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

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 287-287 ◽  
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.

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

Plant Disease ◽  
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.

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

Plant Disease ◽  
2008 ◽  
Vol 92 (8) ◽  
pp. 1249-1249 ◽  
Author(s):  
S. M. Marek ◽  
I. R. Moncrief ◽  
N. R. Walker
Keyword(s):  
Warm Season ◽  
Its Region ◽  
The United States ◽  
Small Subunit ◽  
Its Sequences ◽  
Type I ◽  
Sclerotinia Homoeocarpa ◽  
Dollar Spot ◽  
First Report ◽  
Warm Season Grass

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.

scholarly journals First Report of Dollar Spot Caused by Sclerotinia homoeocarpa on Agrostis stolonifera in North Dakota

Plant Disease ◽  
2012 ◽  
Vol 96 (7) ◽  
pp. 1071-1071
Author(s):  
J. M. LeBoldus ◽  
Q. Zhang ◽  
K. Kinzer
Keyword(s):  
New York ◽  
North Dakota ◽  
Signs And Symptoms ◽  
Creeping Bentgrass ◽  
Agrostis Stolonifera ◽  
Sclerotinia Homoeocarpa ◽  
Dollar Spot ◽  
Full Spectrum ◽  
Spot Disease ◽  
Plastic Bags

Dollar spot disease is a major concern for golf courses nationwide, resulting in poor turf quality and significant damage to playing surfaces. To manage this disease effectively, fungicides need to be applied regularly. This management strategy represents a significant cost to turfgrass managers and may impact the economics of the industry in North Dakota. In the summer of 2011, small, circular, sunken brown patches of dead turf approximately 5 cm in diameter and resembling dollar spot disease were observed on a creeping bentgrass (Agrostis stolonifera L.) variety trial at the North Dakota State University Agricultural Experiment Station in Fargo, ND. Fresh individual leaf specimens with distinct lesions having straw colored centers and reddish brown margins were collected. Leaves were surface disinfected in a 0.05% sodium chloride solution for 60 s, rinsed three times in sterile distilled water, then placed onto potato dextrose agar (PDA). Three isolates were obtained from the disease infested leaves with similar morphology to that described for Sclerotinia homoeocarpa F.T. Bennett (1). Fungal colonies were initially colorless followed by the development of sparse white columnar aerial mycelia with cinnamon colored bases. Hyphae were 5 to 8 μm wide and thin walled with dense granular contents and septations at irregular intervals. Fourteen days after culturing, dark brown to black mycelial stroma developed. A single representative isolate was selected to conduct inoculations. Inoculum was produced by placing six 5-mm-diameter plugs of PDA with actively growing mycelium into an Erlenmeyer flask with 40 g of sterile millet seed. The inoculum was incubated for 14 days at ambient temperature (20 to 25°C). Three creeping bentgrass cultivars, Crenshaw, Declaration, and L-93, were inoculated (two pots per cultivar). Following inoculation, the pots were misted with water, sealed in separate plastic bags, and placed in the dark for 48 h. For the next 5 days, plants were placed for 8 h outside of bags on a bench with full spectrum fluorescent bulbs, followed by 16 h in plastic bags in the dark. Finally, pots were placed on a bench for 48 h. Signs and symptoms of S. homoeocarpa developed on all pots, whereas the controls remained asymptomatic. The same fungus was reisolated from grass leaves, satisfying Koch's postulates. To confirm the identity of the fungus, the internal transcribed spacer was amplified using the ITS4 and ITS5 primers (2). The amplicon was sequenced, generating a 549-bp sequence (Accession No. JQ735942) with 100% similarity to sequences of S. homoeocarpa in GenBank (Accession Nos. GQ924924.1, GQ924923.1, and EU123801.1). To our knowledge, this is the first confirmed report of dollar spot in North Dakota. References: (1) F. T. Bennett. Ann. Appl. Biol. 24:236, 1937. (2) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press Inc., New York, 1990.

scholarly journals First Report of Dollar Spot Caused by Sclerotinia homoeocarpa on Agrostis stolonifera in Argentina

Plant Disease ◽  
2004 ◽  
Vol 88 (12) ◽  
pp. 1384-1384 ◽  
Author(s):  
M. C. Rivera ◽  
E. R. Wright ◽  
L. V. Goldring ◽  
B. A. Pérez ◽  
D. Barreto
Keyword(s):  
Substrate Surface ◽  
Pathogenic Fungi ◽  
Creeping Bentgrass ◽  
Agrostis Stolonifera ◽  
Fluorescent Light ◽  
Infested Soil ◽  
Sclerotinia Homoeocarpa ◽  
Dollar Spot ◽  
First Report ◽  
Agar Plug

During the summer of 2000, circular, yellow-to-brown, blighted, 2- to 4-cm-diameter patches were observed on creeping bentgrass (Agrostis stolonifera) putting greens (cv. Pennlinks) maintained at a 4- to 5-mm height on a golf course in Pilar (Buenos Aires, Argentina). Symptomatic leaves had transverse chlorotic bands that sometimes extended to the tip with brown lesions inside the bands. A fungus was isolated from symptomatic tissue after surface sterilization with 2% bleach for 1 min and plating on 2% potato dextrose agar (PDA). The mycelium was fluffy and white. The culture turned olive to brown and developed black stromata on the lower side of the plate base after 2 weeks. Pathogenicity tests were performed on 2-month-old healthy plants of A. stolonifera (cv. Crenshaw) grown in sterilized sand. Recently cut, 14-mm-diameter plugs of A. stolonifera were placed in 22- × 17-cm plastic trays filled with a sterilized mixture of 50:50 soil/sand (vol/vol). Plants were maintained at a 7-mm height. Two sources of inoculum were prepared; one was cultured on PDA at 22 to 25°C for 10 days and the other was prepared by incubating in sterilized soil at room temperature for 14 days. Twenty pieces of 1-cm-diameter agar blocks containing mycelium were placed in each plug at the base of the plants. In the infested soil inoculation, 25 g of soil were distributed among the plants on the substrate surface. Control plants were treated with either sterile PDA pieces or noninfested soil. The trays were irrigated with sterilized distilled water, covered with polyethylene bags, and kept in a controlled environment chamber at 25°C with 12 h per day of fluorescent light for 30 days. Leaf chlorosis appeared 7 and 10 days after inoculation for the agar-plug and infested-soil methods, respectively. Leaf necrosis was observed at day 23. Controls remained asymptomatic. The inoculated fungus was reisolated from symptomatic leaf tissue. The pathogen was identified as Sclerotinia homoeocarpa (1,2). To our knowledge, this is the first report of Sclerotinia homoeocarpa causing dollar spot disease on Agrostis stolonifera in Argentina and the first report of a disease on golf courses in our country. References: (1) J. E. M. Mordue. Sclerotinia homoeocarpa. No. 618 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1979. (2) R. W. Smiley. Dollar Spot. Pages 14–16 in: Compendium of Turfgrass Diseases. The American Phytopathological Society, St. Paul, MN, 1983.

scholarly journals First report of Colletotrichum siamense Causing Blossom Blight on Thai basil (Ocimum basilicum L.) in Malaysia

Plant Disease ◽  
2020 ◽  
Author(s):  
Siti Izera Ismail ◽  
Nur Adlina Rahim ◽  
Dzarifah Zulperi
Keyword(s):  
Disease Incidence ◽  
Its Region ◽  
Ocimum Basilicum ◽  
Distilled Water ◽  
Single Spore ◽  
First Report ◽  
Plastic Bags ◽  
Colletotrichum Siamense ◽  
Primer Sets ◽  
Acca Sellowiana

Thai basil (Ocimum basilicum L.) is widely cultivated in Malaysia and commonly used for culinary purposes. In March 2019, necrotic lesions were observed on the inflorescences of Thai basil plants with a disease incidence of 60% in Organic Edible Garden Unit, Faculty of Agriculture in the Serdang district (2°59'05.5"N 101°43'59.5"E) of Selangor province, Malaysia. Symptoms appeared as sudden, extensive brown spotting on the inflorescences of Thai basil that coalesced and rapidly expanded to cover the entire inflorescences. Diseased tissues (4×4 mm) were cut from the infected lesions, surface disinfected with 0.5% NaOCl for 1 min, rinsed three times with sterile distilled water, placed onto potato dextrose agar (PDA) plates and incubated at 25°C under 12-h photoperiod for 5 days. A total of 8 single-spore isolates were obtained from all sampled inflorescence tissues. The fungal colonies appeared white, turned grayish black with age and pale yellow on the reverse side. Conidia were one-celled, hyaline, subcylindrical with rounded end and 3 to 4 μm (width) and 13 to 15 μm (length) in size. For fungal identification to species level, genomic DNA of representative isolate (isolate C) was extracted using DNeasy Plant Mini Kit (Qiagen, USA). Internal transcribed spacer (ITS) region, calmodulin (CAL), actin (ACT), and chitin synthase-1 (CHS-1) were amplified using ITS5/ITS4 (White et al. 1990), CL1C/CL2C (Weir et al. 2012), ACT-512F/783R, and CHS-79F/CHS-345R primer sets (Carbone and Kohn 1999), respectively. A BLAST nucleotide search of ITS, CHS-1, CAL and ACT sequences showed 100% similarity to Colletotrichum siamense ex-type cultures strain C1315.2 (GenBank accession nos. ITS: JX010171 and CHS-1: JX009865) and isolate BPDI2 (CAL: FJ917505, ACT: FJ907423). The ITS, CHS-1, CAL and ACT sequences were deposited in GenBank as accession numbers MT571330, MW192791, MW192792 and MW140016. Pathogenicity was confirmed by spraying a spore suspension (1×106 spores/ml) of 7-day-old culture of isolate C onto 10 healthy inflorescences on five healthy Thai basil plants. Ten infloresences from an additional five control plants were only sprayed with sterile distilled water and the inoculated plants were covered with plastic bags for 2 days and maintained in a greenhouse at 28 ± 1°C, 98% relative humidity with a photoperiod of 12-h. Blossom blight symptoms resembling those observed in the field developed after 7 days on all inoculated inflorescences, while inflorescences on control plants remained asymptomatic. The experiment was repeated twice. C. siamense was successfully re-isolated from the infected inflorescences fulfilling Koch’s postulates. C. siamense has been reported causing blossom blight of Uraria in India (Srivastava et al. 2017), anthracnose on dragon fruit in India and fruits of Acca sellowiana in Brazil (Abirami et al. 2019; Fantinel et al. 2017). This pathogen can cause a serious threat to cultivation of Thai basil and there is currently no effective disease management strategy to control this disease. To our knowledge, this is the first report of blossom blight caused by C. siamense on Thai basil in Malaysia.

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

Plant Disease ◽  
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.

scholarly journals First Report of Stem Canker Disease of Pitaya (Hylocereus undatus and H. polyrhizus) Caused by Neoscytalidium dimidiatum in Taiwan

Plant Disease ◽  
2012 ◽  
Vol 96 (6) ◽  
pp. 906-906 ◽  
Author(s):  
M. F. Chuang ◽  
H. F. Ni ◽  
H. R. Yang ◽  
S. L. Shu ◽  
S. Y. Lai ◽  
...  
Keyword(s):  
New York ◽  
Aerial Mycelium ◽  
Its Region ◽  
Stem Canker ◽  
Canker Disease ◽  
First Report ◽  
Neoscytalidium Dimidiatum ◽  
Succulent Plant ◽  
Sterile Medium ◽  
Hylocereus Undatus

Pitaya (Hylocereus undatus and H. polyrhizus Britt. & Rose), a perennial succulent plant grown in the tropics, is becoming an emerging and important fruit plant in Taiwan. In September of 2009 and 2010, a number of pitaya plants were found to have a distinctive canker on stems. The disease expanded quickly to most commercial planting areas in Taiwan (e.g., Pintung, Chiayi, and Chunghua). Symptoms on the stem were small, circular, sunken, orange spots that developed into cankers. Pycnidia were erumpent from the surface of the cankers and the stems subsequently rotted. After surface disinfestation with 0.1% sodium hypochloride, tissues adjacent to cankers were placed on acidified potato dextrose agar (PDA) and incubated at room temperature for 1 week, after which colonies with dark gray-to-black aerial mycelium grew. Hyphae were branched, septate, and brown and disarticulated into 0- to 1-septate arthrospores. Sporulation was induced by culturing on sterile horsetail tree (Casuarina equisetifolia) leaves. Conidia (12.79 ± 0.72 × 5.14 ± 0.30 μm) from pycnidia were one-celled, hyaline, and ovate. The internal transcribed spacer (ITS) region of ribosomal DNA was PCR amplified with primers ITS1 and ITS4 (2) and sequenced. The sequence (GenBank Accession No. HQ439174) showed 99% identity to Neoscytalidium dimidiatum (Penz.) Crous & Slippers (GenBank Accession No. GQ330903). On the basis of morphology and nucleotide-sequence identity, the isolates were identified as N. dimidiatum (1). Pathogenicity tests were conducted in two replicates by inoculating six surface-sterilized detached stems of pitaya with either mycelium or conidia. Mycelial plugs from 2-day-old cultures (incubated at 25°C under near UV) were inoculated to the detached stems after wounding with a sterile needle. Conidial suspensions (103 conidia/ml in 200 μl) were inoculated to nonwounded stems. Noninoculated controls were treated with sterile medium or water. Stems were then incubated in a plastic box at 100% relative humidity and darkness at 30°C for 2 days. The symptoms described above were observed on inoculated stems at 6 to 14 days postinoculation, whereas control stems did not develop any symptoms. N. dimidiatum was reisolated from symptomatic tissues. To our knowledge, this is the first report of N. dimidiatum causing stem canker of pitaya. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, New York, 1990.

scholarly journals First Report of Root and Stem Rot Caused by Phytophthora nicotianae on Peperomia tetraphylla in China

Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1171-1171 ◽  
Author(s):  
D. X. Zeng ◽  
X. L. Wu ◽  
Y. H. Li
Keyword(s):  
Southern China ◽  
Aerial Mycelium ◽  
Its Region ◽  
Soil Extract ◽  
Phytophthora Nicotianae ◽  
Stem Rot ◽  
Molecular Characteristics ◽  
Its Sequence ◽  
First Report ◽  
Pure Cultures

Peperomia tetraphylla, an evergreen herb, is becoming increasingly popular as a potted ornamental plant in southern China. In the summer of 2008, in some commercial flower nurseries in Shenzhen, Guangdong Province, P. tetraphylla showed extensive black stem and root rot, with leaves dropping from the rotten stem. Small pieces (approximately 3 mm2) of stems and leaves were excised from the margins of the black lesions, surface disinfected for 30 s to 1 min in 0.1% HgCl2, plated onto potato dextrose agar (PDA), and incubated at 25°C in the dark. All the plated samples yielded Phytophthora, and microscopic examination of pure cultures grown on PDA plates showed arachnoid colonies with abundant aerial mycelium, chlamydospores, and a few sporangia. Numerous sporangia were formed in sterile soil extract. Sporangia were ovoid or obpyriform, noncaducous, with prominent solitary papillae, and measured 31 to 52 μm (average 38 μm) × 21 to 34 μm (average 27 μm). Chlamydospores were spherical and 21 to 34 μm in diameter (average 28 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS4/ITS5 and sequenced (2). The ITS sequence, when submitted for a BLAST search in the NCBI database, showed 100% homology with the sequences of two reference isolates of Phytophthora nicotianae (Accession Nos. AY833526 and EU433396) and the consensus ITS sequence was deposited in the NCBI as Accession No. GQ499373. The isolate was identified as Phytophthora nicotianae on the basis of morphological and molecular characteristics (1). Pathogenicity of the isolate was confirmed by inoculating 1-year-old plants of P. tetraphylla growing in pots. The isolate was grown for 7 days on PDA plates and mycelial plugs, 5 mm in diameter and taken from the advancing margins of the colonies, were buried approximately 1 cm deep near the base of the stem in such a way that the mycelium on the plugs was in contact with the surface of the stem, which had been wiped earlier with 70% ethanol and gently wounded with a needle. Plants treated the same way but inoculated with sterile PDA plugs served as control plants. Three plants in each pot were inoculated and there were five replications each for the treatment and the control. All plants were kept in a greenhouse at 22 to 32°C. After 6 to 7 days, the inoculated plants showed black lesions around the mycelial plugs; symptoms of root and stem rot developed rapidly thereafter and the plants collapsed within 2 weeks. All symptoms on the inoculated plants were identical to those observed in naturally diseased plants, whereas the control plants remained healthy. The same fungus was consistently reisolated from the inoculated plants. To our knowledge, this is the first report of Phytophthora nicotianae on P. tetraphylla in China. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) J. B. Ristaino et al. Appl. Environ. Microbiol. 64:948, 1998.

scholarly journals Residual Efficacy of Fungicides for Control of Dollar Spot on Creeping Bentgrass

Plant Disease ◽  
2006 ◽  
Vol 90 (5) ◽  
pp. 571-575 ◽  
Author(s):  
Richard Latin
Keyword(s):  
Creeping Bentgrass ◽  
Effective Concentration ◽  
Disease Suppression ◽  
Golf Course ◽  
Sclerotinia Homoeocarpa ◽  
Dollar Spot ◽  
Residual Efficacy ◽  
Bioassay Technique ◽  
Precipitous Decline ◽  
Field Plots

The duration of effective concentrations of fungicides for control of dollar spot on creeping bentgrass fairways was investigated using a bioassay technique. In each of three runs of the experiment, fungicide treatments were applied once to turf in replicated field plots; then, the plots were sampled periodically over 3 weeks by removing turf plugs from the field. The sampled plugs were placed in incubation containers and inoculated with sections from a 4-day-old colony of Sclerotinia homoeocarpa growing on potato dextrose agar. After a 96-h incubation period, the extent of pathogen growth on the turf plugs was measured. Results described a precipitous decline in effective concentration for all fungicide treatments beginning 7 to 10 days after application. The fungicides were only marginally effective at 14 days after application, and none provided any disease suppression at 21 days after application. The efficacy half-life (EHL) associated with four of the fungicides (chlorothalonil, iprodione, propiconazole, and thiophanate methyl) was estimated with two models. EHL estimates ranged from 6.1 to 15.2 days depending on the fungicide and the model. This research contributes to our knowledge of the duration of effective fungicide concentrations on creeping bentgrass and can provide insight for scheduling fungicide sprays for golf course fairways.

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