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

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 Puccinia pelargonii-zonalis Causing Rust on Pelargonium × hortorum in Xinjiang Province, China

Plant Disease ◽

10.1094/pdis-09-21-1965-pdn ◽

2021 ◽

Author(s):

Jin Sun ◽

Guo Qin Wang ◽

Jia Ge Song ◽

Biao Xu

Keyword(s):

Disease Incidence ◽

Large Subunit ◽

Economic Value ◽

The United States ◽

Concentric Circle ◽

Pathogenicity Test ◽

First Report ◽

Potted Plants ◽

Light Yellow ◽

Initial Symptoms

Geranium (Pelargonium × hortorum) is an ornamental plant cultivated throughout world. In May 2019, typical symptoms of rust with nearly 90% disease incidence were observed on leaves of Pel. × hortorum growing in pots in a scenic spot in the ancient city of Kashgar, Xinjiang province, China. Early symptoms of the disease appeared small lesions with pale-yellow halos on the lower leaf surfaces. Infected tissues subsequently formed a concentric circle of sori on lesion reaching a final diameter of 0.5 to 1.5 cm, which resulted in leaf blighting and premature. Two representative specimens were deposited in the Mycological Herbarium of Tarim University (HMUT 8001 and HMUT 8002). Urediniospores were globose or subglobose to ovoid, light brown, echinulate, thin-walled with two conspicuous subequatorial pores, 19.8 to 27.3 × 17.8 to 23.6 μm (24.3 × 21.7 μm average, n=30). Telia and teliospores were not seen. The sizes and characteristics of urediniospores were similar with those of Puccinia pelargonii-zonalis as described by Doidge (1926). To confirm the identification, genomic DNAs were extracted directly from sori on diseased leaves from isolates HMUT 8001 and HMUT 8002. The part of the nuclear large subunit (LSU, 28S) rDNA was amplified and sequenced following Aime et al. (2006), and deposited in GenBank (Accession Nos. MT648851 and MT648852). An NCBI blast search indicated that 99.6% identity of the LSU (772/775 nucleotides) with Puc. pelargonii-zonalis on Pel. hortorum (KX999887) from Australia (Marin-Felix et al. 2017). The phylogenetic analysis based on the LSU sequences using Maximum-Likelihood and Bayesian methods placed the Xinjiang isolates from Pel. × hortorum in the same clade with the reference sequences of Puc. pelargonii-zonalis. Pathogenicity test was confirmed by Koch’s postulates. Leaves of three healthy potted plants were sprayed with a spore suspension (2 × 105 spores/ml). Sterile water was sprayed on three healthy seedlings as controls. Inoculated and control plants were covered with a plastic bag and placed in a moist chamber with 90% relative humidity at 25°C. Initial symptoms were observed with small light-yellow spots on the upper surface after 15 days, but not in the control plants. Puc. pelargonii-zonalis has been previously reported on Pel. × hortorum in Argentina, Australia, Canada, Georgia, Greece, India, Mexico, New Zealand, Norway, Portugal, South Africa, the United States, and Venezuela (Farr and Rossman 2021). Geranium rust was first found in Yunnan province in southwestern China (Zhou and Zhuang 2005). However, this is the first report of geranium rust in Xinjiang province in northwestern China. The occurrence of the disease seriously affected their ornamental and economic value.

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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 Zonate Leaf Spot Caused by Hinomyces moricola on Japanese Hop in Korea

Plant Disease ◽

10.1094/pdis-01-13-0021-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1117-1117 ◽

Cited By ~ 1

Author(s):

S. E. Cho ◽

J. H. Park ◽

S. H. Hong ◽

H. D. Shin

Keyword(s):

Its Region ◽

The United States ◽

Infected Leaf ◽

Invasive Weed ◽

Voucher Specimen ◽

First Report ◽

Leaf Spots ◽

Initial Symptoms ◽

Wide Range ◽

Humulus Japonicus

Japanese hop (Humulus japonicus Siebold & Zucc. = H. scandens (Lour.) Merr.), native to East Asia, is an annual, climbing or trailing vine. The vines can spread to cover large areas of open ground or low vegetation, eventually blanketing the land and vegetation. Pollen of H. japonicus is allergenic, and this species is considered as one of the important causes of pollinosis in Korea and China. It is a notorious invasive weed in the United States and also in France, Hungary, and Italy (1). In September 2012, zonate leaf spots were observed on Japanese hops growing in wetlands in Yeongdong County of Korea. A voucher specimen was preserved in the Korea University Herbarium (KUS-F26901). Initial symptoms included grayish-green to grayish-brown spots without border lines. As the lesions enlarged, they coalesced, leading to leaf blight. Sporophores on the leaf lesions were dominantly hypophyllous, rarely epiphyllous, solitary, erect, easily detachable, and as long as 700 μm. The upper portion of the sporophores consisted of a pyramidal head was ventricose, 320 to 520 μm long and 110 to 150 μm wide. The fungus was isolated from leaf lesions and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 18°C without light, but conidia were not observed in culture. These morphological and cultural characteristics were consistent with those of Hinomyces moricola (I. Hino) Narumi-Saito & Y. Harada (= Cristulariella moricola (I. Hino) Redhead) (3,4). An isolate was preserved in the Korean Agricultural Culture Collection (Accession No. KACC46955). Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The resulting sequence of 452 bp was deposited in GenBank (Accession No. KC460209). A BLAST search in GenBank revealed that the sequence showed an exact match with those of C. moricola (JQ036181 ex Acer negundo and JQ036182 ex Glycine max). To determine the pathogenicity of the fungus, according to the procedure of Cho et al. (2), sporophores with the pyramidal head were carefully detached from a lesion on the naturally infected leaf using a needle. Each sporophore was transferred individually onto five places of four detached healthy leaves. The leaves were placed in dew chambers and incubated at 16°C. Symptoms were observed after 2 days on all inoculated leaves. A number of sporophores and immature sclerotia which were morphologically identical to the ones observed in the field were formed on the abaxial surface of the leaf 2 weeks after inoculation. The pathogen was reisolated from lesions on the inoculated leaves, confirming Koch's postulates. No symptoms were observed on the control leaves kept in humid chambers for 2 weeks. H. moricola was known to cause zonate leaf spots and defoliation on a wide range of woody and annual plants (3). To the best of our knowledge, this is the first report of Hinomyces infection on Japanese hops in Korea. References: (1) Anonymous. Humulus japonicus (Cannabaceae): Japanese hop. Eur. Medit. Plant Prot. Org. (EPPO). 2012. (2) S. E. Cho et al. Plant Dis. 96:906, 2012. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., Online publication, ARS, USDA, Retrieved December 8, 2012. (4) S. A. Redhead. Can. J. Bot. 53:700, 1975.

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

Plant Disease ◽

10.1094/pdis-92-5-0832b ◽

2008 ◽

Vol 92 (5) ◽

pp. 832-832 ◽

Cited By ~ 13

Author(s):

A. Aroca ◽

R. Raposo ◽

D. Gramaje ◽

J. Armengol ◽

S. Martos ◽

...

Keyword(s):

Elongation Factor ◽

Aerial Mycelium ◽

Its Region ◽

Pathogenic Fungi ◽

Vascular Lesions ◽

Malt Extract ◽

Pathogenicity Test ◽

Lasiodiplodia Theobromae ◽

First Report ◽

Mother Plants

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

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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 panicle rot caused by Fusarium asiaticum on foxtail millet in China

Plant Disease ◽

10.1094/pdis-04-21-0856-pdn ◽

2021 ◽

Author(s):

Fanxin Kong ◽

Haijin Zhang ◽

Zhi Liu ◽

Guoqiu Chen ◽

Jing Xu

Keyword(s):

Foxtail Millet ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Its Region ◽

Liaoning Province ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Dark Cycle ◽

First Report ◽

Fusarium Asiaticum

Foxtail millet [ Setaria italica (L.) P. Beauv.] is one of the most important nutritious food crops. It is used for wine and health products in China. In August of 2019, panicle rot symptoms with up to 85% of panicles infected were observed on foxtail millet (cultivar Chaogu 8) in a commercial field located in Chaoyang city of Liaoning Province, China. Typical disease symptoms included brown spots on spikelets at early stages and brown-colored withering and rot of whole panicles at late stages, with the symptoms being more severe at the tip of the panicles. Under high humidity conditions, pink or salmon-colored molds developed on panicles. Symptomatic spikelet pieces were surface-disinfested with 70% ethanol for 1 min followed by 2% NaOCl for 3 min, rinsed with sterilized water for three times, and placed on potato dextrose agar (PDA) medium at 25°C. After 5 days, colonies turned pink to dark red with fluffy aerial mycelium and pigmentation with the age. Ten pure cultures were obtained from single conidia of mycelium grown on carnation leaf agar (CLA) medium at 25°C under a 12-h light-dark cycle using an inoculation needle under stereomicroscope. Macroconidia were hyaline, falcate with foot cells, 3–5 septate and size: 28.5- 44.0 μm × 3.8 - 4.9 μm. Chlamydospores were globose to subglobose (5.4 to 13.8 μm). No microconidia were produced on CLA. Black, ostiolate subglobose perithecia were formed on CLA after one month of incubation at 20°C under a 12-h light-dark cycle. Morphological characteristics of the fungus were in agreement with the description of Fusarium asiaticum (O’Donnell et al. 2004; Leslie and Summerell 2006). To validate this identification, partial translation elongation factor 1 alpha (TEF1-a) gene, and rDNA internal transcribed spacer (ITS) region of five isolates were amplified and sequenced (O’Donnell et al. 2015; White et al.1990). Identical sequences were obtained, and the sequence of one representative isolate (JGF-3) was submitted to GenBank. BLASTn analysis of both TEF sequence (MW685833) and ITS sequence (MW423687), revealed 100% sequence identity with F. asiaticum KT380120 and MT322117, respectively. Pathogenicity test were conducted on cultivar Chaogu 8 of foxtail millet. Inoculum was prepared from the culture of JGF-3 incubated in 2% mung beans juice on a shaker (140 rpm) at 25°C for 48 h. Conidial suspension (5 × 105 conidia per ml) was prepared and sprayed onto the panicles of 20 plants at the initial flowering stage and 20 additional plants that were sprayed with distilled water served as the non-inoculated controls. Treated plants were covered with plastic bags for 48 h and maintained at a greenhouse with day and night temperatures of 26 and 24°C, respectively. Two weeks after inoculation, all inoculated panicles exhibited symptoms similar to the syptoms observed in the field. No symptoms were observed in the non-inoculated control plants. The experiment was repeated twice with similar results. F. asiaticum was reisolated from the inoculated plants and its morphological characteristics matched those of the original isolates; the fungus was not reisolated from the non-inoculated plants. To our knowledge, this is the first report of F. asiaticum causing panicle rot of foxtail millet in China. To date, the disease has been observed to be present in Fuxin and Tieling city of Liaoning Province. Panicle rot can become an important disease in foxtail millet in China. References: O’Donnell, K., et al. 2004. Fungal Genetics and Biology 41: 600. Leslie, J. F., and Summerell, B. A. 2006. The Fusarium laboratory manual. Blackwell Publishing, Ames, pp 176-179. O’ Donnell, K., et al. 2015. Phytoparasitica 43: 583. White, T. J., et al. 1990. Academic Press, San Diego, CA, pp 315-322.

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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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First Report of Sclerotinia sclerotiorum on Calceolaria integrifolia in Italy

Plant Disease ◽

10.1094/pdis-92-7-1133c ◽

2008 ◽

Vol 92 (7) ◽

pp. 1133-1133

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

Sclerotinia Sclerotiorum ◽

Soil Surface ◽

Its Region ◽

The United States ◽

First Report ◽

Potted Plants ◽

Initial Symptoms ◽

Mycelial Plug ◽

Blastn Analysis ◽

White Mycelium

Calceolaria integrifolia L. is an ornamental species grown as a potted plant in Liguria, northern Italy. In the winter of 2006, extensive chlorosis was observed on approximately 10% of the 10-month-old potted plants in a commercial greenhouse. Initial symptoms included stem necrosis and darkening of leaves. As stem and foliar necrosis progressed, infected plants wilted and died. Wilt occurred on young plants within a few days after the initial appearance of symptoms. Infected plants were characterized by the presence of soft, watery tissues that became covered with white mycelium and dark sclerotia. The diseased stem tissue was surface sterilized for 1 min in 1% NaOCl and plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. Sclerotinia sclerotiorum (Lib.) de Bary (3) was consistently recovered from infected stem pieces. Sclerotia observed on infected plants measured 0.7 to 1.0 × 2.8 to 4.4 mm (average 1.6 to 2.1 mm). Sclerotia produced on PDA measured 1.0 to 1.1 × 3.0 to 4.2 mm (average 1.7 to 2.3 mm). The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 522-bp amplicon resulted in 100% homology with the sequence of S. sclerotiorum. The nucleotide sequence has been assigned GenBank Accession No. EU 627004. Pathogenicity of two isolates obtained from infected plants was confirmed by inoculating 10 120-day-old plants grown in individual 14-cm-diameter pots maintained in a greenhouse under partial shade. Inoculum consisted of 1 cm2 of mycelial plugs excised from a 10-day-old PDA culture of each isolate. Plants were inoculated by placing a mycelial plug on the soil surface around the base of each plant. Ten plants were inoculated per isolate and an equal number of noninoculated plants served as controls. The trial was repeated once. All plants were kept at temperatures ranging between 8 and 17°C (average 12.5°C) and watered as needed. All inoculated plants developed leaf yellowing within 8 days after inoculation, soon followed by the appearance of white mycelium and sclerotia, and then by wilt. Control plants remained symptomless. S. sclerotiorum was reisolated from the stems of inoculated plants. S. sclerotiorum was reported previously on a Calceolaria sp. in the United States (2). To our knowledge, this is the first report of white mold on C. integrifolia in Italy. The economic importance of this disease is currently limited. References (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) Anonymous. USDA Agric. Handb. 165:441, 1960. (3) N. F. Buchwald. Den. Kgl. Veterin.er-og Landbohojskoles Aarsskrift 75, 1949.

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