First Report of Spot Form Net Blotch Caused by Pyrenophora teres f. maculata on Barley in the Mon-Dak Area of the United States

Pyrenophora teres Drechs. causes net blotch of barley, a common foliar disease in cultivation zones around the world. The disease occurs in two forms, namely a net form net blotch (NFNB) caused by P. teres f. teres and a spot form net blotch (SFNB) caused by P. teres f. maculata. As in other parts of the northern Great Plains, in the Mon-Dak area (western North Dakota and eastern Montana), NFNB is prevalent. SFNB was first reported in western Montana in 1983 (1) and more recently in eastern North Dakota in 2010 (3) but not in the Mon-Dak area. In the summer of 2011, unusual spot lesions that were surrounded by necrosis or chlorosis were observed on different barley cultivars in fields at Williston, ND, Nesson Valley, ND, and Sidney, MT areas. Diseased leaves from various barley cvs. from the three locations were transferred to water agar and incubated at room temperature for 24 h to induce sporulation. Morphological examination of conidia (45 to 169 × 15 to 21 μm) did not show significant differences from a known isolate of P. teres f. teres 0-1 (provided by Tim Friesen, ARS, Fargo, ND). For pathogenicity testing, six 14-day-old plants of barley cv. Tradition were sprayed until runoff with a 2,000 spore/ml suspension of two isolates from each location and the control P. teres f. maculata isolate DEN2.6 (provided by Tim Friesen). Plants were incubated first in a lighted humidity chamber for 24 h and then in a greenhouse for 7 days at 21°C. Regardless of inoculum source, spot lesions surrounded by necrosis or chlorosis, typical of SFNB, appeared on the inoculated leaves within 7 days. Fungi isolated from symptomatic leaves were identified as P. teres and the morphology of the conidia was undistinguishable from those of P. teres f. teres. All control plants which were sprayed with sterile distilled water were symptomless. The pathogenicity test was repeated. Rapid PCR detection and amplicon sequencing (2) of the internal transcribed spacer (ITS) region of ribosomal genes was performed on field and pathogenicity test leaf lesion samples to confirm the presence of P. teres f. maculata. DNA templates were prepared using the Extract-N-Amp Plant PCR Kits (Sigma Chemical Co., St. Louis, MO) and subjected to PCR using ITS1 and ITS4 primers. Amplicons were then purified and sequenced. The 585-bp nucleotide sequences of P. teres f. maculata from Mon-Dak area were submitted to GenBank under accession nos. PtmNES1 (JX187587), PtmSDY1 (JX187588), PtmSDY2 (JX187589), and PtmWIL1 (JX187590). The sequences from the four locations shared 100% similarity and also with P. teres f. maculata (EF452471) from GenBank while showing 10 nucleotide differences (99% similarity) with P. teres f. teres (EF452472).The results represent first report of SFNB in the Mon-Dak. Barley is one of the most important crops in the area. Resistance of the NFNB and SFNB of barley are controlled by different genes (4). Based on this report, SFNB therefore have to be considered in selection of barley cultivars for cultivation in the area. References: (1) H. E Bockelman et al. Plant Dis. 67:696, 1983. (2) R. T. Lartey et al. J. Sugar Beet Res. 40:1, 2003. (3) Z. H. Liu and T. L. Friesen. Plant Dis. 94:480, 2010. (4) O. M. Manninen et al. Genome. 46:1564, 2006.

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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 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 Botrytis Blight Caused by Botrytis cinerea on Platycodon grandiflorum in Italy

Plant Disease ◽

10.1094/pdis-93-9-0969b ◽

2009 ◽

Vol 93 (9) ◽

pp. 969-969

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Botrytis Cinerea ◽

Its Region ◽

The United States ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Commonwealth Mycological Institute ◽

Platycodon Grandiflorum ◽

First Report ◽

Bedding Plant

Platycodon grandiflorum (balloon flower), a perennial plant belonging to the Campanulaceae family, is widely grown as a bedding plant in temperate gardens. This species is characterized by the ability to bloom profusely throughout the summer into early fall and for its white to blue and pink flowers. In September 2008, symptoms of a previously unknown blight were observed in six gardens located in the Biella Province of northern Italy. When the disease developed, temperatures ranged between 15 and 22°C with frequent rains (149.8 mm of rainfall registered in September 2008 by the meteorological station of Oropa, located in the same area in which the disease appeared). Initially, leaves and petioles appeared chlorotic. Subsequently, lesions developed on the stems and flowers were sometimes affected. In each garden examined, approximately 50% of the plants were affected by the disease. A soft, gray mycelium was observed on symptomatic tissues, especially the stems. Severely infected leaves and stems eventually became completely rotted and later desiccated. Diseased tissue was excised from affected leaves, immersed in a solution containing 1% sodium hypochlorite for 10 s, and then cultured on potato dextrose agar (PDA) medium. A fungus developed that 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, irregular, and measured 1 to 3.5 × 0.9 to 2.5 (average 2.1 × 1.5) mm. Conidia were smooth, ash colored, unicellular, ovoid, and measured 11 to 19 × 7 to 13 (average 15 × 11) μm. These morphological features were typical of those described for Botrytis cinerea (2). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 539-bp segment showed 100% similarity with the sequence of Botryotinia fuckeliana (perfect stage of B. cinerea). The nucleotide sequence has been assigned the GenBank Accession No. GQ149480. Pathogenicity tests were performed by placing 1-cm2 fragments removed from PDA cultures of B. cinerea isolated from balloon flower on leaves of healthy potted P. grandiflorum plants (4-month-old). Five fragments were placed on each plant. Plants inoculated with PDA alone served as controls. Ten plants per treatment were used. Plants were covered with plastic bags for 5 days after inoculation and maintained in a greenhouse at temperatures between 18 and 23°C. The first foliar lesions developed on leaves 3 days after inoculation, and after 5 days, 80% of the leaves were severely infected. As the infection progressed after the inoculation, the stems also became infected. Control plants remained healthy. B. cinerea was consistently reisolated from leaf and stem lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of B. cinerea on P. grandiflorum in Italy, as well as in Europe. Blight on balloon flower attributed to Botrytis spp. was previously reported in the United States (3). References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, England, 1971. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

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First Report of Choanephora Blight Caused by Choanephora infundibulifera on Hibiscus rosa-sinensis in Korea

Plant Disease ◽

10.1094/pdis-02-14-0218-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1275-1275 ◽

Cited By ~ 2

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

S. H. Lee ◽

H. D. Shin

Keyword(s):

Large Subunit ◽

Aerial Mycelium ◽

Its Region ◽

The United States ◽

Pathogenicity Test ◽

Potential Threat ◽

First Report ◽

Hibiscus Rosa Sinensis ◽

Initial Symptoms ◽

Chinese Hibiscus

Hibiscus rosa-sinensis L., commonly known as Chinese hibiscus, is an evergreen flowering shrub belonging to the Malvaceae and is widely cultivated throughout Asia including Korea. In August 2013, blight was observed on Chinese hibiscus in a commercial flower nursery in Seoul, Korea. Initial symptoms began as reddish purple spots at the tip of flowers and expanded to encompass entire flowers. Infected lesions appeared water-soaked, reddish brown, and were followed by rapid rotting of infected tissues. Approximately 50% of the plants surveyed were affected. Monosporous sporangiola formed on infected tissue were transferred to potato dextrose agar (PDA). Fungal colonies were obtained that were at first white with abundant aerial mycelium, and then became yellowish with the appearance of sporangiola. Sporangiophores bearing sporangiola were erect to slightly curved, unbranched, and hyaline. Funnel-shaped secondary vesicles formed on the primary vesicles. Sporangiola were indehiscent, ovoid to subglobose, smooth, non-striated, brown to dark brown, 10 to 27.5 × 8.5 to 17 μm, and sometimes germinated in culture. The fungus was identified as Choanephora infundibulifera (Curr.) D.D. Cunn. based on the morphological and cultural characteristics (2). Voucher specimens were housed in the Korea University Herbarium (KUS). An isolate obtained from KUS-F27535 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47643) and used for a pathogenicity test and molecular analyses. To confirm identity of the fungus, genomic DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The internal transcribed spacer (ITS) region of rDNA and the D1/D2 region of the large subunit (LSU) were amplified with the primers ITS1/ITS4 and NL1/LR3, respectively (3), and sequenced. The resulting 635-bp ITS and 680-bp D1/D2 sequences were deposited in GenBank (Accession Nos. KF486539 and KF486538). A GenBank BLAST search revealed that the ITS sequences showed 100% similarity with that of C. infundibulifera (JN943009) and D1/D2 sequences also showed 100% identity with that of C. infundibulifera (JN939193). A sporangiola suspension (2 × 104 cells/ml) was sprayed over three pots of the shrub, kept in a humid chamber for 2 days, and placed in greenhouse (28°C and 80 to 100% RH). Another three potted plants of the same age were sprayed with sterile water and served as controls. After 4 days, typical blossom blight symptoms, identical to the ones observed in the nursery, developed on the inoculated flowers. No symptoms were observed on controls. C. infundibulifera was re-isolated from inoculated plants. Pathogenicity test was conducted twice with the same results, fulfilling Koch's postulates. Choanephora blight caused by C. infundibulifera on H. rosa-sinenesis has been reported in Japan, Myanmar, Nepal, Guinea, and the United States (1). In Korea, there was one record of this fungus on H. syriacus (1). To our knowledge, this is the first report of C. infundibulifera on H. rosa-sinensis in Korea. This pathogen could be a potential threat to the production of this ornamental shrub over a prolonged period of hot and humid weather. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved February 28, 2014. (2) P. M. Kirk. Mycol. Pap. 152:1, 1984. (3) G. Walther et al. Persoonia 30:11, 2013.

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First Report of Leaf Spot of Fan Columbine (Aquilegia flabellata) Caused by Phoma aquilegiicola in Italy

Plant Disease ◽

10.1094/pdis-05-10-0391 ◽

2011 ◽

Vol 95 (7) ◽

pp. 880-880

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. T. Amatulli ◽

M. L. Gullino

Keyword(s):

United States ◽

Morphological Characteristics ◽

Its Region ◽

Apical Part ◽

The United States ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Perennial Herbaceous Plant ◽

Olive Green

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips that is largely present in gardens. It can also be grown for cut flower production. In September of 2008 and 2009, in a private garden located near Biella (northern Italy), a leaf blight was observed. Leaves of infected plants showed extensive, irregular, brown, necrotic lesions, which were slightly sunken with a well-defined border and surrounded by a violet-brown halo. A hole frequently appeared in the center of dried tissues. Lesions, initially measuring 0.5 mm, later expanded up to 15 mm in diameter and eventually coalesced to cover the entire leaf, which curled without falling. At a later stage, stems were also affected, causing death of the apical part of the plant. The disease affected 90% of the plants in the garden. Dark brown, subglobose pycnidia, 116 to 145 μm, containing light gray, ellipsoid, nonseptate conidia measuring 9.0 to 16.2 × 2.6 to 4.2 (average 12.7 × 3.4) μm were observed on symptomatic tissue. On the basis of these morphological characteristics, the fungus was related to the genus Phoma (2). Diseased tissue was excised from the margin of lesions, rinsed in sterile distilled water, and then cultured on potato dextrose agar (PDA) medium at 23 ± 1°C under alternating daylight and darkness (12-h light and 12-h dark). Fungal colonies produced a pale olive green, lightly floccose mycelium, generating clusters of dark olive green swollen cells. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 504-bp segment showed 100% homology with a sequence of Phoma aquilegiicola (GenBank Accession No. GU237735). The nucleotide sequence of our isolate was assigned GenBank Accession No. HM222537. Pathogenicity tests were performed by spraying a mycelium suspension of a homogenate of mycelium (1 × 105 mycelial fragments per ml) obtained from 15-day-old PDA cultures of the fungus on leaves of six healthy 6-month-old potted A. flabellata plants. Six plants inoculated with a homogenate of PDA served as controls. Plants were maintained in a greenhouse in a high humidity chamber for 7 days after inoculation at 23 ± 1°C and under high relative humidity conditions (70 to 90%). The first foliar lesions developed on leaves 4 days after inoculation. After 15 days, 80% of the leaves were severely infected. Control plants remained healthy. The organism reisolated on PDA from leaf lesions was identical in morphology to the isolate used for inoculation. The pathogenicity test was carried out twice. To our knowledge, this is the first report of the presence of P. aquilegiicola on A. flabellata in Italy. Ascochyta aquilegiae (synonym P. aquilegiicola) has been reported on A. vulgaris in Germany (4) and Aquilegia spp. in the United States (3). Currently, the economic importance of this disease is limited, but may become a more significant problem if the use of A. flabellata in gardens increases. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) G. H. Boerema et al. Phoma Identification Manual. Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing, Wallingford, UK, 2004. (3) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (4) R. Laubert. Gartenwelt 34:621, 1930.

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First Report of Powdery Mildew Incited by Erysiphe heraclei on English Ivy (Hedera helix) in Italy

Plant Disease ◽

10.1094/pdis-92-2-0313a ◽

2008 ◽

Vol 92 (2) ◽

pp. 313-313 ◽

Cited By ~ 1

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

J. Rossi ◽

M. L. Gullino

Keyword(s):

Nucleotide Sequence ◽

Powdery Mildew ◽

Its Region ◽

The United States ◽

Tube Length ◽

Herbarium Specimens ◽

Pathogenicity Test ◽

Hedera Helix ◽

First Report ◽

Germ Tubes

Hedera helix L. (Araliaceae) is a common ornamental species that is able to grow in shaded areas and is often used in parks and gardens. During the fall of 2006, severe outbreaks of a previously unknown powdery mildew were observed in several gardens in Liguria (northern Italy). Both surfaces of young leaves of affected plants were covered with dense, white mycelia and conidia. As the disease progressed, infected leaves turned yellow and dropped. Mycelia and conidia were also observed on young stems. Conidia were hyaline, cylindrical, borne singly, and measured 38 to 51 × 12 to 18 (average 42 × 16) μm. Single germ tubes, moderately long (average 26 μm), developed at the end of conidia. Appressoria of germ tubes and hyphae were lobed (three to four lobes). Conidiophores, 68 to 82 × 7 to 8 (average75 × 8) μm, showed foot cells measuring 39 to 60 × 7 to 8 (average 52 × 8) μm, followed by one shorter cell measuring 19 to 28 × 8 to 9 (average 23 × 9) μm. Fibrosin bodies were absent. Chasmothecia were numerous, spherical, amber-colored then brown at maturity, with diameters ranging from 97 to 140 (average 120) μm, containing four asci shortly stalked, 57 to 72 × 32 to 51 (average 65 × 41 μm). Ascospores were ellipsoid and measured 24 to 34 × 15 to 20 (average 30 × 17) μm. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 613-bp fragment showed an E-value of 0.0 with Erysiphe heraclei. The nucleotide sequence has been assigned GenBank Accession No. EU 010381. In GenBank, our nucleotide sequence shows an E-value of 0.0 also with E. betae. However, the comparison of appressorium shape and germ tube length observed on our microorganism with those described for E. betae by Braun (2) suggests that the causal agent of the powdery mildew reported on ivy is E. heraclei. Furthermore, symptoms described on our host, appressorium shape and the length of conidiophores, are different from those of Oidium araliacearum described by Braun (2) on Araliaceae. Inoculations were made by gently pressing diseased leaves onto leaves of five healthy H. helix plants. Three noninoculated plants served as controls. Inoculated and noninoculated plants were maintained in a greenhouse at temperatures between 21 and 25°C. After 15 days, typical powdery mildew colonies 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 the presence of powdery mildew on H. helix caused by E. heraclei in Italy. A powdery mildew caused by E. cichoracearum was previously reported on H. canariensis var. azorica in Italy (3), while a powdery mildew on H. helix caused by O. araliacearum and Golovinomyces orontii, respectively, were observed in the United States (4) and Germany. Herbarium specimens of this disease are available at AGROINNOVA Collection, University of Torino, Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) U. Braun. A Monograph of the Erysiphaceae (Powdery Mildews). Cramer, Berlin, Germany, 1987. (3) C. Nali. Plant Dis. 83:198, 1999. (4) G. S. Saenz and S. T. Koike. Plant Dis. 82:127, 1998.

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Identification of Pyrenophora teres f. maculata, Causal Agent of Spot Type Net Blotch of Barley in North Dakota

Plant Disease ◽

10.1094/pdis-94-4-0480a ◽

2010 ◽

Vol 94 (4) ◽

pp. 480-480 ◽

Cited By ~ 15

Author(s):

Z. H. Liu ◽

T. L. Friesen

Keyword(s):

North Dakota ◽

Leaf Tissue ◽

Causal Agent ◽

Tan Spot ◽

Pathogenicity Test ◽

Pyrenophora Teres ◽

Net Blotch ◽

Hordeum Vulgare L ◽

Single Spore ◽

Spot Type

Net blotch of barley (Hordeum vulgare L.) caused by the fungus Pyrenophora teres (anamorph Drechslera teres) is found in two forms, net form net blotch (NFNB) and spot form net blotch (SFNB). When inoculated on susceptible varieties, P. teres f. teres produces lesions with a characteristic net-like pattern surrounded by necrosis or chlorosis (NFNB), whereas P. teres f. maculata produces lesions consisting of spots surrounded by necrosis or chlorosis (SFNB). Recently, epidemics of SFNB have occurred throughout the world (4). Currently, net blotch is a significant foliar disease of barley in the North Dakota-Northwestern Minnesota agricultural region, a leading barley-production area. Diseased barley leaf tissue was collected annually from 2004 to 2008 in Fargo and Langdon, ND. Diseased leaves were incubated to promote sporulation. Ten single-spore isolates of P. teres collected from each location each year were tested for virulence by inoculation on 20 commonly used barley net blotch differential lines. Among the 100 isolates collected, one isolate collected in Fargo in 2006 (FGOH06Pt-8) and one isolate collected in Langdon in 2008 (LDNH08Pt-4) were identified as P. teres f. maculata due to their induction of spot-type lesions across the differential set. Conidial morphology of the two isolates was similar to P. teres f. teres isolates. A pathogenicity test of all isolates was performed on regional barley cvs. Tradition, Robust, and Lacey as well as barley lines Rika and Kombar (1) as previously described (3). The net form isolate 0-1 and spot form isolate DEN2.6 (obtained from B. Steffenson, University of Minnesota) were used as controls. The P. teres f. teres isolate 0-1 produced typical net type symptoms on all barley lines except the resistant line Rika, in which only small, dark spots were observed. DEN2.6 produced pin-point spot-like lesions with an extensive yellow halo on Robust, Lacey, Rika, and Kombar, but without chlorosis on Tradition. The two newly identified isolates induced elliptical spot-type lesions measuring 3 × 6 mm, larger than those produced by P. teres f. maculata isolate DEN 2.6, suggesting a higher level of virulence. We constructed a neighbor-joining phylogenetic tree using ClustalW2 ( http://www.ebi.ac.uk/ ) based on sequence identity of the internal transcribed spacer (ITS) region from 0-1 (GenBank No. GU014819), DEN2.6 (GenBank No. GU014820), FGOH06Pt-8 (GenBank No. GU014821), and LDNH08Pt-4 (GenBank No. GU014822) as well as P. teres f. maculata, P. teres f. teres, and P. tritici-repentis (causal agent of tan spot of wheat) accessions obtained from GenBank (2). All P. teres isolates clustered together and were clearly separated from the P. tritici-repentis cluster. Isolates FGOH06Pt-8 and LDNH08Pt-4 had identical ITS sequences and differed from DEN2.6 by only a single nucleotide. To our knowledge, this is the first report of P. teres f. maculata in North Dakota. Resistance to SFNB should now be considered in local barley breeding programs and cultivar releases. Reference: (1) M. Abu Qamar. Theor. Appl. Genet. 117:1261, 2008. (2) R. M. Andrie et al. Fungal Genet. Biol. 45:363, 2008. (3) Z. Lai et al. Fungal Genet. Biol. 44:323, 2007. (4) M. S. McLean et al. Crop Pasture Sci. 60:303, 2009.

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First Report of Leaf Spot of Milky Bellflower (Campanula lactiflora) Caused by a Phoma sp. in Italy

Plant Disease ◽

10.1094/pdis-94-5-0638a ◽

2010 ◽

Vol 94 (5) ◽

pp. 638-638

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

C. Pellegrino ◽

M. L. Gullino

Keyword(s):

United States ◽

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Fluorescent Light ◽

Herbaceous Plant ◽

Pathogenicity Test ◽

First Report ◽

Perennial Herbaceous Plant

Campanula lactiflora (milky bellflower), a perennial herbaceous plant in the Campanulaceae, is used in park and gardens and sometimes cultivated for cut flower production. In June 2008, a previously unknown leaf spot was observed on C. lactiflora ‘New Hybrids’ plants from an experimental nursery located near Carmagnola (Torino, northern Italy). Leaves of infected plants showed extensive and irregular, dark brown, necrotic lesions that were slightly sunken with well-defined borders. Lesions initially ranged from 0.5 to 3 mm, eventually coalesced, and covered the entire leaf. Black pycnidia (107 to 116 μm in diameter) containing hyaline, ellipsoid, nonseptate conidia measuring 3.7 to 4.7 × 1.2 to 2.0 (average 4.3 × 1.6) μm were observed. On the basis of these morphological characteristics, the fungal causal agent of the disease could be related to the genus Phoma. In some cases, the basal leaves turned completely necrotic and the plant died. The disease affected 50% of plants. Diseased tissue was excised, immersed in a solution containing 1% sodium hypochlorite for 2 to 3 s, rinsed in water, and then cultured on potato dextrose agar (PDA) medium. A fungus developed that produced a greenish gray mycelium with a white border when incubated under 12 h/day of fluorescent light at 22 to 25°C. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 459-bp segment showed a 100% similarity with the sequence of a Didymella sp. (synonym Mycosphaerella), anamorphic stage of Phoma spp. The nucleotide sequence has been assigned GenBank Accession No. GU128503. Pathogenicity tests were performed by placing 8-mm-diameter mycelial disks removed from PDA cultures of the fungus isolated from infected plants on leaves of healthy potted 4-month-old C. lactiflora ‘New Hybrids’ plants. Eight disks were placed on each plant. Plants inoculated with PDA alone served as controls. Six plants per treatment were used. Plants were covered with plastic bags for 4 days after inoculation and maintained in a growth chamber with daily average temperatures ranging between 23 and 24°C. The first foliar lesions developed on leaves 5 days after inoculation, and after 8 days, 80% of leaves were severely infected. Control plants remained healthy. A Didymella sp. was consistently reisolated from leaf lesions. The pathogenicity test was completed twice. To our knowledge, this is the first report of the presence of a Didymella sp. on C. lactiflora in Italy. Mycosphaerella campanulae and M. minor were reported on C. americana and C. lasiocarpa in the United States (2). The economic importance of the disease currently is limited, but could become a more significant problem in the future if the cultivation of this species becomes more widespread. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989.

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First Report of Stem Rot on Cereus peruvianus monstruosus Caused by Bipolaris cactivora (Petr.) Alcorn in Italy

Plant Disease ◽

10.1094/pdis-06-13-0614-pdn ◽

2014 ◽

Vol 98 (1) ◽

pp. 159-159 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

United States ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Stem Rot ◽

Pathogenicity Test ◽

First Report ◽

Plastic Bags ◽

Blastn Analysis ◽

Dark Green

Cereus peruvianus monstruosus, known as “monster cactus,” family Cactaceae, is grown as a potted plant. In the winter of 2013, a stem rot was observed on a farm located near Ventimiglia (northern Italy) on 80% of 4,000 9-month-old plants grown in trays in a peat substrate. Symptoms consisted of a rapid rot of the upper portion of the stem. Affected stems at first showed yellowish spots that became brown irregular necrotic lesions with well-defined margins. The tissues below the affected areas were blackened and dry but became soft in the presence of high relative humidity. Fungal sporulation on rotted tissues consisted of caespitose, non-branched, septate conidiophores, olivaceous to brown at the base, paler above, measuring 89.0 to 196.9 × 6.2 to 8.7 (average 124.8 × 7.0) μm. Single conidia were borne on terminal cells. At maturity, conidia with 2 to 5 (average 3) septa were brownish-olivaceous, varying in shape from obclavate, fusiform, ellipsoid or sometimes furcate, and measuring 23.4 to 48.6 × 8.0 to 12.6 (average 38.8 × 10.3) μm. Symptomatic tissues were immersed in 1% sodium hypochlorite for 2 to 3 s and rinsed in sterile distilled water, then fragments excised from the margin of internal lesions were cultured on potato dextrose agar (PDA) medium amended with 25 mg/l of streptomycin sulfate and incubated at 20 to 23°C under alternating daylight and darkness (10 h light, 14 h dark). A fungus that was consistently isolated was subcultured on PDA. At maturity, dark green floccose colonies comprised of light brown septate hyphae, 4.2 to 8.1 (average 5.6) μm in width, produced non-branched, pale to dark brown, septate conidiophores, measuring 99.6 to 176.1 × 4.5 to 6.5 (average 146.7 × 5.4) μm. The conidia produced on PDA were similar to those observed on infected tissues and measured 20.6 to 40.7 × 7.5 to 11.4 (average 32.0 × 9.7) μm, with 1 to 3 septa (average 2). On the basis of the morphological characteristics, the fungus was identified as Bipolaris cactivora (Petr.) Alcorn [Syn.: Drechslera cactivora (Petr.) M. B. Ellis] (4). The internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA) was amplified for one isolate using ITS1/ITS4 primers and sequenced (GenBank Accession No. KF041822). BLASTn analysis (1) of the 557-bp segment showed a 99% similarity with the ITS sequence of Bipolaris cactivora HM598679. For pathogenicity tests, 8 mm diameter mycelial disks removed from 15-day-old PDA cultures of the fungus were placed at the wounded stem apexes of three 7-month-old healthy plants (three disks per plant). Three plants inoculated with non-inoculated PDA disks served as controls. Plants were covered with plastic bags and maintained in a growth chamber at 23 ± 1°C with 12 h light/dark. By 8 days after inoculation, all the inoculated stems were rotted and 10 colonies of B. cactivora were re-isolated from infected tissues. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. Several hosts are listed for B. cactivora including C. peruvianus, and the pathogen has been reported in the United States (2) and in South Korea (3). To our knowledge, this is the first report of B. cactivora on C. peruvianus monstruosus in Italy. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. APS Press, St Paul, MN, 1989. (3) I. H. Hyun et al. Res. Plant Dis. 7:56, 2001. (4) A. Sivanesan. Mycopathologia 111:125, 1990.

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First Report of Fruit Rot in Pear Caused by Botryosphaeria dothidea in Italy

Plant Disease ◽

10.1094/pdis-02-12-0130-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 910-910 ◽

Cited By ~ 9

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Fruit Rot ◽

Pathogenicity Test ◽

Botryosphaeria Dothidea ◽

First Report ◽

Irregular Margin ◽

Pyrus Communis L ◽

Pathogenicity Tests

Pear (Pyrus communis L.) is widely grown in Italy, the leading producer in Europe. In summer 2011, a previously unknown rot was observed on fruit of an old cultivar, Spadoncina, in a garden in Torino Province (northern Italy). The decayed area of the fruit was soft, dark brown, slightly sunken, circular, and surrounded by an irregular margin. The internal decayed area appeared rotten and brown and rotted fruit eventually fell. To isolate the causal agent, fruits were soaked in 1% NaOCl for 30 s and 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 20 and 28°C under alternating light and darkness. Colonies of the fungus initially appeared whitish, then turned dark gray. After about 30 days of growth, unicellular elliptical hyaline conidia were produced in pycnidia. Conidia measured 16 to 24 × 5 to 7 (average 20.1 × 5.7) μm (n = 50). The morphological characteristics are similar to those of the fungus Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not. (4). The internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 473-bp segment showed a 100% similarity with the sequence of the epitype of B. dothidea AY236949. The nucleotide sequence has been assigned the GenBank Accession No. JQ418493. Pathogenicity tests were performed by inoculating six pear fruits of the same cultivar (Spadoncina) after surface disinfesting in 1% sodium hypochlorite and wounding. Mycelial disks (8 mm diameter), obtained from 10-day-old PDA cultures of one strain, were placed on wounds. Six control fruits were inoculated with plain PDA. Fruits were incubated at 25 ± 1°C in plastic boxes. The first symptoms developed 3 days after inoculation. After 5 days, the rot was very evident and B. dothidea was consistently reisolated. Noninoculated fruits remained healthy. The pathogenicity test was performed twice. B. dothidea was identified on decayed pears in the United States (2), South Africa, New Zealand, Japan, and Taiwan (3). To our knowledge, this is the first report of the presence of B. dothidea on pear in Italy, as well as in Europe. In Italy, the economic importance of the disease on pear fruit is at present limited, although the pathogen could represent a risk for this crop. References: (1) S. F. Altschul et al. Nucleic Acids Res., 25:3389, 1997. (2) L. F. Grand. Agr. Res. Serv. Techn. Bull. 240:1, 1985. (3) Y. Ko et al. Plant Prot. Bull. (Taiwan) 35:211, 1993. (4) B. Slippers et al. Mycologia 96:83, 2004.

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