First Report of Leaf Blight on Woodland Sage Caused by Rhizoctonia solani AG 1 in Italy

Woodland sage (Salvia nemorosa L.; Lamiaceae) is a hardy herbaceous perennial plant that is easy to grow and propagate and is used in parks and grown as potted plants. During the summer of 2009 in a nursery near Torino in northern Italy, a leaf blight was observed on 30-day-old plants of cv. Blau Koenigin grown in pots under shade. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along leaf margins. Lesions expanded along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and clung to the shoots. No symptoms were observed on the roots. 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 of streptomycin sulfate. A fungus with morphological characters of Rhizoctonia solani (3) was consistently recovered. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and with radial growth. Sclerotia were irregular and measured between 0.5 and 2 mm. Pairings were made with tester isolates of AG 1, 2, 3, 4, 5, 6, 7, 11, and AG B1. The only successful anastomosis was with tester isolate AG 1 (ATCC 58946). 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 (2). The description of sclerotia of the isolate AG1 was typical for subgroup 1A Type 2 (3). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 688 bp showed a 100% homology with the sequence of R. solani AG-1A and the nucleotide sequence has been assigned (GenBank Accession No. HM044764). For pathogenicity tests, the inoculum of one isolate of R. solani from the nursery was prepared by growing the pathogen on PDA for 7 days. The foliage of 30-day-old potted plants of S. nemorosa cv. Blau Koenigin was artificially inoculated with an aqueous suspension of PDA and mycelium fragments (1 g per mycelium per plant) prepared from cultures with a blender. Plants were covered with plastic bags for 3 days. Plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a glasshouse at 20 to 25°C. The first symptoms, similar to those observed in the nursery, developed 7 days after foliar inoculation. R. solani was consistently reisolated from infected leaves. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. To our knowledge, this is the first report of leaf blight of S. nemorosa caused by R. solani in Italy as well as worldwide. The importance of the disease is still unknown. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. E. Carling. Page 35 in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. Kluwer Academic Publishers, the Netherlands, 1996. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

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First Report of Leaf Blight on Hosta fortunei Caused by Rhizoctonia solani AG 4 in Italy

Plant Disease ◽

10.1094/pdis-93-4-0432c ◽

2009 ◽

Vol 93 (4) ◽

pp. 432-432 ◽

Cited By ~ 3

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Its Region ◽

The United States ◽

Morphological Characters ◽

Leaf Blight ◽

Pathogenicity Test ◽

First Report ◽

Leaf Petiole

Hosta fortunei (Liliaceae) is used in semishaded areas of gardens for its lavender-colored flowers produced in midsummer. In April of 2008, in a greenhouse at the University of Torino, located in Grugliasco (northern Italy), a leaf blight was observed on 15% of potted 60-day-old plants growing at temperatures ranging between 20 and 25°C and relative humidity of 60 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along leaf margins. Lesions expanded for several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and clung to the shoots. 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 characters of Rhizoctonia solani (4) 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 of R. solani obtained from affected plants were 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, but no anastomosis was observed. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 646-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 534556. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Six-month-old plants of H. fortunei were grown in 1-liter pots. Inoculum, which consisted of an aqueous suspension of PDA and mycelium disks (10 g of mycelium per pot), was placed at the collar of plants. Plants inoculated with water and PDA fragments alone served as control treatments. Five plants per treatment were used. Plants were maintained in a growth chamber at 20 ± 1°C. The first symptoms, similar to those observed in the nursery, developed 15 days after 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. R. solani was reported on plants belonging to the genus Hosta in the United States (3). This is, to our knowledge, the first report of leaf blight of H. fortunei 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 Phytopathology Society, St Paul, MN, 1989. (4) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.

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First Report of Leaf Blight on Coral Bells (Heuchera sanguinea) Caused by Rhizoctonia solani AG 1A in Italy

Plant Disease ◽

10.1094/pdis-91-9-1206b ◽

2007 ◽

Vol 91 (9) ◽

pp. 1206-1206

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Morphological Characters ◽

Leaf Blight ◽

Pathogenicity Test ◽

Uniform Size ◽

First Report ◽

Leaf Petiole ◽

Pathogenicity Tests ◽

Hyphal Diameter

Heuchera sanguinea (Saxifragaceae), coral bells or alum root, is an herbaceous perennial used in parks and gardens and sometimes grown in pots for its heart-shaped leaves and upright panicles of bright red, tiny flowers produced in late spring. At the end of fall 2006, a leaf blight was observed on 50% of a crop of potted 45-day-old plants grown in a sphagnum peat/clay/perlite (70:20:10) substrate at temperatures ranging between 20 and 25°C in a nursery. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. For several days, lesions expanded along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Mycelia were often seen on and suspended between leaves. Blight progressed up the plant from the leaves to the shoot tip. Affected plants often died leaving wide empty areas. Diseased tissue was disinfected for 1 min in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 100 μg/liter of streptomycin sulfate. A fungus with the morphological characters of Rhizoctonia solani was consistently and readily recovered, then transferred and maintained in pure culture (3). The isolates of R. solani obtained from affected plants were successfully anastomosed with tester isolate AG 1 (ATCC 58946). 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 (1). Pairings were also made with tester isolates of AG 2, 3, 4, 5, 6, 7, and 11 with no anastomoses observed between the recovered and tester isolates. Sclerotia were of uniform size with a diameter from 0.4 to 4 mm and sometimes joined laterally. The description of sclerotia was typical for subgroup 1A Type 2 (2). For pathogenicity tests, the inoculum of R. solani was prepared by growing three isolates of the pathogen on PDA for 7 days. Plants of 30-day-old H. sanguinea were grown in 10-liter containers (6 plants per container) on a steam disinfested peat/clay/perlite substrate (70:20:10)). Inoculum consisted of an aqueous suspension of PDA and mycelium disks (1 cm2 of mycelium per plant) and was placed at the base of the plant stems and on leaves. Plants inoculated with water and PDA fragments alone served as control treatments. Three replicates were used. Plants were maintained in a growth chamber at 24°C with 12 h of light/dark. The first symptoms, similar to those observed in the nursery, developed 12 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. This is, to our knowledge, the first report of leaf blight of H. sanguinea caused by R. solani in Italy and probably in the world. 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, 1996. (2) R. T. Sherwood. Phytopathology, 59:1924, 1969. (3) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St Paul, MN, 1991.

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Report of Leaf Blight on Washington Lupine (Lupinus polyphyllus) Caused by Rhizoctonia solani AG 4 in Italy

Plant Disease ◽

10.1094/pdis-93-4-0429a ◽

2009 ◽

Vol 93 (4) ◽

pp. 429-429

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

Lupinus Polyphyllus ◽

Leaf Petiole ◽

Sphagnum Peat

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.

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First Report of Leaf Blight on Fan Columbine (Aquilegia flabellata) Caused by Rhizoctonia solani AG 4 in Italy

Plant Disease ◽

10.1094/pdis-93-4-0433a ◽

2009 ◽

Vol 93 (4) ◽

pp. 433-433 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Rhizoctonia Solani ◽

Aqueous Suspension ◽

Morphological Characteristics ◽

The United States ◽

Leaf Blight ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Leaf Petiole

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.

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First Report of Leaf Blight Caused by Rhizoctonia solani AG 1B on Madagascar Periwinkle (Catharanthus roseus) in Italy

Plant Disease ◽

10.1094/pd-90-1361c ◽

2006 ◽

Vol 90 (10) ◽

pp. 1361-1361 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Catharanthus Roseus ◽

The United States ◽

Morphological Characters ◽

Leaf Blight ◽

Pathogenicity Test ◽

Madagascar Periwinkle ◽

Public Park ◽

First Report ◽

Leaf Petiole

Madagascar periwinkle (Catharanthus roseus), a plant belonging to the Apocynaceae family, is used for parks and gardens and sometimes grown in pots. At the end of the summer of 2005, a leaf blight was observed on plants in a public park of Torino. 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. Although lesions were not seen on the stem, affected plants often died leaving wide empty areas. Mycelia of the pathogen were often seen on and suspended between the leaves. Blight progressed from the leaves to the shoot tip. The diseased tissue was disinfected for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 100 μg/l streptomycin sulphate. A fungus with the morphological characters of Rhizoctonia solani was consistently and readily isolated and maintained in pure culture after single-hyphal tipping (4). The isolates of R. solani obtained from affected plants were successfully anastomosed with tester isolate AG 1 (ATCC 58946). The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. These results are consistent with other reports on anastomosis reactions (2). Pairing was also made with AG 2, 3, 4, 5, 7, and 11, with no anastomoses observed between the isolates and testers. Sclerotia were subspheroid in shape and had a size of 1 mm, which indicated that this pathogen was in subgroup 1B (4). For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 7 days. Plants of C. roseus were grown in 3-liter containers (2 plants per pot) on a steam disinfested substrate (peat/ pomix/pine bark/clay). Artificial inoculation was carried out on 7-day-old plants by placing numerous fragments of PDA cultures on the leaves of the plants. Plants inoculated with PDA alone served as control treatments. Three replicates were used. Plants were maintained in a glasshouse at 20 to 25°C. The first symptoms, similar to those observed in the public park, developed 5 days after inoculation, and R. solani was consistently reisolated from infected plants. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of R. solani on periwinkle in Italy. The same disease was reported in India (1) and the United States (3). References: (1) R. Balasubramanian and K. S. Bhama. Indian Phytopathol. 30:556, 1977. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. Pages 37–47 in: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, the Netherlands, 1996. (3) A. K. Hagan and J. M. Mullen Plant Dis. 77:1169, 1993. (4) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

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First Report of Web Blight on Oregano (Origanum vulgare L.) Caused by Rhizoctonia solani AG-1-IB in Italy

Plant Disease ◽

10.1094/pdis-02-13-0138-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1119-1119 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Its Region ◽

Morphological Characters ◽

Anastomosis Group ◽

Morphological Identification ◽

Pathogenicity Test ◽

Origanum Vulgare ◽

First Report ◽

Plastic Bags ◽

Hyphal Diameter

Origanum vulgare L., common name oregano, family Labiatae, is grown for its aromatic and medicinal properties and as ornamental. In the fall of 2012, a blight was observed in a farm located near Albenga (northern Italy) on 6% of 30,000 50-day-old plants, grown in trays in a peat/perlite mix. Semicircular, water soaked lesions appeared on leaves and stems, starting from the basal ones. As the disease progressed, blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Eventually, infected plants died. Leaf and stem fragments taken from the margin of the diseased tissues belonging to 10 plants were disinfected for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA). A fungus with the morphological characters of Rhizoctonia solani was consistently recovered. 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 tester strain. The 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 oregano 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 conducted twice. Mycelium of 10-day-old isolates from oregano appeared reddish brown, coarse, and radiate. Numerous dark brown sclerotia, 0.3 to 1.0 mm diameter (average 0.7) developed within 10 days after transfer of mycelia to PDA in 90 mm diameter petri dishes at 21 to 24°C. The descriptions of mycelium and sclerotia were typical for subgroup IB Type 1 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis (1) of the 538 bp showed a 99% homology with the sequence of R. solani FJ746937, confirming the morphological identification of the species. The nucleotide sequence has been assigned the GenBank Accession KC493638. For pathogenicity tests, one of the isolates assigned to the anastomosis group AG-1-IB was tested by placing 9 mm diameter mycelial disks removed from PDA 10-day-old cultures of the fungus on leaves of 90-day-old oregano plants (n = 35). Thirty-five plants inoculated with non-inoculated PDA disks served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 25 ± 1°C with 12 h light/dark. The first symptoms, similar to those observed in the farm, developed 3 days after inoculation. Nine days after the artificial inoculation, 50% of plants were dead. About 10 colonies of R. solani were reisolated from infected leaves of inoculated plants. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. Symptoms caused by R. solani have been recently observed on O. vulgare in Greece (3). This is, to our knowledge, the first report of blight of O. vulgare caused by R. solani in Italy. 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, pp. 37-47, 1996. (3) C. D. Holevas et al. Benaki Phytopathol. Inst., Kiphissia, Athens, 19:1-96, 2000. (4) R. T. Sherwood. Phytopathology 59:1924, 1969.

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First Report of Web Blight on Rosemary (Rosmarinus officinalis) Caused by Rhizoctonia solani AG-1-IA in Italy

Plant Disease ◽

10.1094/pdis-11-12-1012-pdn ◽

2013 ◽

Vol 97 (6) ◽

pp. 844-844 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Ground Cover ◽

Its Region ◽

Morphological Characters ◽

Anastomosis Group ◽

Rosmarinus Officinalis ◽

Economic Losses ◽

Pathogenicity Test ◽

First Report ◽

Wheat Kernels

Rosmarinus officinalis L., family Labiatae, is an evergreen shrub used in gardens as an aromatic or ground cover plant. In the summer of 2012, a blight was observed in a farm located near Albenga (northern Italy) on 20% of 150,000 70-day-old plants, grown in trays. Water soaked lesions appeared on leaves and stems. As the disease progressed, blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. A light mycelium spread on the substrate. Disease progressed from infected plants to healthy ones and, eventually, infected plants died. Leaf and stem fragments taken from the margin of the diseased tissues belonging to 10 plants were disinfected for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA). A fungus with the morphological characters of Rhizoctonia solani was consistently and readily recovered. Three isolates of R. solani obtained from affected plants were successfully paired with R. solani tester strains AG 1, 2, 3, 4, 6, 7, or 11 and examined microscopically. Three pairings were made for each recovered isolate. The isolates of R. solani from rosemary anastomosed only with tester strain AG 1 (ATCC 58946). Results were consistent with other reports on anastomosis reactions (2). Tests were repeated once. Mycelium of 10-day-old isolates from rosemary appeared light brown, compact, and radiate. Numerous dark brown sclerotia, 0.7 to 2.0 mm diameter (average 1.3), developed within 10 days at 20 to 26°C. The descriptions of mycelium and sclerotia were typical for subgroup IA Type 2 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced (GenBank Accession No. KC005724). BLASTn analysis (1) of the 657-bp showed a 99% similarity with the sequence of R. solani GU596491. For pathogenicity tests, inoculum of R. solani was prepared by growing the pathogen on wheat kernels autoclaved in 1-liter glass flasks for 8 days. One of the isolates assigned to the anastomosis group AG 1 IA was tested. Fifteen 90-day-old rosemary plants were grown in 15-liter pots in a steam disinfested peat:pomice:pine bark:clay mix (50:20:20:10) infested with 3 g/liter of infested wheat kernels, placed at the base of the stem. Fifteen plants inoculated with non-infested wheat kernels served as control treatments. Plants were covered with plastic bags and arranged in a growth chamber at 20 to 24°C with 12 h light/dark for 15 days. The first symptoms, similar to those observed in the farm, developed 10 days after inoculation. About 10 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 with similar results. Symptoms caused by R. solani have been recently observed on R. officinalis in United States (3), India, and Brazil. This is, to our knowledge, the first report of blight of R. officinalis caused by R. solani in Italy. This disease could cause serious economic losses, because rosemary is one of the most cultivated aromatic plants in the Mediterranean region. 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) G. E. Holcomb. Plant Dis. 76:859, 1992. (4) R. T. Sherwood. Phytopathology 59:1924, 1969.

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First Report of Gray Mold Caused by Botrytis cinerea on Stevia rebaudiana in Italy

Plant Disease ◽

10.1094/pdis-93-3-0318a ◽

2009 ◽

Vol 93 (3) ◽

pp. 318-318

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

M. L. Gullino

Keyword(s):

Botrytis Cinerea ◽

Stevia Rebaudiana ◽

Its Region ◽

Morphological Characters ◽

Gray Mold ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Leaf Extracts ◽

First Report ◽

Potted Plants

Stevia rebaudiana (sweetleaf) is a perennial shrub belonging to the Asteraceae family and is widely grown for its sweet leaves. With its extracts having as much as 300 times the sweetness of sugar, this species is used in many countries for the production of sugar substitutes. However, in Italy, as well as in other countries, this species cannot be grown for the use of its leaf extracts. This plant is grown in a few nurseries in the Albenga Region (northern Italy) as potted plants. In February of 2008, 3-month-old plants grown in plastic pots (14-cm diameter) under glasshouse on heated benches started showing symptoms of a previously unknown blight. The temperature in the glasshouse ranged between 16 and 20°C and plants were watered by sprinkle irrigation. Leaves, starting from the basal ones, showed small, brown spots that spread across the entire leaf surface. Subsequently, the crown and stem were infected, and the pathogen developed abundant, soft, gray mycelium on leaves and stems and in the middle of the heads of S. rebaudiana. Flowers were not present when the symptoms appeared. Severely infected leaves dried out and became necrotic. The disease was observed in one nursery in which 5% of the plants were affected. The margins of the lesions were excised from leaves, immersed in a solution containing 1% sodium hypochlorite, and then cultured on potato dextrose agar (PDA) medium. A fungus produced abundant mycelium when incubated under constant fluorescent light at 22 ± 1°C after 10 days. The conidia were smooth, hyaline, ovoid, measuring 15.5 to 8.3 × 11.1 to 7.3 (average 11.6 × 8.6) μm, and were similar to those described for Botrytis cinerea. Conidiophores were slender and branched with enlarged apical cells bearing conidia on short sterigmata. The identity of the fungus was also confirmed by the production of numerous, small, black sclerotia on PDA plates incubated for 20 days at 8 ± 1°C. Sclerotia were dark and irregular with a diameter ranging from 1 to 2 mm. These morphological characters identified the fungus as B. cinerea (2). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 780-bp segment showed a 100% homology with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). The nucleotide sequence has been assigned GenBank Accession No. FJ486270. Pathogenicity tests were performed by spraying leaves of six healthy 6-month-old potted S. rebaudiana plants with a 105 conidia/ml suspension. Six plants sprayed with water only served as controls. Plants were covered with plastic bags for 3 days after inoculation to maintain high relative humidity and were placed in a growth chamber at 20 ± 1°C. The first foliar lesions developed on leaves 4 days after inoculation, whereas control plants remained healthy. B. cinerea was consistently reisolated from these lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of B. cinerea on S. rebaudiana in Italy. The disease has been reported in Ukraine (3) and more recently in Japan (4). The economic importance of this disease is at the moment limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) H. L. Barnett and B. B. Hunter. Illustrated Genera of Imperfect Fungi. Burgess Publishing Company, Minneapolis, MN, 1972. (3) J. Takeuch and H. Horie. Annu. Rep. Kanto-Tosan Plant Prot. Soc. 53:87, 2006. (4) V. F. Zubenko et al. Zash. Rast. 18, 1991.

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First Report of Choanephora Rot Caused by Choanephora cucurbitarum on Hosta plantaginea in Korea

Plant Disease ◽

10.1094/pdis-07-14-0751-pdn ◽

2015 ◽

Vol 99 (1) ◽

pp. 158-158 ◽

Cited By ~ 1

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

B. S. Kim ◽

H. D. Shin

Keyword(s):

Large Subunit ◽

Its Region ◽

Disease Suppression ◽

Representative Specimen ◽

Pathogenicity Test ◽

First Report ◽

Perennial Plant ◽

Potted Plants ◽

Pcr Products ◽

Choanephora Cucurbitarum

Hosta plantaginea (Lam.) Asch. is an herbaceous perennial plant with ornamental value. In August 2013, water-soaked spots and wet rot were found on flowers of H. plantaginea in a garden bedded out for landscaping in Hongcheon County, Korea. Symptoms initially appeared as water-soaked spots at the tips of flowers. Dark gray spots on flower petals often coalesced and led to rotting of flowers, with abundant sporulation. However, no symptoms were found on the leaves. Approximately 30% of the flowers were affected in the landscape bed. A fungal isolate was obtained by plating surface-disinfested diseased flower tissue on potato dextrose agar (PDA). Fungal colonies covering the plate (diam. 90 mm) in 48 h were white at first, with abundant aerial mycelia, but later turned pale yellow with abundant sporangiola. Sporangiophores bearing sporangiola were aseptate, hyaline, and usually arose from infected tissue. Sporangiola were ellipsoid to ovoid, indehiscent, brown to dark brown, pediculate, 7 to 12 μm wide and 9 to 20 μm high, and showed longitudinal striations at high magnification. Sporangia were few-spored to multispored, pale brown to brown, and 50 to 150 μm. Sporangiospores from sporangia were broadly ellipsoid, brown to pale brown, with hyaline polar appendages, 8 to 10 μm wide and 15 to 22 μm high. Zygospores were not observed. The morphological and cultural characteristics, especially based on shape and striation of sporangiola, were identical with those of Choanephora cucurbitarum (Berk. & Ravenel) Thaxt. (2,3). A representative specimen was deposited in the Korea University Herbarium (KUS-F27540). Genomic DNA was extracted using a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). The primers ITS1/ITS4 and NL1/LR3 were used to amplify the internal transcribed spacer (ITS) region of rDNA and the D1/D2 region of the large subunit (LSU), respectively (4). The PCR products were purified and directly sequenced. The resulting 594-bp ITS and 680-bp D1/D2 sequences were submitted to GenBank (Accession Nos. KM200034 and KM200035). A GenBank BLAST search of the fungal database showed that the sequences of ITS and D1/D2 regions matched those of C. cucurbitrarum (JN943006 and JN939195) with 100% similarity. A pathogenicity test was conducted by spraying three healthy potted plants (2 months old) with a sporangiola suspension (2 × 104 conidia/ml). Another three potted plants of the same age were treated with sterile water and served as controls. The plants were kept in humid chambers for 2 days and placed in a greenhouse (28°C and 60 to 80% RH). After 4 to 5 days, water-soaked spots were evident on the flowers of inoculated plants. No symptoms were observed on control plants. A pathogenicity test was conducted twice with the same results, fulfilling Koch's postulates. C. cucurbitarum has a wide host range but has not been previously reported to cause disease on H. plantaginea (1). To our knowledge, this is the first report of C. cucurbitarum on H. plantaginea globally as well as in Korea. Choanephora rot of flowers is an issue under high-moisture conditions, so allowing for adequate airflow and a dry plant canopy should aid in disease suppression. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab. Online publication, ARS, USDA, retrieved July 11, 2014. (2) P. M. Kirk. Mycol. Pap. 152:1, 1984. (3) A. Saroj et al. Plant Dis. 96:293, 2012. (4) G. Walther et al. Persoonia 30:11, 2013.

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First Report of Leaf Blight on Foxglove (Digitalis purpurea) Caused by Rhizoctonia solani AG-1-IA in Italy

Plant Disease ◽

10.1094/pdis-93-3-0318c ◽

2009 ◽

Vol 93 (3) ◽

pp. 318-318

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Morphological Characteristics ◽

Its Region ◽

Leaf Blight ◽

Pathogenicity Test ◽

Uniform Size ◽

First Report ◽

Digitalis Purpurea ◽

Blastn Analysis ◽

Hyphal Diameter

Digitalis purpurea (Scrophulariaceae), foxglove, is used in flower gardens. In the spring of 2008, leaf blight was observed in a nursery near Biella (northern Italy) on 30% of potted 30-day-old plants grown in a peat substrate at temperatures from 20 to 25°C and relative humidity at 75 to 80%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the blade-petiole junction and later along the leaf margins. Lesions expanded for several days along the midvein until the entire leaf was affected. Blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily recovered, then transferred and maintained in pure culture (4). The isolates of R. solani obtained from affected plants successfully anastomosed with tester isolate AG 1 (ATCC 58946). 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 (2). Pairings were also made with tester isolates AG 2, 3, 4, 6, 7, 11, and AG BI and anastomosis was not observed. Ten-day-old colonies grown on PDA appeared light brown, rather compact, and radial. Numerous sclerotia of uniform size (0.5 to 3 mm in diameter) and sometimes joined laterally were formed. Descriptions of mycelium and sclerotia were typical for subgroup IA Type 2 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 724-bp fragment showed a 99% homology with the sequence of R. solani (GenBank Accession No. EU591800). The nucleotide sequence has been assigned GenBank Accession No. FJ467490. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Plants of 30-day-old D. purpurea were grown in 10-liter containers (6 plants per container) in a steam disinfested peat/clay/perlite (70:20:10) substrate. Disks of PDA cultures were placed on leaves (1 cm2 of mycelium per plant). Plants inoculated with PDA alone served as control treatments. Three replicates were used. Plants were maintained in a growth chamber at 24 ± 1°C with 12 h light/dark. First symptoms developed 12 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. R. solani was isolated from a small percentage of infected seeds of D. purpurea in India (3). This is, to our knowledge, the first report of leaf blight of D. purpurea caused by R. solani in Italy as well as in Europe. The spread of R. solani in nurseries might cause a decrease in trade. 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) K. K. Janardhanan and D. Ganguly. Indian Phytopathol. 16:379, 1963. (4) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St Paul, MN, 1991.

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