First Report of Leaf Spot and Root Rot Caused by Phoma betae on Beta vulgaris subsp. vulgaris (Garden Beet Group) in Italy

In the winter of 2007 in Piedmont (northern Italy), symptoms of a previously unknown disease were observed on beet (Beta vulgaris L. subsp. vulgaris) (garden beet group) grown under a tunnel on several commercial farms near Cuneo. First symptoms appeared on 1-month-old plants, occurring as brown, round-to-oval spots as much as 2 cm in diameter with dark concentric rings near the perimeter. Small, dark pycnidia were present throughout the spots in concentric rings. Generally, older, lower leaves were affected more than the younger ones. Ten to fifteen percent of the plants were affected. Symptoms on the roots began near the crown as small, dark, sunken spots that became soft and water soaked. Eventually, spots on the roots turned dark brown to black and black lines separated diseased and healthy tissues. Older infected tissues were black, dry, shrunken, and spongy. Pycnidia were not observed on affected roots. From infected leaves and roots, a fungus was consistently isolated on potato dextrose agar (PDA) amended with 25 mg/l of streptomycin. The fungus was grown on PDA and maintained at 22°C (12 h of light, 12 h of dark). After 10 days, black pycnidia (130 to 328 [204] μm in diameter) developed, releasing abundant hyaline, elliptical, nonseptate conidia measuring 3.9 to 6.7 (5.1) × 2.4 to 5.9 (3.6) μm. On the basis of its morphological characteristics, the fungus was identified as a Phoma sp. (1). The internal transcribed spacer (ITS) region was amplified using primers ITS4/ITS6 (2) and sequenced. BLASTn analysis of the 557 bp obtained showed an E-value of 0.0 with Phoma betae. The nucleotide sequence has been assigned GenBank Accession No. EU003450. Pathogenicity tests were performed by spraying leaves of healthy 20-day-old potted B. vulgaris plants with a spore and mycelial suspension (1 × 106 spores or mycelial fragments per ml). Noninoculated plants sprayed only with water served as controls. Fifteen plants (three per pot) were used for each treatment. Plants were covered with plastic bags for 5 days after inoculation and kept in a growth chamber at 20°C. Symptoms previously described developed on leaves of all inoculated plants 5 days after inoculation, while control plants remained healthy. Later, pycnidia and conidia, with the same dimensions and characteristics previously described, were observed on the infected leaves. The fungus was consistently reisolated from the lesions of the inoculated plants. The pathogenicity test was carried out twice. P. betae on B. vulgaris var. cycla has been reported in Canada (3) as well as in other countries. The same pathogen was reported in Italy on sugar beet (2). References: (1) G. H. Boerema and G. J. Bollen. Persoonia 8:111, 1975. (2) A. Canova. Inf. Fitopatol. 16:207, 1966. (3) D. E L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (4) J. R. Howard et al. Diseases of Vegetable Crops in Canada. Canadian Phytopathological Society, 1994.

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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 Leaf Spot Caused by Phoma multirostrata on Fuchsia × hybrida in Italy

Plant Disease ◽

10.1094/pdis-94-3-0382a ◽

2010 ◽

Vol 94 (3) ◽

pp. 382-382 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

G. Gilardi ◽

M. L. Gullino

Keyword(s):

Leaf Surface ◽

Morphological Characteristics ◽

Its Region ◽

Northern Italy ◽

Pathogenicity Test ◽

First Report ◽

Plastic Bags ◽

Pathogenicity Tests ◽

Necrotic Spots ◽

Mycelial Suspension

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

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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 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 Black Rot Caused by Phomopsis cucurbitae on Cantaloupe (Cucumis melo) in the Piedmont Region of Northern Italy

Plant Disease ◽

10.1094/pdis-06-11-0481 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1317-1317

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Cucumis Melo ◽

Morphological Characteristics ◽

Its Region ◽

Northern Italy ◽

Pathogenicity Test ◽

First Report ◽

Sterile Needle ◽

Pathogenicity Tests ◽

Cucumis Melo L ◽

Piedmont Region

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

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First Confirmed Report of Cercospora Blight of Asparagus Caused by Cercospora asparagi in Korea

Plant Disease ◽

10.1094/pdis-10-12-0984-pdn ◽

2013 ◽

Vol 97 (3) ◽

pp. 428-428 ◽

Cited By ~ 4

Author(s):

K. S. Han ◽

B. S. Kim ◽

J. H. Park ◽

H. D. Shin

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

Culture Collection ◽

Asparagus Officinalis ◽

Organic Farm ◽

Plastic Bags ◽

Japanese Workers ◽

Close Phylogenetic Relationship ◽

Voucher Specimens ◽

Pathogenicity Tests

In September 2011, hundreds of asparagus (Asparagus officinalis L.) showing symptoms of blight with nearly 100% incidence (percentage of plants affected) were found in polyethylene tunnels at an organic farm in Gangneung City of Korea. Lesions on needles and branches of the ferns were small, elliptic to subcircular, pale tan to brown with reddish brown borders, and became gray and cottony due to heavy sporulation under continuous high humidity. Infection caused premature defoliation and weakened plant vigor. The damage purportedly due to this disease has reappeared with confirmation of the causal agent made again in 2012. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were well developed, consisting of brown cells, and were 10 to 30 μm wide. Conidiophores were fasciculate (n = 2 to 12), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones, or 1 to 2 times geniculate in longer ones, 40 to 260 μm long, 3.5 to 5.5 μm wide, and 1- to 6-septate. Conidia were hyaline, cylindric to acicular, straight in shorter ones, curved in longer ones, truncate to obconically truncate at the base with darkened hila, guttulate, 2- to 12-septate, and 40 to 220 × 3 to 5 μm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora asparagi Sacc. (1). Voucher specimens were housed at the Korea University herbarium (KUS). An isolate from KUS-F26046 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC46400). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 497 bp was deposited in GenBank (Accession No. JX964995). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. For pathogenicity tests, conidial suspensions (105 conidia/ml) were prepared by culturing the fungus on V8 juice agar (2) for 3 weeks. Five plants were inoculated with conidial suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain 100% RH for 24 h and then transferred to a greenhouse. Typical symptoms of necrotic lesions appeared on the inoculated plants 6 days after treatment, and were identical to the ones observed in the field. C. asparagi was reisolated from symptomatic tissues, confirming Koch's postulates. No symptoms were observed on control plants. The disease has been reported through the regions of the world where asparagus is grown (3). In Korea, the disease was recorded in 1928 by Japanese workers under Cercosporina asparagicola Speg. (regarded as synonymous with Cercospora asparagi) with brief notes (4). Though one sample of asparagus was sent to the author (KSH) for diagnosis in 2009 summer and determined to be infected with C. asparagi (unpublished data), there has been no additional finding of the disease in Korea for the last 82 years. To our knowledge, this is the first confirmed report of Cercospora blight of asparagus caused by C. asparagi in Korea. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) C. J. Cooperman and S. F. Jenkins. Phytopathology 76:617, 1986. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved October 20, 2012. (4) K. Nakata and S. Takimoto. Bull. Agric. Exp. Stat. Korea 15:1, 1928.

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First Report of Didymella bryoniae Causing Gummy Stem Blight of Chayote in Taiwan

Plant Disease ◽

10.1094/pdis-02-12-0185-pdn ◽

2012 ◽

Vol 96 (10) ◽

pp. 1578-1578 ◽

Cited By ~ 2

Author(s):

Y. C. Tsai ◽

J. F. Chen

Keyword(s):

Morphological Characteristics ◽

Pathogenic Fungi ◽

Molecular Data ◽

Pathogenicity Test ◽

Didymella Bryoniae ◽

Stem Blight ◽

Gummy Stem Blight ◽

Plastic Bags ◽

Pathogenicity Tests ◽

Stems And Leaves

Chayote (Sechium edule (Jacq.) Swartz, Cucurbitaceae), originally native to Mexico, is an important vegetable known as “dragon-whisker vegetable” and is cultivated for its shoots in Ji-an, Hualien County in eastern Taiwan. In June 2010, 70 to 80% of the chayote plants grown in Ji-an developed necrotic spots on stems, leaves, and fruits. The disease was severe during the warm and rainy season from June to August. The symptoms on stems, leaves, and fruits were water-soaked lesions that eventually dried up, cracked, and produced perithecia on necrotic tissues. A single ascospore was isolated from perithecia harvested from diseased stems and cultured on potato dextrose agar (PDA) at 25°C for 1 month. Colonies of three isolates (SE5, SE6, and SE7) were white to olivaceous green bearing unicellular conidia measuring 2 to 5 × 3 to 10 μm, which is consistent with the morphological characteristics of Didymella bryoniae (Auersw.) Rehm (anamorph Phoma cucurbitacearum (Fr.:Fr.) Sacc.) (1,2,3). DNA of SE5, SE6, and SE7 isolates were obtained using microwave-based method (4). The internal transcribed spacer (ITS) rDNA (GenBank accessions AB714984, AB714985, and AB714986), PCR-amplified using primers ITS1 and ITS4, had 98 to 99% nucleotide sequence identity with D. bryoniae (GenBank Accession Nos. GU045304 and GU592001). A pathogenicity test was conducted in a greenhouse with temperature ranging from 20 to 30°C. Three-day-old mycelial plugs (5 × 5 mm) of the three isolates were placed on the needle-pricked wounds of stems and leaves of 36 4-month-old potted chayote plants wrapped in plastic bags to maintain 100% relative humidity for 2 days. Six days after inoculation, water-soaked lesions formed on the stems and leaves. Controls inoculated with sterile water had no symptoms. The fungus reisolated from the lesions of diseased stems and leaves had morphological characteristics of D. bryoniae. Based on the results of morphology, molecular data, and pathogenicity tests, we reported for the first time to our knowledge that gummy stem blight of chayote is caused by D. bryoniae in Taiwan. References: (1) J. W. Huang and W. R. Hsieh. Plant Prot. Bull. 27:325, 1985. (2) A. P. Keinath et al. Phytopathology 85:364, 1995. (3) E. Punithalingam and P. Holliday. P. 332 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1972. (4) S. R. Tendulkar et al. Biotechnol. Lett. 22:1941, 2003.

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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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First Report of Leaf Spot of Saponaria officinalis Caused by Alternaria nobilis in Italy

Plant Disease ◽

10.1094/pdis-09-12-0839-pdn ◽

2013 ◽

Vol 97 (3) ◽

pp. 424-424 ◽

Cited By ~ 2

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Morphological Characteristics ◽

Pathogenicity Test ◽

Academic Press ◽

First Report ◽

Perennial Plant ◽

Plastic Bags ◽

Pcr Product ◽

Pathogenicity Tests ◽

Alternaria Sp ◽

Saponaria Officinalis

Saponaria officinalis (Vize) Simmons (common name bouncingbet) is a low maintenance perennial plant belonging to the Caryophyllaceae family, typically grown in parks and gardens. During the summers of 2011 and 2012, extensive necrosis were observed on leaves of plants grown in private gardens, near Biella (northern Italy). The disease affected 90% of 1- to 2-year-old plants. The first symptoms were usually pale brown lesions 1 to 5 mm in diameter and sometimes coalesced. Lesions were circular to irregular with a dark purple halo, with infected leaves eventually turning chlorotic. The conidia observed on infected leaves were olivaceous brown and obclavate, with a beak. Conidia showed 8 to 15 (average 12) transverse and 4 to 14 (average 11) longitudinal septa, with slight constrictions connected with septa, and were 78.3 to 177.7 (average 135.5) × 19.0 to 34.3 (average 26.5) μm. The beak was 20.0 to 62.2 (average 33.7) μm in length, with 0 to 6 (average 3) transverse septa and no longitudinal septa. The fungus was consistently isolated from infected leaves on potato dextrose agar (PDA). The isolate, grown for 14 days at 20 to 24°C with 10 h of darkness and 14 h of light on sterilized host leaves plated on PDA, produced conidiophores single, unbranched, flexuous, septate with conidia in short chains, similar to those observed on the leaves and previously described. On the basis of its morphological characteristics, the pathogen was identified as Alternaria sp. (3). DNA was extracted using Nucleospin Plant Kit (Macherey Nagel) and PCR carried out using ITS 1/ITS 4 primer (4). A 542-bp PCR product was sequenced and a BLASTn search confirmed that the sequence corresponded to A. dianthi (AY154702), recently renamed A. nobilis (2). The nucleotide sequence has been assigned the GenBank Accession No. JX647848. Pathogenicity tests were performed by spraying leaves of healthy 3-month-old plants of S. officinalis with an aqueous 2 × 105 spore/ml suspension. The inoculum was obtained from cultures of the fungus grown on PDA amended with host leaves for 14 days, in light-dark, at 22 ± 1°C. Plants sprayed only with water served as controls. Four pots (1 plant/pot) were used for each treatment. Plants were covered with plastic bags for 4 days after inoculation and maintained in a glasshouse at 21 ± 1 °C. Lesions developed on leaves 9 days after inoculation with the spore suspension, whereas control plants remained healthy. A. nobilis was consistently reisolated from these lesions. The pathogenicity test was carried out twice. The presence of A. dianthi was reported on S. officinalis in Denmark (1) and Turkey. This is, to our knowledge, the first report of A. nobilis on S. officinalis in Italy. The presence and importance of this disease is, at present, limited. References: (1) P. Neergaard. Danish species of Alternaria and Stemphylium. Oxford University Press, 1945. (2) E. G. Simmons. Mycotaxon 82:7, 2002. (3) E. G. Simmons. Alternaria: An Identification Manual. CBS Biodiversity Series 6, Utrecht, The Netherlands, 2007. (4) T. J. White et al. In: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Leaf Spot of Orange Coneflower (Rudbeckia fulgida) Caused by a Phoma sp. in Italy

Plant Disease ◽

10.1094/pdis-94-6-0788a ◽

2010 ◽

Vol 94 (6) ◽

pp. 788-788

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. T. Amatulli ◽

M. L. Gullino

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Pathogenicity Test ◽

First Report ◽

Humidity Chamber ◽

Pathogenicity Tests ◽

Symptomatic Tissue ◽

White Mycelium

Rudbeckia fulgida (orange coneflower) is an herbaceous species (Asteraceae) grown in full sun in flower beds and borders in gardens. In the summer of 2009, a previously unknown leaf spot was observed on R. fulgida plants in three private gardens located near Biella (northern Italy). Leaves of infected plants showed extensive and irregular, dark brown, necrotic lesions that were slightly sunken with a well-defined border. Lesions initially ranged from 0.5 to 3 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 plant. The disease affected 90% of plants. Dark brown pycnidia, 68 to 195 × 60 to 165 (average 135 × 117) μm in diameter, containing hyaline (light gray in mass), and ellipsoid, nonseptate conidia measuring 4.0 to 7.0 × 2.4 to 3.5 (average 5.4 × 3.0) μm were observed on symptomatic tissue. On the basis of these morphological characteristics, the fungus was related to the genus Phoma. Diseased tissue was excised from the margin of lesions, immersed in a solution containing 1% sodium hypochlorite for 2 to 3 s, rinsed in sterile distilled water, and then cultured on potato dextrose agar (PDA) medium. Fungal colonies initially produced a white mycelium that became greenish gray when incubated at temperatures ranging between 22 and 25°C under alternating daylight and darkness (13 h of light and 11 h of dark). After 14 days of incubation, unicellular, cylindrical or truncated cone-shaped, light brown chlamydospores measuring 6 to 12 μm in diameter developed in long chains. The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 498-bp segment showed 100% homology with a sequence of a Phoma sp. (EF585395). The nucleotide sequence of our isolate was assigned GenBank Accession No. GU573979. Pathogenicity tests were performed by placing 100 ml of a water homogenate of mycelium (1 × 105 mycelial fragments/ml) obtained from 15-day-old PDA cultures of the fungus on leaves of three healthy 4-month-old potted R. fulgida plants. Three 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 temperatures ranging from 18 to 22°C and under high relative humidity conditions (70 to 90%). The first foliar lesions developed on leaves 7 days after inoculation, and after 10 to 12 days, 80% of 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 a Phoma sp. on R. fulgida in Italy. Mycosphaerella ligulicola was reported on Rudbeckia sp. (2), while M. rudbeckiae and Phoma exigua have been reported on R. hirta (3). Currently, the economic importance of this disease is limited, but may become a more significant problem if the cultivation of this species increases. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) C. G. C. Chesters and J. P. Blakeman. Ann. Appl. Biol. 60:385, 1967. (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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