scholarly journals First Report of Rhizoctonia solani AG 4 on Lamb's Lettuce in Italy

Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1109-1109 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce or corn salad (Valerianella olitoria) is increasingly grown in Italy and used primarily in the preparation of mixed processed salad. In the fall of 2005, plants of lamb's lettuce, cv Trophy, exhibiting a basal rot were observed in some commercial greenhouses near Bergamo in northern Italy. The crown of diseased plants showed extensive necrosis, progressing to the basal leaves, with plants eventually dying. The first symptoms, consisting of water-soaked zonate lesions on basal leaves, were observed on 30-day-old plants during the month of October when temperatures ranged between 15 and 22°C. Disease was uniformly distributed in the greenhouses, progressed rapidly in circles, and 50% of the plants were affected. Diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 μg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily isolated and maintained in pure culture after single-hyphal tipping (3). The five isolates of R. solani, obtained from affected plants successfully anastomosed with tester isolate AG 4, no. RT 31, received from R. Nicoletti of the Istituto Sperimentale per il Tabacco, Scafati, Italy (2). The hyphal diameter at the point of anastomosis was reduced, and cell death of adjacent cells occurred (1). Pairings were also made with AG 1, 2, 3, 5, 7, and 11 with no anastomoses observed between the five isolates and testers. For pathogenicity tests, the inoculum of R. solani (no. Rh. Vale 1) was grown on autoclaved wheat kernels at 25°C for 10 days. Plants of cv. Trophy were grown in 10-liter containers (20 × 50 cm, 15 plants per container) on a steam disinfested substrate (equal volume of peat and sand). Inoculations were made on 20-day-old plants by placing 2 g of infected wheat kernels at each corner of the container with 3 cm as the distance to the nearest plant. Plants inoculated with clean wheat kernels served as controls. Three replicates (containers) were used. Plants were maintained at 25°C in a growth chamber programmed for 12 h of irradiation at a relative humidity of 80%. The first symptoms, consisting of water-soaked lesions on the basal leaves, developed 5 days after inoculation with crown rot and plant kill in 2 weeks. Control plants remained healthy. R. solani was consistently reisolated from infected plants. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of R. solani on lamb's lettuce in Italy as well as worldwide. The isolates were deposited at the AGROINNOVA fungal collection. The disease continues to spread in other greenhouses in northern Italy. References: (1) D. Carling. Rhizoctonia Species: Pages 37–47 in: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, the Netherlands, 1996. (2) J. Parmeter et al. Phytopathology, 59:1270, 1969. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN, 1996.

Plant Disease ◽  
2007 ◽  
Vol 91 (9) ◽  
pp. 1206-1206
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

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.


Plant Disease ◽  
2011 ◽  
Vol 95 (10) ◽  
pp. 1317-1317
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
A. Poli ◽  
M. L. Gullino

Cucumis melo L., belonging to the Cucurbitaceae family, is cultivated on more than 23,000 ha in Italy. Cantaloupe (C. melo L. var. cantalupensis Naudin) is the most popular variety. In summer 2010, a previously unknown rot was observed on fruits produced in Italy and marketed in the Piedmont Region of northern Italy. Early symptoms on fruit consisted of irregular, brown, soft, sunken lesions up to 10 cm in diameter. No surface mold was visible and pycnidia were not present. Internally, the decay is adjacent to the sunken area of the fruit's surface and is soft, water soaked, spongy, with a nearly circular margin, and easily separated from healthy tissues. Fragments (approximately 3 mm3) were taken from the margin of the internal diseased tissues, cultured on potato dextrose agar (PDA), and incubated at 24 ± 1°C, (16 h of light and 8 h of darkness). Fungal colonies initially appeared coarse, at first whitish then buff brown, and produced dark pycnidia 0.5 mm in diameter, which exuded numerous conidia belonging to two types. Type A conidia were hyaline, unicellular, ellipsoidal to fusiform, sometimes slightly constricted in the middle, and measured 5.6 to 10.3 × 1.7 to 2.6 (average 8.0 × 2.1) μm. Type B conidia were hyaline, long, slender, curved, and measured 17.1 to 26.6 × 0.7 to 1.4 (average 22.0 × 1.0) μm. Sclerotia were not produced. The morphological characteristics of the fungus corresponded to those of the genus Phomopsis (1). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis of the 543-bp segment showed a 99% similarity with the sequence of a Phomopsis sp. (GenBank Accession No. HM999947). The nucleotide sequence has been assigned the GenBank Accession No. JN032733. Both Phomopsis cucurbitae and P. sclerotioides are pathogenic to Cucurbitaceae, however P. cucurbitae is identifiable by the production of B conidia and the absence of sclerotia. Therefore, P. cucurbitae has been considered the causal agent of the disease. Pathogenicity tests were performed by inoculating three wounded cantaloupe fruits after surface disinfesting in 1% sodium hypochlorite. Six wounds per fruit, 1 cm deep, were made with a sterile needle. Mycelial disks (10 mm in diameter), obtained from PDA cultures of one strain, were placed on each wound. Three control fruits were inoculated with PDA. Fruits were incubated at 16 ± 1°C in the dark. The first symptoms developed 4 days after the artificial inoculation. Two days later, the rot developed at all inoculation points and the pathogen was consistently reisolated. Noninoculated fruit remained healthy. The pathogenicity test was performed twice with similar results. P. cucurbitae has been reported on melon in many countries (2,3). To our knowledge, this is the first report of the disease in Italy. Currently, the relevance of the disease in the country is not yet well known. However, attention must be paid considering that the pathogen can be transmitted through seeds. References: (1) H. L. Barnett and B. B. Hunter. Illustrated Genera of Imperfect Fungi. Burgess Publishing Company, Minneapolis, MN, 1972. (2) L. Beraha and M. J. O'Brien. Phytopathol. Z. 94:199, 1979. (3) E. Punithalingam and P. Holliday. Phomopsis cucurbitae. IMI Descriptions of Fungi and Bacteria. 47, Sheet 469, 1975.


Plant Disease ◽  
2005 ◽  
Vol 89 (2) ◽  
pp. 205-205 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce (Valerianella olitoria) is increasingly grown in Italy and used in the preparation of processed salad. In the fall of 2003, plants of lamb's lettuce cvs. Trophy and Palmares showing symptoms of severe stunting were observed in several commercial plastic greenhouses near Bergamo in northern Italy. The distribution of the disease was generally uniform in the greenhouses and 10 to 30% of the plants were affected. The first symptoms, consisting of reduced growth of the plants and extensive chlorosis, developed in October at temperatures ranging between 10 and 20°C on 30-day-old plants. The roots of diseased plants showed extensive necrosis that extended to the crown of the plants. The diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 µg/l streptomycin sulfate. A fungus with the morphological characteristics of Thielaviopsis basicola was consistently and readily isolated from symptomatic tissues (1). Catenulate, cylindrical, hyaline endoconidia and catenulate, subrectangular, thick-walled chlamydospores (aleuriospores) were observed. Ten-day-old plants of cvs. Trophy and Palmares were artificially inoculated by dipping three isolates of T. basicola obtained from infected plants for 15 min in a spore suspension (1 × 106 CFU/ml). Noninoculated plants served as control treatments. Each treatment consisted of 30 plants. Plants were maintained at 20°C in a growth chamber, with 12 h of light/day. Symptoms developed 25 days after the artificial inoculation on both cultivars, and T. basicola was consistently reisolated from diseased plants. The noninoculated plants remained healthy. The pathogenicity test was carried out twice. To our knowledge, this is the first report of T. basicola on lamb's lettuce in Italy as well as in the world. The same disease was described on corn salad (Valerianella locusta) in the United States (2). References: (1) D. E. Mathre and A.V. Ravenscroft. Phytopathology 56:337, 1966. (2) M. E. Stanghellini et al. Plant Dis. 74:81, 1990.


Plant Disease ◽  
2010 ◽  
Vol 94 (3) ◽  
pp. 382-382 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Fuchsia × hybrida (Onagraceae) is widely used in gardens and very much appreciated as a potted plant. During the summer of 2008, a severe foliar disease was observed on 1- to 2-year-old plants in several gardens located near Biella (northern Italy). Small necrotic spots were observed on the upper and lower sides of infected leaves. Spots enlarged to form round areas of 2 to 12 mm in diameter and were well defined by a brown-purple margin at temperatures between 15 and 25°C. Severely infected leaves wilted and abscised as disease progressed. The disease occurred on 100% of the plants and at least 30% of the leaf surface was affected. Stems and flowers were not affected by the disease. A fungus was consistently isolated from infected leaves on potato dextrose agar amended with 25 mg/liter of streptomycin. The fungus was grown on leaf extract agar, including 30 g of autoclaved fuchsia leaves per liter, and maintained at 22°C (12-h light, 12-h dark). After 30 days, black pycnidia 150 to 450 μm in diameter developed, releasing abundant hyaline, elliptical, nonseptate conidia measuring 5.6 to 14.3 (10.3) × 1.9 to 5.6 (3.5) μm. On the basis of these morphological characteristics, the fungus was identified as a Phoma sp. (2). The internal transcribed spacer (ITS) region of rDNA of the isolate coded FuHy1 was amplified using primers ITS4/ITS6 (3) and sequenced. BLAST analysis (1) of the 488-bp segment obtained showed an E-value of 0.0 with Phoma multirostrata. The nucleotide sequence has been assigned GenBank Accession No. GU220539. Pathogenicity tests were performed by spraying leaves of healthy 6-month-old potted Fuchsia × hybrida plants with a spore and mycelial suspension (1 × 106 spores or mycelial fragments per milliliter). Noninoculated plants sprayed with water served as controls. Five plants were used for each treatment. Plants were covered with plastic bags for 5 days after inoculation and kept under greenhouse conditions at 20 to 24°C. Symptoms previously described developed on leaves 12 days after inoculation, whereas control plants remained healthy. The fungus was consistently reisolated from the lesions of the inoculated plants. The pathogenicity test was carried out twice. To our knowledge, this is the first report of the presence of P. multirostrata on fuchsia in Italy as well as worldwide. The importance of the disease is still limited in Italy. References: (1) S. F. Altschud et al. Nucleic Acids Res. 25:3389, 1997. (2) G. H. Boerema and G. J. Bollen. Persoonia 8:111, 1975. (3) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997.


Plant Disease ◽  
2011 ◽  
Vol 95 (10) ◽  
pp. 1316-1316 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
C. Bertoldo ◽  
M. L. Gullino

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.


Plant Disease ◽  
2009 ◽  
Vol 93 (3) ◽  
pp. 318-318
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

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.


Plant Disease ◽  
2012 ◽  
Vol 96 (4) ◽  
pp. 585-585 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
P. Pensa ◽  
M. T. Amatulli ◽  
M. L. Gullino

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.


Plant Disease ◽  
2008 ◽  
Vol 92 (5) ◽  
pp. 836-836 ◽  
Author(s):  
D. Aiello ◽  
G. Parlavecchio ◽  
A. Vitale ◽  
E. Lahoz ◽  
R. Nicoletti ◽  
...  

Lagunaria patersonii (Adr.) G. Don (cow itch tree) is native to Australia and tolerates salted winds. During July 2007, damping-off of cow itch tree was observed on 4-month-old seedlings growing in a commercial nursery in eastern Sicily, Italy. More than 20% of the seedlings showed disease symptoms. First symptoms consisting of water-soaked lesions at the seedling base that expand rapidly girdle the stem and collapse the seedling in a few days. Diseased tissues were disinfested for 1 min in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with streptomycin sulphate at 100 mg/l, and then incubated at 25°C. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently yielded. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Microscopic examination revealed that hyphae had a right-angle branching pattern, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. Basidia were not observed in the greenhouses or on the plates. Hyphal cells were determined to be multinucleate when stained with 0.5% aniline blue solution and examined at ×400 magnification with a microscope. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (3). Pairings were made with tester strains of AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, AG-11. Anastomosis was observed only with tester isolates of AG-4 producing both C2 and C3 reactions. 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 were consistent with other reports on anastomosis reactions (1). The identification of group AG-4 within R. solani has been confirmed by electrophoretic patterns of pectic enzymes (polygalacturonases) in vertical pectin-acrylamide gel stained with ruthenium red (2). Pathogenicity tests were conducted on potted, healthy, 3-month-old seedlings of cow itch tree. Twenty plants were inoculated by placing plugs of PDA from 5-day-old mycelial cultures near the base of the stem. The same number of plants was treated with 1 cm2 PDA plugs as control. Plants were kept at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Wilt symptoms due to basal stem rot, identical to ones observed in the nursery, appeared 10 days after inoculation and all inoculated plants showed symptoms within 1 month. Control plants remained healthy. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report in the world of R. solani causing disease on L. patersonii. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (2) R. H. Cruickshank and G. C. Wade. Anal. Biochem. 107:177, 1980. (3) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.


Plant Disease ◽  
2006 ◽  
Vol 90 (9) ◽  
pp. 1260-1260 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
D. Minerdi ◽  
M. L. Gullino

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.


Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 433-433 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

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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