing me to apply for a scholarship abroad. I could not believe it! Their advice was evidently not to assume I would win a position on the first try, since I was still young and had few academic titles, but it was worth taking a shot.

The importance and sad fame of Claviceps purpurea, a fungus that attacks rye, are linked to the disastrous consequences for human and animal health caused by the presence of its sclerotia in the flours and fodder.

Few of us know that the first great economic crisis—a real financial crash—was unleashed by tulips in the Dutch Republic of late 1600s. Who would have said that the fondness for this flower, made popular by the Ottoman Turks, could have led to such speculative mania? Let’s try to retrace this curious and fascinating story.

Global food security depends to a substantial degree on the rice crop. Rice production is threatened by many pathogens, requiring a range of prevention and control strategies. These need to be integrated with efficient monitoring and detection. This chapter focuses on diagnostic tools developed for three of the most important rice pathogens: Pyricularia oryzae, the agent of rice blast; Bipolaris oryzae, the agent of rice brown spot; and Fusarium fujikuroi, the agent of bakanae disease. Accurate, effective, sensitive and specific diagnostic techniques are required to forecast and manage these diseases. Observation of symptoms in field crops is not always accurate. Serological and, especially, molecular techniques present advantages in terms of speed, specificity and ease of use. Whole-genome sequencing has revealed novel genomic regions and genes and can be used to distinguish between clonal lineages of pathogens. High-throughput sequencing applied to the aerial mycobiome of rice crops has revealed substantial biodiversity that can affect the airborne inoculum and may play a key role in survival and dispersal of fungal spores. The LAMP (Loop-mediated isothermal AMPlification) assay has been identified as a suitable solution for on-site detection. The application of LAMP combined with alkaline DNA extraction has an enormous potential to assess airborne and other inoculum. The use of molecular field testing is revolutionising disease diagnostics, giving results in a few hours. There are exciting prospects of further advances through machine learning. Improved detection should lead to better disease control and hence improved food security. However, pathogens are genetically dynamic so plant disease management will remain challenging.

Once upon a time there was methyl bromide… If it were a fairy tale it would start like this. But it is not a fairy tale. Or maybe it is. Because this topic has really involved twenty years of my life and, a bit like it had been, in the eighties, fifteen years before with grey mould, it left its mark.

Today Ceylon (now Sri Lanka) is worldwide known for its tea production, but few of us know that in 1870 this island was the main international coffee producer.

Twelve trials, in closed soilless culture under controlled conditions, were carried out to evaluate the efficacy of resistance inducers (based on K-phosphite and K-silicate used alone or in combination), and of experimental biocontrol agents (Trichoderma sp. TW2, a mixture of Pseudomonas FC 7B, FC 8B, and FC 9B, Fusarium solani FUS25, Pseudomonas sp. PB26), and a commercial formulation of Trichoderma gamsii + T. asperellum, against diseases caused by Phytophthora capsici of sweet pepper. The products were applied using three different protocols, and effects on incidence of Phytophthora crown, stem and root rots (% dead plants), disease development (area under the disease progress curve; AUDPC), and plant fresh weights were evaluated. Potassium phosphite, applied directly at standard P2O5:K2O, 1.30 + 1.05 g L-1) and at half standard rates, onto growing media, or via nutrient solution, and before infestation of peat plant growing medium with P. capsici, provided the best disease management in a dose-dependent manner, with an 80% reduction of Phytophthora crown, stem and root rots for the standard dosage and for both types of application. These treatments also reduced proportions of dead plants by 47% from the standard rate and by 62% at the half standard rate, when applied via the nutrient solution or directly to the substrate. K-silicate alone partially reduced the percentage of dead plants, with efficacy of 20–23%. No improvement in disease control was observed when K-silicate was applied in combination with phosphite, while K-silicate alone or combined with K-phosphite reduced disease development, compared to untreated controls. Biocontrol agents (BCAs), applied preventively, reduced Phytophthora crown, stem and root rots, with similar or better results than those from the commercial mixture of Trichoderma asperellum + T. gamsii. Among the tested BCAs, Fusarium solani FUS25 provided the most consistent disease reduction (60–65%) and gave increased plant fresh weights. All the tested BCAs reduced disease development, with a similar trend for different disease pressures. The least AUDPC values, compared to the non-treated controls, were from Fusarium solani FUS25, followed by the tested Pseudomonas strains and Trichoderma sp. TW2. These results indicate the potential for potassium phosphite and biocontrol agents in management of Phytophthora crown, stem and root rots of pepper grown in soilless systems.

Very often the most interesting results in the agricultural sector come from acute observations by farmers. It was the mid-1970s when an Israeli farmer noticed that the fields in which plastic films, used to reduce weed growth during the summer, were kept on the ground longer presented, subsequently, more limited attacks than some soil-borne fungal pathogens (the same descripted in the last chapter).

Paradoxically, protecting crops from pests is more difficult today than it was in the past, despite the availability of effective and sophisticated technologies, and it will be even more so in the future. In a context of general crisis that requires to lower production costs, and with the need to limit the environmental impact of interventions, the role of research will be increasingly important.

The “story” of Phytophthora ramorum is another example of the economic and environmental impact of introducing an exotic pathogen on forest plants.

Consumers’ negative attitude to pesticides is well-known. These chemical products used for plant disease management are often considered “poisonous” for human health. Yet they are not poisons, just agrochemicals (see box page 167), which, after many years of rigorous studies, as it happens with pharmaceuticals, are registered and made available to farmers as control means against pests threatening agricultural crops (see box page 168).