First Report of Septoria pistaciae Causing Leaf Spot of Pistachio in Egypt

In May 2006, leaf spots were observed on approximately 60% of 8-year-old pistachio (Pistacia vera L.) trees in an orchard located at Rhafah, north of Sinai, Egypt. These spots were typically circular, or occasionally irregular, with white-to-light tan or gray centers and a purple or brown border measuring 1 to 5 mm in diameter, or occasionally larger on the upper leaf surface. A narrow, brown border surrounded the spot, and with age, the lesion cracked. Spots occasionally turned brown and closed to form blotches. Fungal isolates from the leaf spots were identified as Septoria pistaciae on the basis of characteristics of pycnidia and conidia. From the leaf spots, numerous black pycnidia were found that produced hyaline conidia, 3 to 7 septate, generally filiform although tapering at one end, and measuring 46 to 75 × 3 to 4 μm. Pycnidia were dark, separate, globe shaped with an ostiole from which conidia were extruded, and erupted through the surface of infected plant tissue. Conidia were produced on short conidiophores. Single conidial isolations onto 2% malt agar consistently formed slow-growing, dark green colonies. To confirm the pathogenicity of the isolate of S. pistaciae, a suspension of 5 × 105 conidia per ml in water was applied at 1 ml per leaf to 20 leaves of 10 rooted cuttings of pistachio trees in 30-cm pots. Ten controls were misted with water only. All plants were covered with plastic bags for 48 h on a greenhouse bench. Greenhouse temperatures ranged from 15 to 20°C with a 16-h photoperiod. After 6 days, all inoculated plants developed symptoms, and the fungus was reisolated from lesions. No symptoms were observed on control plants. S. pistaciae was previously reported on pistachio in Texas and Arizona (2,3). Reports included mention of its occurrence in the United States (California), Asia (Armenia Republic of Georgia, India, Israel, Kazakhstan, Kirgizstan, Syria, Turkey, Turkmenistan, and Uzbekistan), and Europe (Albania, France, Greece, Italy, and Portugal) (1). To our knowledge, this is the first report of Septoria leaf spots of pistachio in Egypt. References: (1) T. Andrianova and D. Minler. Septoria pistaciae. Page 159 in: IMI Descriptions of Fungi and Bacteria. CAB International, Wallingford, UK. 2004. (2) A. Chitzanidis. Ann. Inst. Phytopathol. Benaki 10:29, 1956. (3) D. J. Young and T. Michailides. Plant Dis. 73:775, 1989.

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First Report of Stigmina palmivora Causing Leaf Spots on Phoenix roebelenii in Brazil

Plant Disease ◽

10.1094/pdis-10-13-1030-pdn ◽

2014 ◽

Vol 98 (6) ◽

pp. 849-849 ◽

Cited By ~ 1

Author(s):

A. Colmán ◽

R. A. da Silva ◽

R. Alves ◽

M. Silva ◽

R. W. Barreto

Keyword(s):

Pure Culture ◽

Leaf Surface ◽

The United States ◽

Culture Collection ◽

Commonwealth Mycological Institute ◽

First Report ◽

Leaf Spots ◽

Nucleotide Homology ◽

Query Coverage ◽

Representative Samples

Phoenix roebelenii (Arecaceae), known as dwarf date (tamareira-anã in Brazil), is a palm native to Southeast Asia and widely cultivated worldwide because of its ornamental value and ease of adaptation to a broad range of climates and soil types (4). In June 2012, some individuals were observed in a private garden in the municipality of Viçosa (state of Minas Gerais, Brazil) bearing numerous necrotic lesions on its leaves. Representative samples were taken, dried in a plant press, and brought to the laboratory for examination. A fungus was regularly associated with the leaf spots. Fungal structures were mounted in lactophenol and slides were examined under a microscope (Olympus BX 51). Spores were taken from sporulating colonies with a sterile fine needle and plated on PDA for isolation. A pure culture was deposited in the culture collection of the Universidade Federal de Viçosa (accession COAD1338). A dried herbarium sample was deposited in the local herbarium (VIC39741). The fungus had the following morphology: conidiophores grouped on sporodochia, cylindrical, 12 to 29 × 5 to 6 μm, dark brown; conidiogenous cells, terminal, proliferating percurrently (annellidic), 8 to 20 × 5 to 6 μm, pale to dark brown; conidia obclavate to subcylindrical, straight, 58 to 147 × 5 to 6 μm, 6 to 16 septate, hila thickened and darkened with a thin-walled projecting papilla, dark brown, and verrucose. The morphology of the Brazilian collections agrees well with the description of Stigmina palmivora (2), a species known to cause leaf spots on P. roebelenii in the United States (Florida) and Japan (3). Pathogenicity was demonstrated through inoculation of leaves of healthy plants by placing 6 mm diameter cuture disks of COAD1338 on the leaf surface followed by incubation in a moist chamber for 48 h and then transferred to a greenhouse bench at 21 ± 3°C. Typical leaf spots were observed 15 days after inoculation. DNA was extracted from the isolate growing in pure culture and ITS and LSU sequences were generated and deposited in GenBank under the accession numbers KF656785 and KF656786, respectively. These were compared by BLASTn with other entries in GenBank, and the closest match for each region were Mycosphaerella colombiensis strain X215 and M. irregulariamosa strain CPC 1362 (EU514231, GU2114441) with 93% of nucleotide homology (over 100% query coverage) for ITS and 98% of nucleotide homology (over 100% query coverage) for LSU. There are no sequences for S. palmivora deposited in public databases for comparison, but for Stigmina platani, the type species in this genus, 86% and 96% nucleotide homology for ITS and LSU with S. palmivora were found. The genus Stigmina is regarded as being polyphyletic (1) and this is probably reflected by these low homology levels found in the BLASTn search. To our knowledge, this is the first report of Stigmina palmivora in Brazil. References: (1) P. W. Crous et al. Stud. Mycol. 75:37, 2012. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, UK, 1971. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 2013. (4) H. Lorenzi et al. Palmeira no Brasil: Exóticas e Nativas, 2nd ed. Editora Plantarum, Nova Odessa, Brazil, 2005.

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First Report of Volutella Blight on Pachysandra Caused by Volutella pachysandricola in China

Plant Disease ◽

10.1094/pdis-11-11-0997 ◽

2012 ◽

Vol 96 (4) ◽

pp. 584-584

Author(s):

Q. Bai ◽

Y. Xie ◽

R. Dong ◽

J. Gao ◽

Y. Li

Keyword(s):

Morphological Characteristics ◽

Stem Canker ◽

Botanical Garden ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

Conidial Suspension ◽

First Report ◽

Leaf Spots ◽

Plastic Bags

Pachysandra (Pachysandra terminalis, Buxaceae) and Japanese Pachysandra, also called Japanese Spurge, is a woody ornamental groundcover plant distributed mostly in Zhejiang, Guizhou, Henan, Hubei, Sichuan, Shanxi, and Gansu provinces in China. In April 2010, P. terminalis asymptomatic plants were shipped from Beijing Botanical Garden Institute of Botany Chinese Academy of Science to the garden nursery of Jilin Agricultural University (43°48′N, 125°23′E), Jilin Province. In June 2011, Volutella blight (sometimes called leaf blight and stem canker) of P. terminalis was observed on these plants. Infected leaves showed circular or irregular, tan-to-brown spots often with concentric rings and dark margins. The spots eventually grew and coalesced until the entire leaf died. Cankers appeared as greenish brown and water-soaked diseased areas, subsequently turning brown or black, and shriveled and often girdled the stems and stolons. During wet, humid weather in autumn, reddish orange, cushion-like fruiting structures of the fungus appeared on the stem cankers and undersides of leaf spots. Symptoms of the disease were consistent with previous descriptions (2–4). Five isolates were obtained from necrotic tissue of leaf spots and cankers of stems and stolons and cultured on potato dextrose agar. The colony surface was salmon colored and slimy. Conidia were hyaline, one celled, spindle shaped, and 12.57 to 22.23 × 3.33 to 4.15 μm with rounded ends. Morphological characteristics of the fungus were consistent with the description by Dodge (2), and the fungus was identified as Volutella pachysandricola (telemorph Pseudonectria pachysandricola). The internal transcribed spacer (ITS) regions of the nuclear rDNA were amplified using primers ITS4/ITS5 (1). The ITS sequences were 553 bp long and identical among these five isolates (GenBank Accession No. HE612114). They were 100% identical to Pseudonectria pachysandricola voucher KUS-F25663 (Accession No. JN797821) and 99% identical to P. pachysandricola culture-collection DAOM (Accession No. HQ897807). Pathogenicity was confirmed by spraying leaves of clonally propagated cuttings of P. terminalis with a conidial suspension (1 × 106 conidia/ml) of the isolated V. pachysandricola. Control leaves were sprayed with sterile water. Plants were covered with plastic bags and kept in a greenhouse at 20 to 25°C for 72 h. After 5 to 8 days, typical disease symptoms appeared on leaves, while the control plants remained healthy. V. pachysandricola was reisolated from the leaf spots of inoculated plants. Pachysandra leaf blight and stem canker also called Volutella blight, is the most destructive disease of P. terminalis and previously reported in the northern humid areas of the United States (Illinois, Connecticut, Ohio, Indiana, Iowa, Massachusetts, Missouri, Kentucky, and Wisconsin), northern Europe (Britain, Germany, and Poland), and the Czech Republic. To our knowledge, this is the first report of the disease caused by V. pachysandricola in China. The disease may become a more significant problem in P. terminalis cultivation areas if the disease spreads on P. terminalis in nursery beds. References: (1) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (2) B. O. Dodge. Mycologia 36:532, 1944. (3) S. M. Douglas. Online publication. Volutella Blight of Pachysandra. The Connecticut Agricultural Experiment Station, 2008. (4) I. Safrankova. Plant Protect. Sci.43:10, 2007.

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First Report of Daylily Leaf Streak Caused by Kabatiella microsticta in China

Plant Disease ◽

10.1094/pdis-04-12-0381-pdn ◽

2012 ◽

Vol 96 (10) ◽

pp. 1579-1579 ◽

Cited By ~ 1

Author(s):

Q. R. Bai ◽

S. Han ◽

Y. Y. Xie ◽

R. Dong ◽

J. Gao ◽

...

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

The United States ◽

Control Treatment ◽

Conidial Suspension ◽

First Report ◽

Perennial Plant ◽

Leaf Spots ◽

Plastic Bags ◽

Hemerocallis Citrina

Daylily (Hemerocallis spp.) is an herbaceous, perennial plant, cultivated for its flowers. Daylily is sold in Asian markets as fresh or dried flowers (the flowers of some species, e.g., Hemerocallis citrina, are edible) or as the corm, which is used for medicinal purposes. In June 2011, daylily leaf streak was found in a nursery of Jilin Agricultural University, Jilin Province, China. Symptoms included water-soaked, irregular spots along the leaf midvein that turned orange to reddish brown and eventually enlarged to coalesce into extensive, necrotic streaks along the length of the leaf, as previously reported (2). Heavily infected leaves often withered and died. Four isolates were recovered from necrotic tissue of leaf spots and cultured on potato dextrose agar (PDA) at 25°C. All colonies were initially cream to peach colored and appeared slimy. With the maturation of the culture, the colonies became dark brown to black with sparse aerial hyphae. Blastic conidia formed simultaneously on intercalary or terminal, undifferentiated conidiogenous cells, and were scattered in dense sections on culture surface. When the conidia dropped from conidiogenous cell, an indistinct scar or a denticle remained. Conidia were hyaline, one-celled, smooth, ellipsoidal, and variable in size (2.73 to 6.01 × 8.45 to 19.36 μm), and all morphological characteristics were consistent with Kabatiella microsticta Bubak (syn. Aureobasidium microstictum; 2,4). The internal transcribed spacer (ITS) region of the nuclear rDNA was amplified using primers ITS4/ITS5 (1). ITS (534 bp) was identical among all four isolates (GenBank Accession No. HE798117) and 100% identical to that of K. microsticta CBS 114.64 (FJ150873). Pathogenicity was confirmed by spraying 20 seedlings of daylily, propagated in tissue-culture medium, with a conidial suspension (106 conidia/ml) of each isolate. A second set of 20 seedlings was sprayed with the same volume of sterile water as the noninoculated control treatment. Plants were grown in the greenhouse at 20 to 25°C and were covered with plastic bags to maintain humidity on the foliage for 72 h. After 5 days, the foliar symptoms described earlier for the field plants appeared on the leaves, whereas the control plants remained healthy. K. microsticta was reisolated from the leaf spots of all 20 inoculated plants. Leaf streak is the most destructive disease of daylily, and was previously reported in Japan and the United States (Illinois, Kentucky, Mississippi, Louisiana, Pennsylvania, Maryland, Virginia, Florida, North Carolina, and Georgia) (3). To our knowledge, this is the first report of the disease caused by K. microsticta in China. References: (1) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (2) E. J. Hermanides-Nijhof. Stud. Mycol. 15:153, 1977. (3) R. M. Leahy et al. Plant Pathology Circular No. 376, 1996. (4) P. Zalar et al. Stud. Mycol. 61:21, 2008.

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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 Impatiens Downy Mildew Outbreaks Caused by Plasmopara obducens Throughout the Hawai'ian Islands

Plant Disease ◽

10.1094/pdis-10-13-1017-pdn ◽

2014 ◽

Vol 98 (5) ◽

pp. 696-696 ◽

Cited By ~ 7

Author(s):

J. A. Crouch ◽

M. P. Ko ◽

J. M. McKemy

Keyword(s):

United States ◽

Downy Mildew ◽

The United States ◽

Molecular Data ◽

Economic Losses ◽

Voucher Specimen ◽

First Report ◽

Plastic Bags ◽

Leaf Surfaces ◽

Impatiens Walleriana

Downy mildew of impatiens (Impatiens walleriana Hook.f.) was first reported from the continental United States in 2004. In 2011 to 2012, severe and widespread outbreaks were documented across the United States mainland, resulting in considerable economic losses. On May 5, 2013, downy mildew disease symptoms were observed from I. walleriana ‘Super Elfin’ at a retail nursery in Mililani, on the Hawai'ian island of Oahu. Throughout May and June 2013, additional sightings of the disease were documented from the islands of Oahu, Kauai, Maui, and Hawai'i from nurseries, home gardens, and botanical park and landscape plantings. Symptoms of infected plants initially showed downward leaf curl, followed by a stippled chlorotic appearance on the adaxial leaf surfaces. Abaxial leaf surfaces were covered with a layer of white mycelia. Affected plants exhibited defoliation, flower drop, and stem rot as the disease progressed. Based on morphological and molecular data, the organism was identified as Plasmopara obducens (J. Schröt.) J. Schröt. Microscopic observation disclosed coenocytic mycelium and hyaline, thin-walled, tree-like (monopodial branches), straight, 94.0 to 300.0 × 3.2 to 10.8 μm sporangiophores. Ovoid, hyaline sporangia measuring 11.0 to 14.6 × 12.2 to 16.2 (average 13.2 × 14.7) μm were borne on sterigma tips of rigid branchlets (8.0 to 15.0 μm) at right angle to the main axis of the sporangiophores (1,3). Molecular identification of the pathogen was conducted by removing hyphae from the surface of three heavily infected leaves using sterile tweezers, then extracting DNA using the QIAGEN Plant DNA kit (QIAGEN, Gaithersburg, MD). The nuclear rDNA internal transcribed spacer was sequenced from each of the three samples bidirectionally from Illustra EXOStar (GE Healthcare, Piscataway, NJ) purified amplicon generated from primers ITS1-O and LR-0R (4). Resultant sequences (GenBank KF366378 to 80) shared 99 to 100% nucleotide identity with P. obducens accession DQ665666 (4). A voucher specimen (BPI892676) was deposited in the U.S. National Fungus Collections, Beltsville, MD. Pathogenicity tests were performed by spraying 6-week-old impatiens plants (I. walleriana var. Super Elfin) grown singly in 4-inch pots with a suspension of 1 × 104 P. obducens sporangia/ml until runoff using a handheld atomizer. Control plants were sprayed with distilled water. The plants were kept in high humidity by covering with black plastic bags for 48 h at 20°C, and then maintained in the greenhouse (night/day temperature of 20/24°C). The first symptoms (downward curling and chlorotic stippling of leaves) and sporulation of the pathogen on under-leaf surfaces of the inoculated plants appeared at 10 days and 21 days after inoculation, respectively. Control plants remained healthy. Morphological features and measurements matched those of the original inoculum, thus fulfilling Koch's postulates. To our knowledge, this is the first report of downy mildew on I. walleriana in Hawai'i (2). The disease appears to be widespread throughout the islands and is likely to cause considerable losses in Hawai'ian landscapes and production settings. References: (1) O. Constantinescu. Mycologia 83:473, 1991. (2) D. F. Farr and A. Y. Rossman. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ July 16, 2013. (3) P. A. Saccardo. Syllogue Fungorum 7:242, 1888. (4) M. Thines. Fungal Genet Biol 44:199, 2007.

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First Report of Leaf Blight and Stem Canker of Pachysandra terminalis Caused by Pseudonectria pachysandricola in Korea

Plant Disease ◽

10.1094/pdis-09-11-0813 ◽

2012 ◽

Vol 96 (2) ◽

pp. 287-287

Author(s):

K. S. Han ◽

J. H. Park ◽

S. E. Cho ◽

H. D. Shin

Keyword(s):

United States ◽

Its Region ◽

Stem Canker ◽

The United States ◽

Culture Collection ◽

Leaf Blight ◽

First Report ◽

Cornell University ◽

Plastic Bags ◽

Young Leaves

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.

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First Report of Alternaria Leaf Spot of Banana Caused by Alternaria alternata in the United States

Plant Disease ◽

10.1094/pdis-01-13-0007-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1116-1116 ◽

Cited By ~ 3

Author(s):

V. Parkunan ◽

S. Li ◽

E. G. Fonsah ◽

P. Ji

Keyword(s):

United States ◽

Alternaria Alternata ◽

Leaf Spot ◽

Morphological Characteristics ◽

The United States ◽

Its Sequencing ◽

Single Spore ◽

First Report ◽

Alternaria Leaf Spot ◽

Leaf Spots

Research efforts were initiated in 2003 to identify and introduce banana (Musa spp.) cultivars suitable for production in Georgia (1). Selected cultivars have been evaluated since 2009 in Tifton Banana Garden, Tifton, GA, comprising of cold hardy, short cycle, and ornamental types. In spring and summer of 2012, 7 out of 13 cultivars (African Red, Blue Torres Island, Cacambou, Chinese Cavendish, Novaria, Raja Puri, and Veinte Cohol) showed tiny, oval (0.5 to 1.0 mm long and 0.3 to 0.9 mm wide), light to dark brown spots on the adaxial surface of the leaves. Spots were more concentrated along the midrib than the rest of the leaf and occurred on all except the newly emerged leaves. Leaf spots did not expand much in size, but the numbers approximately doubled during the season. Disease incidences on the seven cultivars ranged from 10 to 63% (10% on Blue Torres Island and 63% on Novaria), with an average of 35% when a total of 52 plants were evaluated. Six cultivars including Belle, Ice Cream, Dwarf Namwah, Kandarian, Praying Hands, and Saba did not show any spots. Tissue from infected leaves of the seven cultivars were surface sterilized with 0.5% NaOCl, plated onto potato dextrose agar (PDA) media and incubated at 25°C in the dark for 5 days. The plates were then incubated at room temperature (23 ± 2°C) under a 12-hour photoperiod for 3 days. Grayish black colonies developed from all the samples, which were further identified as Alternaria spp. based on the dark, brown, obclavate to obpyriform catenulate conidia with longitudinal and transverse septa tapering to a prominent beak attached in chains on a simple and short conidiophore (2). Conidia were 23 to 73 μm long and 15 to 35 μm wide, with a beak length of 5 to 10 μm, and had 3 to 6 transverse and 0 to 5 longitudinal septa. Single spore cultures of four isolates from four different cultivars were obtained and genomic DNA was extracted and the internal transcribed spacer (ITS1-5.8S-ITS2) regions of rDNA (562 bp) were amplified and sequenced with primers ITS1 and ITS4. MegaBLAST analysis of the four sequences showed that they were 100% identical to two Alternaria alternata isolates (GQ916545 and GQ169766). ITS sequence of a representative isolate VCT1FT1 from cv. Veinte Cohol was submitted to GenBank (JX985742). Pathogenicity assay was conducted using 1-month-old banana plants (cv. Veinte Cohol) grown in pots under greenhouse conditions (25 to 27°C). Three plants were spray inoculated with the isolate VCT1FT1 (100 ml suspension per plant containing 105 spores per ml) and incubated under 100% humidity for 2 days and then kept in the greenhouse. Three plants sprayed with water were used as a control. Leaf spots identical to those observed in the field were developed in a week on the inoculated plants but not on the non-inoculated control. The fungus was reisolated from the inoculated plants and the identity was confirmed by morphological characteristics and ITS sequencing. To our knowledge, this is the first report of Alternaria leaf spot caused by A. alternata on banana in the United States. Occurrence of the disease on some banana cultivars in Georgia provides useful information to potential producers, and the cultivars that were observed to be resistant to the disease may be more suitable for production. References: (1) E. G. Fonsah et al. J. Food Distrib. Res. 37:2, 2006. (2) E. G. Simmons. Alternaria: An identification manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

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First Report of a Leaf Spot Caused by Alternaria brassicae on the Invasive Weed Lepidium draba in North America

Plant Disease ◽

10.1094/pdis-93-8-0846a ◽

2009 ◽

Vol 93 (8) ◽

pp. 846-846 ◽

Cited By ~ 1

Author(s):

A. J. Caesar ◽

R. T. Lartey

Keyword(s):

North America ◽

Chinese Cabbage ◽

Leaf Spot ◽

The United States ◽

Alternaria Brassicae ◽

Voucher Specimen ◽

First Report ◽

Leaf Spots ◽

Black Spots ◽

Lepidium Draba

The exotic, rangeland weed Lepidium draba L., a brassicaceous perennial, is widely distributed in the United States. For example, Oregon contains 100,000 ha of land infested with L. draba (2). Because it is capable of aggressive spread and has the potential to reduce the value of wheat-growing land (4), it is the target of biological control research. The application of multiple pathogens has been advocated for control of other brassicaceous weeds, including the simultaneous application of biotrophic and necrotrophic pathogens (3). In pursuit of this approach, in 2007, we discovered the occurrence of leaf spots on approximately 90% of L. draba plants near Shepherd, MT, which were distinct from leaf lesions caused by Cercospora bizzozeriana (1). The lesions were initially tiny, black spots enlarging over time to become circular to irregular and cream-colored around the initial black spots and sometimes with dark brown borders or chlorotic halos. Conidia from the lesions were light brown, elongate and obclavate, produced singly from short conidia, with 8 to 12 transverse septa, and 2 to 6 longitudinal septa. The spore body measured 25 to 35 × 200 to 250 μm with a beak cell 42 to 100 μm long. On the basis of conidial and cultural characteristics, the fungus was identified as Alternaria brassicae (Berk.) Sacc. Leaf tissues bordering lesions were plated on acidified potato dextrose agar. Colonies on V8 and alfalfa seed agar were black with concentric rings, eventually appearing uniformly black after 10 to 14 days. The internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 and sequenced. BLAST analysis of the 575-bp fragment showed a 100% homology with a sequence of A. brassicae Strain B from mustard (GenBank Accession No. DQ156344). The nucleotide sequence has been assigned GenBank Accession No. FJ869872. For pathogenicity tests, aqueous spore suspensions approximately 105/ml were prepared from cultures grown at 20 to 25°C for 10 to 14 days on V8 agar and sprayed on leaves of three L. draba plants. Inoculated plants were enclosed in plastic bags and incubated at 20 to 22°C for 72 to 80 h. In addition, three plants of the following reported hosts of A. brassicae were inoculated: broccoli, canola, Chinese cabbage, collards, broccoli raab, kale, mustard greens, radish, rape kale, and turnip. Within 10 days, leaf spots similar to those described above developed on plants of radish, canola, Chinese cabbage, and turnip and A. brassicae was reisolated and identified. Control plants sprayed with distilled water remained symptomless. These inoculations were repeated and results were the same. To our knowledge, this is the first report of a leaf spot disease caused by A. brassicae on L. draba in North America. A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) A. J. Caesar et al. Plant Dis. 93:108, 2009. (2) G. L. Kiemnec and M. L. McInnis. Weed Technol. 16:231, 2002. (3) A. Maxwell and J. K. Scott. Adv. Bot. Res. 43:143, 2005. (4) G. A. Mulligan and J. N. Findlay. Can. J. Plant Sci. 54:149, 1974.

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First Report of Phoma exigua as a Pathogen of Salal (Gaultheria shallon) in British Columbia, Canada

Plant Disease ◽

10.1094/pd-89-0685b ◽

2005 ◽

Vol 89 (6) ◽

pp. 685-685 ◽

Cited By ~ 4

Author(s):

S. F. Shamoun ◽

S. Zhao

Keyword(s):

British Columbia ◽

The United States ◽

Gaultheria Shallon ◽

Centraalbureau Voor ◽

Potato Dextrose Broth ◽

Voucher Specimen ◽

Disease Symptoms ◽

Phoma Exigua ◽

First Report ◽

Plastic Bags

Salal (Gaultheria shallon Pursh.) is an ericaceous, evergreen, and rhizomatous shrub that competes for nutrients and moisture with young conifers in low elevation, coastal British Columbia (BC). A survey was conducted on southern Vancouver Island, BC during the summer of 1999 to find fungal pathogens of salal that might serve as biocontrol organisms (3). Phoma exigua Desmaz. (isolate PFC2705) near Parksville, BC proved to be pathogenic on salal. Identification of PFC2705 at the Centraalbureau voor Schimmelcultures was based on morphology and ITS sequences (GenBank Accession No. AY927784). Pathogenicity was determined with 24 salal seedlings (3-month-old) by inoculating with mycelial suspensions (20% v/v) or conidial suspensions (1 × 106 conidia per ml in 0.5% potato dextrose broth). Inoculated seedlings were placed in plastic bags and incubated in a greenhouse (16 to 23°C with natural light). Plastic bags were removed after 2 days. Initial disease symptoms were observed 2 days after inoculation. Brown, sunken lesions appeared on the surface of young leaves and stems and extended quickly. All seedlings were killed within 14 days. Twelve control plants showed no disease symptoms. With diseased salal leaves incubated at 23°C with 12-h fluorescent light/dark and 100% relative humidity, pycnidia appeared on leaf surfaces within 5 days. Conidia were hyaline, ellipsoid, one-celled, sometimes two- to three-celled, 2.5 to 3.8 × 5 to 12.5 μm, with a rounded base; the colony was gray or dark gray on potato dextrose agar after 5 to 7 days. Reisolation from the inoculated diseased leaves produced a mycelial colony that shared the same growth and morphological characteristics as the initial isolate. Phyllosticta gaultheriae Ellis & Everh., a widely reported foliar pathogen of salal, is distinct morphologically from P. exigua (1). To our knowledge, this is the first report of P. exigua as a pathogen of salal in Canada (2). A voucher specimen has been deposited at the Pacific Forestry Center Herbarium (DAVFP No. 28735). References: (1) J. Bissett and S. J. Darbyshire. No. 275 in: Fungi Canadenses, 1984. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society. St. Paul. MN, 1989. (3) S. F. Shamoun et al. Can. J. Plant Pathol. 22:192, 2000.

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First Report of Stem and Foliar Blight of Sunflower Caused by Alternariaster helianthi in Louisiana

Plant Disease ◽

10.1094/pdis-01-12-0107-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 761-761 ◽

Cited By ~ 1

Author(s):

R. Singh ◽

D. M. Ferrin

Keyword(s):

Disease Incidence ◽

Fluorescent Light ◽

Infected Leaf ◽

Stem Tissue ◽

First Report ◽

Foliar Blight ◽

Leaf Spots ◽

Plastic Bags ◽

Alternaria Helianthi ◽

Stem Lesions

During the fall of 2009, sunflower (Helianthus annuus L.) planted at the LSU AgCenter's Burden Center in Baton Rouge, LA exhibited severe stem and foliar blight symptoms. Symptoms on stems and petioles included elongated, slightly sunken lesions with dark brown margins. Leaf symptoms included irregular to circular, dark brown lesions with white centers and surrounded by a yellow halo. Several spots often coalesced to form large, blighted areas, and severely affected leaves turned yellow, followed by defoliation. The corolla and calyx exhibited similar lesions except for the yellow halo. Disease developed rapidly and the whole (100% disease incidence) field was blighted within a week following a rain (4 mm). Infected leaf and stem tissue was surface disinfested and plated on ¼-strength potato dextrose agar (PDA). Both leaf and stem tissue consistently produced dark olivaceous-to-black fungal colonies at room temperature under 12 h of fluorescent light per day. Conidia were 53 to 128 × 10 to 26 μm, borne singly on the conidiophores, hyaline to dark olivaceous, cylindrical, rounded at both ends, and with 6 to 10 transverse and 0 to 2 longitudinal septa. Conidiophores were single, unbranched, septate, hyaline to dark olivaceous, and measured 77 to 128 × 7 to 13 μm. Morphologically, the fungus was identified as Alternariaster helianthi (Hansf.) E.G. Simmons (= Alternaria helianthi [Hansf.] Tubaki & Nishih) (1). A single-spore isolate (PDC-4291) was obtained from the original culture and DNA from this isolate was extracted with a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Primers ITS1 and ITS4 were used to amplify and sequence the internal transcribed spacer regions 1 and 2, and NCBI blast analysis of the 552-bp sequence (GenBank Accession No. JN208925) resulted in 100% homology with Alternaria helianthi isolated from sunflower infected with leaf spot and blight disease in India (GenBank Accession No. DQ156343). Pathogenicity was determined by inoculating 20 potted sunflower plants (Full Sun Improved TD, Fred C. Gloeckner and Company, Inc., Harrison, NY) with conidia from a 2-week-old culture of isolate PDC-4291. Each plant was sprayed with 25 ml of suspension containing 106 conidia/ml. Twenty control plants were sprayed with 25 ml of sterile distilled water. Inoculated and control plants were covered with plastic bags and maintained in a greenhouse at 28 ± 2°C. Plastic bags were removed 72 h after inoculation. Leaf spots similar to the original symptoms appeared on all 20 inoculated plants 5 days after inoculation. A few stem lesions were observed on 13 plants. Two weeks after inoculation, infected leaves turned yellow and blighted. Alternariaster helianthi (= Alternaria helianthi) was reisolated from the leaf spots and stem lesions. No symptoms developed on any of the 20 control plants. On the basis of morphology and sequence data, this pathogen was identified as A. helianthi, and to our knowledge, this is the first report of sunflower stem and foliar blight caused by A. helianthi in Louisiana. In Louisiana, sunflower is a popular ornamental that is grown in landscapes and gardens and by commercial flower growers who grow it for cut flower arrangements. Louisiana's hot, humid weather is ideal for disease development, which may discourage gardeners and commercial growers from planting sunflower. Reference: (1) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Center, Utrecht, the Netherlands, 2007.

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