First Report of a Leaf Spot Caused by Alternaria compacta on Hydrangea anomala subsp. petiolaris in Italy

Hydrangea anomala subsp. petiolaris (synonym H. petiolaris and H. scandens), also known as climbing hydrangea, is cultivated as an ornamental for landscaping in parks and gardens. This species, belonging to the Hydrangeaceae and native to the woodlands of Japan and coastal China, is widely appreciated for its abundant, creamy white flowers with a sweet aroma, particularly in shade gardens. During the summer of 2006, extensive necroses were observed on leaves and young stems of 3-year-old plants grown outdoors in several gardens of Piedmont of northern Italy. In many cases, on the upper side of the leaves, necrotic spots (4 to 10 mm in diameter) turned progressively black. Lesions often coalesced, generating larger (2 to 6 cm in diameter) necrotic areas. Necroses initially developed mainly at leaf margins and near petioles, and severely affected plants were defoliated. Infected plants rarely died, but the presence of lesions reduced the aesthetic quality and subsequently the commercial value. The disease occurred on 50 of 100 plants. A fungus was consistently isolated from infected leaves on potato dextrose agar (PDA) and identified on the basis of its morphological characteristics as an Alternaria sp. Conidia were dark gray, multicellular, clavate to pear shaped, measuring 23 to 54 × 10 to 13 μm (average 38 × 12 μm), with five longitudinal crosswalls and a relatively short apical beak. DNA was extracted with a Nucleospin Plant Kit (Macherey Nagel, Brockville, ON, Canada) and PCR was carried out with ITS 6/ITS 4 primer (2). A 557-bp PCR product was sequenced, and a BLASTn search (1) confirmed that the sequence corresponded to Alternaria compacta (99% homology). The nucleotide sequence has been assigned GenBank Accession No. EU 128529. Pathogenicity tests were performed by spraying leaves of healthy 1-year-old potted H. anomala plants with an aqueous 105 CFU/ml spore suspension. The inoculum was obtained from cultures of the fungus grown on sterilized host leaves placed on PDA for 20 days in light/dark at 23 ± 1°C. Plants sprayed only with water served as controls. Five plants were used for each treatment. Plants were covered with plastic bags for 3 days after inoculation and maintained between 12 and 22°C. Lesions developed on leaves 8 days after inoculation with the spore suspension, whereas control plants remained healthy. A. compacta was consistently reisolated from these lesions. The pathogenicity test was repeated twice. The presence of an Alternaria sp. on H. macrophylla was reported in the United States (3), whereas A. hortensiae was observed in Spain on H. hortensis. Recently, A. alternata belonging to the alternata group was reported on H. macrophylla in Italy (4). This is, to our knowledge, the first report of A. compacta on H anomala subsp. petiolaris in Italy. References: (1) S. F. Altschud et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (3) M. L. Daughtrey et al. Page 9 in: Compendium of Flowering Potted Plant Diseases. American Phytopathological Society. St. Paul, MN, 1995. (4) A. Garibaldi et al. Plant Dis. 91:767, 2007.

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First Report of Leaf Spot Caused by Alternaria alternata on Hydrangea macrophylla in Italy

Plant Disease ◽

10.1094/pdis-91-6-0767a ◽

2007 ◽

Vol 91 (6) ◽

pp. 767-767 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

S. Frati ◽

M. L. Gullino

Keyword(s):

Single Cells ◽

Morphological Characteristics ◽

The United States ◽

Major Allergen ◽

Plant Diseases ◽

Pathogenicity Test ◽

Aesthetic Quality ◽

First Report ◽

Hydrangea Macrophylla ◽

Alternaria Sp

Hydrangea macrophylla is cultivated as an ornamental and also used in the landscape. During the fall of 2005, leaves and young stems on 12-month-old plants (cvs. Hanabi, Nigra, and Zaffiro) grown in pots in several gardens and commercial nurseries in the Piedmont (northern Italy) had extensive necrosis. In many cases, 4-mm-diameter spots on the upper side of the leaves were surrounded by a chlorotic halo, which turned progressively black. Lesions often coalesced into 3- to 8-cm-diameter necrotic areas. Initial necrosis developed mainly on the leaf margins and near the petioles. Severely affected plants were defoliated. Infected plants rarely died, but the presence of lesions reduced the aesthetic quality and subsequently the commercial value. The disease occurred on 30 to 50% of the plants. Leaf spots contained dark brown, multicellular, pear-shaped conidia. Conidia were 19.2 to 36.5 μm (average 26.3 μm) long and 7.7 to 11.5 μm (average 8.9 μm) wide, with 3 to 4 longitudinal cross walls and an average of 4.4 single cells. A fungus identified on the basis of its morphological characteristics as an Alternaria sp. was consistently isolated from symptomatic leaves onto potato dextrose agar. DNA was extracted from mycelium (Nucleospin Plant Kit, Macherey Nagel, Brockville, ON, Canada) and PCR was completed using Alt-for/Alt-rev primers (3), which amplified a part of the gene that encodes for the protein Alt a 1, the major allergen produced by the genus Alternaria. A 305-bp fragment was amplified, sequenced, and the sequence was subjected to BLASTn analysis (1), which confirmed that the isolate belonged to the genus Alternaria and to the alternata group (3). The nucleotide sequence has been deposited in GenBank (Accession No. EF446670). Pathogenicity tests were performed by spraying leaves of healthy potted H. macrophylla plants, cvs. Zaffiro (6-month-old) and Hanabi (12-month-old) with a spore suspension (105 conidia/ml). Plants sprayed with water only served as a control. Ten plants per cultivar were used for each treatment. Plants were covered with plastic bags for 5 days after inoculation and maintained at 20°C for an additional 7 days. Plants were transferred outdoors and kept at temperatures ranging from 19 to 25°C. The first foliar lesions developed on leaves 15 days after inoculation, whereas control plants remained healthy. Alternaria sp. was consistently reisolated from these lesions. The pathogenicity test was completed twice. The presence of Alternaria sp. on Hydrangea spp. was reported in the United States (2), whereas A. hortensiae was observed in Spain (4). To our knowledge, this is the first report of Alternaria sp. belonging to the alternata group infecting H. macrophylla in Italy. The disease is currently spreading in other Italian areas. References: (1) S. F. Altschud et al. Nucleic Acids Res. 25:3389, 1997. (2) M. L. Daughtrey et al. Page 9 in: Compendium of Flowering Potted Plant Diseases. The American Phytopathological Society, St. Paul, MN, 1995. (3) S. Gyu Hong et al. Fungal Genet. Biol. 42:119, 2005. (4) L. M. Unamuno. An. Jard. Bot. Madr. 4:145, 1944.

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Leaf Spot Caused by Alternaria sp. on Iberis sempervirens in Italy

Plant Disease ◽

10.1094/pd-89-1243b ◽

2005 ◽

Vol 89 (11) ◽

pp. 1243-1243 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

Leaf Spot ◽

Single Cells ◽

Morphological Characteristics ◽

Weather Conditions ◽

The United States ◽

Plant Diseases ◽

Pathogenicity Test ◽

Aesthetic Quality ◽

Leaf Spots ◽

Alternaria Sp

Iberis sempervirens (candytuft) is increasingly grown in Liguria (northern Italy) as a potted plant for ornamental purposes, particularly under cool-weather conditions. At the end of the summer of 2003, extensive necrosis was observed on leaves and young stems of 4-month-old plants grown in 14-cm diameter pots outdoors at a commercial farm. In many cases, on the upper side of the leaves, necrotic spots were surrounded by a chlorotic halo that turned progressively black. The necrotic areas often coalesced, generating larger and irregularly shaped spots. On the lower side of the leaves, no chlorotic areas were observed. Severely affected plants were defoliated. Infected plants rarely died, but the presence of lesions on mature plants decreased aesthetic quality and subsequently market value. The disease occurred on 40% of plants at each of the two farms. Leaf spots contained dark brown, multicellular pear-shaped conidia. Conidia were 22.5 to 50.0 μm (average 32.8 μm) long and 7.5 to 15.0 μm (average 12.3 μm) wide, with 5 to 7 longitudinal cross walls and an average of 6 to 7 single cells. From infected leaves, a fungus identified on the basis of its morphological characteristics as Alternaria sp. was consistently isolated on potato dextrose agar. Pathogenicity tests were performed by spraying leaves of healthy 12-month-old potted I. sempervirens plants with a spore and mycelial suspension (105 CFU/ml). Plants without inoculation served as control. Ten plants were used for each treatment. Plants were covered with plastic bags for 10 days after inoculation and kept outdoors for 60 days at temperatures ranging from 0 to 32°C (average 12°C). The first lesions developed on leaves 45 days after inoculation, while control plants remained healthy. From such lesions, Alternaria sp. was consistently reisolated. The pathogenicity test was carried out twice. The presence of A. brassicae was reported in Tanganica on Iberis sp., I. umbellata in Denmark (2), and I. amara in the United States (4); A. matthiolae was observed on seeds of I. amara and I. umbellata (3). A leaf spot incited by Alternaria sp. on I. amara was observed in Florida (1). This is, to our knowledge, the first report of Alternaria sp. on I. sempervirens in Italy as well as worldwide. References: (1) S. A. Alfieri et al. Index of Plant Diseases in Florida. Bull. 11, 1984. (2) P. Neergaard. Rev. Appl. Micol. 18:572, 1939. (3) P. Neergaard. Rev. Appl. Micol. 25:382, 1946). (4) R. D. Raabe. Comb. Proc. Int. Plant Propagators Soc. 40:160, 1991.

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First Report of Alternaria Leaf Spot on Camellia in Italy

Plant Disease ◽

10.1094/pdis-91-3-0324c ◽

2007 ◽

Vol 91 (3) ◽

pp. 324-324 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

M. L. Gullino

Keyword(s):

Single Cells ◽

Morphological Characteristics ◽

The United States ◽

Tea Plant ◽

Northern Italy ◽

Plant Diseases ◽

Pathogenicity Test ◽

Camellia Japonica ◽

Aesthetic Quality ◽

First Report

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.

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First Report of Alternaria Leaf Blight of Aralia japonica Caused by Alternaria panax in Europe

Plant Disease ◽

10.1094/pdis.2004.88.1.82b ◽

2004 ◽

Vol 88 (1) ◽

pp. 82-82 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

M. L. Gullino

Keyword(s):

Central Italy ◽

The United States ◽

Leaf Blight ◽

Plant Diseases ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Single Spore ◽

Aesthetic Quality ◽

First Report ◽

Alternaria Panax

Aralia japonica (synonym Fatsia japonica), belonging to the Araliaceae family, is a foliage plant highly valued in Italy for landscape and interior decoration. In the fall of 2002, a leaf blight disease was observed on plants grown in pots that were maintained under shade at a density of 15 to 20 pots per m2 at a nursery located in central Italy (Teramo Province). Typical symptoms were tan-to-dark brown leaf spots and rapid blighting of foliage under moist conditions. Chlorotic zones around necrotic lesions were common, and considerable leaf drop was associated with the disease. Affected plants were rarely killed, but the presence of lesions on mature plants reduced aesthetic quality and market value. The disease occurred on 70% of the plants. A fungus identified morphologically as Alternaria panax (2) was consistently isolated from infected leaves on potato dextrose agar (PDA). The fungus grows slowly and sparsely on PDA and produces a light brown mycelium, a characteristic red diffusible pigment in the agar medium, and rare conidia under 12-hr photoperiods. Measurements were carried out on conidia formed from single-spore isolates grown on autoclavated host tissue on water agar (LWA) at 24°C for 10 days. In LWA culture, conidia were borne singly or in chains of two to four conidia. Conidia produced in culture were smaller than those formed on the host and were highly variable in shape. They appeared obclavate, ellipsoidal, and obpyriform and pale to dark brown with relatively short or false beaks. Conidial bodies were 14.4 to 48.0 μm long (average 30.5 μm) and 7.2 to 12.0 μm wide (average 9.9 μm) with 3 to 10 transverse and a few longitudinal septa. Length of appendages was 9.6 to 26.0 μm (average 16.0 μm). Pathogenicity tests were performed by inoculating leaves of healthy Aralia japonica and Schefflera actinophylla plants by placing mycelial disks (5 mm in diameter) directly on wounded leaf tissues. Uninoculated, wounded plants served as controls. Four plants of each species were used. Plants were covered for 72 h with plastic bags and maintained in a growth chamber at 20°C (12 hours per day of fluorescent light). Control plants were maintained similarly. The first lesions developed on leaves of inoculated plants of both species after 7 days. A. panax was consistently reisolated from the lesions. The pathogenicity test was carried out twice. The presence of A. panax on Aralia japonica has been reported in Japan, Korea (2), and the United States (1) but to our knowledge, this is the first report of A. panax on Aralia japonica in Europe. References: (1) S. Alfieri et al. Index of plant diseases in Florida. Bull. 11:52, Florida Department of Agriculture and Consumer Services, 1984 (2) S. H. Yu et al. Ann. Phytopathol. Soc. Jpn. 50:313, 1984.

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First Report of Downy Mildew Caused by Peronospora parasitica on Iberis sempervirens in Italy

Plant Disease ◽

10.1094/pd-90-1261c ◽

2006 ◽

Vol 90 (9) ◽

pp. 1261-1261

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

New York ◽

Downy Mildew ◽

Morphological Characteristics ◽

Sprinkler Irrigation ◽

The United States ◽

Plant Diseases ◽

Pathogenicity Test ◽

Peronospora Parasitica ◽

First Report ◽

Downy Mildews

Iberis sempervirens (evergreen candytuft) is a garden species belonging to the Brassicaceae family. During June 2004, a damaging foliar disease was observed in several commercial farms near Albenga (northern Italy) on I. sempervirens plants grown outdoors in containers. More than 30% of the plants were affected. Symptoms appeared on both sides of leaves, buds, flowers, and fruits. Initially, leaves were slightly chlorotic, but within 5 to 7 days a characteristic whitish furry growth developed on the lower and upper leaf surfaces. The efflorescence was particularly evident on the lower surfaces of leaves and consisted of sporangiophores and sporangia. The appearance and severity of the disease increased because of overhead sprinkler irrigation. Microscopic observations revealed dichotomously branched sporangiophores with slender curved tips. Sporangiophores with a length of 115 to 410 μm (average 295 μm) ended with sterigmata bearing single sporangia. Sporangia were ovoid and measured 18 to 28 × 25 to 45 μm (average 22 × 35 μm). The pathogen was identified as Peronospora parasitica on the basis of its morphological characteristics (3). Pathogenicity was confirmed by inoculating leaves of 10 45-day-old healthy plants grown in 14-cm-diameter pots with a sporangial suspension (1 × 103 conidia/ml). Ten noninoculated plants served as controls. Plants were maintained outdoors at 50% light intensity with temperatures ranging between 16 and 25°C (average 18°C) and 85 to 100% relative humidity. The pathogenicity test was carried out twice. After 18 days, typical symptoms of downy mildew developed on the inoculated plants and P. parasitica was observed on the leaves. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of P. parasitica on evergreen candytuft in Italy. P. parasitica was previously reported on I. sempervirens in the United Kingdom (1) and on I. amara in California (2). Voucher specimens are available at the AGROINNOVA Collection, University of Torino, Italy. References: (1) S. Francis and G. Waterhouse. Trans. Br. Mycol. Soc. 91:1, 1988. (2) P. R. Muller et al. Index of Plant Diseases in the United States. USDA Handbook No. 165, 1960. (3) D. M. Spencer. The Downy Mildews. Academic Press, New York, 1981.

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First Report of Rust Caused by Pucciniastrum circaeae on Fuchsia × hybrida in Italy

Plant Disease ◽

10.1094/pdis-11-11-0976-pdn ◽

2012 ◽

Vol 96 (4) ◽

pp. 588-588 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

G. Ortu ◽

M. L. Gullino

Keyword(s):

Phylogenetic Trees ◽

Aqueous Suspension ◽

Morphological Characteristics ◽

Pcr Amplification ◽

Signs And Symptoms ◽

The United States ◽

Pathogenicity Test ◽

First Report ◽

Potted Plants ◽

Plastic Bags

Fuchsia is a genus of flowering plants that is native to South America and New Zealand and belongs to the family Onagraceae. In September 2011, 2-year-old potted plants of Fuchsia × hybrida, cv. Citation, in a garden located near Biella (northern Italy) showed signs and symptoms of a previously unknown disease. Typically, infected plants showed leaf chlorosis followed by the appearance of necrosis on the adaxial leaf surfaces, while the abaxial surfaces showed orange uredinia irregularly distributed. As the disease progressed, infected leaves turned yellow and wilted. Affected plants showed a progressive phylloptosis and also flowering was negatively affected. Urediniospores were globose, yellow to orange, and measured 14.6 to 25.9 (average 19.6) μm. Teliospores were not observed. Morphological characteristics of the fungus corresponded to those of the genus Pucciniastrum. DNA extraction and PCR amplification were carried out with Terra PCR Direct Polymerase Mix (Clontech, Saint Germain-en-Laye, France) and primers ITS1/ITS4 (4). A 700-bp PCR product was sequenced and a BLASTn search (1) confirmed that the sequence corresponded with a 96% identity to Pucciniastrum circaeae. The nucleotide sequence has been assigned the GenBank Accession No. JQ029688. Pathogenicity tests were performed by spraying leaves of healthy 1-year-old potted Fuchsia × hybrida plants with an aqueous suspension of 1 × 103 urediniospores ml–1. The inoculum was obtained from infected leaves. Plants sprayed only with water served as controls. Three plants were used for each treatment. Plants were covered with plastic bags for 4 days after inoculation and maintained outdoors at temperatures ranging between 18 and 25°C. Lesions developed on leaves 20 days after inoculation with the urediniospore suspension, showing the same symptoms as the original plants, whereas control plants remained healthy. The organism that was recovered from the lesions after inoculation was the same as the one obtained from the diseased plants. The pathogenicity test was carried out twice with similar results. The presence of P. fuchsiae, later identified as P. epilobii, was repeatedly reported in the United States (3). P. epilobii and P. circaeae have closely related hosts and morphologically similar urediniospores. These species were reported to form a single group in molecular phylogenetic trees (2). This is, to our knowledge, the first report of P. circaeae on Fuchsia × hybrida in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) Y. M. Liang et al. Mycoscience 47:137, 2006. (3) L. B. Loring and L. F. Roth. Plant Dis. Rep. 48:99, 1964. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Stem Rot on Cereus peruvianus monstruosus Caused by Bipolaris cactivora (Petr.) Alcorn in Italy

Plant Disease ◽

10.1094/pdis-06-13-0614-pdn ◽

2014 ◽

Vol 98 (1) ◽

pp. 159-159 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

United States ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Stem Rot ◽

Pathogenicity Test ◽

First Report ◽

Plastic Bags ◽

Blastn Analysis ◽

Dark Green

Cereus peruvianus monstruosus, known as “monster cactus,” family Cactaceae, is grown as a potted plant. In the winter of 2013, a stem rot was observed on a farm located near Ventimiglia (northern Italy) on 80% of 4,000 9-month-old plants grown in trays in a peat substrate. Symptoms consisted of a rapid rot of the upper portion of the stem. Affected stems at first showed yellowish spots that became brown irregular necrotic lesions with well-defined margins. The tissues below the affected areas were blackened and dry but became soft in the presence of high relative humidity. Fungal sporulation on rotted tissues consisted of caespitose, non-branched, septate conidiophores, olivaceous to brown at the base, paler above, measuring 89.0 to 196.9 × 6.2 to 8.7 (average 124.8 × 7.0) μm. Single conidia were borne on terminal cells. At maturity, conidia with 2 to 5 (average 3) septa were brownish-olivaceous, varying in shape from obclavate, fusiform, ellipsoid or sometimes furcate, and measuring 23.4 to 48.6 × 8.0 to 12.6 (average 38.8 × 10.3) μm. Symptomatic tissues were immersed in 1% sodium hypochlorite for 2 to 3 s and rinsed in sterile distilled water, then fragments excised from the margin of internal lesions were cultured on potato dextrose agar (PDA) medium amended with 25 mg/l of streptomycin sulfate and incubated at 20 to 23°C under alternating daylight and darkness (10 h light, 14 h dark). A fungus that was consistently isolated was subcultured on PDA. At maturity, dark green floccose colonies comprised of light brown septate hyphae, 4.2 to 8.1 (average 5.6) μm in width, produced non-branched, pale to dark brown, septate conidiophores, measuring 99.6 to 176.1 × 4.5 to 6.5 (average 146.7 × 5.4) μm. The conidia produced on PDA were similar to those observed on infected tissues and measured 20.6 to 40.7 × 7.5 to 11.4 (average 32.0 × 9.7) μm, with 1 to 3 septa (average 2). On the basis of the morphological characteristics, the fungus was identified as Bipolaris cactivora (Petr.) Alcorn [Syn.: Drechslera cactivora (Petr.) M. B. Ellis] (4). The internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA) was amplified for one isolate using ITS1/ITS4 primers and sequenced (GenBank Accession No. KF041822). BLASTn analysis (1) of the 557-bp segment showed a 99% similarity with the ITS sequence of Bipolaris cactivora HM598679. For pathogenicity tests, 8 mm diameter mycelial disks removed from 15-day-old PDA cultures of the fungus were placed at the wounded stem apexes of three 7-month-old healthy plants (three disks per plant). Three plants inoculated with non-inoculated PDA disks served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 23 ± 1°C with 12 h light/dark. By 8 days after inoculation, all the inoculated stems were rotted and 10 colonies of B. cactivora were re-isolated from infected tissues. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. Several hosts are listed for B. cactivora including C. peruvianus, and the pathogen has been reported in the United States (2) and in South Korea (3). To our knowledge, this is the first report of B. cactivora on C. peruvianus monstruosus in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. APS Press, St Paul, MN, 1989. (3) I. H. Hyun et al. Res. Plant Dis. 7:56, 2001. (4) A. Sivanesan. Mycopathologia 111:125, 1990.

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First Report of Black Rot of Carrots Caused by Alternaria radicina in Michigan

Plant Disease ◽

10.1094/pd-90-0684b ◽

2006 ◽

Vol 90 (5) ◽

pp. 684-684

Author(s):

C. Saude ◽

M. K. Hausbeck

Keyword(s):

United States ◽

New York ◽

Sequence Similarity ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Distilled Water ◽

First Report ◽

Plastic Bags ◽

Alternaria Sp

In April 2005, an Alternaria sp. was isolated from carrot (Daucus carota) roots harvested in the fall of 2004 and held at 1 to 3°C in a storage facility in Newaygo County, MI. The pathogen was readily isolated on water agar from root tissue exhibiting grayish black, sunken lesions. Morphological characteristics were noted 5 to 7 days after single-conidium cultures were established on potato dextrose agar (3). Sixteen Alternaria sp. isolates were recovered. Cultures were dark olive brown, and conidia were pigmented, ellipsoidal, and produced singly or in chains of two. Conidia were 35 to 45 μm long and 15 to18 μm in diameter, usually with three to eight transverse and one to four longitudinal septa. Pathogenicity of isolates was tested on carrot roots in the laboratory and carrot seedlings (cv. Goliath) in the greenhouse. In the laboratory, four surface-sterilized, whole carrot roots were sprayed until runoff with 2 × 106 conidia/ml of each isolate and incubated at 23 to 25°C in a moist chamber for 10 days. Controls were sprayed with sterile distilled water. Ten to fifteen days after inoculation, inoculated carrots exhibited grayish black, sunken lesions, and an Alternaria sp. was reisolated from the margin of the lesions. Controls remained healthy. In the greenhouse, seven pots containing one 2-week-old carrot seedling were watered to saturation and plants were sprayed until runoff with 2 × 106 conidia/ml for each isolate. Control plants were sprayed with sterile distilled water. After inoculation, plants were enclosed in clear plastic bags, placed under 63% woven shade cloth and watered regularly. Black lesions were observed on the foliage 7 days after inoculation, and wilt and death of plants were observed 15 to 30 days after inoculation. Alternaria sp. was reisolated from the foliage of symptomatic plants. Control plants remained healthy. DNA was extracted from all isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region amplified with primers ITS4 and ITS5 and sequenced. A portion of the ITS sequence has been deposited in the NCBI database (GenBank Accession No. DQ394073). A BLAST search of the NCBI database with the ITS sequences revealed A. radicina, Accession No AY154704, as the closest match with 100% sequence similarity. In September 2005, an Alternaria sp. was isolated from black lesions on carrot roots, crowns, and foliage that were collected from fields in Newaygo and Oceana counties, MI. The recovered isolates were morphologically similar to A. radicina isolates obtained from stored carrots in April 2005. First isolated and identified on stored carrots in New York (3), A. radicina is also present in other carrot-producing areas of the United States (1) and was isolated not only from stored carrots but also from carrots in the field (2) and carrot seeds (4). To our knowledge, this is the first report of A. radicina on stored and field carrots in Michigan, which signifies a serious risk to a carrot industry that ranks among the top five in the United States. References: (1) D. F. Farr et al. Fungi on Plants and Plant Produce in the United States.The American Phytopathological Society, St. Paul, MN, 1989. (2) R. G. Grogan and W. C. Snyder. Phytopathology 42:215, 1952. (3) F. C. Meier and E. D. Eddy. Phytopathology 12:157, 1922. (4) B. M. Pryor and R. L. Gilbertson. Plant Dis. 85:18, 2001.

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First Report of Leaf Spot of Saponaria officinalis Caused by Alternaria nobilis in Italy

Plant Disease ◽

10.1094/pdis-09-12-0839-pdn ◽

2013 ◽

Vol 97 (3) ◽

pp. 424-424 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Morphological Characteristics ◽

Pathogenicity Test ◽

Academic Press ◽

First Report ◽

Perennial Plant ◽

Plastic Bags ◽

Pcr Product ◽

Pathogenicity Tests ◽

Alternaria Sp ◽

Saponaria Officinalis

Saponaria officinalis (Vize) Simmons (common name bouncingbet) is a low maintenance perennial plant belonging to the Caryophyllaceae family, typically grown in parks and gardens. During the summers of 2011 and 2012, extensive necrosis were observed on leaves of plants grown in private gardens, near Biella (northern Italy). The disease affected 90% of 1- to 2-year-old plants. The first symptoms were usually pale brown lesions 1 to 5 mm in diameter and sometimes coalesced. Lesions were circular to irregular with a dark purple halo, with infected leaves eventually turning chlorotic. The conidia observed on infected leaves were olivaceous brown and obclavate, with a beak. Conidia showed 8 to 15 (average 12) transverse and 4 to 14 (average 11) longitudinal septa, with slight constrictions connected with septa, and were 78.3 to 177.7 (average 135.5) × 19.0 to 34.3 (average 26.5) μm. The beak was 20.0 to 62.2 (average 33.7) μm in length, with 0 to 6 (average 3) transverse septa and no longitudinal septa. The fungus was consistently isolated from infected leaves on potato dextrose agar (PDA). The isolate, grown for 14 days at 20 to 24°C with 10 h of darkness and 14 h of light on sterilized host leaves plated on PDA, produced conidiophores single, unbranched, flexuous, septate with conidia in short chains, similar to those observed on the leaves and previously described. On the basis of its morphological characteristics, the pathogen was identified as Alternaria sp. (3). DNA was extracted using Nucleospin Plant Kit (Macherey Nagel) and PCR carried out using ITS 1/ITS 4 primer (4). A 542-bp PCR product was sequenced and a BLASTn search confirmed that the sequence corresponded to A. dianthi (AY154702), recently renamed A. nobilis (2). The nucleotide sequence has been assigned the GenBank Accession No. JX647848. Pathogenicity tests were performed by spraying leaves of healthy 3-month-old plants of S. officinalis with an aqueous 2 × 105 spore/ml suspension. The inoculum was obtained from cultures of the fungus grown on PDA amended with host leaves for 14 days, in light-dark, at 22 ± 1°C. Plants sprayed only with water served as controls. Four pots (1 plant/pot) were used for each treatment. Plants were covered with plastic bags for 4 days after inoculation and maintained in a glasshouse at 21 ± 1 °C. Lesions developed on leaves 9 days after inoculation with the spore suspension, whereas control plants remained healthy. A. nobilis was consistently reisolated from these lesions. The pathogenicity test was carried out twice. The presence of A. dianthi was reported on S. officinalis in Denmark (1) and Turkey. This is, to our knowledge, the first report of A. nobilis on S. officinalis in Italy. The presence and importance of this disease is, at present, limited. References: (1) P. Neergaard. Danish species of Alternaria and Stemphylium. Oxford University Press, 1945. (2) E. G. Simmons. Mycotaxon 82:7, 2002. (3) E. G. Simmons. Alternaria: An Identification Manual. CBS Biodiversity Series 6, Utrecht, The Netherlands, 2007. (4) T. J. White et al. In: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Leaf Spot of Wild (Diplotaxis tenuifolia) and Cultivated (Eruca vesicaria) Rocket Caused by Alternaria japonica in Italy

Plant Disease ◽

10.1094/pdis-04-11-0280 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1316-1316 ◽

Cited By ~ 13

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

C. Bertoldo ◽

M. L. Gullino

Keyword(s):

Morphological Characteristics ◽

Northern Italy ◽

Pathogenicity Test ◽

Academic Press ◽

First Report ◽

Plastic Bags ◽

Pcr Product ◽

Pathogenicity Tests ◽

Alternaria Sp ◽

Potential Impact

Wild (Diplotaxis tenuifolia) and cultivated (Eruca vesicaria) rocket, popular crops in Italy as well as in many Mediterranean areas, are grown for fresh consumption as well as for dish decoration. During fall and winter of 2010 to 2011, extensive necroses were observed on leaves of D. tenuifolia and E. vesicaria that were grown in commercial greenhouses in Piedmont and Liguria (northern Italy). The disease affected 30 to 40% of 60-day-old plants. First symptoms were usually black-brown lesions, 1 to 30 mm in diameter, which progressively turned black. Lesions usually started on the upper side of older leaves at the leaf margins and tips and developed a yellow halo. Eventually, lesions also affected leaf veins and stems. A fungus was consistently isolated from infected leaves on potato dextrose agar and was grown on water agar (15 g/liter) amended with autoclaved rocket tissues (100 g/liter). After 12 days of growth at 22°C and 12-h dark/12-h light, conidia that were produced were dark brown, obclavate, obpyriform, ovoid or ellipsoid, with beaks. Round conidia without beaks were also present. Conidia showed two to seven (average three to four) transverse and one to three longitudinal septa, and measured 17.7 to 56.2 (average 30.9) × 6.6 to 17.8 (average 10.8) μm. Conidia were produced singly or in short chains (two to three elements) and mostly presented a conical or cylindrical beak, 1.8 to 7.3 (average 3.6) μm, pale light brown to brown. On the basis of its morphological characteristics, the pathogen was identified as an Alternaria sp. (3). DNA was extracted with Terra PCR Direct Polymerase Mix (Clontech, Mountain View, CA) and PCR was carried out with ITS 1/ ITS 4 primer (4). A 553-bp PCR product was sequenced and a BLASTn search (1) confirmed that the sequence corresponded to Alternaria japonica. The nucleotide sequence has been assigned the GenBank Accession No. JP 742643. Pathogenicity tests were performed by spraying leaves of healthy 30-day-old wild and cultivated rocket plants with an aqueous 1 × 105 spore/ml suspension. The inoculum was obtained from cultures of the fungus grown on sterilized host leaves placed on water agar for 20 days in light/dark at 22 ± 1°C. Plants sprayed only with water served as controls. Three pots (four plants per pot) were used for each treatment. Plants were covered with plastic bags for 4 days after inoculation and maintained in a glasshouse at 22 ± 1°C. Lesions developed on leaves 7 days after inoculation with the spore suspension, whereas control plants remained healthy. A. japonica was consistently reisolated from these lesions. The pathogenicity test was carried out twice. The presence of A. japonica has been reported on several brassica hosts, such as Brassica napus, B. nigra, B. oleracea, and B. rapa (2). This is, to our knowledge, the first report of A. japonica on wild and cultivated rocket in Italy as well as in Europe. Because of the importance of rocket in many countries, the potential impact of this disease is high. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) J. C. David, IMI Description of Fungi and Bacteria. 144:1432, 2000. (3) E. G. Simmons. Alternaria. An Identification Manual. CBS Biodiversity Series 6, Utrecht, The Netherlands, 2007. (4) T. J. White et al. In: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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