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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.

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.

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.

A TaqMan quantitative PCR (qPCR) assay based on the translation elongation factor 1-α gene was developed for the quantification of

Serious outbreaks of Alternaria leaf spot and plant decay have recently been recorded on several ornamental plants in the Biella Province (Northern Italy). Twenty-two fungal isolates were obtained from

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 (Vasic 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 (Petroczy 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 (Vasic 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.

The ready-to-eat salad sector, also called fresh-cut or bagged salads, is a fast-growing segment of the fresh-food industry. The dynamism and specialization of this sector, together with the lack of adequate crop rotation, the globalization of the seed market, and climate change, are the main causes of the development of many new diseases that cause severe production losses. Newly detected diseases of the most important crops grown (lettuce, wild and cultivated rocket, lamb’s lettuce, chicory, endive, basil, spinach, and Swiss chard) are critically discussed. The management of these diseases represents a formidable challenge, since few fungicides are registered on these minor-use crops. An interesting feature of the ready-to-eat salad sector is that most crops are grown under protection, often in soilless systems, which provide an environment helpful to the implementation of innovative control methods. Current trends in disease management are discussed, with special focus on the most sustainable practices.

Cineraria maritima L. (synonym Senecio cineraria DC.), commonly known as dusty-miller, is grown in Italy for landscape use in parks and gardens. In the spring of 2001, severe outbreaks of a previously unknown disease were observed in commercial farms located in northern Italy. Leaves of infected plants showed several sori on the abaxial surface, progressing to the adaxial surface, and often in the interveinal areas. On the adaxial surface of leaves, chlorotic areas developed and eventually turned brown. Severely infected leaves wilted, but remained attached to the stem. Signs of the fungus were present as whitish and catenulate sporangia emerging from the sori. Sporangia, organized in chains, had an average diameter of 20.5 × 26.5 μm. On the basis of the microscopic observations, the causal agent of the disease was identified as Albugo tragopogonis. Pathogenicity was confirmed by inoculating leaves of healthy C. maritima plants with a sporangial suspension (5 × 102 sporangia per ml) obtained from infected plants. Noninoculated plants served as a control. Plants were kept covered with plastic bags for 72 h and maintained at 15°C. After 10 days, typical symptoms of white rust developed on inoculated plants starting from the basal leaves. Within 30 days, affected leaves were completely wilted. Microscopic examination of sporangia within sori verified the pathogen to be A. tragopogonis. No symptoms developed on the control plants. A. tragopogonis has been reported as the causal agent of white rust on several species belonging to the genus Senecio in the United States (1). In New Zealand, the presence of A. tragopogonis was reported on the genus Cineraria in 1959 (2). To our knowledge, this is the first report of the presence of white rust on Cineraria maritima in Italy. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (2) J. M. Dingley. N. Z. J. Agric. Res. 2:380, 1959.

Thymus × citriodorus is well known for the citrus aroma released by its leaves and is grown as a potted plant in northern Italy. This species is widely used in gardens and landscapes and for culinary purposes. In the Liguria Region alone, 1.5 million plants are grown. In the winter of 2002, extensive chlorosis was observed on potted plants of Thymus × citriodorus cv. Silver Queen grown outdoors on commercial farms near Albenga. Initial symptoms included stem necrosis at the soil level and darkening of leaves. As stem necrosis progressed, infected plants wilted and died. Wilt, characterized by the presence of soft and watery tissues, occurred within a few days on young plants. Necrotic tissues became covered with whitish mycelium that produced dark sclerotia. Sclerotinia sclerotiorum (Lib.) de Bary (1) was consistently recovered from infected stem pieces of Thymus × citriodorus. The diseased stem tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 100 ppm of streptomycin sulfate. Sclerotia produced on PDA were ellipsoid and measured 5.2 to 4.4 × 2.1 to 1.5 mm (average 3.5 × 3.0 mm). Pathogenicity of three isolates obtained from infected plants was confirmed by inoculating 30-day-old plants grown in 14-cm-diameter pots in a screenhouse. Inoculum that consisted of wheat kernels infested with mycelium and sclerotia of each isolate was placed on the soil surface around the base of each of 10 plants. Noninoculated plants served as controls. The inoculation trial was repeated once. All plants were kept at temperatures ranging between 5 and 26°C and watered as needed. Inoculated plants developed symptoms of leaf yellowing within 13 days, soon followed by the appearance of white mycelium, and eventually wilted. Control plants remained symptomless. White mycelium and sclerotia developed on infected tissues and S. sclerotiorum was reisolated from inoculated plants. To our knowledge, this is the first report of white mold of Thymus × citriodorus caused by S. sclerotiorum. The economic importance of this disease for the crop can be considered low. Reference: (1) N. F. Buchwald. Den. Kgl. Veterin.er-og Landbohojskoles Aarsskrift, 1949.

Lamium galeobdolon L. (Labiatae) is a common ornamental species that grows in shade areas and often used as a ground cover in gardens. During the summer of 2006, severe outbreaks of a previously unknown powdery mildew were observed on all Lamium spp. plants in some gardens near Biella (northern Italy). Both surfaces of the leaves of affected plants were covered with dense, white mycelia and conidia. As the disease progressed, infected leaves turned yellow and died. Mycelia and conidia also were observed on stems and flowers. Conidia were hyaline, ellipsoid, borne in short chains (with a maximum of five conidia per chain), and measured 29 to 37 × 16 to 20 μm (average 33 × 18 μm). Conidiophores, 91 to 104 μm (average 96 μm) long, showed the foot cell measuring 28 to 49 × 9 to 11 μm (average 38 × 10 μm), followed by three shorter cells measuring 14 to 26 × 9 to 15 μm (average 21 × 11 μm). Fibrosin bodies were absent. Chasmothecia were not observed in the collected samples. The internal transcribed spacer region (ITS) of rDNA was amplified using the primers ITS4/ITS6 (4) and sequenced. BLASTn analysis (1) of the 436 bp obtained showed an E-value of 0.0 with Golovinomyces orontii (Erysiphe orontii.) (3). The nucleotide sequence has been assigned GenBank Accession No. EF 121871. Inoculations were made by gently pressing diseased leaves onto leaves of five healthy L. galeobdolon plants. Five noninoculated plants served as controls. Inoculated and noninoculated plants were maintained in a greenhouse at temperatures between 15 and 28°C. After 10 days, typical powdery mildew colonies developed on inoculated plants. Noninoculated plants did not show symptoms. The pathogenicity test was carried out twice. To our knowledge, this is the first report of the presence of powdery mildew on L. galeobdolon caused by G. orontii in Italy. Blumer (2) was able to reproduce powdery mildew symptoms on L. galeobdolon using populations from cucumber, while Braun (3) reported L. galeobdolon as a possible host of E. orontii. Herbarium specimens of this disease are available at AGROINNOVA Collection, University of Torino, Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) S. Blumer. Ber. Schweiz. Bot. Ges. 62:384, 1952. (3) U. Braun. A Monograph of the Erysiphaceae (Powdery Mildews). Cramer, Berlin, GDR, 1987. (4) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997.

Penstemon barbatus (Cav.) Roth (synonym Chelone barbata), used in parks and gardens and sometimes grown in pots, is a plant belonging to the Scrophulariaceae family. During the summers of 2004 and 2005, symptoms of a root rot were observed in some private gardens located in Biella Province (northern Italy). The first symptoms resulted in stunting, leaf discoloration followed by wilt, root and crown rot, and eventually, plant death. The diseased tissue was disinfested for 1 min in 1% NaOCl and plated on a semiselective medium for Oomycetes (4). The microorganism consistently isolated from infected tissues, grown on V8 agar at 22°C, produced hyphae with a diameter ranging from 4.7 to 5.2 μm. Sporangia were papillate, hyaline, measuring 43.3 to 54.4 × 26.7 to 27.7 μm (average 47.8 × 27.4 μm). The papilla measured from 8.8 to 10.9 μm. These characteristics were indicative of a Phytophthora species. The ITS region (internal transcribed spacer) of rDNA was amplified using primers ITS4/ITS6 (3) and sequenced. BLASTn analysis (1) of the 800 bp obtained showed a 100% homology with Phytophthora citrophthora (R. & E. Sm.) Leonian. The nucleotide sequence has been assigned GenBank Accession No. DQ384611. For pathogenicity tests, the inoculum of P. citrophthora was prepared by growing the pathogen on autoclaved wheat and hemp kernels (2:1) at 25°C for 20 days. Healthy plants of P. barbatus cv. Nano Rondo, 6 months old, were grown in 3-liter pots (one plant per pot) using a steam disinfested substrate (peat/pomix/pine bark/clay 5:2:2:1) in which 200 g of kernels per liter of substrate were mixed. Noninoculated plants served as control treatments. Three replicates were used. Plants were maintained at 15 to 20°C in a glasshouse. The first symptoms, similar to those observed in the gardens, developed 21 days after inoculation, and P. citrophthora was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was carried out twice with similar results. A nonspecified root and crown rot of Penstemon spp. has been reported in the United States. (2). To our knowledge, this is the first report of P. citrophthora on P. barbatus in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) F. E. Brooks and D. M. Ferrin. Plant Dis. 79:212, 1995. (3) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (4) H. Masago et al. Phytopathology 67:425, 1977.

Lettuce (Lactuca sativa L.) is an important crop used for fresh and processing markets in Italy and is grown on more than 21,000 ha. During October and November of 2006, wilt symptoms were observed on field-grown lettuce, cv. Estelle, in Forlì, Emila Romagna (northeastern Italy) and on cv. Ballerina grown under plastichouses in Piedmont (northwestern Italy). Both lettuce cultivars were of a butterhead type. Affected plants were stunted and developed yellow leaves with brown or black streaks in the vascular tissue. Yellowing started from the external leaves. Discoloration was observed in the vascular tissue of roots, crown, and leaves. A fungus was consistently and readily isolated from symptomatic vascular tissue, previously disinfested in 1% sodium hypochlorite, when cultured on potato dextrose agar (PDA). Microscopic observations revealed hyaline hyphae with many ovoid, dark microsclerotia measuring 32 to 43 × 16 to 26 μm developing after 15 days of growth at 18°C in the dark. Conidiophores showed two verticils of three elements. Conidia were hyaline, elliptical, single celled, and measured 3.5 to 8.5 × 1.8 to 4.3 μm (average 5.5 × 2.5 μm). According to its morphological characteristics, the fungus was identified as Verticillium dahliae (2). Healthy, 20-day-old lettuce plants, cvs. Principessa and Maxima, both belonging to the butterhead type, were separately inoculated by root dip with a conidial suspension (106/ml) of two isolates of V. dahliae isolated, respectively, at Forlì and Torino. Noninoculated lettuce plants served as control treatments. Plants (10 per treatment) were grown in pots (10-liter vol.) in a steam-disinfested peat/perlite/sand (3:1:1 vol/vol) substrate and were maintained in a glasshouse at temperatures ranging between 17 and 22°C and relative humidity ranging between 60 and 70%. First wilt symptoms and vascular discoloration in the roots, crown, and veins developed 40 days after the artificial inoculation. Forty percent of the plants were affected in the case of cv. Maxima and 30% for cv. Principessa. Noninoculated plants remained healthy. The pathogenicity tests were repeated twice. To our knowledge, this is the first report in Italy of Verticillium wilt on lettuce. The disease has been previously reported in Greece (1) and the United States (3). Currently, Verticillium wilt of lettuce seems restricted in Italy to very few farms in the two locations; moreover, its incidence is very low (0.05%). References: (1) E. K. Ligoxigakis et al. Phytoparasitica 30:141, 2002. (2) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (3) G. E. Vallad et al. Plant Dis. 89:317, 2005.

Lupinus polyphyllus Lindl., a perennial ornamental belonging to the Leguminosae family, is grown in gardens for flower beds and borders. During the summer of 2006, in several gardens located in the Biella Province (northern Italy), a new wilt of Lupine was observed in 20 to 30% of the plants. The vascular tissue in stems of affected plants appeared brown. These plants were stunted and developed yellow leaves with brown or black streaks in the vascular tissue. Verticillium dahliae was consistently isolated from symptomatic vascular tissue and leaves when cultured on potato dextrose agar (PDA) (3). Microscopic observations revealed hyaline hyphae, with many irregular, dark microsclerotia, ranging from 17 to 61 μm. Conidiophores showed two verticils of three elements. Conidia were hyaline, elliptical, single-celled, measuring 3.4 to 6.0 × 1.8 to 3.1 μm (average 4.5 × 2.4 μm). The ITS region (internal transcribed spacer) of rDNA was amplified using the primers ITS4/ITS6 (2) and sequenced. BLASTn analysis (1) of the 521 bp obtained showed an E-value of 0.0 with V. dahliae. The nucleotide sequence has been assigned GenBank Accession No. EF015891. Healthy 30-day-old plants (10 per treatment) of L. polyphyllus were inoculated by root dip with a conidial suspension (0.5 × 106 CFU/ml) of V. dahliae isolated from infected plants. Ten noninoculated plants served as control treatments. All plants were transplanted into pots filled with a mix of sphagnum peat/pomix/pine bark/clay (50:20:20:10) and grown outdoors at temperatures ranging from 15 to 25°C. First wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 20 days on each inoculated plant and become evident after 50 days. V. dahliae was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of V. dahliae on L. polyphyllus in Italy. A wilt caused by V. dahliae on L. polyphyllus was observed in the Netherlands in 1925 (4). References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (3) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (4) J. H. H. Van der Meer. Meded. Landbouwhogesch. Wagening. 28, 1925.

In the winter of 2007 in Piedmont (northern Italy), symptoms of a previously unknown disease were observed on beet (Beta vulgaris L. subsp. vulgaris) (garden beet group) grown under a tunnel on several commercial farms near Cuneo. First symptoms appeared on 1-month-old plants, occurring as brown, round-to-oval spots as much as 2 cm in diameter with dark concentric rings near the perimeter. Small, dark pycnidia were present throughout the spots in concentric rings. Generally, older, lower leaves were affected more than the younger ones. Ten to fifteen percent of the plants were affected. Symptoms on the roots began near the crown as small, dark, sunken spots that became soft and water soaked. Eventually, spots on the roots turned dark brown to black and black lines separated diseased and healthy tissues. Older infected tissues were black, dry, shrunken, and spongy. Pycnidia were not observed on affected roots. From infected leaves and roots, a fungus was consistently isolated on potato dextrose agar (PDA) amended with 25 mg/l of streptomycin. The fungus was grown on PDA and maintained at 22°C (12 h of light, 12 h of dark). After 10 days, black pycnidia (130 to 328 [204] μm in diameter) developed, releasing abundant hyaline, elliptical, nonseptate conidia measuring 3.9 to 6.7 (5.1) × 2.4 to 5.9 (3.6) μm. On the basis of its morphological characteristics, the fungus was identified as a Phoma sp. (1). The internal transcribed spacer (ITS) region was amplified using primers ITS4/ITS6 (2) and sequenced. BLASTn analysis of the 557 bp obtained showed an E-value of 0.0 with Phoma betae. The nucleotide sequence has been assigned GenBank Accession No. EU003450. Pathogenicity tests were performed by spraying leaves of healthy 20-day-old potted B. vulgaris plants with a spore and mycelial suspension (1 × 106 spores or mycelial fragments per ml). Noninoculated plants sprayed only with water served as controls. Fifteen plants (three per pot) were used for each treatment. Plants were covered with plastic bags for 5 days after inoculation and kept in a growth chamber at 20°C. Symptoms previously described developed on leaves of all inoculated plants 5 days after inoculation, while control plants remained healthy. Later, pycnidia and conidia, with the same dimensions and characteristics previously described, were observed on the infected leaves. The fungus was consistently reisolated from the lesions of the inoculated plants. The pathogenicity test was carried out twice. P. betae on B. vulgaris var. cycla has been reported in Canada (3) as well as in other countries. The same pathogen was reported in Italy on sugar beet (2). References: (1) G. H. Boerema and G. J. Bollen. Persoonia 8:111, 1975. (2) A. Canova. Inf. Fitopatol. 16:207, 1966. (3) D. E L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (4) J. R. Howard et al. Diseases of Vegetable Crops in Canada. Canadian Phytopathological Society, 1994.

Camellia cultivation has a long history in the Lake Maggiore area of northern Italy where a wide selection of varieties is present. Camellias are appreciated for their large, colorful flowers that bloom from late fall through early spring. In July 2005, a previously unknown foliar disease was observed on a collection of 2- to 12-month-old camellia cultivars (Camellia japonica) grown in several nurseries located in the Verbania Province (northern Italy). The disease was observed on plants grown in pots (10 to 24 cm in diameter) that were maintained either in the open or in a greenhouse and was present for the entire growing season. However, symptoms were more severe during the summer with temperatures ranging between 25 and 30°C with high relative humidity values. During the months of June and July of 2005, severe attacks involving as much as 70% of plants were observed on C. japonica cvs. Mrs. Tingley, Burnside, Hagoromo (synonym Magnoliaeflora), and Giuseppe Traverso. The disease was again observed in 2006. On the upper side of the younger leaves, small necrotic spots (3 to 8 mm in diameter) initially developed mainly at the margin of the leaves and near the petioles. Necrotic areas were surrounded by a chlorotic halo that turned progressively black. The necrotic areas often coalesced, generating larger spots with a diameter ranging from 15 to 30 mm. Severely affected plants were defoliated. Infected plants sometimes died. The presence of lesions on mature plants decreased aesthetic quality and market value. Leaf spots contained dark brown, multicellular, pyriform conidia. Conidia, generally in short chains, were 20.5 to 34.8 μm (average 29.3 μm) long, 6.9 to 12.2 μm (average 9.9 μm) wide, with 3 to 4 longitudinal cross walls, and an average of 5.7 single cells. From 15 samples of infected leaves, several isolates of a fungus identified on the basis of its morphological characteristics as belonging to the Alternaria alternata complex (2) were consistently isolated on potato dextrose agar containing 25 mg/l of streptomycin sulfate. Pathogenicity tests were performed by spraying leaves of healthy 6-month-old potted C. japonica cv. Burnside plants with a spore and mycelial suspension (1 × 105 CFU/ml) prepared by using a mixture of three isolates obtained in 2005 grown on PDA for 30 days at 23 ± 2°C in a growth chamber (12 h of light per day). Plants without inoculation served as a control. Five plants were used for each treatment. Plants were covered with plastic bags for 3 days after inoculation and maintained at 25°C in growth chambers. The first lesions developed on leaves 3 days after inoculation, while control plants remained healthy. Sixty days after artificial inoculation, 25% of the inoculated plants were dead, while the control plants remained healthy. From lesions of infected plants, a fungus belonging to the A. alternata complex was consistently reisolated. The pathogenicity test was carried out twice. The presence of A. alternata on C. sinensis, the commercial tea plant, was reported in India (1). Previously, a flower blight caused by A. tenuis was reported in the United States (3). This is, to our knowledge, the first report of A. alternata on C. japonica in Italy and probably in the world. The disease was present in 2005 and 2006 in several commercial nurseries affecting 50% of plants of susceptible cultivars. References: (1) B. N. Chakraborty et al. Plant Pathol. 55:303, 2006. (2) E. G. Simmons. Pages 1–35 in: Alternaria Biology, Plant Diseases and Metabolites. J. Chelchowski and A. Visconti, eds. Elsevier, Amsterdam, 1992. (3) A. J. Watson. Plant Dis. Rep. 34:186, 1950.

Paris daisy (Argyranthemum frutescens), also known as Marguerite daisy, is an economically important crop in the Riviera Ligure (northern Italy) where approximately 18 million potted plants per year are produced for export. During the fall and winter of 2007, Paris daisy ‘Bright Carmen’ plants, started in a greenhouse and growing outside in a commercial nursery at Albenga, showed a previously unknown powdery mildew. Young stems, particularly in the interior portions of the plant, were covered with a white mycelium. As the disease progressed, leaves became covered with the mycelium, resulting in smaller, chlorotic leaves. Conidia were hyaline, cylindrical, borne in chains (two to three conidia per chain) and measured 30 × 12 μm (20 to 34 × 10 to 15 μm). Conidia were generated by conidiophores represented by a foot cell measuring 55 to 101 × 11 to 12 μm followed by two shorter cells measuring 19 to 29 × 11 to 14 and 24 to 33 × 12 to 14 μm. Fibrosin bodies were absent. Chasmothecia were not observed in the collected samples. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 441 bp showed a 100% homology with the sequence of Golovinomyces cichoracearum (= Erysiphe cichoracearum) (3). The nucleotide sequence has been assigned GenBank Accession No. EU486992. Pathogenicity was confirmed through inoculation by gently pressing diseased leaves onto leaves of healthy Paris daisy plants of cvs. Blazer Rose, Bright Carmine, Cherry Harmony, Crowned Rose, Fulvia, Sole Mio, Stella 2000, Summit Pink, and Sun Light. Three plants per cultivar were inoculated, while the same number served as noninoculated controls. The pathogenicity test was carried out twice. Plants were maintained in a greenhouse at temperatures ranging from 15 to 21°C. Fifteen days after inoculation, typical symptoms of powdery mildew developed on inoculated plants of all cultivars, with the exception of Stella 2000. The fungus observed on inoculated plants was morphologically identical to that originally observed. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of powdery mildew on A. frutescens in Italy. G. cichoracearum has been reported on Chrysanthemum frutescens in Switzerland (2). The economic impact of this disease is limited but can easily increase because of the intensive cultivation of this crop. The availability of resistant or partially resistant cultivars will help reduce the impact of this new disease. Voucher specimens are available at the AGROINNOVA Collection, University of Torino. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) A. Bolay, Cryptogam. Helv. 20:1, 2005. (3) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000.

Rudbeckia fulgida (common name orange coneflower) is an herbaceous perennial (Asteraceae) grown in full sun in perennial borders in gardens. At the end of the summer of 2007, in a public garden located in Turin (northern Italy), symptoms of vascular wilt and stunting were observed on approximately 80% of the plants grown in a mixed border. Initial symptoms were yellowing of external leaves and brown or black streaks in the vascular tissue of roots, crown, and leaves. A fungus was consistently and readily isolated on potato dextrose agar from symptomatic vascular tissue previously disinfested in 1% sodium hypochloride. Ovoid, dark microsclerotia, 41 to 108 μm, developed in hyaline hyphae after 10 days of growth at 23°C (12 h of light and 12 h of dark). Hyaline, elliptical, single-celled conidia, 3.2 to 7.3 × 2.1 to 3.7 (average 4.7 × 2.8) μm, developed on verticillate conidiophores. On the basis of these morphological characteristics, the fungus was identified as Verticillium dahliae (4). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 530 bp showed a 100% homology with the sequence of V. dahliae. The nucleotide sequence has been assigned GenBank Accession No. EU 627007. Healthy, 30-day-old R. fulgida plants were grown in a steam-disinfested mix of sphagnum peat:pomix:pine bark:clay (50:20:20:10) infested with a conidial suspension (1.5 × 106/ml) of three isolates of V. dahliae isolated from infected plants. Noninoculated plants served as controls. Plants (16 per treatment) were grown in pots (3 liter vol) and maintained in a glasshouse at temperatures between 22 and 25°C and relative humidity between 50 and 70%. First wilt symptoms and vascular discoloration in the roots, crown, and veins developed 17 days after inoculation. Noninoculated plants remained healthy. The pathogenicity tests were carried out twice. To our knowledge, this is the first report in Italy of Verticillium wilt on R. fulgida. Wilts caused by V. dahliae on R. laciniata in Poland (3) and V. albo-atrum on R. hirta in the United States (2) were previously reported. The importance and economic impact of this disease is currently limited but may increase because of the popularity of Rudbeckia spp. in private and public parks. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Their Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (3) B. Leski. Rocz. Nauk Roln. 253, 1974. (4) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002.

Paris daisy (Argyranthemum frutescens (L.) Sch. Bip.) is an economically important crop on the Riviera Ligure (northern Italy), where approximately 10 million plants per year are produced for export. In the winter of 2007, extensive wilting was observed on 5-month-old potted plants of A. frutescens grown in a commercial greenhouse near Albenga. First symptoms included stem necrosis, darkening and withering of leaves, and wilting of young buds. As stem and foliar necrosis progressed, infected plants wilted and died. Wilt occurred a few days after the appearance of the first symptoms. Infected plants were characterized by the presence of soft, watery tissues. Lesions became covered with a whitish mycelium and dark sclerotia were produced on the mycelium. Diseased stem tissue was surface sterilized for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 100 mg/l streptomycin sulfate. Sclerotinia sclerotiorum (Lib.) de Bary (2) was consistently recovered from infected stem pieces. Sclerotia produced on PDA measured 1.4 to 5.2 × 2.3 to 6.7 (average 3.3 to 4.2) mm. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and then sequenced. BLAST analysis (1) of the 531-bp segment showed a 100% homology with the sequence of S. sclerotiorum. The nucleotide sequence has been assigned GenBank Accession No. EU 556701. Pathogenicity of two isolates obtained from infected plants was confirmed by inoculating 10 90-day-old plants for each isolate. Plants were grown singly in 18-cm-diameter pots maintained in a greenhouse under shade and were regularly irrigated and fertilized. Mycelium plugs 1 cm2 were excised from a 10-day-old PDA culture of both isolates and placed on the soil surface around the base of each plant. Ten noninoculated plants served as controls. Plants were maintained in a greenhouse under shade at temperatures ranging between 6 and 22°C (average 18°C) and relative humidity at >90%. The inoculation trial was carried out twice. All inoculated plants developed leaf yellowing by 22 days after inoculation. White, cottony mycelium and black sclerotia developed on stems and at the base of all inoculated plants. Eventually, infected plants wilted. Control plants remained symptomless. S. sclerotiorum was reisolated from the stems of inoculated plants. To our knowledge, this is the first report of S. sclerotiorum causing white mold on A. frutescens in Italy as well as worldwide. The economic importance of this disease can be considered limited at the moment. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) N. F. Buchwald. Kongl. Veterisk Landb. Aarssk. 75, 1949.

Lampranthus sp. N.B. Brown (figmarigold) of the Aizoaceae family is used as groundcover in gardens. In October of 2008, severe outbreaks of a previously unknown rot were observed in a nursery located in Liguria, near Savona (northern Italy), on 35-day-old rooted cuttings grown in a peat substrate. Approximately 50% of rooted cuttings of red-flowered cultivars were affected. Lesions on collars and young stems were brown, water soaked, and soft. Plants eventually collapsed as roots rotted. Thin, aerial hyphae were visible on the surface of the stems and substrate. Later, a thick, light yellow, mycelial mat surrounded infected plants. Tissue fragments were excised from the margins of the lesions, dipped in a solution containing 1% sodium hypochlorite, and plated on potato dextrose agar and a medium selective for Oomycetes (4). Plates were incubated under constant fluorescent light at 23 ± 1°C for 4 to 5 days. Hyphae of five isolates grown on V8 medium were aseptate and 4.2 to 7.9 (average 6.2) μm wide. Sporangia consisted of complexes of swollen hyphal branches. Oogonia were globose, smooth, and 23.5 to 28.0 (average 25.9) μm in diameter. Antheridia were barrel shaped, intercalary, and diclinous. Oospores were globose and 19.4 to 23.6 (average 21.4) μm in diameter. The internal transcribed spacer (ITS) region of rDNA of a single isolate (DB24112008) was amplified with primers ITS4/ITS6 and sequenced. A BLAST analysis (1) in GenBank of the 1,074-bp segment showed a 100% homology with the sequence of Pythium aphanidermatum (Accession No. EU245039). The nucleotide sequence has been assigned the GenBank Accession No. FJ492745. Pathogenicity tests were performed twice on a red-flower cultivar of a Lampranthus sp. grown in 1-liter pots containing a peat moss substrate infested with wheat and hemp kernels colonized with one isolate of P. aphanidermatum at a rate of 20 g/liter. Ten plants were grown in infested media and 10 plants were grown in noninfested media. Greenhouse temperatures were 18 to 24°C. The first symptoms of stem and root rot developed 15 days later, while control plants remained healthy. P. aphanidermatum was consistently reisolated from the lesions. To our knowledge, this is the first report of P. aphanidermatum on a Lampranthus sp. in Italy. The disease has been reported in Japan (3) in 2008, while in the United States, a Pythium sp. was reported on L. aureus and L. glomeratus (2). Currently, the economic importance of Pythium rot on figmarigold in Italy is still limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (3) H. Kawarazachi et al. J. Gen. Plant Pathol. 74:94, 2008. (4) H. Masago et al. Phytopathology, 67, 425, 1977.

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips. It can also be grown for cut flower production. In April of 2008, in several nurseries located near Biella (northern Italy), a leaf blight was observed on 10 to 15% of potted 30-day-old plants grown on a sphagnum peat substrate at 15 to 20°C and relative humidity of 80 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded over several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and abscised. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and had radial growth. Sclerotia were not present. Isolates obtained from affected plants successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates of AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 648-bp fragment showed a 100% homology with the sequence of R. solani AG-4 AB000018. The nucleotide sequence has been assigned GenBank Accession No. FJ 534555. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Five plants of 30-day-old A. flabellata were grown in 3-liter pots. Inoculum consisting of an aqueous suspension of PDA and mycelium disks (5 g of mycelium + agar per plant) was placed at the collar of plants. Five plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a greenhouse at temperatures between 20 and 24°C. The first symptoms, similar to those observed in the nursery, developed 7 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. The presence of R. solani AG1-IB on A. flabellata has been reported in Japan (4), while in the United States, Rhizoctonia sp. is described on Aquilegia sp. (3). This is, to our knowledge, the first report of leaf blight of A. flabellata caused by R. solani in Italy as well as in Europe. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) E. Imaizumi et al. J. Gen. Plant Pathol. 66:210, 2000.

Lupinus polyphyllus (Leguminosae), Washington lupine, is a perennial herbaceous plant. In March 2008, in a campus greenhouse at the University of Torino, Grugliasco (northern Italy), a leaf blight was observed on 20% of potted 30-day-old plants. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Severely infected plants died. Plants were grown in a sphagnum peat/perlite/clay (70:20:10) substrate at temperatures between 18 and 25°C and relative humidity of 60 to 80%. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani (4) was consistently and readily recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 20 ± 1°C appeared light brown, rather compact, and exhibited radial growth. The isolates of R. solani successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (3). Pairings were also made with tester isolates AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 660-bp fragment showed 100% homology with the sequence of R. solani. The nucleotide sequence has been assigned GenBank Accession No. FJ486272. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old L. polyphyllus were grown in 10-liter containers (10 plants per container) on a steam disinfested sphagnum peat/perlite/clay (70:20:10) medium. Inoculum, consisting of an aqueous suspension of mycelium disks prepared from PDA cultures (5 g of mycelium per plant), was placed at the collar of plants. Plants inoculated with water and PDA fragments alone served as control treatments. Three replicates were used. Plants were maintained in a greenhouse at temperatures between 18 and 23°C. First symptoms, similar to those observed in the nursery, developed 10 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was repeated twice. The susceptibility of L. polyphyllus to R. solani was reported in Poland (2). This is, to our knowledge, the first report of leaf blight of L. polyphyllus caused by R. solani in Italy. The importance of the disease is at the moment limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) W. Blaszczak. Rocz. Nauk. Roln. Ser A 85:705, 1962. (3) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (4) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.

Spider flower (Cleome hassleriana L. [synonym = C. spinosa]) is used as a tall border in parks and gardens worldwide. During July 2008, severe outbreaks of a powdery mildew were observed in a public park in Torino (northern Italy). Leaves were covered with dense, white hyphae and conidia, especially on the adaxial surface. Hyphae were also present on petioles and fruits, but not on petals and stems. As the disease progressed, infected leaves turned chlorotic, curled, and wilted. Conidia were hyaline, cylindrical, single, and measured 31.1 to 48.2 × 12.9 to 17.6 μm (average 37.1 × 15.6 μm). Germ tubes terminating in a moderately lobed appressorium were produced terminally. The cylindrical foot cells of the erect condiophores were 19.2 to 27.8 × 6.5 to 8.6 μm (average 23.3 × 7.7 μm). Fibrosin bodies were absent. Chasmothecia were observed mostly on the lower surfaces of leaves. At maturity, they were dark amber and spherical with a diameter of 92.9 to 151.0 μm (average 121.4 μm). Each chasmothecium contained six stalked asci (average size 63.7 × 35.9 μm). Each ascus contained four ellipsoid ascospores that measured 17.3 to 26.4 × 10.9 to 15.6 μm (average 23.3 × 12.8 μm). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced (1). The 602-bp sequence was deposited in GenBank under the Accession No. GQ149478 and was 99% similar to that of Erysiphe cruciferarum (Accession No. EU140958). As proof of pathogenicity, diseased leaves of C. hassleriana were pressed against leaves of three healthy 4-month-old potted plants of the same species for 10 min. Three noninoculated plants served as controls. Inoculated and noninoculated plants were maintained in a greenhouse at 22 to 25°C in isolation. After 11 days, typical powdery mildew colonies developed on inoculated plants. Noninoculated plants did not develop symptoms. The pathogenicity test was repeated once. Powdery mildew on C. hassleriana caused by E. cruciferarum was reported in Italy (2) but the pathogen was not characterized. Herbarium specimens are deposited at AGROINNOVA Collection, University of Torino, Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) U. Braun. The Powdery Mildews (Erysiphales) of Europe. Gustav Fischer Verlag, Jena, Germany, 1995.

In the summer of 2007, a wilt of endive (Cichorium endivia) cv. Myrna plants was observed on several commercial farms near Alessandria in northern Italy. Approximately 40% of the plants on each farm were symptomatic. Affected plants were stunted and yellow and their root systems were poorly developed. Basal leaves eventually wilted. Black streaks were observed in the proximal part of the leaf vascular system. Fusarium oxysporum was consistently isolated from symptomatic vascular tissue on a Fusarium-selective medium (1). To confirm identification of the pathogen, the translation elongation factor 1α and the ribosomal DNA intergenic spacer (IGS) region were partially sequenced (2). Genomic DNA was extracted from mycelia growing on potato dextrose agar. Amplification of the EF-1α region generated a sequence of 648 bp; the IGS amplicon was 2,500 bp. The EF-1α sequence (GenBank Accession No GQ398152) was 99% similar to the sequence of a F. oxysporum strain isolated from soil and a strain pathogenic on cotton plants (GenBank Accession No. EU246574). The IGS sequence (GenBank Accession No GQ398153) was 97% similar to the sequence of a F. oxysporum strain (GenBank Accession No. EF661647). Pathogenicity tests were conducted on 15-day-old endive plants (cv. Myrna). Ten plants were inoculated by dipping their roots in a 1 × 106 CFU/ml suspension of one of the isolates recovered from a wilted endive plant. Inoculated and noninoculated plants were transplanted into pots filled with steamed soil and maintained in a glasshouse at 23 to 28°C. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 60 days after inoculation. F. oxysporum was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of wilt caused by F. oxysporum on endive in Italy. References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) G. Mbofung et al. Phytopathology 97:87, 2007.

In northern Italy, blue mold can occur generally on apples after 3 months of storage under controlled atmospheres. The mold can be caused by Penicillium griseofulvum Dierckx (synonym P. urticae Bainier). During 2008, several postharvest fruit rots were observed on apples (cv. Golden Delicious) after 180 to 240 days of storage at 1°C. Approximately 8% of the fruits showed blue mold. Apples had been cultivated in Aosta (Aosta Valley Region) and Lagnasco (Piedmont Region). Infected fruits showed soft, watery, brown spots enlarging rapidly at 20°C. There was a distinct margin between soft rotted flesh and firm healthy tissues. Under high humidity, masses of blue-green spores formed on the surface of the lesion. Apple fruit excisions from the margin between the healthy and diseased tissues were plated on potato dextrose agar (PDA), pH 5.6. The recovered fungus produced abundant mycelium and conidia, with the colonies attaining a diameter of 2.0 to 2.4 cm after 7 days at 20 ± 2°C on PDA. Colonies were mostly yellow-green, with a yellowish-to-orange brown underside. Conidiophores were mononematous or loosely synnematous, hyaline, with branches strongly divergent. Phialides were cylindrical with a very short neck. Conidia were ellipsoidal, sometimes subglobose, 2.5 to 3.5 × 2.2 to 2.5 μm, hyaline to greenish. Preliminary morphological identification of the fungus (2) was confirmed by PCR using genomic DNA extracted from mycelia of pure cultures. Two sequences, obtained through the amplification of ribosomal region ITS1-5.8S-ITS2 (1), were blast searched in GenBank and showed 99% sequence coverage and 99% similarity to ribosomal sequences of P. griseofulvum. Two sequences were deposited in GenBank with Accession Nos. HQ012498 (a strain from Aosta Valley) and HQ012499 (a strain from the Piedmont Region). Pathogenicity was tested on 20 ripe fruits each of four apple cultivars (Golden Delicious, Red Chief, Granny Smith, and Royal Gala). Fruits were surface sterilized with 1% sodium hypochlorite. Conidial suspensions (30 μl of 105 conidia/ml) of the fungus were placed on artificial wounds generated on the apple surface. Control fruits were treated with sterile water. Seven days after inoculation, the symptoms were reproduced on the four cultivars and P. griseofulvum was reisolated on PDA from the inoculated fruits of all four cultivars. Control fruits were symptomless. An analysis using high-performance liquid chromatography with diode array of the rotting tissues associated with inoculated fruits of all four cultivars (4) confirmed, as in the case of other strains of P. griseofulvum, the production of the mycotoxin patulin (12.1 to 44.4 mg kg–1). Previously, P. griseofulvum was reported on apple in other countries such as the United States (3), Japan, Egypt, and Brazil. To our knowledge, this is the first report of P. griseofulvum on apples during storage in Italy. References: (1) R. Nilsson et al. FEMS Microbiol. Lett. 296:97, 2009. (2) R. A. Samson and J. L. Pitt. Integration of Modern Taxonomic Methods for Penicillium and Aspergillus Classification. Harwood Academic Publishers, Singapore, 2001. (3) P. G. Sanderson and R. A. Spotts. Phytopathology 85:103, 1995. (4) D. Spadaro et al. Food Addit. Contam. B 1:134, 2008.

Spearmint (Mentha spicata) is grown for its aromatic and carminative oil and as an ornamental. During the fall of 2009, 4-month-old plants grown on a commercial farm located near Albenga (northern Italy) showed signs and symptoms of an unknown powdery mildew. The adaxial leaf surfaces were covered with white mycelia and conidia, while the abaxial surfaces were less infected. As the disease progressed, infected leaves turned yellow and wilted. Mycelia were also observed on stems. Conidia were hyaline, elliptical to doliform, borne in short chains (two to three conidia per chain), and measured 35 × 21 μm (30 to 43 × 18 to 26 μm). Conidiophores measured 86 × 11 μm (76 to 97 × 9 to 13 μm) followed by one to three shorter cells, measuring, respectively, 22 × 11 μm (13 to 28 × 9 to 15 μm). Fibrosin bodies were absent. Chasmothecia were not observed in the collected samples. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced (1) (GenBank Accession No. HM053470). The 567-bp amplicon had 100% homology with the sequence of Golovinomyces biocellatus (GenBank Accession No. AB307675). Pathogenicity was confirmed through inoculation by gently pressing diseased leaves onto leaves of healthy M. spicata plants. Three plants were inoculated, while the same number of noninoculated plants served as a control. Plants were maintained at temperatures from 18 to 25°C. Twelve days after inoculation, typical symptoms of powdery mildew developed on inoculated plants. The fungus observed on inoculated plants was morphologically identical to that originally observed. Noninoculated plants did not show symptoms. The pathogenicity test was carried out twice. G. biocellatus has been reported on Mentha spp. in Australia (3) and the United States (4) and the pathogen is present on other plant genera in Italy. A similar powdery mildew of M. spicata was attributed to Erysiphe orontii in the United States (2). The economic importance of this disease is currently limited in Italy because of the limited planting of this host, but potted aromatic plants represent a steadily increasing crop in Italy. Voucher specimens are available at the Agroinnova Collection, University of Torino. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) S. T. Koike and G. S. Saenz. Plant Dis. 83:399, 1999. (3) J. R. Liberato and J. H. Cunnington. Australas. Plant Dis. Notes 2:83, 2007. (4) D. B. Marcum et al. Plant Dis. 94:276, 2010.

Euphorbia susannae (common name Suzane's spurge) and E. inermis (Euphorbiaceae family) are grown in pots and commercialized in northern Italy. In March 2012, plants of these species grown on the same commercial farm showed signs of powdery mildew. On E. susannae, apexes of stems were covered with white mycelia and conidia, with the ultimate development of necrosis on symptomatic tissues more interested by the disease. Of 5,000 plants, 5% were affected. Conidia were hyaline, elliptical, borne in short chains (two to three conidia per chain), and 27.7 (24.4 to 30.6) μm long and 16.1 (13.6 to 19.1) μm wide. Conidiophores were erect, with a foot cell straight or slightly flexuous, 82.8 (52.7 to 117.1) μm long and 10.0 (9.2 to 11.2) μm wide, followed by two to three shorter cells 19.8 (14.7 to 28.9) μm long and 10.7 (8.7 to 13.0) μm wide. On E. inermis, a white mycelium covered the stems starting from the apexes, causing yellowing and necrosis on leaves finally killing infected tissues. Of 500 plants, 80% were affected. Conidia were 33.2 (25.7 to 42.0) μm long and 17.3 (12.6 to 21.6) μm wide. Conidiophores were erect, with a foot cell straight or slightly flexuous, 96.7 (67.0 to 138.6) μm long and 9.5 (7.7 to 11.7) μm wide, followed by two to three shorter cells 26.3 (17.5 to 42.2) μm long and 11.8 (8.3 to 16.2) μm wide. Fibrosin bodies were present on both hosts, while chasmothecia were not observed in samples from either host. The internal transcribed spacer (ITS) region of rDNA of samples of mycelium and conidia collected from the two hosts was amplified using the primers ITS1-ITS4 (3) and directly sequenced (GenBank Accession Nos. JX179221 and JX179219 for E. susannae and E. inermis, respectively). By performing BLAST analysis, the 692-bp amplicon from E. susannae and the 541-bp amplicon from E. inermis showed 100% homology with several sequences of Podosphaera spp. On the basis of morphological characteristics of the imperfect state and the ITS analysis, the causal agent of powdery mildew on E. susannae and E. inermis was identified as Podosphaera sp. Pathogenicity was confirmed by gently brushing healthy 20-month-old potted plants of E. susannae and E. inermis with mycelia and conidia from diseased plants. Four plants/treatment/species were used. Non-inoculated plants belonging to the two species served as control. Plants were maintained in a glasshouse at temperatures ranging from 18 to 25°C and relative humidity from 65 to 80%. About 20 days after inoculation, typical symptoms of powdery mildew developed on inoculated plants. The fungus observed on inoculated plants was morphologically identical to the original isolate. Non-inoculated plants did not exhibit symptoms. The pathogenicity test was performed in duplicate. Several agents of powdery mildew have been reported on Euphorbia spp. Among others, Podosphaera euphorbiae was reported on numerous Euphorbiaceae (1), P. euphorbiae-hirtae was observed on E. hirta (1) and P. euphorbiae-helioscopiae on E. pekinensis (2). To our knowledge, this is the first report of Podosphaera sp. on E. susannae and E. inermis in Italy. This disease is not presently of economic importance. Specimens are available at the Agroinnova Collection. References: (1) U. Braun. A Monograph of the Erysiphales (Powdery Mildews). J. Cramer, Berlin-Stuttgard, German Democratic Republic, 1987. (2) S. Y. Liu. Plant Dis. 95:1314, 2011. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

During fall 2011, symptoms of a wilt disease were observed in a commercial nursery near Ventimiglia as well as in the Research Center of Floriculture of Sanremo (northern Italy) on plants of Papaver nudicaule (Iceland poppy) of a local unnamed cultivar. In the commercial nursery, 15 to 20% of plants were affected, while about 3% of plants were affected at the Research Center. Symptoms consisted of chlorosis, premature leaf drop, and foliar wilting, followed by the stem wilting, bending, and eventually rotting from the base. Brown discoloration was observed in the stem vascular tissue. Using Komada's Fusarium-selective agar medium (2), a fungus was consistently and readily isolated from symptomatic vascular tissue of plants collected from both sites. The isolates were purified and subcultured on potato dextrose agar (PDA), on which medium both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with pale purple pigments in the agar medium. The isolate generated short monophialides with unicellular, ovoid-elliptical microconidia of 3.9 to 6.7 × 1.4 to 3.0 (average 5.4 × 2.3) μm. On carnation leaf agar (CLA) (1), isolates produced pale orange sporodochia with macroconidia that were 3-septate, slightly falcate with a foot-shaped basal cell and a short apical cell, and 26.0 to 43.5 × 3.1 to 4.4 (average 35.3 × 3.7) μm. Chlamydospores were abundant, terminal, and intercalary, rough walled, mostly singles but sometime in short chains or clusters, and 5.2 to 10.1 μm in diameter. Such characteristics are typical of Fusarium oxysporum (3). The internal transcribed spacer (ITS) region of rDNA was amplified from the isolates using the primers ITS1/ITS4 (4), and sequenced. BLASTn analysis of the 507-bp ITS sequence of one isolate from P. nudicaule collected from the commercial nursery (GenBank Accession No. JX103564) showed an E-value of 0.0 and 100% identity with the ITS sequence of F. oxysporum (HQ649820). To confirm pathogenicity of one of the Iceland poppy isolates, tests were conducted on 2-month-old plants of the same cultivar on which symptoms were first observed. Plants (n = 14) were inoculated by dipping roots in a 1 × 107 CFU/ml conidial suspension of the isolate of F. oxysporum prepared from 10-day-old cultures grown in potato dextrose broth (PDB) on a shaker (90 rpm) for 10 days at 22 ± 1°C (12-h fluorescent light, 12-h dark). Non-inoculated control plants (n = 14) were dipped in sterilized water. All the plants were transplanted into pots filled with steamed potting mix (sphagnum peat/perlite/pine bark/clay at 50:20:20:10), and maintained in a glasshouse at 24 to 28°C. Inoculated plants showed typical symptoms of Fusarium wilt after 10 days. The stems then wilted and plants died. Non-inoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants but not from control plants. The pathogenicity test was conducted twice with the same results. Since Fusarium wilt has not previously been described on Iceland poppy at any location, this is first report of F. oxysporum on P. nudicaule in Italy and anywhere in the world. References: (1) N. L. Fisher et al. Phytopathology 72:151, 1982. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) J. F. Leslie and B.A. Summerell. The Fusarium Laboratory Manual, Blackwell Professional, IA, 2006. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

Satureja montana L. (winter savory “Repandens”) is an evergreen shrub. In late summer 2010, blight was observed on a farm near Albenga (northern Italy) on 3% of 500 potted 2-month-old plants. Semicircular, water-soaked lesions appeared first on stems then on leaves. As the disease progressed, blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage within 5 to 6 days. Stem fragments taken from the margin of the diseased tissues of 10 plants were disinfected for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 100 μg/liter streptomycin sulfate. A fungus with morphological characters of Rhizoctonia solani was consistently isolated. Three isolates of R. solani obtained from affected plants were successfully anastomosed with R. solani isolate AG 1 (ATCC 58946). Three pairings were made for each tested strain. Hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (2). Isolates from winter savory were paired with R. solani isolates AG 2, 3, 4, 6, 7, or 11 and examined microscopically. Anastomosis was not observed in any of the pairings. Tests were repeated once. Mycelium of 10-day-old isolates from winter savory appeared light brown, compact, and radiate. Numerous, dark brown sclerotia, 1 to 4 mm in diameter (average 1.7), developed within 20 days after transfer of mycelia to PDA in 90-mm-diameter petri dishes and incubated (11-h daylight, 13-h dark) at 21 to 24°C. Descriptions of mycelium and sclerotia were typical for subgroup IA Type 2 (3). The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and sequenced. BLASTn analysis (1) of the 696 bp showed a 99% homology with the sequence of R. solani. The nucleotide sequence has been assigned GenBank No. JQ313811. For pathogenicity tests, inoculum of R. solani was prepared by growing the pathogen on wheat kernels autoclaved in 1-liter glass flasks (30 min at 121°C and 1 atm) for 15 days. One of the isolates assigned to the anastomosis group AG 1 IA was tested. Five 90-day-old plants of S. montana were inoculated. Each plant grown in 2-liter pots in a steam disinfested peat/pumice/pine bark/clay mix (50:20:20:20:10) was inoculated with 10 g of infested wheat kernels placed at the base of the stem. Five plants inoculated with noninfested wheat kernels served as the control. Plants covered with plastic bags were arranged randomly in a growth chamber at 20 ± 1°C with 12-h light/dark for 5 days. Symptoms, similar to those observed in the farm, developed 4 days after inoculation. Ten colonies of R. solani were reisolated from infected leaves and stems of each inoculated plant. Control plants remained healthy. The pathogenicity test was carried out twice. Symptoms caused by R. solani have been recently observed on S. hortensis in Poland (4). This is, to our knowledge, the first report of blight of S. montana caused by R. solani in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, The Netherlands, 1996. (3) R. T. Sherwood. Phytopathology 59:1924, 1969. (4) B. Zimowska. Herba Polonica 56:29, 2010.

During an extensive survey carried out in Piedmont (northern Italy) aimed at identifying the emerging soilborne diseases affecting tomato in commercial fields where alternatives to methyl bromide have been implemented in response to national and international regulations, sudden collapse of tomato plants, cv. Tomahawk, grafted on cv. Beaufort, were repeatedly observed in a commercial plastic tunnel operation. Affected plants suddenly collapsed 60 days after transplant during the month of May 2010. Symptoms included chlorosis, stunting, and severe root and crown rot, leading to sudden collapse of approximately 25% of the plants within 60 days of transplant. Symptomatic tissues from the root and collar of infected plants were surface disinfested for 1 min in a 1% NaOCl solution, rinsed for 5 min in water, and submerged in selective medium based on corn meal agar. A Phytophthora-like organism (2) with characteristic coenocytic hyphae was consistently isolated and transferred to V8 agar. The sporangia were spherical to ovoid, papillate, and 40 to 77 × 23 to 34 (average 55.1 × 30.3) μm. Oospores were globose and 22.2 to 30.8 μm. The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 750-bp segment showed a 100% homology with the sequence of Phytophthora capsici JN382543.1. The nucleotide sequence has been assigned the GenBank Accession No. JX090306. Pathogenicity tests were performed on healthy 30-day-old tomato plants cv. Beaufort by using one strain of P. capsici grown for 15 days at 22 to 25°C on a mixture of 2:1 wheat/hemp kernels, and then 1 g per L of the inoculum was mixed into a substrate based on peat blonde/peat black (15:85 v/v). Two plants were transplanted into 3-L pots, with five replicates. Ten non-inoculated plants represented the control treatment; the trial was repeated once. All plants were kept in a greenhouse at temperatures ranging from 22 to 25°C. Inoculated plants became chlorotic 7 days after inoculation and root and crown rot developed 30 days after inoculation. Control plants remained symptomless. P. capsici consistently was reisolated from inoculated plants. In Italy, the presence of P. nicotianae on hybrids of Solanum lycopersicum × S. hirsutum is known (3), while, to the best of our knowledge, this is the first report of P. capsici on the hybrid S. lycopersicum × S. hirsutum in Italy. The economic importance of the disease can increase due to the expanding use of grafted tomato plants. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. APS Press, St Paul, MN, 1996. (3) A. Garibaldi and M. L. Gullino. Acta Hortic. 833:35, 2010. (4) H. M. Masago et al. Phytopathology 67:425, 1977.

During spring 2012, symptoms of an unusual leaf spot disease were observed in several commercial greenhouses near Salerno (southern Italy) on plants of Diplotaxis tenuifolia (cv Selvatica). The first symptoms on leaves of affected plants consisted of small (1 mm) black-brown spots of irregular shape, later coalescing into larger spots, 1 cm in diameter. Spots were surrounded by a yellow halo, and were mostly located on the foliar limb, rib, and petiole. Affected leaves were often distorted, appearing hook-like. The disease was severe under 75 to 90% RH, at air temperature of 20 to 26°C, and caused severe production losses on about 50 ha. Particularly, affected tissues rotted quickly after packaging and during transit and commercialization of processed rocket. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 60 s, rinsed in water, then placed on potato dextrose agar (PDA) medium, containing 25 mg/l of streptomycin sulphate. After 5 days, a fungus developed producing a whitish-orange mycelium when incubated under 12 h/day of fluorescent light at 22°C. The isolates obtained were purified on PDA. On this medium, they produced hyaline elliptical and ovoid conidia, sometimes one-septate, measuring 4.5 to 9.2 × 1.7 to 3.5 (average 6.8 × 2.6) μm. Conidia were born on phialides, measuring 6.8 to 20.2 × 1.3 to 3.1 (average 16.5 × 2.1) μm. Such characteristics are typical of Plectosphaerella sp. (2). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (3) and sequenced. BLAST analysis of the 519-bp segment showed a 98% similarity with the sequence of Plectosphaerella cucumerina (GenBank Accession No. AB469880). The nucleotide sequence has been assigned the GenBank Accession JX185769. To confirm pathogenicity, tests were conducted on 45-day-old D. tenuifolia plants. Plants (21/treatment), grown in 15 liter pots (7 plants/pot) were inoculated by spraying a 1 × 106 CFU/ml conidial suspension of one isolate of P. cucumerina, prepared from 10-day-old cultures, grown on PDA. Inoculated plants were maintained in a growth chamber at 23 ± 1°C, at 90% RH for 4 days. Non-inoculated plants served as control. Inoculated plants showed the typical first leaf spots 6 days after the artificial inoculation. Four days later, spots enlarged and leaves became distorted, showing chlorosis. Non-inoculated plants remained healthy. P. cucumerina was reisolated from inoculated plants. The pathogenicity test was conducted twice with identical results. This is, to our knowledge, the first report of P. cucumerina on D. tenuifolia in Italy as well as worldwide. P. cucumerina has been described as associated with root and collar rots of other horticultural crops in southern Italy (1). Due to the importance of the crop in Italy, this disease can cause serious economic losses. References: (1) A. Carlucci et al., Persoonia, 28:34, 2012. (2) M. E. Palm et al. Mycologia, 87:397, 1995. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

During summer 2012, symptoms of a new leaf spot disease were observed in several commercial fields in Treviglio (Bergamo, northern Italy) on plants of curly (Cichorium endivia var. crispum) and Bavarian (C. endivia var. latifolium) endive (Asteraceae). This crop is widely grown in the region for fresh market. The first symptoms on leaves of affected plants consisted of small (1 mm) black-brown spots of irregular shape, later coalescing into larger spots, up to 10 to 15 mm diameter. Eventually, spots were surrounded by a yellow halo. Particularly, affected tissues rotted quickly under high moisture. Disease severity was greatest at 75 to 90% RH and air temperature between 23 and 30°C, where affected tissues rotted quickly. This disease resulted in severe production losses. On one farm in particular, three different fields totaling 2 ha, 5 to 13% of the plants were affected. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 60 s, rinsed in water, then placed on potato dextrose agar (PDA) medium, containing 25 mg/liter of streptomycin sulphate. After 5 days, a fungus developed producing a whitish-orange mycelium when incubated under 12 h/day of fluorescent light at 23°C. The isolates obtained were purified on PDA. On this medium, they produced hyaline elliptical and ovoid conidia, rarely septate, measuring 5.0 to 9.0 × 1.7 to 3.9 (average 6.0 × 2.9) μm. Conidia were born on phialides, single, clavate, and 2.8 × 1.4 μm. Such characteristics are typical of Plectosphaerella sp. (1,2). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (3) and sequenced. BLAST analysis of the 530-bp segment obtained from C. endivia var. crispum isolate PLC28 and of the 527-bp from C. endivia var. latifolium isolate PLC 30, respectively, showed 99% similarity with the sequence of Plectosphaerella cucumerina (anamorph Plectosporium tabacinum), GenBank EU5945566. The nucleotide sequences of isolates PLC 28 and PLC 30 have been assigned the GenBank accession numbers KC293994 and KC293993, respectively. To confirm pathogenicity, tests were conducted on 30-day-old C. endivia plants. C. endivia var. crispum cv Myrna and C. endivia var. latifolium cv. Sardana plants, grown in 2-liter pots (1 plant per pot, 10 plants per treatment) were inoculated by spraying a 106 CFU/ml conidial suspension of the two isolates of P. cucumerina, prepared from 10-day-old cultures, grown on PDA. Inoculated plants were maintained in a growth chamber at 25 ± 1°C and 90% RH for 5 days. Non-inoculated plants, only sprayed with water, served as controls. All plants inoculated with the two isolates, showed typical leaf spots 7 days after the artificial inoculation, similar to those observed in the field. Later, spots enlarged and leaves rotted. Non-inoculated plants remained healthy. P. cucumerina was reisolated from inoculated plants. The pathogenicity tests were conducted twice with identical results. This is, to our knowledge, the first report of P. cucumerina on endive n Italy, as well as worldwide. Due to the importance of the crop in Italy, this disease can cause serious economic losses. References: (1) A. Carlucci et al. Persoonia 28:34, 2012. (2) M. E. Palm et al. Mycologia 87:397, 1995. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

Rebutia perplexa, Cactaceae family, is a clumping fine thorny cactus, producing several flushes of pink flowers. In the spring of 2013, a blight was observed in a farm located near Imperia (northern Italy) on 2% of 2,000 3-year-old plants, grown in plastic pots. Affected plants showed pale brown discoloration of stems, starting from the base, and eventually collapsed. Flowers also rotted and wilted. In the presence of high relative humidity, a rare, whitish mycelium developed on the surface of the substrate. Eventually, infected plants died. Symptomatic tissues of the stem were taken from 10 plants and plated on potato dextrose agar (PDA). A fungus with the morphological characters of Rhizoctonia solani (3) was consistently recovered. Three representative isolates obtained from affected plants were successfully paired with tester strains of R. solani (AG 1, AG 2-2-IIIB, AG 2-2-IV, AG 4, AG 7, AG 11) (2) and examined microscopically. Three replicated pairings were made for each tester strain. The Rebutia isolates anastomosed only with AG 2-2-IIIB tester strain with high hyphal fusion frequency. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and death of adjacent cells was observed, indicating anastomosis reactions (1). Tests were performed twice. Mycelium of 15-day-old isolates maintained at 27 to 30°C, appeared whitish or pale buff in color, coarse, with a concentric zonation, scarce aerial mycelium, and without sclerotia. The optimum temperature for mycelium growth was 30°C (daily growth rate: 24.6 mm) and isolates grew also at 35°C. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis of the 523-bp amplicon (GenBank Accession No. KF719318) showed a 99% homology with the sequence of the R. solani AG 2-2-IIIB isolate GU811672. The nucleotide sequence has been assigned the GenBank Accession No. KF719318. Therefore, on the basis of molecular characteristics, anastomosis tests, temperature growth, and cultural characteristics, the isolates from R. perplexa were identified as R. solani AG 2-2-IIIB. For pathogenicity tests, 3 g of colonized wheat kernel from 10-day-old cultures of a representative isolate of the fungus was added per 1 l of substrate in 12 potted healthy plants of R. perplexa. The inoculum was prepared by inoculating wheat kernels with the mycelium of 10-day-old cultures of the fungus and incubating at 25 ± 1°C (12 h fluorescent light, 12 h dark). Twelve plants inoculated with non-infested wheat kernels served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 25 ± 1°C. The first symptoms, similar to those observed in the farm, developed 5 days after inoculation. Fifteen days after the artificial inoculation, all inoculated plants were dead. R. solani was re-isolated only from the stems of symptomatic plants. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of blight of R. perplexa caused by R. solani in Italy as well as worldwide. References: (1) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease control. Kluwer Academic Publishers, The Netherlands, pp. 37-47, 1996. (2) A. Ogoshi. Ann. Rev. Phytopathol. 25:125, 1987. (3) B. Sneh et al. Identification of Rhizoctonia species. APS Press, St Paul, MN, 1991.

Nettle-leaved bellflower (Campanula trachelium) is a hardy plant belonging to the Campanulaceae, cultivated in gardens for its large and showy flowers. During September 2020, symptoms of a rust were noted on about 90% out of around thirty 7- to 8-month-old plants of C. trachelium growing in a private garden located in the Biella province (northern Italy; 45°36′00″N, 8°03′00″E). Symptoms consisted of leaf chlorosis and round pustules typical of rusts on the abaxial leaf surfaces of infected plants. The percentage of infected leaves was about 70%, whereas the disease severity on affected leaves ranged from 5 to 20%. Orange uredinia were circular to elliptical and were 0.2 to 0.5 mm in diameter. Urediniospores were orange in color, ovoid to ellipsoid, and measured 16.2 to 30.0 × 14.1 to 23.2 (average 23.7 × 18.8) µm (n = 50). Teleutospores were not observed. On the basis of its morphology, the causal agent of the disease was determined to belong to the genus Coleosporium (Cummins 1978). The urediniospores of the pathogen were gently collected from affected leaves, and the DNA of the fungus was extracted using the E.Z.N.A. Fungal DNA Mini Kit (Omega Bio-Tek, Darmstadt, Germany). PCR was performed with primers ITS1/ITS4 (White et al. 1990), obtaining a 578-bp sequence (GenBank accession no. MW056179). The BLASTn analysis of this sequence showed 99.48% similarity with Coleosporium campanulae (KY810468). Pathogenicity was confirmed through inoculations by gently pressing diseased leaves onto leaves of three healthy 6-month-old plants of C. trachelium. Three noninoculated plants served as a control. All plants were covered by plastic bags for 7 days and maintained at temperatures ranging from 15 to 25°C. About 10 days after inoculation, typical rust symptoms developed on inoculated plants, whereas controls did not show symptoms. The pathogenicity test was carried out twice, obtaining the same results. The rust DNA on newly affected plants was identified as C. campanulae by using primers ITS1/ITS4, obtaining a 625-bp sequence (GenBank accession no. MW337297). C. campanulae is reported in several countries, on many host plants including C. trachelium (Farr and Rossman 2020). In Italy, this pathogen has been recently detected on C. rapunculoides (Garibaldi et al. 2017). To our knowledge, this is the first report of rust caused by C. campanulae on C. trachelium in Italy. The spread of C. campanulae to Campanula spp. complicates the management of this disease, discouraging the use of these ornamentals in public gardens, where their cultivation is increasing, especially for mixed borders in low-maintenance gardens. Voucher specimens are available at the Agroinnova Collection, University of Torino.

Phlox maculata (common name spotted phlox), Polemoniaceae family, is an herbaceous perennial plant with showy panicle inflorescences, commonly grown in mixed borders. During summer 2019, about twenty 5-year-old plants of P. maculata growing in a garden located in the Biella province (northern Italy) showed symptoms of a leaf spot, starting from the basal part of the foliage. Symptoms consisted in brown spots with not well-defined margins, irregularly circular to polygonal in shape, up to 2 to 3 mm in diameter, sometimes confluent. The internal part of spots dried. Chlorosis and yellowing surrounded the spots, involving the most part of affected leaf surfaces, due to the spread of the disease. The ornamental value of affected plants was significantly reduced. Several symptomatic leaves were washed in sterile water for 1 min. Isolations were carried out, taking several small fragments of tissues from the margins of the spots and placing them on potato dextrose agar. Fungal colonies that developed were dark green at the end of development. Successively, single-spore colonies were obtained, and pure cultures were transferred on potato carrot agar (PCA, Simmons 2007) under a light/dark regime of 11 h/13 h. On this medium, colonies formed branched conidiophores with brown, multicellular conidia, ovoid to obclavate in shape, containing one to four (average 2.4, n = 50) transverse septa and zero to one (average 0.1, n = 50) longitudinal septa. Conidia measured 10.8 to 29.3 × 5.3 to 11.2 µm (average 18.7 × 7.6 µm, n = 50). If present, the beak was 2.1 to 11.0 µm (average 5.5 µm) long. These morphological characteristics permitted identifying the fungus isolated from P. maculata as Alternaria section alternata (Woudenberg et al. 2015). The fungal DNA from the 19/33 isolate was extracted using the E.Z.N.A. Fungal DNA Mini Kit (Omega Bio-Tek, Darmstadt, Germany), and PCR was performed using the ITS (White et al. 1990), the rpb2, and the endoPG primers (Woudenberg et al. 2015). Three sequences with 529 bp (ITS), 889 bp (rpb2), and 452 bp (endoPG) (GenBank accession nos. MN871825, MN871979, and MN871978, respectively) were produced. A BLASTn analysis of these sequences exhibited 100% identity with the reference strain CBS 117.44 of A. alternata (Fries) Keissler in rpb2 and endoPG portions (accession nos. KP124772 and KP124001, respectively) and 99.6% identity in ITS region (accession no. KP124303). One P. maculata fungal isolate (19/33) was grown on PCA. A spore suspension obtained from 15-day-old cultures was sprayed at the concentration of 10⁵ CFU/ml onto leaves of three healthy plants of P. maculata. Three control plants were treated with sterile water. All plants were closed in moist plastic bags for 7 days and maintained at temperatures ranging from 15 to 28°C. The first symptoms of leaf spot appeared 10 days after the inoculation, and A. alternata was reisolated from affected leaves. Control plants remained symptomless. The pathogenicity test was repeated once using the isolate 19/40, obtaining the same results. To our knowledge, this is the first report of A. alternata on P. maculata in Italy as well as worldwide. The spread of A. alternata on P. maculata and the difficulty to manage this disease, particularly in public gardens, may discourage the use of this ornamental in Italy.

Bakanae disease (caused by Fusarium fujikuroi) and rice blast (caused by Magnaporthe oryzae) are two of the most important seedborne pathogens of rice. The detection of both pathogens in rice seed is necessary to maintain high quality standards and avoid production losses. Currently, blotter tests are used followed by morphological identification of the developing pathogens to provide an incidence of infection in seed lots. Two loop-mediated isothermal amplification assays were developed with primers designed to target the elongation factor 1-α sequence of F. fujikuroi and the calmodulin sequence of M. oryzae. The specificity, sensitivity, selectivity, repeatability, and reproducibility for each assay was assessed in line with the international validation standard published by the European and Mediterranean Plant Protection Organization (PM7/98). The results showed a limit of detection of 100 to 999 fg of DNA of F. fujikuroi and 10 to 99 pg of M. oryzae DNA. When combined with a commercial DNA extraction kit, the assays were demonstrated to be effective for use in detection of the pathogens in commercial batches of infected rice seed of different cultivars, giving results equivalent to the blotter method, thus demonstrating the reliability of the method for the surveillance of F. fujikuroi and M. oryzae in seed-testing laboratories.

Tradescantia virginiana, also known as Virginia spiderwort, belonging to Commelinaceae, is a rustic perennial plant, generally used in low-maintenance gardens. During the summer of 2019 plants grown in mixed planting borders and pots in several gardens located in the Biella province (northern Italy) showed extensive foliar disease. Sixty percent of about 100 plants grown in one garden were severely diseased. Early symptoms were small, light brown, necrotic spots with a reddish border, later reaching 10 mm diameter with an irregular shape. In some cases, the internal part of the necrotic areas dried with the appearance of holes. The disease progressed from the base to the apex of plants, and most of the leaves turned completely necrotic. For identification of the causal agent, symptomatic tissues were immersed in 1% sodium hypochlorite for 2 s, rinsed with sterile distilled water, and dried. Small fragments were excised from the margin of lesions and plated on potato dextrose agar (PDA) amended with 25 mg/liter of streptomycin sulfate. A single fungus with felty, white cream to gray colonies consistently developed. A 14-day-old culture of the isolate 19-29, selected among those collected as representative based on morphological characteristics grown on PDA at 21 to 23°C under alternating daylight and darkness (12 h photoperiod), produced black globose or subglobose pycnidia measuring 89 to 211 μm in diameter, with masses of hyaline, elliptical conidia, mainly nonseptate, measuring 8.9 to 15.3 × 5.6 to 7.9 (average 11.4 × 7.1) μm. On the basis of the morphological characteristics, the fungus was identified as Phoma sp. (Boerema et al. 2004). The internal transcribed spacer of rDNA was amplified using the primers ITS1/ITS4 (White et al. 1990) and then sequenced. BLAST analysis (Altschul et al. 1997) of a 488-bp segment of isolate 19-29 (GenBank accession no. MN526484) and a 541-bp segment of isolate 19-30 previously isolated in the same area (GenBank accession no. KJ443356.1) were identical to the sequence of Phoma commelinicola strain CBS100409 (GenBank accession no. GU237712). Pathogenicity tests were performed by spraying five T. virginiana plants grown in 3-liter pots with 1 × 10⁵ conidia/ml obtained from the 19-29 isolate, and the same number of healthy plants inoculated with sterile water served as controls. Plants were maintained in the garden at 18 to 25°C and covered with plastic bags for 10 days to maintain 90% relative humidity. The first foliar lesions, similar to those observed in the field, developed on leaves 10 days after inoculation, and after 30 days all inoculated plants were severely damaged. Control plants remained healthy. The pathogenicity test was carried out twice. The fungus reisolated with 100% frequency from leaf lesions of six samples obtained from the inoculated plants was identical in morphology to the isolate used for inoculation. P. commelinicola has been reported on Commelina nudiflora in Puerto Rico (Young 1915) and on Tradescantia sp. in New Zealand (Boerema et al. 2004). To our knowledge, this is the first report of the presence of P. commelinicola on T. virginiana in Italy and Europe (Farr and Rossman 2019). At present the economic importance of this disease is unknown, but it may become a more significant problem if the cultivation of Virginia spiderwort in gardens increases.

During the spring and autumn of 2017, 40-day-old spinach (Spinacia oleracea L.) plants, cultivar Crocodile, grown in a commercial field in the Asti province in Piedmont (Northern Italy), showed symptoms of a previously unknown foliar disease. The first symptoms developed at air temperatures ranging from 10 to 22°C. Five to 25% of plants showed small circular, yellow to pale brown leaf spots (1 to 2 mm in diameter), with a well-defined border, generally surrounded with a yellow halo. As the lesions expanded, they changed from circular to elliptical or irregular, becoming necrotic and broken in the center. Small tissue fragments were excised from symptomatic leaves, dipped in a solution containing 1% sodium hypochlorite for 5 s, washed in sterile water, dried on sterile paper, and plated on potato dextrose agar. After 7 days of incubation under 12-h fluorescent light at 22°C, 80% of fragments were colonized by the same fungus. Examination of the isolates IT27, IT28, and IT50 grown on V8 agar medium (200 ml of V8, 15 g of agar, 0.5 g of CaCO₃, and 1 liter of distilled water) revealed multicellular, light brown pigmented, obclavate or subspherical conidia, measuring 19.2 to 31.9 μm long × 10.7 to 21.4 μm wide (length/width ratio 1.1 to 1.4), with two to four transverse and three to four longitudinal septa. Conidia developed on a distinctive brown conidiophore with a terminal enlargement. These morphological characters permitted identifying the fungus as Stemphylium sp. (Simmons 1969). Genomic DNA from 7-day-old pure cultures of IT27, IT28, and IT50 were extracted with E.Z.N.A. Plant DNA Kit (Omega Bio-Tek). Polymerase chain reaction (PCR) was performed using primers ITS1/ITS4 to amplify the internal transcribed spacer, the intergenic region between 28 S and 18 S sequences of the ribosomal RNA, including the 5.8 S sequence. The PCR products were purified and sent for sequencing to BMR Genomics (Padova, Italy). BLASTn analysis (Altschul et al. 1997) of the three isolates (483 bp) showed 100% similarity with Stemphylium beticola CBS 141024 (GenBank no. NR_154925.1). Additionally, alignment with the CBS isolates used by Woudenberg et al. (2017) and the isolates from spinach was carried out using Clustal W with Mega 6.0.6 (Tamura et al. 2013). Sequence analysis of the same length sequences was performed using the maximum likelihood method with the Kimura two-parameter model plus gamma distribution. The sequences from IT27, IT28, and IT50 isolates clustered with the S. beticola strains CBS 141024, CBS 141025, and CBS 141026 (GenBank nos. KU850520, KU850521, and KU850522, respectively), allowing confirmation of the identity of isolates from spinach as S. beticola. The sequences from the isolates IT50, IT27, and IT28 were deposited in GenBank with the accession numbers MG913379, MG913380, and MG91338, respectively. The pathogenicity test was carried out by spraying a conidial suspension of 10⁵conidia/ml from a single-spore culture of strain IT50 onto leaves of 30-day-old spinach cultivar Crocodile plants, maintained under >90% relative humidity conditions for 5 days after inoculation. Pots (five per treatment, 10 plants/pot) were kept in a growth chamber at an average temperature of 20°C. The first lesions developed on leaves 8 days after inoculation, whereas control plants remained healthy. The pathogenicity test was conducted three times, providing the same results. Based on morphological and molecular analysis, the same fungus was consistently reisolated from the lesions. This is to our knowledge the first report of S. beticola on spinach in Italy and probably in Europe (Farr and Rossman 2018), although the same species has been reported on spinach in the United States (Woudenberg et al. 2017). The importance of the disease is at present limited but could rapidly increase in the cultivation areas where spinach is intensively grown.