First Report of Dollar Spot of Buffalograss Caused by Sclerotinia homoeocarpa in Oklahoma

Buffalograss (Buchloe dactyloides (Nutt.) Engelm.) is a perennial, warm-season grass native to the central plains of North America and a dominant plant over much of the shortgrass prairie ecosystem. Its prostrate growth habit and excellent drought tolerance make it a commercially promising turfgrass species, and numerous turf-type cultivars have been released. In the spring of 2007, the southern plains states experienced prolonged periods of excessive precipitation during which numerous buffalograss swards throughout north-central Oklahoma exhibited symptoms of dollar spot (1). A fungus morphologically identical to Sclerotinia homoeocarpa Bennett was consistently isolated from diseased buffalograss leaves collected from three locations in Oklahoma, two from Payne County and one from Logan County. Thirty-day-old seedlings of B. dactyloides (‘Cody’ and ‘Topgun’) and Agrostis stolonifera (‘SR1020’) were inoculated by placing potato dextrose agar (PDA) plugs, colonized by mycelia of each S. homoeocarpa isolate, onto the seedlings' leaves. Sterile PDA plugs were placed on plants as controls. Leaf lesions developed after 4 days only on inoculated plants, and S. homoeocarpa was reisolated from lesions, satisfying Koch's postulates. The nuclear ribosomal internal transcribed spacer (ITS) region was amplified from DNA extracted from cultures of the three buffalograss isolates and a bentgrass isolate using primers ITS4 and ITS5 (2) and sequenced. Sequences were similar to one another (97 to 99% identical), however, two isolates shared a 420-bp, type I intron in the 18S small subunit rDNA. A search of GenBank at NCBI found the ITS sequences were most similar to the ITS regions of other S. homoeocarpa accessions (97% identical). The ITS sequences from the four isolates were deposited in GenBank (Accession Nos. EU123800–EU123803). To our knowledge, this is the first report of dollar spot on a native, warm-season grass in the United States and the disease appears to be endemic to buffalograss in Oklahoma and Kansas (N. A. Tisserat, personal communication). References: (1) R. W. Smiley et al. Page 22 in: Compendium of Turfgrass Diseases. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2005. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press Inc., New York, 1990.

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First Report of Dollar Spot of Sandbur Caused by Sclerotinia homoeocarpa in Oklahoma

Plant Disease ◽

10.1094/pdis-03-14-0244-pdn ◽

2014 ◽

Vol 98 (8) ◽

pp. 1160-1160

Author(s):

F. Flores ◽

N. R. Walker

Keyword(s):

Warm Season ◽

Its Region ◽

Small Subunit ◽

Type I ◽

Academic Press ◽

Sclerotinia Homoeocarpa ◽

Dollar Spot ◽

First Report ◽

Small Subunit Rdna ◽

Disturbed Soils

Sandbur (Cenchrus incertus Curtis) is a warm-season, annual, noxious, grassy weed native to southern North America. It is common in sandy, disturbed soils and can also be found in home lawns and sport fields where low turf density facilitates its establishment. In July 2013, after a period of frequent rainfall and heavy dew, symptoms of dollar spot-like lesions (1) were observed on sandbur plants growing in a mixed stand of turf-type and native warm-season grasses in Logan County, Oklahoma. Lesions, frequently associated with leaf sheaths, were tan and surrounded by a dark margin. Symptomatic leaves were surface sterilized and plated on potato dextrose agar amended with 10 ppm rifampicin, 250 ppm ampicillin, and 5 ppm fenpropathrin. After incubation, a fungus morphologically identical to Sclerotinia homoeocarpa Bennett was consistently isolated. The nuclear ribosomal internal transcribed spacer (ITS) region of two different isolates, SCL2 and SCL3, were amplified using primers ITS4 and ITS5 (2). The DNA products were sequenced and BLAST analyses were used to compare sequences with those in GenBank. The sequence for isolate SLC2 was 869 bp, contained a type I intron in the 18S small subunit rDNA, and was identical to accession EU123803. The ITS sequence for isolate SLC3 was 535 bp and identical to accession EU123802. Twenty-five-day-old seedlings of C. incertus were inoculated by placing 5-mm-diameter agar plugs, colonized by mycelia of each S. homoeocarpa isolate, onto two of the plants' leaves. Plugs were held in place with Parafilm. Two plants were inoculated with each isolate and sterile agar plugs were placed on two leaves of another seedling as control. Plants were incubated in a dew chamber at 20°C and a 12-h photoperiod. After 3 days of incubation, water-soaked lesions surrounded by a dark margin appeared on inoculated plants only. Fungi that were later identified as S. homoeocarpa isolates SLC2 and SLC3 by sequencing of the ITS region were re-isolated from symptomatic leaves, fulfilling Koch's postulates. To our knowledge, this is the first report of dollar spot on sandbur. References: (1) R. W. Smiley et al. Page 22 in: Compendium of Turfgrass Diseases. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2005. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Dollar Spot Caused by Sclerotinia homoeocarpa on Switchgrass in the United States

Plant Disease ◽

10.1094/pdis-04-11-0332 ◽

2011 ◽

Vol 95 (12) ◽

pp. 1585-1585 ◽

Cited By ~ 2

Author(s):

A. L. Vu ◽

K. D. Gwinn ◽

B. H. Ownley

Keyword(s):

Panicum Virgatum ◽

Morphological Characteristics ◽

Leaf Tissue ◽

Leading Edge ◽

The United States ◽

Small Subunit ◽

Grass Species ◽

Sclerotinia Homoeocarpa ◽

Dollar Spot ◽

Leaf Spots

Sclerotinia homoeocarpa causes dollar spot on many grass species; however, it has not been described on switchgrass (Panicum virgatum L.) as a host. In August 2010, bleached, tan-to-straw-colored leaf spots with dark brown-to-reddish brown margins were found in patchy distribution in small field plots of ‘Alamo’ switchgrass at the East Tennessee Research and Education Center, Knoxville, TN. The plots had been planted to switchgrass for the past 21 years. Disease lesions covered 75 to 80% of leaf tissue per patch and were also evident on stems. To identify the pathogen, center portions of diseased leaves were cut into 20- to 30-cm segments, surface disinfested (95% ethanol for 30 s, 10% bleach for 1 min, and 95% ethanol for 30 s), and dried. Disinfested leaves (5-cm sections that included a leading edge of a lesion) were plated on potato dextrose agar (PDA). Plates were incubated at 22°C. Within 12 h, white, fluffy, aerial mycelium developed. Viewed from above, colonies were tan to cinnamon in color with a dark brown-to-black substratal stroma on and in the agar, which appeared brown as viewed from below the petri dish. No spores were observed. Morphological characteristics of colony and hyphal growth were identical to those of S. homoeocarpa F.T. Bennett (1). Pathogenicity studies were conducted with 6-week-old ‘Alamo’ switchgrass grown from scarified (2), surface-disinfested seed. Nine (9 × 9-cm2) pots with 18 plants each were inoculated with 20 mycelial plugs (6-mm diameter) per pot, taken from 3-to-5-day-old fungal cultures. Two control pots were inoculated with sterile PDA plugs and subjected to the same conditions. Plugs were placed on leaf surfaces and around the plant crowns. Plants were subjected to high humidity by enclosure in a plastic bag and incubated in a growth chamber at 25/20°C with a 12-h photoperiod. Plastic bags were removed after 48 h. Leaf spots appeared as early as 2 days postinoculation, with full symptoms after 2 weeks for eight of nine replicates. Control plants had no symptoms. The fungus was cultured from leaf spots and stem lesions of inoculated plants as described above. The same disease and fungus were observed, completing Koch's postulates. The internal transcribed spacer (ITS) regions of ribosomal DNA from the original isolate used for inoculation and from the isolate recovered from plants in the pathogenicity assay were amplified with PCR with primers ITS4 and ITS5 (4). PCR amplicons of ~565 bp were sequenced; sequences of amplicons from the original isolate and reisolate were identical and submitted to GenBank (Accession No. HQ850151). The sequence had 99% homology with several S. homoeocarpa isolates in GenBank, including three isolates from buffalograss in Oklahoma (Accession Nos. EU123800, EU123802, and EU123803). The mitochondrial small subunit region was amplified from the original isolate with primers NMS1 and NMS2 (3). The resultant 536-bp fragment was sequenced and submitted to GenBank (Accession No. HQ850152), but no S. homoeocarpa sequences were available for comparison. To our knowledge, this is the first confirmed report of switchgrass as a natural host for S. homoeocarpa, extending the known host range for the pathogen. References: (1) F. T. Bennett. Ann. Appl. Biol. 24:236, 1937. (2) K. D. Gwinn et al. Crop Sci. 31:1369, 1991. (3) K. N. Li et al. Appl. Environ. Microbiol. 60:4324, 1994. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, NY, 1990.

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First Report of Dollar Spot, Caused by Sclerotinia homoeocarpa, of Creeping Bentgrass in Norway

Plant Disease ◽

10.1094/pdis-07-14-0775-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 287-287 ◽

Cited By ~ 4

Author(s):

T. Espevig ◽

M. B. Brurberg ◽

A. Kvalbein

Keyword(s):

Room Temperature ◽

Aerial Mycelium ◽

Its Region ◽

Creeping Bentgrass ◽

Grass Species ◽

Sclerotinia Homoeocarpa ◽

Dollar Spot ◽

Running Water ◽

First Report ◽

Plastic Bags

In September 2013, symptoms similar to dollar spot caused by Sclerotinia homoeocarpa F.T. Benn., were observed on creeping bentgrass (Agrostis stolonifera L.) fairways at Losby Golf Course, Lørenskog, Akershus County, in Norway (59.8864′ N, 10.9862′ E). There were small, circular spots and larger irregular patches of sunken, bleached, straw-colored turf. Affected leaves had light-tan lesions with light reddish-brown margins (2). Abundant aerial mycelium was observed in the diseased turf after incubation for 24 h at room temperature in a moist chamber. The mycelium was septate with y-shaped branches. No spores were observed. Diseased leaf segments were washed 30 min in cold running water, surface-sterilized for 60 s using 70% ethanol, placed on water agar, and incubated at room temperature. After 4 days, water agar plugs containing the fungus were transferred to 50% potato dextrose agar (PDA; 19.5 g PDA and 7.5 g agar per 1 liter of media). The fungus colonized the entire 9-cm PDA plates in 6 days. The diameter of the hyphae varied from 2.5 to 12.5 μm. The white, floccose mycelium turned olive green after 7 days and cinnamon brown after 21 days. The cultures became brown from the bottom, forming flat, dark-brown stroma of 0.5 to 5.0 mm in diameter. DNA was extracted from three isolates (from different plants) using a DNeasy Plant Mini Kit (Qiagen). The ribosomal internal transcribed spacer (ITS) region was PCR-amplified using primers ITS1 and ITS4 (3). All three isolates were identical in sequence (GenBank Accession No. KJ775860) and showed up to 97.6% similarity with isolates of S. homoeocarpa of the common type (C-type; e.g., GenBank Accession No. HQ449691) (1). This similarity is considered quite low within a species and indicates that the Norwegian isolates are distinct from other S. homoeocarpa. For Koch's postulates, the fungus was scraped off 21-day-old PDA cultures and chopped, using a sterile scalpel. All three sequenced isolates were pooled and mixed with 200 ml of autoclaved water. Four mature, healthy sod plugs of creeping bentgrass cv. Independence (10-cm-diameter and 10-cm-depth) were taken from an experimental golf green at Landvik, inserted into pots, and inoculated by even distribution of 50 ml of the fungal suspension. Two control pots with creeping bentgrass received 50 ml of sterile water only. All six pots were incubated individually in plastic bags at room temperature and 16-h daylight. After 14 days, 30 to 90% of the inoculated pots of turfgrass exhibited dollar spot symptoms and controls remained healthy. The fungus was recovered from inoculated turf and identified morphologically. This is the first report of dollar spot on any grass species in Norway. For climatic reasons, dollar spot has been considered to be nonexistent in Scandinavia. However, during recent years, symptoms resembling dollar spot have been observed on more than 15 golf courses in Sweden, Denmark, and Norway, and the damage has varied from low to severe. The disease has been given the Norwegian name myntflekk (i.e., coin spot). References: (1) D. Liberti et al. Phytopathology 102:506, 2012. (2) J. D. Smith et al. Fungal diseases of amenity turf grasses. E. & F. E. Spon, London. 1989. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.

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First Report of Botrytis Blight Caused by Botrytis cinerea on Periwinkle (Catharanthus roseus) in Italy

Plant Disease ◽

10.1094/pdis-93-5-0554b ◽

2009 ◽

Vol 93 (5) ◽

pp. 554-554 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

Botrytis Cinerea ◽

Catharanthus Roseus ◽

Warm Season ◽

Its Region ◽

The United States ◽

Plant Diseases ◽

Fluorescent Light ◽

Pathogenicity Test ◽

First Report ◽

Bedding Plant

Catharanthus roseus (periwinkle), a perennial plant belonging to the Apocynaceae family, is grown as a warm-season bedding plant in temperate gardens. This species is characterized by a long flowering period and prized for its white-to-dark pink flowers. In October of 2008, 15% of C. roseus plants in a public garden located in Torino (northern Italy) showed symptoms of a previously unknown blight. When the disease developed, temperatures ranged between 10 and 24°C (average 17.3°C) and plants were being watered through sprinkle irrigation. Necrosis developed on the stems first, eventually spreading to leaf stalks, and the the entire leaf. Subsequently, the pathogen developed a scant, delicate, gray mycelium on affected tissues, particularly diffused on the stems. Severely infected leaves and stems eventually became completely rotted and desiccated. Tissues were excised from diseased leaves, immersed in a solution containing 1% sodium hypochlorite for 10 s, and cultured on potato dextrose agar (PDA) medium. The fungus produced abundant mycelium on PDA medium when incubated under constant fluorescent light at 22 ± 1°C. Numerous sclerotia were produced on PDA plates incubated for 20 days at 8 ± 1°C. Sclerotia were dark and irregular, measuring 0.5 to 2.8 × 0.5 to 2.2 (average 1.4 × 1.1) mm. Conidia were smooth, ash colored, ovoid, measuring 8 to 16 × 6 to 10 (average 10 × 7) μm, and similar to those described for Botrytis cinerea (2). The internal transcribed spacer (ITS) region of rDNA was amplified with 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. FJ486271. Pathogenicity tests were performed by placing numerous fragments of PDA cultures on leaves of healthy, potted, 8-month-old C. roseus plants. Plants inoculated with PDA alone served as controls. Three plants per treatment were used. Plants were covered with plastic bags for 5 days after inoculation and maintained in a greenhouse at temperatures ranging between 18 and 25°C. The first foliar lesions developed on leaves 5 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 C. roseus in Italy. The same disease was previously reported in many countries including the United States (3) and Taiwan (4). 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) M. L. Daughtrey et al. Compendium of Flowering Potted Plant Diseases. The American Phytopathological Society, St Paul, MN, 1995. (4) W. Ou-Yang and W. S. Wu. Plant Pathol. Bull. 7:147, 1998.

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First Report of Leaf Blight and Stem Canker of Pachysandra terminalis Caused by Pseudonectria pachysandricola in Korea

Plant Disease ◽

10.1094/pdis-09-11-0813 ◽

2012 ◽

Vol 96 (2) ◽

pp. 287-287

Author(s):

K. S. Han ◽

J. H. Park ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

United States ◽

Its Region ◽

Stem Canker ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

First Report ◽

Cornell University ◽

Plastic Bags ◽

Young Leaves

Pachysandra terminalis Siebold & Zucc., known as Japanese pachysandra, is a creeping evergreen perennial belonging to the family Buxaceae. In April 2011, hundreds of plants showing symptoms of leaf blight and stem canker with nearly 100% incidence were found in a private garden in Suwon, Korea. Plants with the same symptoms were found in Seoul in May and Hongcheon in August. Affected leaves contained tan-to-yellow brown blotches. Stem and stolon cankers first appeared as water soaked and developed into necrotic lesions. Sporodochia were solitary, erumpent, circular, 50 to 150 μm in diameter, salmon-colored, pink-orange when wet, and with or without setae. Setae were hyaline, acicular, 60 to 100 μm long, and had a base that was 4 to 6 μm wide. Conidiophores were in a dense fascicle, not branched, hyaline, aseptate or uniseptate, and 8 to 20 × 2 to 3.5 μm. Conidia were long, ellipsoid to cylindric, fusiform, rounded at the apex, subtruncate at the base, straight to slightly bent, guttulate, hyaline, aseptate, 11 to 26 × 2.5 to 4.0 μm. A single-conidial isolate formed cream-colored colonies that turned into salmon-colored colonies on potato dextrose agar (PDA). Morphological and cultural characteristics of the fungus were consistent with previous reports of Pseudonectria pachysandricola B.O. Dodge (1,3,4). Voucher specimens were housed at Korea University (KUS). Two isolates, KACC46110 (ex KUS-F25663) and KACC46111 (ex KUS-F25683), were accessioned in the Korean Agricultural Culture Collection. Fungal DNA was extracted with DNeasy Plant Mini DNA Extraction Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced using ABI Prism 337 automatic DNA sequencer (Applied Biosystems, Foster, CA). The resulting sequence of 487 bp was deposited in GenBank (Accession No. JN797821). This showed 100% similarity with a sequence of P. pachysandricola from the United States (HQ897807). Isolate KACC46110 was used in pathogenicity tests. Inoculum was prepared by harvesting conidia from 2-week-old cultures on PDA. Ten young leaves wounded with needles were sprayed with conidial suspensions (~1 × 106 conidia/ml). Ten young leaves that served as the control were treated with sterile distilled water. Plants were covered with plastic bags to maintain a relative humidity of 100% at 25 ± 2°C for 24 h. Typical symptoms of brown spots appeared on the inoculated leaves 4 days after inoculation and were identical to the ones observed in the field. P. pachysandricola was reisolated from 10 symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in the United States, Britain, Japan, and the Czech Republic (2,3), but not in Korea. To our knowledge, this is the first report of P. pachysandricola on Pachysandra terminalis in Korea. Since this plant is popular and widely planted in Korea, this disease could cause significant damage to nurseries and the landscape. References: (1) B. O. Dodge. Mycologia 36:532, 1944. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , September 24, 2011. (3) I. Safrankova. Plant Prot. Sci. 43:10, 2007. (4) W. A. Sinclair and H. H. Lyon. Disease of Trees and Shrubs. 2nd ed. Cornell University Press, Ithaca, NY, 2005.

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First Report of Anthracnose of Onion Caused by Colletotrichum coccodes in Ohio

Plant Disease ◽

10.1094/pdis-01-14-0032-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1271-1271 ◽

Cited By ~ 2

Author(s):

F. Baysal-Gurel ◽

N. Subedi ◽

D. P. Mamiro ◽

S. A. Miller

Keyword(s):

Leaf Tissue ◽

Its Region ◽

The United States ◽

Tween 20 ◽

Colletotrichum Coccodes ◽

Academic Press ◽

Conidial Suspension ◽

First Report ◽

Allium Cepa L ◽

Total Dna

Dry bulb onion (Allium cepa L. cvs. Pulsar, Bradley, and Livingston) plants with symptoms of anthracnose were observed in three commercial fields totaling 76.5 ha in Huron Co., Ohio, in July 2013. Symptoms were oval leaf lesions and yellowing, curling, twisting, chlorosis, and death of leaves. Nearly half of the plants in a 32.8-ha field of the cv. Pulsar were symptomatic. Concentric rings of acervuli with salmon-colored conidial masses were observed in the lesions. Conidia were straight with tapered ends and 16 to 23 × 3 to 6 μm (2). Colletotrichum coccodes (Wallr.) S. Hughes was regularly isolated from infected plants (2). Culturing diseased leaf tissue on potato dextrose agar (PDA) amended with 30 ppm rifampicin and 100 ppm ampicillin at room temperature yielded white aerial mycelia and salmon-colored conidial masses in acervuli. Numerous spherical, black microsclerotia were produced on the surface of colonies after 10 to 14 days. To confirm pathogen identity, total DNA was extracted directly from a 7-day-old culture of isolate SAM30-13 grown on PDA, using the Wizard SV Genomic DNA Purification System (Promega, Madison, WI) following the manufacturer's instructions. The ribosomal DNA internal transcribed spacer (ITS) region was amplified by PCR using the primer pair ITS1 and ITS4 (2), and sequenced. The sequence, deposited in GenBank (KF894404), was 99% identical to that of a C. coccodes isolate from Michigan (JQ682644) (1). Ten onion seedlings cv. Ebenezer White at the two- to three-leaf stage of growth were spray-inoculated with a conidial suspension (1 × 105 conidia/ml containing 0.01% Tween 20, with 10 ml applied/plant). Plants were maintained in a greenhouse (21 to 23°C) until symptoms appeared. Control plants were sprayed with sterilized water containing 0.01% Tween 20, and maintained in the same environment. After 30 days, sunken, oval lesions each with a salmon-colored center developed on the inoculated plants, and microscopic examination revealed the same pathogen morphology as the original isolates. C. coccodes was re-isolated consistently from leaf lesions. All non-inoculated control plants remained disease-free, and C. coccodes was not re-isolated from leaves of control plants. C. coccodes was reported infecting onions in the United States for the first time in Michigan in 2012 (1). This is the first report of anthracnose of onion caused by C. coccodes in Ohio. Unusually wet, warm conditions in Ohio in 2013 likely contributed to the outbreak of this disease. Timely fungicide applications will be necessary to manage this disease in affected areas. References: (1) A. K. Lees and A. J. Hilton. Plant Pathol. 52:3. 2003. (2) L. M. Rodriguez-Salamanca et al. Plant Dis. 96:769. 2012. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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Genetic Diversity of Sclerotinia homoeocarpa Isolates from Turfgrasses from Various Regions in North America

Plant Disease ◽

10.1094/pdis.2004.88.11.1269 ◽

2004 ◽

Vol 88 (11) ◽

pp. 1269-1276 ◽

Cited By ~ 19

Author(s):

G. Viji ◽

W. Uddin ◽

N. R. O'Neill ◽

S. Mischke ◽

J. A. Saunders

Keyword(s):

United States ◽

Genetic Diversity ◽

Creeping Bentgrass ◽

The United States ◽

Vegetative Compatibility ◽

Major Group ◽

Golf Courses ◽

Sclerotinia Homoeocarpa ◽

Dollar Spot ◽

Geographical Regions

Sixty-seven isolates of Sclerotinia homoeocarpa, causing dollar spot disease in creeping bentgrass, annual bluegrass, Bermudagrass, and perennial ryegrass turf, collected from 23 golf courses in various geographical regions of the United States and Canada between 1972 and 2001, were characterized by vegetative compatibility, genetic diversity, and pathogenicity. Eleven vegetative compatibility groups (VCGs A to K) were identified among the isolates tested in this study, and five of them (VCGs G to K) were new. VCG B was the most predominant group, typifying 33 isolates (51%) tested. S. homoeocarpa isolates collected from golf courses in Pennsylvania belonged to seven VCGs (A, B, E, F, G, I, and K), whereas three groups were observed in those collected from New York (B, E, and G) and New Jersey (E, H, and I). Two isolates, one each from Pennsylvania and Canada, were incompatible when paired with the tester isolates in all possible combinations, and did not fall into any known VCG. An isolate collected from Canada was compatible with tester isolates from two VCGs (C and D). Genetic analyses using amplified fragment length polymorphism (AFLP) showed the presence of two genetically distinct groups, designated as major group and the minor group. The major group included 36 isolates collected from various golf courses in the United States and Canada. Two isolates collected from bermudagrass in Florida formed a separate cluster, the minor group. Isolates that belonged to the major group were further divided into two subgroups (1 and 2). Subgroup 1 consisted of all the isolates that belonged to VCGs A, E, G, H, and I. Three of the four isolates that belonged to VCG K also were clustered with isolates of subgroup 1. Subgroup 2 consisted of all the isolates from VCG B, and one each from VCGs F and K. Pathogenicity assays on Penncross creeping bentgrass showed significant differences (P = 0.05) in virulence among the isolates. Overall, a relationship between virulence and VCGs was observed, in which certain virulence groups corresponded to specific VCGs; however, such a relationship was not observed between virulence and AFLPs. Close similarity among isolates of S. homoeocarpa collected from different locations in the United States and Canada suggests that isolates of the same genotype could be involved in outbreaks of dollar spot epidemics at multiple locations.

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First Report of Phytophthora citrophthora on Penstemon barbatus in Italy

Plant Disease ◽

10.1094/pd-90-1260a ◽

2006 ◽

Vol 90 (9) ◽

pp. 1260-1260 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

D. Minerdi ◽

M. L. Gullino

Keyword(s):

Its Region ◽

The United States ◽

Phytophthora Species ◽

Crown Rot ◽

Pathogenicity Test ◽

Phytophthora Citrophthora ◽

First Report ◽

Blastn Analysis ◽

Root And Crown Rot ◽

Pathogenicity Tests

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.

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First Report of Zonate Leaf Spot Caused by Hinomyces moricola on Japanese Hop in Korea

Plant Disease ◽

10.1094/pdis-01-13-0021-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1117-1117 ◽

Cited By ~ 1

Author(s):

S. E. Cho ◽

J. H. Park ◽

S. H. Hong ◽

H. D. Shin

Keyword(s):

Its Region ◽

The United States ◽

Infected Leaf ◽

Invasive Weed ◽

Voucher Specimen ◽

First Report ◽

Leaf Spots ◽

Initial Symptoms ◽

Wide Range ◽

Humulus Japonicus

Japanese hop (Humulus japonicus Siebold & Zucc. = H. scandens (Lour.) Merr.), native to East Asia, is an annual, climbing or trailing vine. The vines can spread to cover large areas of open ground or low vegetation, eventually blanketing the land and vegetation. Pollen of H. japonicus is allergenic, and this species is considered as one of the important causes of pollinosis in Korea and China. It is a notorious invasive weed in the United States and also in France, Hungary, and Italy (1). In September 2012, zonate leaf spots were observed on Japanese hops growing in wetlands in Yeongdong County of Korea. A voucher specimen was preserved in the Korea University Herbarium (KUS-F26901). Initial symptoms included grayish-green to grayish-brown spots without border lines. As the lesions enlarged, they coalesced, leading to leaf blight. Sporophores on the leaf lesions were dominantly hypophyllous, rarely epiphyllous, solitary, erect, easily detachable, and as long as 700 μm. The upper portion of the sporophores consisted of a pyramidal head was ventricose, 320 to 520 μm long and 110 to 150 μm wide. The fungus was isolated from leaf lesions and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 18°C without light, but conidia were not observed in culture. These morphological and cultural characteristics were consistent with those of Hinomyces moricola (I. Hino) Narumi-Saito & Y. Harada (= Cristulariella moricola (I. Hino) Redhead) (3,4). An isolate was preserved in the Korean Agricultural Culture Collection (Accession No. KACC46955). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 452 bp was deposited in GenBank (Accession No. KC460209). A BLAST search in GenBank revealed that the sequence showed an exact match with those of C. moricola (JQ036181 ex Acer negundo and JQ036182 ex Glycine max). To determine the pathogenicity of the fungus, according to the procedure of Cho et al. (2), sporophores with the pyramidal head were carefully detached from a lesion on the naturally infected leaf using a needle. Each sporophore was transferred individually onto five places of four detached healthy leaves. The leaves were placed in dew chambers and incubated at 16°C. Symptoms were observed after 2 days on all inoculated leaves. A number of sporophores and immature sclerotia which were morphologically identical to the ones observed in the field were formed on the abaxial surface of the leaf 2 weeks after inoculation. The pathogen was reisolated from lesions on the inoculated leaves, confirming Koch's postulates. No symptoms were observed on the control leaves kept in humid chambers for 2 weeks. H. moricola was known to cause zonate leaf spots and defoliation on a wide range of woody and annual plants (3). To the best of our knowledge, this is the first report of Hinomyces infection on Japanese hops in Korea. References: (1) Anonymous. Humulus japonicus (Cannabaceae): Japanese hop. Eur. Medit. Plant Prot. Org. (EPPO). 2012. (2) S. E. Cho et al. Plant Dis. 96:906, 2012. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved December 8, 2012. (4) S. A. Redhead. Can. J. Bot. 53:700, 1975.

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First Report of Postharvest Fruit Rot in Persimmon Caused by Phacidiopycnis washingtonensis in Italy

Plant Disease ◽

10.1094/pdis-94-6-0788b ◽

2010 ◽

Vol 94 (6) ◽

pp. 788-788 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. T. Amatulli ◽

M. L. Gullino

Keyword(s):

United States ◽

Its Region ◽

The United States ◽

Fruit Rot ◽

Diospyros Kaki ◽

Pathogenicity Test ◽

Conidial Suspension ◽

First Report ◽

Pathogenicity Tests ◽

Phacidiopycnis Washingtonensis

Persimmon (Diospyros kaki L.) is widely grown in Italy, the leading producer in Europe. In the fall of 2009, a previously unknown rot was observed on 3% of fruit stored at temperatures between 5 and 15°C in Torino Province (northern Italy). The decayed area was elliptical, firm, and appeared light brown to dark olive-green. It was surrounded by a soft margin. The internal decayed area appeared rotten, brown, and surrounded by bleached tissue. On the decayed tissue, black pycnidia that were partially immersed and up to 0.5 mm in diameter were observed. Light gray conidia produced in the pycnidia were unicellular, ovoid or lacriform, and measured 3.9 to 6.7 × 2.3 to 3.5 (average 5.0 × 2.9) μm. Fragments (approximately 2 mm) were taken from the margin of the internal diseased tissues, cultured on potato dextrose agar (PDA), and incubated at temperatures between 23 and 26°C under alternating light and darkness. Colonies of the fungus initially appeared ash colored and then turned to dark greenish gray. After 14 days of growth, pycnidia and conidia similar to those described on fruit were produced. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 502-bp segment showed a 100% similarity with the sequence of Phacidiopycnis washingtonensis Xiao & J.D. Rogers (GenBank Accession No. AY608648). The nucleotide sequence has been assigned the GenBank Accession No. GU949537. Pathogenicity tests were performed by inoculating three persimmon fruits after surface disinfesting in 1% sodium hypochlorite and wounding. Mycelial disks (10 mm in diameter), obtained from PDA cultures of one strain were placed on wounds. Three control fruits were inoculated with plain PDA. Fruits were incubated at 10 ± 1°C. The first symptoms developed 6 days after the artificial inoculation. After 15 days, the rot was very evident and P. washingtonensis was consistently reisolated. Noninoculated fruit remained healthy. The pathogenicity test was performed twice. Since P. washingtonensis was first identified in the United States on decayed apples (2), ‘Fuji’, ‘Gala’, ‘Golden Delicious’, ‘Granny Smith’, ‘Red Chief’, and ‘Stark Delicious’, apple fruits also were artificially inoculated with a conidial suspension (1 × 106 CFU/ml) of the pathogen obtained from PDA cultures. For each cultivar, three surface-disinfested fruit were wounded and inoculated, while three others served as mock-inoculated (sterile water) controls. Fruits were stored at temperatures ranging from 10 to 15°C. First symptoms appeared after 7 days on all the inoculated apples. After 14 days, rot was evident on all fruit inoculated with the fungus, and P. washingtonensis was consistently reisolated. Controls remained symptomless. To our knowledge, this is the first report of the presence of P. washingtonensis on persimmon in Italy, as well as worldwide. The occurrence of postharvest fruit rot on apple caused by P. washingtonensis was recently described in the United States (3). In Italy, the economic importance of the disease on persimmon fruit is currently limited, although the pathogen could represent a risk for apple. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) Y. K. Kim and C. L. Xiao. Plant Dis. 90:1376, 2006. (3) C. L. Xiao et al. Mycologia 97:473, 2005.

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