Thlaspi arvense, a New Host for Alternaria brassicicola

A. C. Cobb; H. R. Dillard

doi:10.1094/pdis.1998.82.8.960b

Thlaspi arvense, a New Host for Alternaria brassicicola

Plant Disease ◽

10.1094/pdis.1998.82.8.960b ◽

1998 ◽

Vol 82 (8) ◽

pp. 960-960 ◽

Cited By ~ 2

Author(s):

A. C. Cobb ◽

H. R. Dillard

Keyword(s):

Leaf Spot ◽

Production Systems ◽

Geographic Location ◽

Alternaria Brassicicola ◽

New Host ◽

Cornell University ◽

Thlaspi Arvense ◽

Leaf Spots ◽

Plastic Bags ◽

Field Pennycress

A leaf spot was observed on cruciferous weeds growing in a cabbage field located in Geneva, NY, on 1 August 1996. The leaf spots on the weeds were dark gray to black in color and varied in size from pinpoints to 1 mm in diameter. The cabbage (Brassica oleracea L. var. capitata L.) was infected with Alternaria brassicicola (Schwein.) Wiltshire, the cause of Alternaria leaf spot. The weeds were identified as Thlaspi arvense L., a winter annual commonly referred to as field pennycress, stinkweed, or fanweed depending on geographic location. Isolations from the diseased weed tissue yielded A. brassicicola (2). The numerous conidia occurred in chains of 10 or more, ranged in size from 14 to 53 μm in length, were 5 to 18 μm wide, contained from 1 to 6 transverse septa with rare longitudinal septa, and were olivaceous in color. An apical beak was absent. On potato dextrose agar (PDA) the colony was dark olive-green to black in color and velvety. Seed was collected from the T. arvense plannts in the spring of 1997. One hundred seeds were placed in petri plates containing PDA amended with 0.01% of chloramphenicol and streptomycin sulfate. A. brassicicola was not isolated from the seeds. A different area of the field was planted to cabbage in 1997 and the cruciferous weeds were allowed to grow. The 1997 population of T. arvense consisted of plants from the previous season that flowered early and plants from seeds that germinated late in the season but did not flower. A. brassicicola was isolated from nonflowering weeds in September and from flowering weeds in October. Nonflowering plants were removed from the field in November, planted in pots, and placed in the greenhouse to induce flowering. Identity of both plant populations was confirmed as T. arvense (Warren Lamboy, Cornell University, Geneva, NY). Pathogencity of A. brassicicola isolates from T. arvense was demonstrated on cabbage and T. arvense by following Koch's postulates. Conidia (105) from a 5-day-old culture isolated from T. arvense grown on PDA were atomized onto field pennycress and cabbage plants with a Preval sprayer. The plants were enclosed in plastic bags and put under lathe shading in the greenhouse. The pathogen was reisolated from symptomatic tissue of both plants after 5 days. This weed could serve as a potential source of A. brassicicola inoculum because it is not controlled by herbicides used in crucifer production systems. Alternaria raphani has been reported on T. arvense in Canada (1). This is believed to be the first report of A. brassicicola on T. arvense. References: (1) K. Mortensen et al. Can. Plant Dis. Surv. 73:129, 1993. (2) P. Neergaard. 1945. Danish Species of Alternaria and Stemphylium. Oxford University Press, London. pp. 137–138.

Download Full-text

First Report of Leaf Spot of Spinach Caused by Pleospora betae in Spain

Plant Disease ◽

10.1094/pdis-04-14-0367-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1583-1583 ◽

Cited By ~ 4

Author(s):

D. D. M. Bassimba ◽

J. L. Mira ◽

A. Vicent

Keyword(s):

Leaf Spot ◽

Spinacia Oleracea ◽

Disease Incidence ◽

Control Measures ◽

Conidial Suspension ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Swiss Chard ◽

Spinach Leaves

The production of spinach (Spinacia oleracea L.) in Spain has increased 50% since 2009, mainly due to the commercialization of fresh-cut spinach leaves packaged in modified atmosphere containers. In October 2012, light brown leaf spots 1 to 2 cm in diameter with dark concentric rings were observed in a commercial spinach production area in Valencia Province, Spain. The initial outbreak comprised an area of about 3 ha with a 20% disease incidence. Symptomatic leaves from spinach cv. Apollo were collected in the affected area and were surface disinfected with 0.5% NaOCl for 2 min. Small fragments from lesions were placed onto potato dextrose agar (PDA) amended with 0.5 g streptomycin sulfate/liter. Fungal colonies developed after 3 days of incubation at 23°C from about 90% of the infected tissues plated. Isolates were transferred to oatmeal agar (OA) (1) and water agar (WA) amended with autoclaved pea seeds (2). Plates were incubated for 30 days at 24°C with 13 h of fluorescent light and 11 h of dark for morphological examination. Colonies were olivaceous grey in OA and pycnidia developed in WA were globose to subglobose, olivaceous black, and 100 to 200 μm in diameter. Conidia were globose to ellipsoidal, hyaline, aseptate, and 3.8 to 7.7 × 2.4 to 3.9 μm. Swollen cells were observed. Isolates showed a positive reaction to NaOH (1). Partial 18S, ITS1, 5.8S, ITS2, and partial 28S ribosomal RNA (rRNA) regions were amplified using the primers ITS1 and ITS4 (4) and sequenced from DNA extracted from the isolate designated as IVIA-V004 (GenBank Accession No. KF321782). The sequence had 100% identity (e-value 0.0) with that of Pleospora betae (Berl.) Nevod. (syn. Phoma betae A.B. Frank) representative strain CBS 523.66 (1). Pathogenicity tests were performed twice by inoculating 4-month-old plants of spinach cv. Apollo, table beet (Beta vulgaris L.) cv. Detroit, and Swiss chard (B. vulgaris subsp. cicla) cv. Verde de Penca Blanca. Plants were inoculated by spraying a conidial suspension of isolate IVIA-V004 (10 ml/plant, 105 conidia/ml water) using a manual pressure sprayer. Plants were immediately covered with black plastic bags and incubated in a growth chamber at 23°C. In each experiment, four plants of each host were inoculated with the fungus and four additional plants sprayed with sterile distilled water were used as controls. Plastic bags were removed after 48 h and leaf spots similar to those observed in affected spinach plants in the field were visible on all spinach, table beets, and Swiss chard plants 3 to 5 days after inoculation. No symptoms were observed on control plants. Fungal colonies morphologically identified as P. betae were re-isolated from leaf lesions on inoculated plants, but not from asymptomatic leaves of control plants. To our knowledge, this is the first report of leaf spot caused by P. betae on spinach in Spain, where it was previously described affecting sugar beet (3). The disease reduces the quality of spinach leaves and proper control measures should be implemented. References: (1) G. H. Boerema et al. Phoma Identification Manual, Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing, Wallingford, UK, 2004. (2) O. D. Dhingra and J. B. Sinclair. Basic Plant Pathology Methods, 2nd ed. CRC Press, Boca Raton, FL, 1995. (3) P. Melgarejo et al. Patógenos de Plantas Descritos en España. MARM-SEF, Madrid, 2010. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

First Report of Pestalotiopsis microspora Causing Leaf Spot of Hidcote (Hypericum patulum) in Japan

Plant Disease ◽

10.1094/pdis-94-8-1064b ◽

2010 ◽

Vol 94 (8) ◽

pp. 1064-1064 ◽

Cited By ~ 3

Author(s):

M. Zhang ◽

H. Y. Wu ◽

T. Tsukiboshi ◽

I. Okabe

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

Basal Cells ◽

Single Spore ◽

Management Options ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Pestalotiopsis Microspora

Hidcote, Hypericum patulum Thunb. ex Murray, is a deciduous shrub that is cultivated as an ornamental in landscape gardens and courtyards in Japan. In early August 2008, severe leaf spotting was observed on plants growing in a courtyard in Nasushiobara, Tochigi, Japan. More than 30% of the leaves on five shrubs exhibited leaf spot symptoms. Small, round, pale brown lesions were initially observed. Later, they expanded to 5 to 12 mm in diameter, round to irregular-shaped with pale brown centers and dark brown margins. Under continuously wet or humid conditions, black acervuli developed on the leaf lesions. Conidia were straight or slightly curved, fusiform to clavate, and five-celled with constrictions at the septa. Conidia ranged from 17 to 21 × 5 to 8 μm with hyaline apical and basal cells. Fifteen percent of apical cells had two and the rest had three appendages (setulae) ranging from 10 to 21 μm long. The basal hyaline cell tapered into a 2 to 4 μm pedicel. The three median cells ranged from light or dark brown to olive green. These morphological characteristics matched those of Pestalotiopsis microspora (Speg.) G.C. Zhao & N. Li (1,2). The identity of the fungus was confirmed by DNA sequencing of the internal transcribed spacer (ITS) region (GenBank Accession No. GU908473) from single-spore isolates, which revealed 100% homology with those of other P. microspora isolates (e.g., GenBank Accession Nos. FJ459950 and DQ456865). Koch's postulates were confirmed using leaves of three detached branches of a field-grown asymptomatic plant of H. patulum. Thirty leaves of each branch were inoculated by placing mycelial plugs obtained from the periphery of 7-day-old single-spore cultures on the leaf surface. Potato dextrose agar plugs without mycelium served as controls. Leaves on branches were covered with plastic bags for 24 h to maintain high relative humidity in a greenhouse (approximately 24 to 28°C). After 5 days, all inoculated leaves showed symptoms identical to those described above, whereas control leaves remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was P. microspora. To our knowledge, this is the first report of leaf spots on H. patulum caused by P. microspora in Japan. Management options may have to be developed and implemented to protect Hidcote plants in areas where leaf spot cannot be tolerated. References: (1) P. A. Saccardo. Sylloge Fungorum III:789, 1884. (2) G. C. Zhao and N. Li. J. Northeast For. Univ. 23(4):21, 1995.

Download Full-text

First Report of Bipolaris peregianensis Causing Leaf Spot of Cynodon dactylon in China

Plant Disease ◽

10.1094/pdis-12-11-1066-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 917-917 ◽

Cited By ~ 2

Author(s):

Z. Y. Wang ◽

S. N. Xie ◽

Y. Wang ◽

H. Y. Wu ◽

M. Zhang

Keyword(s):

Leaf Spot ◽

Cynodon Dactylon ◽

Control Strategies ◽

Tap Water ◽

Morphological Characteristics ◽

Its Region ◽

Basal Cells ◽

First Report ◽

Leaf Spots ◽

Plastic Bags

Cynodon dactylon is widely cultivated as a sod crop in warm climates worldwide. In early September 2009, heavy leaf spot infection was observed on C. dactylon from Zhengzhou, Henan, China. Early symptoms appeared as small, elliptical, pale brown lesions on the leaves. Lesions later expanded to 5 to 10 mm long and 1 to 2 mm wide, becoming brown to dark brown. A fungus was consistently isolated from leaf spots on potato dextrose agar (PDA), but with poor sporulation. Morphological characteristics were observed from single-conidium cultures on tap water agar + wheat straw (TWA+W) after 5 to 7 days. Conidiophores were light to medium brown, cylindrical, solitary or clustered, unbranched, usually with basal cells enlarged, and 94.5 to 147.0 × 4.0 to 9.0 μm. Conidia were fusoid, strongly curved, end cells broadly hemiellipsoidal, brown, 58.5 to 84.5 × 13.5 to 18.5 μm, with 6 to 10 septa. These morphological characteristics are similar to those of Bipolaris peregianensis (2). The identity of our fungus was confirmed to be B. peregianensis by DNA sequencing of the internal transcribed spacer (ITS) region (GenBank Accession No. JQ316121), which was 99% homologous to those of other B. peregianensis isolates (= Cochliobolus peregianensis; Accession Nos AF071328 and AF158111) (1). Koch's postulates were performed with the leaves of three pots of C. dactylon. Leaves were sprayed with 1 × 105 conidia/ml of B. peregianensis; an equal number of leaves in the pots of the same plant sprayed with sterile distilled water served as the control. All test plants were covered with plastic bags for 24 h to maintain high relative humidity at 23 to 25°C. After 7 days, more than 50% of inoculated leaves showed symptoms identical to those observed in natural condition, whereas controls remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was B. peregianensis. To our knowledge, this is the first report of leaf spots caused by B. peregianensis on C. dactylon in China. The disease cycle and the control strategies in the regions are being further studied. References: (1) M. L. Berbee et al. Mycologia 91:964, 1999. (2) A. Sivanesan. Mycol. Pap. 158:1, 1987.

Download Full-text

First Report of a Leaf Spot on Basella alba Caused by a Bipolaris sp. in Florida

Plant Disease ◽

10.1094/pdis-04-11-0276 ◽

2011 ◽

Vol 95 (7) ◽

pp. 880-880 ◽

Cited By ~ 3

Author(s):

S. Zhang ◽

Y. Fu ◽

Z. Mersha ◽

X. Mo ◽

A. J. Palmateer

Keyword(s):

Leaf Spot ◽

Its Region ◽

Plant Diseases ◽

Home Garden ◽

Pathogenicity Test ◽

Ambient Conditions ◽

First Report ◽

Basella Alba ◽

Leaf Spots ◽

Plastic Bags

Malabar spinach (Basella alba L.) is a fast-growing, perennial vegetable crop grown largely in the tropics of Asia and Africa. This crop is widely used in the cuisine of different regions for its thick, semisucculent leaves, mild flavor, and mucilaginous texture. Leaf spots were observed on both surfaces of symptomatic leaf samples received from a home garden in Homestead, FL in November 2009. The necrotic lesions (up to 2 mm in diameter) were round, semicircular, or irregular-shaped with grayish centers surrounded by dark brown borders. A fungus was consistently isolated from symptomatic tissues on clarified V8 (CV8) agar. Fungal colonies on CV8 agar were black and velvet-like with minimal mycelial growth and conidiophores were dark brown, simple, borne singly or in groups upon the substrate. Conidia were straight, pale to medium golden brown, rounded at the ends with three to six septa, and on average measured 75 × 15 μm (48 to 97 × 9 to 18 μm). Cultural and conidial characteristics of the isolates were closely similar to those of a Bipolaris sp. (1). The internal transcribed spacer (ITS) region (~570 bp) of rDNA was amplified using the primers ITS1/ITS4 and sequenced bidirectionally (GenBank Accession No. JF506092). Subsequent database searches by the BLASTN program indicated that the resulting sequence had a 95% identity over 531 bp with the corresponding gene sequence of Bipolaris portulacae (GenBank Accession No. AY004778.1), a fungal pathogen reported to cause leaf spot on purslane (Portulaca oleracea) (2,3). However, our isolate has consistently smaller conidia and does not match descriptions of B. portulacae (BPI 871173, U.S. National Fungus Collections). The pathogenicity was confirmed through inoculation of healthy Malabar spinach plants with conidia of the isolate reproduced on CV8. Six Malabar spinach plants were inoculated with a suspension containing 1 × 106 conidia per ml and sprayed until runoff (approximately 15 ml per plant) with a handheld pressurized canister. Another six noninoculated plants served as a control. Immediately after inoculation, plants were covered with plastic bags for 24 h to maintain high relative humidity and maintained in a greenhouse under ambient conditions. Ten days after inoculation, the symptoms described above were observed on leaves of all inoculated plants, whereas symptoms did not develop on the control plants. A Bipolaris sp. was reisolated and identified by the above methods, fulfilling Koch's postulates. This pathogenicity test was carried out three times. To our knowledge, this is the first report of a Bipolaris sp. affecting Malabar spinach in Florida. Further work should be conducted to confirm identity of these isolates. Because of limited plantings of Malabar spinach, the economic importance of this disease in Florida is currently not known. Nevertheless, this pathogen poses a threat to the growing market of continuously produced oriental vegetables in Florida. References: (1) J. L. Alcorn. Mycotaxon 39:361, 1990. (2) S. A. Alfieri, Jr. et al. Bull. 14. Index of Plant Diseases in Florida (Revised). Florida Dep. Agric. Consumer Serv., Div. Plant Ind., 1984. (3) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory. ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 25 January 2010.

Download Full-text

Morphology, Molecular Phylogeny, and Pathogenicity of Neofusicoccum parvum, Associated with Leaf Spot Disease of a New Host, the Japanese Bay Tree (Machilus thunbergii)

Forests ◽

10.3390/f12040440 ◽

2021 ◽

Vol 12 (4) ◽

pp. 440

Author(s):

Sungyu Choi ◽

Narayan Chandra Paul ◽

Kye-Han Lee ◽

Hyun-Jun Kim ◽

Hyunkyu Sang

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Its Region ◽

Leaf Spot Disease ◽

New Host ◽

Neofusicoccum Parvum ◽

Spot Disease ◽

Leaf Spots ◽

Machilus Thunbergii

During a survey of diseased plants on Wando Island, Korea from May to June 2020, a severe leaf spot disease was observed in the upper leaves of Japanese bay tree (Machilus thunbergii). Early symptoms were light blackish spots on the leaf surface and enlargement of older spots. Dry leaf spots surrounded with deep black margins were common throughout the plants. Symptomatic leaf samples were collected, and the causal pathogen was isolated on potato dextrose agar (PDA). Three fungal isolates (CMML20-1, CMML20-3, and CMML20-4) were cultured on PDA for morphological characterization at 25 °C in the darkness. Fungal colonies were circular, fast-growing, olivaceous to dark grey, and with abundant aerial mycelium. Sporulation was induced in 14 h-10 h light-dark conditions, and the conidia were single-celled, thin-walled with a smooth surface, ellipsoid with round apices, and measuring 17.5–20.5 (avg. 17.5) μm × 7.5–10.0 (7.9) μm. The morphological characteristics resembled those typical for Neofusicoccum parvum. Molecular identification was confirmed by partially sequencing the internal transcribed spacer (ITS) region and the translation elongation factor 1-α (EF1-α) genes. Pathogenicity tests were conducted on detached leaves and whole plants of M. thunbergii. High disease prevalence was observed, and Koch postulates were fulfilled. This is the first worldwide report of N. parvum causing leaf spots on Machilus thunbergii.

Download Full-text

First Report of Alternaria alternata Causing Black Leaf Spot of Rubber Tree in China

Plant Disease ◽

10.1094/pdis-09-14-0954-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 290-290 ◽

Cited By ~ 11

Author(s):

Z. Y. Cai ◽

Y. X. Liu ◽

G. H. Li ◽

Y. F. Wang ◽

M. Zhou

Keyword(s):

Alternaria Alternata ◽

Leaf Spot ◽

Rubber Tree ◽

Morphological Characteristics ◽

Correct Identification ◽

Post Inoculation ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

Black Leaf Spot

We first reported Alternaria heveae (E.G. Simmons ) to be the pathogen that caused black leaf spot of rubber tree (Hevea brasiliensis Muell. Arg) in Heikou county in July 2014 (1). Black leaf spots that resembled the symptoms caused by A. heveae were observed on the leaves of rubber trees of the whole propagule collection nursery in Jingping County (22°68′ N and 103°05′ E) of Yunnan Province. Black foliar spots (0.1 to 2 mm in diameter) surrounded by a yellow halo with lesions slightly sunken on the leaf surface were observed. To confirm whether the disease was caused by the same pathogen, 5-mm2 sections were removed from the leading edge of the lesion and were surface-sterilized in 75% ethanol, air-dried, plated on potato carrot agar (PCA), and incubated at 28°C in the dark. Colonies of the fungus on PCA had round margins and little aerial mycelia with gray-black coloration after 6 days of growth on PCA (2). Medium brown conidia were found to be in short chains of two to eight spores, ovoid, obclavate, and obpyriform, with or without a short conical or cylindrical-shaped apical beak. Conidia ranged from 22.5 to 67.5 μm long (mean 39.9 μm) × 10 to 15 μm wide (mean 12.5 μm; 100 colodia were measured), with three to six transverse septa and zero to three longitudinal or oblique septa. Morphological characteristics matched the descriptions of A. alternata [(Fries) Keissler] (4).The ITS1-5.8S-ITS2 region of one single-spore isolate, Ah02JP1, was amplified with primers ITS1 and ITS4. The PCR product was sequenced directly and deposited in GenBank (Accession No. KM111289). A BLAST search of the GenBank database revealed 100% similarity with A. alternata isolates KJ829535.1, KJ677246.1, and KF813070.1. Therefore, the pathogen was identified as A. alternata on the basis of its morphological characteristics and ITS sequence. Pathogenicity of a representative isolate, Ah02JP1 was confirmed using a field rubber tree inoculation method. Three rubber plants (the clone of rubber tree Yunyan77-4) were grown to the copper-colored leaf stage. Leaves were spray-inoculated (104 conidia per milliliter spore suspension) until drops were equally distributed using a manual pressure sprayer. Three rubber plants sprayed with sterile distilled water were used as controls. After inoculation, the plants were covered with plastic bags to maintain high relative humidity. The plastic bags were removed 2 days post-inoculation (dpi), and the plants were monitored daily for symptom development. Five days post-inoculation, spots similar to the original ones seen on the field trees developed on all inoculated leaves, while control leaves remained symptomless. A. alternata was re-isolated from spray-inoculated leaves, confirming Koch's postulates. A. alternata has been reported as the causal agent of leaf blight of rubber tree in India, which initially appeared as minute spots on leaves and enlarged with the growth of the leaves (3). However, in the present study, the symptoms (black leaf spots) remained small over time after inoculation. To our knowledge, this is the first report of A. alternata on rubber tree in China. Correct identification of pathogens is essential for disease management strategies. This report will establish a foundation for the further study of Alternaria alternata to address the disease effectively. References: (1) Z. Y. Cai et al. Plant Dis. 98:1011, 2014. (2) E. Mirkova. J. Phytopathol. 151:323, 2003. (3) C. B. Roy et al. J. Plantation Crops 34:499, 2006. (4) T. Y. Zhang. Page 32 in: Flora Fungorum Sinicorum, Vol. 16: Alternaria. Science Press, Beijing, 2003.

Download Full-text

Alternaria brassicicola Causes a Leaf Spot on Isatis indigotica in China

Plant Disease ◽

10.1094/pdis-03-14-0242-pdn ◽

2014 ◽

Vol 98 (10) ◽

pp. 1431-1431 ◽

Cited By ~ 3

Author(s):

J. Gao ◽

Y. N. Liu ◽

N. Nan ◽

B. H. Lu ◽

W. Y. Xia ◽

...

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Petri Dish ◽

Alternaria Brassicicola ◽

Control Treatment ◽

Conidial Suspension ◽

Isatis Indigotica ◽

Plastic Bags ◽

Dark Green ◽

Branched Chains

Chinese woad (Isatis indigotica) is a biennial herb in the Brassicaceae that is widely cultivated in China. Extracts from the roots and leaves have potential pharmaceutical use for treatment of flu, encephalitis, measles, hepatitis, and mumps (2). In June 2012, a leaf spot was observed on 1-year-old plants of I. indigotica in the medicinal garden of Jilin Agricultural University, Changchun, Jilin Province, China. More than 50% of the leaves and 100% of the plants in the garden were symptomatic. In the initial stage of infection, irregular to circular, dark gray spots, each surrounded by a chlorotic halo, appeared on leaves. The spots ranged from pinpoint to 5 mm in diameter. Some spots enlarged and coalesced, forming concentric rings. Black, sunken, fusiform lesions were observed on the petioles. Lesions gradually dried and exhibited a shot-hole appearance, and entire infected leaves desiccated. Small pieces of infected leaves and petioles were surface-disinfested in 75% ethanol for 60 s, rinsed thrice in sterilized distilled water, dried, and plated on potato dextrose agar. Olive-green mycelium developed after 2 days of incubation at 25°C, turned dark green, and covered the petri dish 10 days later. The periphery of each colony was gray and velvety. On potato carrot agar medium, conidia formed on branched chains. Conidiophores arose singly or in clusters, were straight or flexuous, separated, and measured 6.8 to 26.7 × 3.1 to 11.9 μm Conidia on host plant tissues were olivaceous, cylindrical or inverted clavate, and 25.8 to 65.2 × 10.9 to 18.3 μm Larger conidia were cylindrical or obclavate, and smaller conidia were oval. Transverse and longitudinal septa of conidia ranged from 3 to 10 and from 0 to 7 μm, respectively. A very small conidial beak or no beak was observed on each conidium. On the basis of these morphological characteristics,the fungus was identified as Alternaria brassicicola (3). A PCR assay with the ITS4 and ITS5 primers was used to amplify DNA extracted from each of four isolates (1). The sequence (567 bp) of isolate Sl-8 was submitted to GenBank (Accession No. KF531832), and showed 100% similarity to that of an A. brassicicola isolate (AF392985.1), confirming the species identification. Pathogenicity assays with 10 single-conidium isolates were done by spraying a conidial suspension (1 × 106 conidia/ml) of each isolate, or sterilized water for the control treatment, onto healthy leaves and petioles of five 3-month-old plants of I. indigotica. Inoculated and control plants were enclosed in plastic bags for 48 h. After 7 days, symptoms on inoculated plants were similar to those on the original diseased plants, while control plants remained symptomless. Re-isolation from inoculated plants produced mycelial colonies with morphological characteristics of A. brassicicola, fulfilling Koch's postulates. No fungus was isolated from control plants. A. napiformis and A. brassicae have been reported as causal agents of Alternaria leaf spot on I. indigotica in China (3). To our knowledge, however, this is the first report of A. brassicicola as a pathogen on I. indigotica in China. References: (1) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (2) A. J. Li et al. Flora Reipublicae Popularis Sinicae Tomus 33, 1998. (3) T. Y. Zhang. Alternaria. Pages 99-100 in: Flora Fungorum Sinicorum, 2003.

Download Full-text

First Report of Leaf Spot Caused by Cercospora fukushiana on New Guinea Impatiens in Korea

Plant Disease ◽

10.1094/pdis-04-14-0363-pdn ◽

2014 ◽

Vol 98 (9) ◽

pp. 1280-1280

Author(s):

B. S. Kim ◽

K. S. Baek ◽

C. H. Pak ◽

J. H. Park ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Its Region ◽

New Guinea ◽

Culture Collection ◽

Distilled Water ◽

First Report ◽

Leaf Spots ◽

Plastic Bags ◽

New Guinea Impatiens

New Guinea impatiens, Impatiens hawkeri W. Bull, is widely cultivated as a potted plant and garden plant. In July 2013, hundreds of young plants (cv. Fanfare) showing symptoms of leaf spot with approximately 50% incidence were found in polyethylene tunnels in Yongin City, Korea. Leaf spots were circular to oblong, reaching 6 mm or more in diameter. The spots were initially uniformly brown to reddish brown, turning gray with reddish brown margin. Diseased plants defoliated prematurely and were abandoned without marketing due to signs of discoloration and yellowing on leaves. A cercosporoid fungus was consistently observed in association with disease symptoms. Stromata were brown, small, and composed of a few swollen hyphal cells. Conidiophores were emerging through the cuticle, fasciculate (n = 2 to 20), olivaceous to brown, paler toward the apex, straight to mildly curved, geniculate, 30 to 260 μm long, 3.5 to 5 μm wide, 1- to 6-septate, and with conspicuous conidial scars. Conidia were hyaline and acicular. Smaller conidia were straight and longer conidia were mildly curved. Conidia were subacute to obtuse at the apex, truncate to obconically truncate at the base, 2- to 18-septate, 30 to 320 × 3.5 to 5.5 μm, and with thickened, darkened hila at the base. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora fukushiana (Matsuura) W. Yamam. (1). Voucher specimens were housed in the Korea University herbarium (KUS). An isolate from KUS-F27438 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC47640). Fungal DNA was extracted with DNeasy Plant Mini Kits (Qiagen Inc., Valencia, CA). The complete internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 (4) and sequenced. The resulting sequence of 497 bp was deposited in GenBank (Accession No. KJ620981). This showed >99% similarity with sequence of C. fukushiana (EF600954) on I. balsamina from Korea. Isolate of KACC47640 was used in the pathogenicity tests. Hyphal suspensions were prepared by grinding 3-week-old colonies grown on PDA with distilled water using a mortar and pestle. Five plants were inoculated with hyphal suspensions and five plants were sprayed with sterile distilled water. The plants were covered with plastic bags to maintain a relative humidity of 100% for 24 h and then transferred to a 25 ± 2°C greenhouse with a 12-h photoperiod. Typical symptoms of necrotic spots appeared on the inoculated leaves 10 days after inoculation, and were identical to the symptoms observed in the field. C. fukushiana was re-isolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on water-inoculated control plants. Previously, leaf spots of Impatiens spp. associated with C. apii, C. balsaminae, and C. fukushiana have been reported (1,2,3). To our knowledge, this is the first report of C. fukushiana on I. hawkeri in Korea. Our observations in the nurseries of I. hawkeri suggest that low humidity with good ventilation as well as plant hygiene in greenhouses might be main strategies for preventing this disease. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, ARS, USDA, retrieved March 25, 2014. (3) J. M. Soares et al. Plant Dis. 93:1214, 2009. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

First Report of Leaf Spot and Shoot Blight Caused by Cylindrocladium scoparium on Mallee Honeymyrtle in Italy

Plant Disease ◽

10.1094/pdis-93-10-1078a ◽

2009 ◽

Vol 93 (10) ◽

pp. 1078-1078 ◽

Cited By ~ 1

Author(s):

G. Polizzi ◽

D. Aiello ◽

V. Guarnaccia ◽

G. Parlavecchio ◽

A. Vitale

Keyword(s):

South African ◽

Leaf Spot ◽

First Record ◽

Crown Rot ◽

Single Spore ◽

Centraalbureau Voor ◽

Leaf Spots ◽

Plastic Bags ◽

Shoot Blight ◽

Twig Blight

In December of 2008, a widespread disease was observed on several blocks of approximately 15,000 plants (6-month to 2-year-old) of mallee honeymyrtle (Melaleuca acuminata F. Muell.). The plants were grown in two nurseries in eastern Sicily where high diffusion of diseases caused by Cylindrocladium pauciramosum and C. scoparium was previously detected. The plants exhibited leaf spots, defoliation, and apical blight of shoots. Crown rot and root rot were not present. Leaf spots were detected on all plants, whereas shoot blight was observed on approximately 3% of the plants. A Cylindrocladium sp. was consistently isolated from the diseased portions of plants onto potato dextrose agar. To determine the species, 20 single-conidia isolates of the fungus obtained from symptomatic tissues from different blocks and nurseries were cultured on carnation leaf agar (CLA) for 7 days at 25°C under 12-h light/dark conditions. Mycelia and spores growing on the carnation leaves were examined with a light microscope and the isolates were identified as C scoparium Morgan (teleomorph Calonectria morganii Crous, Alfenas & M.J. Wingf.) on the basis of their pyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, and conidia (1). In addition, the ability of the isolates to mate with South African and Italian opposite tester strains of C. scoparium (2,4) confirmed the identification of all the isolates. Koch's postulates were fulfilled by inoculating 30 6-month-old plants of mallee honeymyrtle with a spore suspension (105 conidia per ml) of one isolate of the pathogen (DISTEF-MA1) obtained from 14-day-old single-spore colonies grown on CLA at 24°C under fluorescent cool white lights on a 12-h light/dark regimen. Following inoculation, all plants were maintained in plastic bags in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. The same number of mallee honeymyrtle plants was used as uninoculated controls. Leaf spots, defoliation, and apical shoot blight identical to those observed in the nurseries appeared within 4 to 25 days. No symptoms were detected on the control plants. C. scoparium was reisolated from the artificially infected tissues and was identified as previously described. The isolate used in the pathogenicity proof was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (Accession No. CBS 124658). In Italy, C. scoparium was detected for the first time on Pistacia lentiscus in 2005 (3). Another report confirmed the spread of the pathogen in Sicilian ornamental nurseries (4). To our knowledge, this is the first record in the world of C. scoparium causing disease on mallee honeymyrtle. The data demonstrate the high susceptibility of this species to the C. scoparium leaf spot and twig blight especially when environmental conditions (heavy rains and mild temperatures) are conductive to the infections. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) G. Polizzi et al. Plant Dis. 90:1110, 2006. (4) G. Polizzi et al. Plant Dis. 91:769, 2007.

Download Full-text

First Report of Zonate Leaf Spot of Artocarpus altilis Caused by Cristulariella moricola in Taiwan

Plant Disease ◽

10.1094/pdis.2002.86.10.1179a ◽

2002 ◽

Vol 86 (10) ◽

pp. 1179-1179 ◽

Cited By ~ 1

Author(s):

B. Y. Hu ◽

W. W. Hsiao ◽

C. H. Fu

Keyword(s):

Leaf Spot ◽

Morphological Characteristics ◽

Eastern United States ◽

First Report ◽

Artocarpus Altilis ◽

Leaf Spots ◽

Plastic Bags ◽

Basal Septum ◽

Bull's Eye ◽

Northern Taiwan

Breadfruit (Artocarpus altilis (Parkinson) Fosberg) is an important landscape and garden tree in Taiwan. During the spring of 2002, zonate leaf spots of breadfruit were observed at a Taipei nursery in northern Taiwan. Initially, several small, brown, zonate lesions developed on leaves. As lesions enlarged, they coalesced, leading to blighting of leaves and premature defoliation. Sporophores on the host were generally hypophyllous but sometimes amphigenous, solitary, erect, easily detachable, and as much as 850 μm long. The upper portion of the sporophore is considered an individual conidium and consisted of a pyramidal head that was fusiform to ventricose and cristulate, 495 to 534 μm long and 210 to 290 μm wide at the broadest point. Branches within the pyramidal head were short and compact, and dichotomously or trichotomously branched. The conidia were hyaline, broad, septate, tapering toward an acute apex, and sometimes constricted at the basal septum. Conidiophores were 400 to 680 × 20 to 100 μm. The fungus was isolated from infected tissue and maintained on potato dextrose agar (PDA). Sclerotia were produced on PDA after 4 to 5 weeks at 20°C without light, but conidia were not observed in culture. The fungus was identified as Cristulariella moricola (Hino) Redhead based on morphological characteristics (1,2). To complete Koch's postulates, three sporophores from infected leaves or three sclerotia from cultures were placed individually on each of 10 breadfruit leaves. The plants were placed in plastic bags and incubated at 16 to 20°C. Symptoms were observed after 2 to 3 days on 100% of plants inoculated with sporophores and after 6 days on 50% of plants inoculated with sclerotia. The pathogen was reisolated from lesions on plants inoculated with sporophores and sclerotia. No symptoms were observed on the control plants. C. moricola has been known to cause a bull's eye or zonate leaf spot and defoliation on woody and annual plants, including at least 51 host species and 36 families distributed in the central and eastern United States (1). To our knowledge, this is the first report of zonate leaf spot and defoliation of breadfruit caused by C. moricola. References: (1) T. T. Chang. Bull. Taiwan For. Res. Inst. New Ser. 10 (2):235, 1995. (2) S. A. Redhead. Can. J. Bot. 53:700, 1975.

Download Full-text