First Report of Phytophthora palmivora, Causal Agent of Black Pod, on Cacao in Puerto Rico

Black pod or Phytophthora pod rot is the most economically important and widespread disease of cacao, Theobroma cacao L. Total losses due to Phytophthora exceed $400 million worldwide (1), and several species are known to attack cacao with P. palmivora (E.J. Butler) E. J. Butler as the most common. All plant parts are infected, but pod infections are particularly damaging. Symptoms resembling those of black pod disease were observed at the National Plant Germplasm Collection System of cacao at the USDA-ARS Tropical Agriculture Research Station (TARS) in Mayaguez, Puerto Rico for a number of years. During May of 2005, to determine the etiology of the disease, small, surface disinfested sections of pod lesions were placed on water agar and incubated for 4 days. The formation of papillate, deciduous, ellipsoidal to ovoid sporangia produced on sympodial sporangiophores on fruits, fit the description of P. palmivora and the identification was confirmed on cultures on water agar (2). Chlamydospores were readily observed in diseased pods and observed in pure cultures on V8 agar (2). Eight, single hyphal tips were transferred to potato dextrose agar (PDA) (Sigma-Aldrich, St. Louis, MO) and maintained as stock cultures. For pathogenicity tests, healthy mature pods were surface disinfested and placed in a humidity chamber lined with moist paper towels. Eight isolates were tested on four fruits per isolate and the pathogenicity test was repeated once. Inoculum was prepared by growing each isolate on PDA for 5 days with irradiation at 24°C, adding approximately 3.0 ml of water to each plate, dislodging the sporangia with a glass rod, mixing the suspension, estimating spore numbers with a hemacytometer, and adjusting to 104 sporangia per ml. A small, sterile scalpel was used to make an approximately 20.0 mm cut on the fruit epidermis, and approximately 0.2 ml of inoculum was placed on the wound. Pods were evaluated daily for 2 weeks. For molecular analysis, each of the eight cultures were grown in 50% potato dextrose broth to produce mycelia for DNA extraction using the FastDNA kit (Q-Biogen1, Irvine, CA). The internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster was amplified, purified, and sequenced for all eight isolates. The ITS sequences of GenBank Accession Nos. DQ987915 to DQ987922 were identical and exhibited strong similarity (>99% identity) to that of three previously described isolates of P. palmivora from cacao (GenBank Accession Nos. AF 228097, AF467093, and AF467089). P. palmivora has been reported on citrus, coconut, black pepper, and Arracacia xanthorrhiza in Puerto Rico (2,3) and inoculum may have originated from these host or imported on cacao planted into the cacao collection before 2000. USDA-ARS-TARS is the official site for the cacao germplasm collection, thus, a detailed integrated pest management plan that includes the evaluation for resistance, sanitation measures, and use of fungicides to reduce disease levels has been implemented. Decreasing incidence and severity of this disease is a top priority. To our knowledge, this is the first report of P. palmivora on cacao in Puerto Rico. References: (1) M. C. T. Braga et al. Agrotropica 1:108, 1989. (2) D. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996. (3) E. Rosa-Marquez. J. Agric. Univ. P. R. 84:53, 2000.

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First Report of Neofusicoccum mangiferae Causing Rachis Necrosis and Inflorescence Blight of Mango (Mangifera indica) in Puerto Rico

Plant Disease ◽

10.1094/pdis-08-13-0878-pdn ◽

2014 ◽

Vol 98 (4) ◽

pp. 570-570 ◽

Cited By ~ 2

Author(s):

L. M. Serrato-Diaz ◽

L. I. Rivera-Vargas ◽

R. D. French-Monar

Keyword(s):

Puerto Rico ◽

Mycelial Growth ◽

Mangifera Indica ◽

Elongation Factor ◽

Germplasm Collection ◽

Its Region ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Reference Isolate ◽

Β Tubulin

Inflorescence blight is a major disease in mango production (2,3). During a disease survey of mango in Puerto Rico conducted from February to April in 2009, 20% of the inflorescences were affected with inflorescence blight showing rachis and flower necrosis. Symptoms were observed in 70% of samples at the Mango Germplasm Collection of the University of Puerto Rico's Experiment Station in Juana Diaz. Blighted inflorescence tissue (necrotic and the interface between necrotic and healthy tissue) from mango cultivars ‘Haden’ and ‘Irwin’ were disinfested with 70% ethanol, rinsed with sterile water and transferred to acidified potato dextrose agar (APDA). Isolations (40%) produced fungi in the Botryosphaeriaceae. Isolates 90LY, 94LY, and 89LY were purified and identified morphologically using taxonomic keys (1,4) and by DNA sequence analyses as Neofusicoccum mangiferae (Syd. & P. Syd.) Crous, Slippers & A.J.L. Phillips. On APDA, colonies were gray with aerial mycelia that turned dark gray with age. Pycnidia were globose to pyriform and dark brown to black. Conidia (n = 50) were hyaline, ovoid, one-celled, and averaged 13.2 × 6.3 μm in size. PCR amplifications of the internal transcribed spacer region of rDNA using ITS5-ITS4 primers, and fragments of both β-tubulin and translation elongation factor 1-alpha (EF1-α) genes using Bt2a-Bt2b and EF1728F-EF1986R primers, respectively, were sequenced and analyzed using BLASTn query. Accession numbers of gene sequences submitted to GenBank were KF479465 to 67 for ITS region; KF479468 to 70 for β-tubulin; and KF479471 to 73 for EF1-α. All sequences were 99 to 100% identical to reference isolate CMW7024 (4) of N. mangiferae (GenBank Accession Nos. AY615185, AY615172, and DQ093221). For each fungal isolate, pathogenicity tests were conducted on mango trees using six randomly selected, healthy mango inflorescences at full bloom using two trees per cultivar. Both needle-wounded and unwounded inflorescences were inoculated with 5-mm diameter mycelial disks from 8-day-old cultures grown on APDA. Inflorescences were incubated in clear plastic bags for 8 days under field conditions. Controls were treated with APDA disks only. Inflorescences on ‘Irwin’ turned brown with necrosis extending from the rachis to flowers. Mycelial growth and inflorescence blight was observed with lesions ranging from 2 to 5 cm in length. On ‘Haden,’ the rachis tissues turned brown and necrotic with lesions ranging from 1.5 to 2 cm long and without mycelial growth. N. mangiferae was re-isolated from all diseased inflorescences, and no symptoms developed on controls, which fulfilled Koch's postulates. The test was repeated once. N. mangiferae was associated with blossom blight in Australia and South Africa (2,3). This is the first report of N. mangiferae causing rachis necrosis and inflorescence blight on mango in Puerto Rico. N. mangiferae belongs to a complex of pathogens causing inflorescence blight and rachis necrosis and, therefore, effective management of this important disease complex must involve control of this pathogen. References: (1) P. W. Crous et al. Stud. Mycol. 55:235, 2006. (2) G. I. Johnson et al. Ann. Appl. Biol. 119:465, 1991. (3) J. H. Lonsdale and J. M. Kotzé. Acta Hortic. 341:345, 1993. (4) A. J. L. Phillips. Key to the various lineages in “Botryosphaeria” Version 01 2007. Last retrieved 5 February 2014 from http://www.crem.fct.unl.pt/botryosphaeria_site/key.htm .

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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 Dolabra nepheliae Associated with Corky Bark Disease of Rambutan and Pulasan in Honduras

Plant Disease ◽

10.1094/pdis-01-12-0081-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 765-765 ◽

Cited By ~ 2

Author(s):

A. Rossman ◽

J. Melgar ◽

D. Walker ◽

A. Gonzales ◽

T. Ramirez ◽

...

Keyword(s):

Puerto Rico ◽

Sequence Data ◽

Management Strategies ◽

Pairwise Comparison ◽

Its Region ◽

Its Sequence ◽

Centraalbureau Voor ◽

First Report ◽

Intergenic Spacers ◽

Old Trees

In the last decade, rambutan (Nephelium lappaceum L., Sapindaceae) and pulasan (N. mutabile Blume) have been cultivated in Honduras to produce exotic fruits for export to North America (2). Recently, a disease was observed that produces dark brown to black fissured cankers from 1 to 3 cm long and 1 to 4 cm wide. The infected bark tissue becomes swollen with the middle region 3 to 8 mm thick. Symptoms appear when the trees are approximately 3 years old. As the trees mature, the cankers increase in size and weaken the branches, often resulting in breakage with the weight of the fruit causing substantial plant damage and fruit loss. In August 2010, fissured branch samples of rambutan and pulasan were collected from 6- to 8-year-old trees from the Humid Tropical Demonstrative Agroforestry Center in Honduras, Atlantida, La Masica (15°33′47.4″N, 87°05′2.5″W, elevation 106 m). A fungus associated with the cankers was identified as Dolabra nepheliae. It produces black, stipitate, elongate ascomata, 312 to 482 × 250 to 281 μm with broadly cylindric, bitunicate asci, 120 to 138 × 11.2 to 15.0 μm, and filiform, hyaline ascospores, 128 to 135 × 2.8 to 3.2 μm. Fungi from rambutan and pulasan were isolated on cornmeal agar plus 0.5% dextrose and antibiotics. On potato dextrose agar, the ascospores produced slow-growing colonies, 5 mm per week. In culture, isolates from both hosts produced pycnidia with elongated, slightly to strongly curved or S-shaped, hyaline conidia, 22.8 to 46.4 × 2.8 to 3.7 μm. This fungus was first reported on rambutan and pulasan from Malaysia (1,4), and later reported on rambutan and litchi in Hawaii and Puerto Rico (3). To our knowledge, this is the first report of D. nepheliae on pulasan and rambutan from Honduras. Specimens have been deposited at the U.S. National Fungus Collections (BPI 882442 on N. lappaceum and BPI 882443 on N. mutabile). Cultures were deposited at the Centraalbureau voor Schimmelcultures (CBS) as CBS 131490 on N. lappaceum and CBS 131491 on N. mutabile. Sequences of the internal transcribed spacer (ITS) region including ITS1, 5.8S, and ITS2 intergenic spacers were deposited in GenBank (Accession No. JQ004281 on N. lappaceum and Accession No. JQ004280 on N. mutabile). A BLAST search and pairwise comparison using the GenBank web server were used to compare ITS sequence data and recovered the following results: (i) CBS 131490 on N. lappaceum is 99% (538 of 544) identical to D. nepheliae CBS 123297 on Litchi chinensis from Puerto Rico; and (ii) CBS 131491 on N. mutabile is 99% (527 of 533) identical to the same strain of D. nepheliae. On the basis of the ITS sequence data, the isolates from Honduras were confirmed as the same species, D. nepheliae from Puerto Rico. Efforts to develop resistant germplasm and management strategies to control this disease have been initiated. References: (1) C. Booth and W. P. Ting. Trans. Brit. Mycol. Soc. 47:235, 1964. (2) T. Ramírez et al. Manual Para el Cultivo de Rambutan en Honduras. Fundación Hondureña de Investigación Agrícola. La Lima, Cortes, Honduras, 2003. (3) A. Y. Rossman et al. Plant Dis. 91:1685, 2007. (4) H. Zalasky et al. Can. J. Bot. 49:559, 1971.

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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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First Report of Typhula Blight on Agrostis stolonifera and Poa annua in Italy

Plant Disease ◽

10.1094/pdis.2003.87.7.875c ◽

2003 ◽

Vol 87 (7) ◽

pp. 875-875 ◽

Cited By ~ 1

Author(s):

P. Titone ◽

M. Mocioni ◽

A. Garibaldi ◽

M. L. Gullino

Keyword(s):

Agrostis Stolonifera ◽

Northern Italy ◽

Potato Dextrose Agar ◽

Poa Annua ◽

Golf Course ◽

Pathogenicity Test ◽

Potato Dextrose Broth ◽

First Report ◽

Pathogenicity Tests ◽

Typhula Incarnata

During January 2002, Agrostis stolonifera and Poa annua turfgrasses on a golf course in Avigliana (northern Italy) exhibited 10- to 45-cm-diameter circular patches when the snow melted from the greens, tees, and fairways. Many patches coalesced to form large areas of strawcolored blighted turfgrass. At the patch margin, infected plants were covered with white-to-gray mycelium. Plants within patches were matted and appeared slimy with mycelium and sclerotia that were light pink, irregularly shaped, and less than 5 mm in diameter. Isolation from infected leaves on potato dextrose agar, supplemented with 100 mg/l of streptomycin sulfate, consistently yielded a fungus with mycelial, sclerotia, and cultural characteristics of Typhula incarnata (1). Pathogenicity tests were performed by spraying a suspension of mycelium and sclerotia, prepared by chopping mycelium and sclerotia produced in potato dextrose broth, onto 8-week-old A. stolonifera plants grown in plastic trays (45 × 30 cm). Trays were maintained at 0°C for 8 weeks in the dark. Blight symptoms developed on inoculated plants after 6 weeks. Non-inoculated plants remained healthy. The pathogen was reisolated from inoculated plants, and the pathogenicity test was repeated once. Typhula blight incited by T. incarnata was reported in Scandinavian countries and in several European countries including Holland, Germany, Austria, and Switzerland (1). To our knowledge, this is the first report of Typhula blight on turfgrass in Italy. Reference: (1) J. D. Smith et al. 1989. Fungal Diseases of Amenity Turf Grasses. E & FN Spong Ltd, London.

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

Plant Disease ◽

10.1094/pdis-02-13-0138-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1119-1119 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Rhizoctonia Solani ◽

Its Region ◽

Morphological Characters ◽

Anastomosis Group ◽

Morphological Identification ◽

Pathogenicity Test ◽

Origanum Vulgare ◽

First Report ◽

Plastic Bags ◽

Hyphal Diameter

Origanum vulgare L., common name oregano, family Labiatae, is grown for its aromatic and medicinal properties and as ornamental. In the fall of 2012, a blight was observed in a farm located near Albenga (northern Italy) on 6% of 30,000 50-day-old plants, grown in trays in a peat/perlite mix. Semicircular, water soaked lesions appeared on leaves and stems, starting from the basal ones. As the disease progressed, blighted leaves turned brown, withered, clung to the shoots, and matted on the surrounding foliage. Eventually, infected plants died. Leaf and stem fragments taken from the margin of the diseased tissues belonging to 10 plants were disinfected for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA). A fungus with the morphological characters of Rhizoctonia solani was consistently recovered. Three isolates of R. solani obtained from affected plants were successfully anastomosed with R. solani isolate AG 1 (ATCC 58946). Three pairings were made for each tester strain. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and death of adjacent cells was observed. Results were consistent with other reports on anastomosis reactions (2). Isolates from oregano were paired with R. solani isolates AG 2, 3, 4, 6, 7, or 11 and examined microscopically. Anastomosis was not observed in any of the pairings. Tests were conducted twice. Mycelium of 10-day-old isolates from oregano appeared reddish brown, coarse, and radiate. Numerous dark brown sclerotia, 0.3 to 1.0 mm diameter (average 0.7) developed within 10 days after transfer of mycelia to PDA in 90 mm diameter petri dishes at 21 to 24°C. The descriptions of mycelium and sclerotia were typical for subgroup IB Type 1 (4). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis (1) of the 538 bp showed a 99% homology with the sequence of R. solani FJ746937, confirming the morphological identification of the species. The nucleotide sequence has been assigned the GenBank Accession KC493638. For pathogenicity tests, one of the isolates assigned to the anastomosis group AG-1-IB was tested by placing 9 mm diameter mycelial disks removed from PDA 10-day-old cultures of the fungus on leaves of 90-day-old oregano plants (n = 35). Thirty-five plants inoculated with non-inoculated PDA disks served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 25 ± 1°C with 12 h light/dark. The first symptoms, similar to those observed in the farm, developed 3 days after inoculation. Nine days after the artificial inoculation, 50% of plants were dead. About 10 colonies of R. solani were reisolated from infected leaves of inoculated plants. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. Symptoms caused by R. solani have been recently observed on O. vulgare in Greece (3). This is, to our knowledge, the first report of blight of O. vulgare caused by R. solani in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res., 25:3389, 1997. (2) D. E. Carling. Grouping in Rhizoctonia solani by hyphal anastomosis reactions. In: Rhizoctonia Species: Taxonomy, Molecular Biology, Ecology, Pathology and Disease control. Kluwer Academic Publishers, The Netherlands, pp. 37-47, 1996. (3) C. D. Holevas et al. Benaki Phytopathol. Inst., Kiphissia, Athens, 19:1-96, 2000. (4) R. T. Sherwood. Phytopathology 59:1924, 1969.

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First Report of Powdery Mildew Caused by Podosphaera xanthii on Calendula officinalis in Italy

Plant Disease ◽

10.1094/pdis-92-1-0174c ◽

2008 ◽

Vol 92 (1) ◽

pp. 174-174 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

M. L. Gullino

Keyword(s):

Powdery Mildew ◽

Its Region ◽

Pathogenicity Test ◽

Podosphaera Xanthii ◽

Calendula Officinalis ◽

First Report ◽

Potted Plants ◽

Blastn Analysis ◽

Voucher Specimens ◽

Pot Marigold

Calendula officinalis L. (Asteraceae) (pot marigold or English marigold) is an ornamental species grown in gardens and as potted plants for the production of cut flower. It was also used in ancient Greek, Roman, Arabic, and Indian cultures as a medicinal herb as well as a dye for fabrics, foods, and cosmetics. During the summer of 2007, severe outbreaks of a previously unknown powdery mildew were observed on plants in several gardens near Biella (northern Italy). Both surfaces of leaves of infected plants were covered with dense, white mycelia and conidia. As the disease progressed, infected leaves turned yellow and died. Mycelia and conidia also were observed on stems and flower calyxes. Conidia were hyaline, ellipsoid, born in short chains (four to six conidia per chain), and measured 27.0 to 32.1 (31.4) × 12.9 to 18.4 (18.2) μm. Conidiophores measured 49 to 77.3 (67.2) × 8 to 13.3 (10.8) μm and showed a foot cell measuring 44 to 59 (51.9) × 9.3 to 12.6 (11.3) μm followed by one shorter cell measuring 15.6 to 18.9 (17.6) × 10.4 to 13.6 (12.2) μm. Fibrosin bodies were present. Chasmothecia were spherical, amber colored, with a diameter of 89 to 100 (94.5) μm. Each chasmothecium contained one ascus with eight ascospores. On the basis of its morphology, the causal agent was determined to be a Podosphaera sp. (2). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 588 bp showed a 100% homology with the sequence of Podosphaera xanthii (2). The nucleotide sequence has been assigned GenBank Accession No. EU100973. Pathogenicity was confirmed through inoculations by gently pressing diseased leaves onto leaves of healthy C. officinalis plants. Five plants were inoculated. Five noninoculated plants served as control. Plants were maintained in a greenhouse at temperatures ranging from 20 to 26°C. Eleven days after inoculation, typical symptoms of powdery mildew developed on inoculated plants. Noninoculated plants did not show symptoms. The pathogenicity test was carried out twice. To our knowledge, this is the first report of powdery mildew on C. officinalis in Italy. C. officinalis was previously described as a host to Sphaerotheca fuliginea (synonym S. fusca) in Great Britain (4) as well as in Romania (3). Voucher specimens are available at the AGROINNOVA Collection, University of Torino. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (3) E. Eliade. Rev. Appl. Mycol. 39:710, 1960. (4) F. J. Moore. Rev. Appl. Mycol. 32:380, 1953.

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First Report of Powdery Mildew on Spanish Needles (Bidens bipinnata) Caused by Podosphaera xanthii in Korea

Plant Disease ◽

10.1094/pdis-10-12-0966-pdn ◽

2013 ◽

Vol 97 (10) ◽

pp. 1385-1385

Author(s):

H. B. Lee ◽

C. J. Kim ◽

H. Y. Mun

Keyword(s):

Powdery Mildew ◽

Its Region ◽

Broad Host Range ◽

Pathogenicity Test ◽

Podosphaera Xanthii ◽

Tropical Regions ◽

First Report ◽

Rdna Its ◽

Humid Condition ◽

Primer Sets

Spanish needles (Bidens bipinnata L.) is an annual herb that belongs to a genus of flowering plants in family Asteraceae native to United States, and tropical regions around world. The plant produces important flavonoid compounds quercitin and hyperoside that function as anti-allergens, anti-inflammatories, anti-microbials, and anti-cancer agents. Between July and October 2011 and 2012, white superficial mycelia were observed initially on leaf and stem portions, but later progressed to the flower head. Surveys showed that the disease was widespread in Gwangju and most areas of South Korea. Abundant, necrotic, dark brown spots showing chasmothecia were frequently observed in October and were abundant on the adaxial surface of leaves. Chasmothecia were blackish brown to yellow without typical appendages. They ranged from 51.2 to 71.1 (mean 66.8) μm in diameter. Conidia were formed singly and the primary conidia were ellipsoid, rounded at the apex, truncated base, and ranged from 25.4 to 33.2 (mean 27.3) μm long × 10.2 to 12.2 (mean 11.3) μm wide. Conidiophores were erect, 60.1 to 101.3 (mean 98.3) μm long × 6.2 to 9.2 (mean 7.3) μm wide. From extracted genomic DNA, the internal transcribed spacer (ITS) region inclusive of 5.8S and 28S rDNA was amplified with ITS1F (5′-TCCGTAGGTGAACCTGCGG-3′) and LR5F (5′-GCTATCCTGAGGGAAAC-3′), and LROR (5′-ACCCGCTGAACTTAAGC-3′) and LR5F primer sets, respectively. rDNA ITS (GenBank Accession No. JX512555) and 28S (JX512556) homologies of the fungus (EML-BBPW1) represented 99.6% (532/534) and 100% (661/661) identity values with Podosphaera xanthii (syn. P. fusca) AB040349 and P. xanthii (syn. P. fusca) AB462798, respectively. The rDNA sequence analysis revealed that the causal fungus matched P. xanthii (syn. P. fusca), forming a xanthii/fusca group (3,4). A pathogenicity test was performed on three plants in a greenhouse. The treated leaves were sealed in vinyl pack in humid condition for 2 days. Seven days after inoculation, similar symptoms were observed on the inoculated Spanish needles plant leaves. No symptoms were observed on control plants treated with distilled water. Koch's postulates were fulfilled by re-observing the fungal pathogen on the inoculated leaves. Podosphaera (syn. Sphaerotheca) xanthii (or fusca) has been known as an ubiquitous species with a broad host range. So far, five records regarding P. xanthii (=P. fusca) have been found in plants of genus Bidens. P. xanthii has been reported to occur on B. cernua in Belarus and Switzerland. In addition, the powdery mildew species was reported to occur on B. frondosa and B. tripartita in Korea, Russia, and Switzerland (2). To our knowledge, this is the first report of powdery mildew caused by P. xanthii on Spanish needles (B. bipinnata) in Korea. References: (1) U. Braun et al. Schlechtendalia 10:91, 2003. (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/ , 2012. (3) H. B. Lee. J. Microbiol. 51:1075, 2012. (4) S. Takamatsu, et al. Persoonia 24:38, 2010.

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First Report of Powdery Mildew Caused by Golovinomyces cichoracearum on English Daisy (Bellis perennis) in Italy

Plant Disease ◽

10.1094/pdis-92-3-0484a ◽

2008 ◽

Vol 92 (3) ◽

pp. 484-484 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

Powdery Mildew ◽

Its Region ◽

Northern Italy ◽

Flowering Plant ◽

Pathogenicity Test ◽

First Report ◽

Golovinomyces Cichoracearum ◽

Blastn Analysis ◽

Commercial Farms ◽

Voucher Specimens

Bellis perennis (English daisy) is a flowering plant belonging to the Asteraceae and is increasingly grown as a potted plant in Liguria (northern Italy). In February 2007, severe outbreaks of a previously unknown powdery mildew were observed on plants in commercial farms at Albenga (northern Italy). Both surfaces of leaves of affected plants were covered with white mycelia and conidia. As the disease progressed, infected leaves turned yellow. Mycelia and conidia also were observed on stems and flower calyxes. Conidia were hyaline, ellipsoid, borne in chains (as many as three conidia per chain), and measured 27.7 × 16.9 (15.0 to 45.0 × 10.0 to 30.0) μm. Conidiophores measured 114.0 × 12.0 (109.0 to 117.0 × 11.0 to 13.0) μm and showed a foot cell measuring 78.0 × 11.0 (72.0 to 80.0 × 11.0 to 12.0) μm followed by two shorter cells. Fibrosin bodies were absent. Chasmothecia were not observed in the collected samples. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 415 bp obtained showed an E-value of 7e–155 with Golovinomyces cichoracearum (3). The nucleotide sequence has been assigned the GenBank Accession No. AB077627.1 Pathogenicity was confirmed through inoculations by gently pressing diseased leaves onto leaves of healthy B. perennis plants. Twenty plants were inoculated. Fifteen noninoculated plants served as a control. Plants were maintained in a greenhouse at temperatures ranging from 10 to 30°C. Seven days after inoculation, typical symptoms of powdery mildew developed on inoculated plants. The fungus observed on inoculated plants was morphologically identical to that originally observed. Noninoculated plants did not show symptoms. The pathogenicity test was carried out twice. To our knowledge, this is the first report of powdery mildew on B. perennis in Italy. The disease was already reported in other European countries (2). Voucher specimens are available at the AGROINNOVA Collection, University of Torino. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) U. Braun The Powdery Mildews (Erysiphales) of Europe. Gustav Fischer Verlag, Jena, Germany, 1995. (3) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000.

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First Report of Lasiodiplodia theobromae Associated with Decline of Grapevine Rootstock Mother Plants in Spain

Plant Disease ◽

10.1094/pdis-92-5-0832b ◽

2008 ◽

Vol 92 (5) ◽

pp. 832-832 ◽

Cited By ~ 13

Author(s):

A. Aroca ◽

R. Raposo ◽

D. Gramaje ◽

J. Armengol ◽

S. Martos ◽

...

Keyword(s):

Elongation Factor ◽

Aerial Mycelium ◽

Its Region ◽

Pathogenic Fungi ◽

Vascular Lesions ◽

Malt Extract ◽

Pathogenicity Test ◽

Lasiodiplodia Theobromae ◽

First Report ◽

Mother Plants

A field of Richter 110 rootstock mother plants in Valencia Province (eastern Spain) was surveyed during November 2006 to study the mycoflora of declining plants. Two canes with stunted leaves were collected from a plant with a reduced number of shoots. No cankers or vascular lesions were observed in the collected canes. Six wood chips (1 to 2 mm thick) were taken from one basal fragment (3 to 4 cm long) of each cane, surface sterilized in 70% ethanol for 1 min, and plated on malt extract agar supplemented with 0.5 g L–1 of streptomycin sulfate. Petri dishes were incubated for 7 days at 25°C. A fungus was consistently isolated from all samples that showed the following characteristics: colonies grown on potato dextrose agar (PDA) at 25°C developed a white, aerial mycelium that turned gray after 4 to 6 days and produced pycnidia after 1 month on sterile grapevine slivers of twigs placed on the PDA surface; conidia from culture were ellipsoidal, thick walled, initially hyaline, nonseptate, and measuring 20 to 25 (22.5) × 12 to 14 (13) μm; aged conidia were brown, 1-septate with longitudinal striations in the wall; and pseudoparaphyses variable in form and length were interspersed within the fertile tissue. The fungus was identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. from the above characteristics (2). Identity was confirmed by analysis of the nucleotide sequences of the internal transcribed spacer (ITS) region from the rRNA repeat and part of the translation elongation factor 1-alpha (EF1-α) and the β-tubulin (B-tub) genes, as done elsewhere (1,3). BLAST searches at GenBank showed a high identity with reference sequences (ITS: 100%, EF1-α: 97%; B-tub: 99%). Representative sequences of the studied DNA regions were deposited at GenBank (Accession Nos.: ITS: EU254718; EF1-α: EU254719; and B-tub: EU254720). A pathogenicity test was conducted on 1-year-old grapevine plants cv. Macabeo grafted onto Richter 110 rootstocks maintained in a greenhouse. A superficial wound was made on the bark of 10 plants with a sterilized scalpel, ≈10 cm above the graft union. A mycelial plug obtained from the margin of an actively growing fungal colony (isolate JL664) was placed in the wound and the wound was wrapped with Parafilm. Ten additional control plants were inoculated with sterile PDA plugs. All control plants grew normally, and the inoculation wound healed 3 months after inoculation. Plants inoculated with L. theobromae showed no foliar symptoms in the same period, but developed cankers variable in size surrounding the inoculation sites. Vascular necroses measuring 8.4 ± 1.5 cm (mean ± standard error) developed in the inoculated plants that were significantly longer than the controls (0.3 ± 0.2 cm). The pathogen was reisolated from all inoculated plants and no fungus was reisolated from the controls. These results confirmed the pathogenicity of L. theobromae to grapevine and points to a possible involvement of L. theobromae in the aetiology of grapevine decline as previously reported (3,4). To our knowledge, this is the first report of L. theobromae isolated from grapevine in Spain. References: (1) J. Luque et al. Mycologia 97:1111, 2005. (2) E. Punithalingam. No. 519 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1976. (3) J. R. Úrbez-Torres et al. Plant Dis. 90:1490, 2006. (4) J. M. van Niekerk et al. Phytopathol. Mediterr. 45(suppl.):S43, 2006.

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