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

Plant Disease ◽  
2014 ◽  
Vol 98 (1) ◽  
pp. 159-159 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
P. Pensa ◽  
A. Poli ◽  
M. L. Gullino

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.

Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 880-880
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. T. Amatulli ◽  
M. L. Gullino

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.


Plant Disease ◽  
2010 ◽  
Vol 94 (5) ◽  
pp. 638-638
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
C. Pellegrino ◽  
M. L. Gullino

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.


Plant Disease ◽  
2006 ◽  
Vol 90 (5) ◽  
pp. 684-684
Author(s):  
C. Saude ◽  
M. K. Hausbeck

In April 2005, an Alternaria sp. was isolated from carrot (Daucus carota) roots harvested in the fall of 2004 and held at 1 to 3°C in a storage facility in Newaygo County, MI. The pathogen was readily isolated on water agar from root tissue exhibiting grayish black, sunken lesions. Morphological characteristics were noted 5 to 7 days after single-conidium cultures were established on potato dextrose agar (3). Sixteen Alternaria sp. isolates were recovered. Cultures were dark olive brown, and conidia were pigmented, ellipsoidal, and produced singly or in chains of two. Conidia were 35 to 45 μm long and 15 to18 μm in diameter, usually with three to eight transverse and one to four longitudinal septa. Pathogenicity of isolates was tested on carrot roots in the laboratory and carrot seedlings (cv. Goliath) in the greenhouse. In the laboratory, four surface-sterilized, whole carrot roots were sprayed until runoff with 2 × 106 conidia/ml of each isolate and incubated at 23 to 25°C in a moist chamber for 10 days. Controls were sprayed with sterile distilled water. Ten to fifteen days after inoculation, inoculated carrots exhibited grayish black, sunken lesions, and an Alternaria sp. was reisolated from the margin of the lesions. Controls remained healthy. In the greenhouse, seven pots containing one 2-week-old carrot seedling were watered to saturation and plants were sprayed until runoff with 2 × 106 conidia/ml for each isolate. Control plants were sprayed with sterile distilled water. After inoculation, plants were enclosed in clear plastic bags, placed under 63% woven shade cloth and watered regularly. Black lesions were observed on the foliage 7 days after inoculation, and wilt and death of plants were observed 15 to 30 days after inoculation. Alternaria sp. was reisolated from the foliage of symptomatic plants. Control plants remained healthy. DNA was extracted from all isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region amplified with primers ITS4 and ITS5 and sequenced. A portion of the ITS sequence has been deposited in the NCBI database (GenBank Accession No. DQ394073). A BLAST search of the NCBI database with the ITS sequences revealed A. radicina, Accession No AY154704, as the closest match with 100% sequence similarity. In September 2005, an Alternaria sp. was isolated from black lesions on carrot roots, crowns, and foliage that were collected from fields in Newaygo and Oceana counties, MI. The recovered isolates were morphologically similar to A. radicina isolates obtained from stored carrots in April 2005. First isolated and identified on stored carrots in New York (3), A. radicina is also present in other carrot-producing areas of the United States (1) and was isolated not only from stored carrots but also from carrots in the field (2) and carrot seeds (4). To our knowledge, this is the first report of A. radicina on stored and field carrots in Michigan, which signifies a serious risk to a carrot industry that ranks among the top five in the United States. References: (1) D. F. Farr et al. Fungi on Plants and Plant Produce in the United States.The American Phytopathological Society, St. Paul, MN, 1989. (2) R. G. Grogan and W. C. Snyder. Phytopathology 42:215, 1952. (3) F. C. Meier and E. D. Eddy. Phytopathology 12:157, 1922. (4) B. M. Pryor and R. L. Gilbertson. Plant Dis. 85:18, 2001.


Plant Disease ◽  
2008 ◽  
Vol 92 (6) ◽  
pp. 975-975 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
S. Frati ◽  
M. L. Gullino

Rudbeckia fulgida (orange coneflower), a flowering plant belonging to the Asteraceae, is increasingly used as a border in parks and gardens. In September 2007, severe outbreaks of a previously unknown powdery mildew were observed on plants in a public park in Torino (northern Italy). More than 90% of the plants were affected by the disease. Both surfaces of leaves of affected plants were covered with white mycelia and conidia. As the disease progressed, infected leaves turned yellow and wilted. Mycelia and conidia also were observed on stems and flower calyxes. Conidia were hyaline, ellipsoid, borne in chains (as many as three to four conidia per chain) and measured 34 × 23 (30 to 39 × 21 to 25) μm. Conidiophores measured 129 × 12 (89 to 181 × 11 to 13) μm and showed a foot cell measuring 88 × 12 (48 to 129 × 11 to 13) 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 the primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 619 bp showed a 100% homology with the sequence of Golovinomyces cichoracearum (3). The nucleotide sequence has been assigned GenBank Accession No. EU 233820. Pathogenicity was confirmed through inoculations by gently pressing diseased leaves onto leaves of healthy R. fulgida plants. Twenty plants were inoculated. Fifteen noninoculated plants served as the control. Plants were maintained in a greenhouse at temperatures ranging from 18 to 22°C. Eight 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 R. fulgida in Italy. Powdery mildew on Rudbeckia spp. was previously reported in the United States (4), Poland, and more recently, India and Switzerland. Particularly, in Switzerland the disease has been observed on R. laciniata and R. nitida (2). The economic importance of this disease is currently limited. 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) A. Bolay. Cryptogam. Helv. 20:1, 2005. (3) U. Braun and S. Takamatsu. Schlechtendalia 4:1, 2000. (4) D. F. Farr et al. Page 82 in: Fungi on Plants and Plants Products in the United States. The American Phytopathological Society, St Paul, MN, 1989.


Plant Disease ◽  
2021 ◽  
Author(s):  
Charles Krasnow ◽  
Nancy Rechcigl ◽  
Jennifer Olson ◽  
Linus Schmitz ◽  
Steven N. Jeffers

Chrysanthemum (Chrysanthemum × morifolium) plants exhibiting stem and foliage blight were observed in a commercial nursery in eastern Oklahoma in June 2019. Disease symptoms were observed on ~10% of plants during a period of frequent rain and high temperatures (26-36°C). Dark brown lesions girdled the stems of symptomatic plants and leaves were wilted and necrotic. The crown and roots were asymptomatic and not discolored. A species of Phytophthora was consistently isolated from the stems of diseased plants on selective V8 agar (Lamour and Hausbeck 2000). The Phytophthora sp. produced ellipsoid to obpyriform sporangia that were non-papillate and persistent on V8 agar plugs submerged in distilled water for 8 h. Sporangia formed on long sporangiophores and measured 50.5 (45-60) × 29.8 (25-35) µm. Oospores and chlamydospores were not formed by individual isolates. Mycelium growth was present at 35°C. Isolates were tentatively identified as P. drechsleri using morphological characteristics and growth at 35°C (Erwin and Ribeiro 1996). DNA was extracted from mycelium of four isolates, and the internal transcribed spacer (ITS) region was amplified using universal primers ITS 4 and ITS 6. The PCR product was sequenced and a BLASTn search showed 100% sequence similarity to P. drechsleri (GenBank Accession Nos. KJ755118 and GU111625), a common species of Phytophthora that has been observed on ornamental and vegetable crops in the U.S. (Erwin and Ribeiro 1996). The gene sequences for each isolate were deposited in GenBank (accession Nos. MW315961, MW315962, MW315963, and MW315964). These four isolates were paired with known A1 and A2 isolates on super clarified V8 agar (Jeffers 2015), and all four were mating type A1. They also were sensitive to the fungicide mefenoxam at 100 ppm (Olson et al. 2013). To confirm pathogenicity, 4-week-old ‘Brandi Burgundy’ chrysanthemum plants were grown in 10-cm pots containing a peat potting medium. Plants (n = 7) were atomized with 1 ml of zoospore suspension containing 5 × 103 zoospores of each isolate. Control plants received sterile water. Plants were maintained at 100% RH for 24 h and then placed in a protected shade-structure where temperatures ranged from 19-32°C. All plants displayed symptoms of stem and foliage blight in 2-3 days. Symptoms that developed on infected plants were similar to those observed in the nursery. Several inoculated plants died, but stem blight, dieback, and foliar wilt were primarily observed. Disease severity averaged 50-60% on inoculated plants 15 days after inoculation. Control plants did not develop symptoms. The pathogen was consistently isolated from stems of symptomatic plants and verified as P. drechsleri based on morphology. The pathogenicity test was repeated with similar results. P. drechsleri has a broad host range (Erwin and Ribeiro 1996; Farr et al. 2021), including green beans (Phaseolus vulgaris), which are susceptible to seedling blight and pod rot in eastern Oklahoma. Previously, P. drechsleri has been reported on chrysanthemums in Argentina (Frezzi 1950), Pennsylvania (Molnar et al. 2020), and South Carolina (Camacho 2009). Chrysanthemums are widely grown in nurseries in the Midwest and other regions of the USA for local and national markets. This is the first report of P. drechsleri causing stem and foliage blight on chrysanthemum species in the United States. Identifying sources of primary inoculum may be necessary to limit economic loss from P. drechsleri.


Plant Disease ◽  
2012 ◽  
Vol 96 (2) ◽  
pp. 287-287
Author(s):  
K. S. Han ◽  
J. H. Park ◽  
S. E. Cho ◽  
H. D. Shin

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


Plant Disease ◽  
2007 ◽  
Vol 91 (11) ◽  
pp. 1515-1515 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

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.


Plant Disease ◽  
2006 ◽  
Vol 90 (9) ◽  
pp. 1260-1260 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
D. Minerdi ◽  
M. L. Gullino

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.


Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 433-433 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
D. Bertetti ◽  
M. L. Gullino

Aquilegia flabellata (Ranunculaceae), fan columbine, is a perennial herbaceous plant with brilliant blue-purple flowers with white petal tips. It can also be grown for cut flower production. In April of 2008, in several nurseries located near Biella (northern Italy), a leaf blight was observed on 10 to 15% of potted 30-day-old plants grown on a sphagnum peat substrate at 15 to 20°C and relative humidity of 80 to 90%. Semicircular, water-soaked lesions developed on leaves just above the soil line at the leaf-petiole junction and later along the leaf margins. Lesions expanded over several days along the midvein until the entire leaf was destroyed. Blighted leaves turned brown, withered, and abscised. Severely infected plants died. Diseased tissue was disinfested for 10 s in 1% NaOCl, rinsed with sterile water, and plated on potato dextrose agar (PDA) amended with 25 mg/liter streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently recovered, then transferred and maintained in pure culture. Ten-day-old mycelium grown on PDA at 22 ± 1°C appeared light brown, rather compact, and had radial growth. Sclerotia were not present. Isolates obtained from affected plants successfully anastomosed with tester isolate AG 4 (AG 4 RT 31, obtained from tobacco plants). Results were consistent with other reports on anastomosis reactions (2). Pairings were also made with tester isolates of AG 1, 2.1, 2.2, 3, 6, 7, 11, and BI with no anastomoses observed between the recovered and tester isolates. The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 648-bp fragment showed a 100% homology with the sequence of R. solani AG-4 AB000018. The nucleotide sequence has been assigned GenBank Accession No. FJ 534555. For pathogenicity tests, the inoculum of R. solani was prepared by growing the pathogen on PDA for 10 days. Five plants of 30-day-old A. flabellata were grown in 3-liter pots. Inoculum consisting of an aqueous suspension of PDA and mycelium disks (5 g of mycelium + agar per plant) was placed at the collar of plants. Five plants inoculated with water and PDA fragments alone served as control treatments. Plants were maintained in a greenhouse at temperatures between 20 and 24°C. The first symptoms, similar to those observed in the nursery, developed 7 days after the artificial inoculation. R. solani was consistently reisolated from infected leaves and stems. Control plants remained healthy. The pathogenicity test was carried out twice with similar results. The presence of R. solani AG1-IB on A. flabellata has been reported in Japan (4), while in the United States, Rhizoctonia sp. is described on Aquilegia sp. (3). This is, to our knowledge, the first report of leaf blight of A. flabellata caused by R. solani in Italy as well as in Europe. 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, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) E. Imaizumi et al. J. Gen. Plant Pathol. 66:210, 2000.


Plant Disease ◽  
2010 ◽  
Vol 94 (6) ◽  
pp. 788-788 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. T. Amatulli ◽  
M. L. Gullino

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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