Yellow Stunt, a Tobacco Disease Caused by Pythium dissotocum, in Southern Parts of Brazil

Yellow stunt, an emerging disease of tobacco (Nicotiana tabacum), has become increasingly prevalent in tobacco-growing regions in southern Brazil. Major symptoms are moderate to severe stunting, yellowing of leaves, severe wilting, darkened roots, necrosis of stem tissue directly above the soil line, and plant death. Phytophthora glovera was first proposed in 1999 as the primary causal agent of yellow stunt (1), but since then, there has been no data or completion of Koch's postulates to support this. Fifty-six isolates of fungi and fungus-like organisms were obtained from stem and root samples of tobacco plants with typical symptoms of yellow stunt in the Brazilian States of Rio Grande do Sul, Santa Catarina, and Paraná during the growing seasons of 2004/05 to 2006/07. They were identified to species level by analysis of the morphological characteristics (2) and sequence of rDNA internal transcribed spacer regions 1 and 2 (ITS1 and ITS2) (3). The isolates were identified as Pythium dissotocum (29), Fusarium oxysporum (10), P. graminicola (5), Rhizoctonia solani (4), F. solani (3), P. ultimum (3), P. deliense (1), and P. inflatum (1). The ITS sequences of the 29 isolates identified as P. dissotocum were identical. The nucleotide sequence of one isolate, LFM27/2005, has been deposited in GenBank (GQ495982). Analysis of ITS sequences alone was not sufficient to differentiate this isolate from other species in the Pythium subclade B2, such as P. coloratum, P. lutarium, P. marinum, or P. diclinum. However, the combination of morphological and cultural characteristics (2) and sequence data support our identification of LFM27/2005 and similar isolates as P. dissotocum. Colonies of LFM27/2005 on cornmeal agar had filamentous sporangia and formed slightly inflated, dendroid structures. Zoospores formed at 5°C. Daily growth rate on potato carrot agar was 13 mm at 25°C. The oogonia (22 μm in diameter) were nonornamented and either intercalary or terminal. Antheridia, commonly 1 to 2 per oogonium, were sessile, born on unbranched stalks, and either monoclinous or diclinous. Aplerotic or nearly plerotic oospores measured 20 μm in diameter with a smooth wall 2.5 μm thick. Pathogenicity tests for each pathogen were performed in a greenhouse at ~24°C in pots filled with pine bark substrate infested with inoculum at the time Burley tobacco plants showed five expanded leaves. Each test consisted of five plants and was repeated three times. Inoculum for one to three isolates representative of each pathogen was prepared by growing 2-month-old cultures at 28°C in the dark for 7 days on potato dextrose agar medium overlaid with three sterile oat kernels. Noninfested oat kernels were used for control plants. Forty days after inoculation, only plants inoculated with isolates of P. dissotocum exhibited all symptoms associated with yellow stunt. P. inflatum and R. solani did not induce yellow stunt symptoms and the others induced only wilting and root rot. P. dissotocum was recovered from an inoculated, symptomatic plant, fulfilling Koch's postulates. Its morphology was identical to isolates obtained from original field samples. The results demonstrate the association of isolates of P. dissotocum with tobacco yellow stunt in Brazil. References: (1) H. D. Shew et al. Phytopathology (Abstr.) 89(suppl):S72, 1999. (2) A. J. van der Plaats-Niterink. Stud. Mycol. 21:1, 1981. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Stemphylium botryosum Causing Leaf Blight of Kiwi in the Province Imathia, Northern Greece

Plant Disease ◽

10.1094/pdis-92-4-0650b ◽

2008 ◽

Vol 92 (4) ◽

pp. 650-650 ◽

Cited By ~ 1

Author(s):

T. Thomidis ◽

T. J. Michailides

Keyword(s):

Apical Cell ◽

Morphological Characteristics ◽

Leaf Blight ◽

Width Ratio ◽

Northern Greece ◽

Cell Width ◽

Koch’S Postulates ◽

First Report ◽

Stemphylium Botryosum ◽

Koch's Postulates

In Greece, kiwi (Actinidia deliciosa) is mostly found in the northern part of the country where approximately 440,000 ha are grown. In the summer of 2006, a Stemphylium sp. was frequently isolated from leaves of kiwi (cv. Hayward) grown in the province of Imathia. Symptomatic leaves were covered with irregular, necrotic, brown areas. Lesions had a distinct margin that, in some cases, covered a wide part of the diseased leaves. Intense symptoms were frequently observed and associated with defoliation. This Stemphylium sp. was consistently isolated from diseased leaves onto potato dextrose agar (PDA) after surface sterilization with 0.1% chlorine solution. On the basis of morphological characteristics of mycelia, dimensions (length 20 to 29 μm and width 14 to 21 μm) and mean length/width ratio (1.42 μm) of conidia, and width and apical cell width of condiophores, the fungus was identified as Stemphylium botryosum (Wallr.) (2,3) Koch's postulates were completed in the laboratory by inoculating leaves of kiwi (cv. Hayward) with an isolate of S. botryosum originated from a symptomatic leaf of a Hayward kiwi. Twenty leaves were surface sterilized by dipping them into 0.1% chlorine solution for 2 to 3 min, washing in sterile distilled water, and allowing them to dry in a laminar flow hood. A leaf was then placed into a petri plate containing a wet, sterilized paper towel. Inoculation was made by transferring a 5-mm-diameter mycelial disc from the margins of a 7-day-old culture onto the center of each leaf surface. Petri plates were closed and incubated at 25°C with 12 h of light for 6 days. Koch's postulates were satisfied when the same S. botryosum was reisolated from 100% of inoculated leaves that developed symptoms similar to those observed in the vineyards. Leaves inoculated with a PDA plug alone (with no S. botryosum) did not develop any symptoms. Previously, Alternaria alternata was reported as the causal agent of a leaf spot pathogen of kiwi (1,4). To our knowledge, this is the first report of the occurrence of S. botryosum causing leaf blight of kiwi in Greece and worldwide. This pathogen can cause a high level of defoliation in diseased plants. References: (1) L. Corazza et al. Plant Dis. 83:487, 1999. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Mycology Institute. London, England, 1971. (3) E. G. Simmons. Mycologia 61:1, 1969. (4) C. Tsahouridou and C. C. Thanassoulopoulos. Plant Dis. 84:371, 2000

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Candida albicans and Staphylococcus aureus Pathogenicity and Polymicrobial Interactions: Lessons beyond Koch’s Postulates

Journal of Fungi ◽

10.3390/jof5030081 ◽

2019 ◽

Vol 5 (3) ◽

pp. 81 ◽

Cited By ~ 8

Author(s):

Olivia A. Todd ◽

Brian M Peters

Keyword(s):

Staphylococcus Aureus ◽

Candida Albicans ◽

Sequence Data ◽

Bacterial Pathogen ◽

Toxin Production ◽

Microbial Interactions ◽

Human Pathogens ◽

Koch’S Postulates ◽

Bacterial Interaction ◽

Koch's Postulates

While Koch’s Postulates have established rules for microbial pathogenesis that have been extremely beneficial for monomicrobial infections, new studies regarding polymicrobial pathogenesis defy these standards. The explosion of phylogenetic sequence data has revolutionized concepts of microbial interactions on and within the host. However, there remains a paucity of functional follow-up studies to delineate mechanisms driven by such interactions and how they shape health or disease. That said, one particular microbial pairing, the fungal opportunist Candida albicans and the bacterial pathogen Staphylococcus aureus, has received much attention over the last decade. Therefore, the objective of this review is to discuss the multi-faceted mechanisms employed by these two ubiquitous human pathogens during polymicrobial growth, including how they: establish and persist in inter-Kingdom biofilms, tolerate antimicrobial therapy, co-invade host tissue, exacerbate quorum sensing and staphylococcal toxin production, and elicit infectious synergism. Commentary regarding new challenges and remaining questions related to future discovery of this fascinating fungal–bacterial interaction is also provided.

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Two new Lycoperdon species collected from Korea: L. albiperidium and L. subperlatum spp. nov.

Phytotaxa ◽

10.11646/phytotaxa.260.2.1 ◽

2016 ◽

Vol 260 (2) ◽

pp. 101 ◽

Cited By ~ 1

Author(s):

CHANG SUN KIM ◽

JONG WON JO ◽

YOUNG-NAM KWAG ◽

GI-HO SUNG ◽

JAE-GU HAN ◽

...

Keyword(s):

Sequence Data ◽

Elongation Factor ◽

Morphological Characteristics ◽

Its Region ◽

Its Sequences ◽

Translation Elongation ◽

Protein Coding ◽

Dna Sequence Data ◽

Coding Regions ◽

Elongation Factor 1 Alpha

Thirty-four Lycoperdon specimens from Korea were examined with the internal transcribed spacer (ITS) region of ribosomal DNA sequence data. The result of the ITS sequences phylogenetic analysis indicated that the Korean specimens represented nine different species. To confirm the taxonomic position of these species, we conducted an intensive morphological investigation, and additional phylogenetic investigation of the protein coding regions RNA polymerase subunit II (RPB2) and translation elongation factor 1-alpha (TEF1). We discovered two new species (L. albiperidium and L. subperlatum) and one (L. ericaeum) newly discovered in Korea. Lycoperdon albiperidium is closely related to L. ericaeum based on ITS, RPB2 and TEF1 sequence data, but these species were distinguishable by morphological characteristics, especially the shape of the basidiocarps, the diameter of the eucapillitial threads and the size of the basidospores. Lycoperdon subperlatum is quite similar to the European and American L. perlatum based on morphological characteristics. However, L. subperlatum is clearly distinct from European and American L. perlatum based on ITS, RPB2 and TEF1 sequence data, and somewhat differs from them in macro- and microscopic characteristics. Based on morphological characteristics, L. ericaeum is related to L. subumbrinum and L. lividum but it is distinguishable by the presence of fragile, eucapillitial threads, the diameters of the threads and ITS sequences. Here, we describe four Lycoperdon species collected in Korea.

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First Report of Leaf Spot Disease on Walnut Caused by Colletotrichum fioriniae in China

Plant Disease ◽

10.1094/pdis-09-14-0938-pdn ◽

2015 ◽

Vol 99 (2) ◽

pp. 289-289 ◽

Cited By ~ 9

Author(s):

Y. Z. Zhu ◽

W. J. Liao ◽

D. X. Zou ◽

Y. J. Wu ◽

Y. Zhou

Keyword(s):

Dna Sequences ◽

Leaf Spot ◽

Juglans Regia ◽

Morphological Characteristics ◽

Leaf Spot Disease ◽

Koch’S Postulates ◽

First Report ◽

Spot Disease ◽

Leaf Spots ◽

Koch's Postulates

In May 2014, a severe leaf spot disease was observed on walnut tree (Juglans regia L.) in Hechi, Guangxi, China. Leaf spots were circular to semicircular in shape, water-soaked, later becoming grayish white in the center with a dark brown margin and bordered by a tan halo. Necrotic lesions were approximately 3 to 4 mm in diameter. Diseased leaves were collected from 10 trees in each of five commercial orchards. The diseased leaves were cut into 5 × 5 mm slices, dipped in 75% ethanol for 30 s, washed three times in sterilized water, sterilized with 0.1% (w/v) HgCl2 for 3 min, and then rinsed five times with sterile distilled water. These slices were placed on potato dextrose agar (PDA), followed by incubating at 28°C for about 3 to 4 days. Fungal isolates were obtained from these diseased tissues, transferred onto PDA plates, and incubated at 28°C. These isolates produced gray aerial mycelium and then became pinkish gray with age. Moreover, the reverse of the colony was pink. The growth rate was 8.21 to 8.41 mm per day (average = 8.29 ± 0.11, n = 3) at 28°C. The colonies produced pale orange conidial masses and were fusiform with acute ends, hyaline, sometimes guttulate, 4.02 to 5.25 × 13.71 to 15.72 μm (average = 4.56 ± 0.31 × 14.87 ± 1.14 μm, n = 25). The morphological characteristics and measurements of this fungal isolate matched the previous descriptions of Colletotrichum fioriniae (Marcelino & Gouli) R.G. Shivas & Y.P. Tan (2). Meanwhile, these characterizations were further confirmed by analysis of the partial sequence of five genes: the internal transcribed spacer (ITS) of the ribosomal DNA, beta-tubulin (β-tub) gene, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene, chitin synthase 3(CHS-1) gene, and actin (ACT) gene, with universal primers ITS4/ITS5, T1/βt2b, GDF1/GDR1, CHS1-79F/CHS1-354R, and ACT-512F/ACT-783R, respectively (1). BLAST of these DNA sequences using the nucleotide database of GenBank showed a high identify (ITS, 99%; β-tub, 99%; GAPDH, 99%; CHS-1, 99%; and ACT, 100%) with the previously deposited sequences of C. fioriniae (ITS, KF278459.1, NR111747.1; β-tub, AB744079.1, AB690809.1; GAPDH, KF944355.1, KF944354.1; CHS-1, JQ948987.1, JQ949005.1; and ACT, JQ949625.1, JQ949626.1). Koch's postulates were fulfilled by inoculating six healthy 1-year-old walnut trees in July 2014 with maximum and minimum temperatures of 33 and 26°C. The 6-mm mycelial plug, which was cut from the margin of a 5-day-old colony of the fungus on PDA, was placed onto each pin-wounded leaf, ensuring good contact between the mycelium and the wound. Non-colonized PDA plugs were placed onto pin-wounds as negative controls. Following inoculation, both inoculated and control plants were covered with plastic bags. Leaf spots, similar to those on naturally infected plants, were observed on the leaves inoculated with C. fioriniae within 5 days. No symptoms were observed on the negative control leaves. Finally, C. fioriniae was re-isolated from symptomatic leaves; in contrast, no fungus was isolated from the control, which confirmed Koch's postulates. To our knowledge, this is the first report of leaf disease on walnut caused by C. fioriniae. References: (1) L. Cai et al. Fungal Divers. 39:183, 2009. (2) R. G. Shivas and Y. P. Tan. Fungal Divers. 39:111, 2009.

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First Report of Aspergillus niger Causing Postharvest Fruit Rot of Cherry in the Prefectures of Imathia and Pella, Northern Greece

Plant Disease ◽

10.1094/pdis-07-11-0620 ◽

2012 ◽

Vol 96 (3) ◽

pp. 458-458 ◽

Cited By ~ 1

Author(s):

T. Thomidis ◽

E. Exadaktylou

Keyword(s):

Aspergillus Niger ◽

Fungal Spores ◽

Morphological Characteristics ◽

Fruit Rot ◽

Northern Greece ◽

Centraalbureau Voor ◽

Koch’S Postulates ◽

First Report ◽

Postharvest Rot ◽

Koch's Postulates

In June 2011, symptoms of postharvest rot were observed on approximately 3% of all cherries collected from commercial orchards of cultivars Lapen and Ferrovia in the prefectures of Imathia and Pella (northern Greece). Fruit were harvested in a timely manner to avoid overripeness. No wounds or other predisposing injuries were observed on the infected fruits. Lesions enlarged rapidly and separated easily from healthy tissue when pressure was applied. Infected tissues were pale and water soaked and the associated fungal spores were dark and powdery and easily liberated when mature. The fungus grew rapidly and produced black colonies on acidified potato dextrose agar (2.5 ml of 85% lactic acid per liter of nutrient medium) after 5 days at 24°C. Identification of the pathogen was based on morphological characteristics (1). The conidial head was radiate, vesicles were nearly spherical and covered with metulae and phialides (biseriate). Conidia were globose (3 to 5 μm in diameter) and usually very rough with irregular ridges, bars, and verrucae. Koch's postulates were completed in the laboratory by inoculating mature cherry fruits (cv. Lapen). The fruits were surface sterilized by dipping in 10% chloride bleach solution, allowed to dry in a laminar flow hood, and wounded with a sharp glass rod that was 2 mm in diameter. A 40-μl drop of a suspension containing 20,000 conidia per ml of water was placed on each wound. There were 20 inoculated and 20 control fruits (similarly wounded and inoculated with a 40-μl drop of sterile distilled water) in a randomized design and incubated at 24 to 26°C for 6 days. Koch's postulates were satisfied after reisolating the fungus from inoculated fruit that developed symptoms similar to those observed on fruit collected from orchards. Control fruits did not show any symptom of the disease. To our knowledge, this is the first report of the occurrence of Aspergillus niger as the causal agent of postharvest rots of cherries in Greece. Postharvest fruit rots caused by A. niger have been reported in cherry orchards of other countries around the world (2). Because this disease causes postharvest rots of cherry fruits, measures may need to be implemented to manage the pathogen. References: (1) M. A. Klich. Page 12 in: Identification of Common Aspergillus Species. Centraalbureau Voor Schimmelcultures, Utrecht, the Netherlands, 2002. (2) A. Valiuskaite et al. Phytopathol. Pol. 35:197, 2005.

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Occurrence of a Fruit Spot Disease of Pomegranates Caused by Colletotrichum gloeosporiodes in the Prefecture of Komotini, Greece

Plant Disease ◽

10.1094/pdis-04-11-0294 ◽

2011 ◽

Vol 95 (7) ◽

pp. 872-872 ◽

Cited By ~ 3

Author(s):

T. Thomidis ◽

E. Exadaktylou

Keyword(s):

Room Temperature ◽

Morphological Characteristics ◽

Punica Granatum ◽

Lesion Development ◽

Koch’S Postulates ◽

Randomized Design ◽

Fruit Spot ◽

Koch's Postulates ◽

Punica Granatum L ◽

Wounded Area

Pomegranate (Punica granatum L.) is an important crop in Greece. In 2010, fruits of the pomegranate cv. Wonderful, in commercial fields located in the Prefecture of Komotini in eastern Greece, were observed to have symptoms of distinct dark brown spots. The waxy acervuli observed in infected tissue were subepidermal, typically with setae and simple, short, erect conidiophores. Conidia were hyaline, one celled, ovoid to oblong, slightly curved or dumbbell shaped, and 10 to 15 μm long and 5 to 7 μm wide. The pathogen was isolated on acidified potato dextrose agar (PDA) (2.5 ml of 85% lactic acid per liter of nutrient medium) and incubated at 23°C for 7 days. The pathogen was identified as Colletotrichum gloeosporiodes (Penz) Sacc. on the basis of morphological characteristics and internal transcribed spacer (ITS) sequence (100% identity to AJ301912, C. gloeosporioides species complex) (3) by CBS Fungal Biodiversity Centre, Identification Service, Utrecht, the Netherlands, (CG-1 isolate deposited in CBS Collection; Accession No. CBS 129372). Koch's postulates were completed in the laboratory by placing a 40-μl drop of suspension (4 × 105 conidia ml–1 of water) on a wounded area of healthy fruits of cv. Wonderful. Fruits were surface sterilized by dipping in 0.1% chlorine solution and allowed to dry in a laminar flow hood. There were 15 inoculated and 15 control fruits (similarly sprayed with sterile distilled water) in a randomized design. Fruits were covered with perforated polythene bags to maintain a high humidity necessary for infection that were removed 48 h after inoculation and the fruits were maintained at room temperature (23 ± 2°C). Lesion development was recorded daily for each fruit. Koch's postulates were satisfied after reisolating the fungus from inoculated fruit that developed symptoms similar to those observed on fruits collected from fields. To our knowledge, this is the first report of the occurrence of C. gloeosporioides as a causal agent of fruit spot of pomegranates in Greece. Fruit spots caused by C. gloeosporioides have been reported in pomegranate fields of other countries around the world (1,2). References: (1) B. K. M. Lakshmi et al. Trop. Agric. Res. 22:183, 2011. (2) D. S. Patel. Indian Phytopathol. 62:252, 2009. (3) B. S. Weir and P. R. Johnston. Mycotaxon 111:209, 2010.

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First Report of Botryosphaeria dothidea Causing Shoot Blight of Kiwifruit (Actinidia deliciosa) in Greece

Plant Disease ◽

10.1094/pdis-07-10-0528 ◽

2010 ◽

Vol 94 (12) ◽

pp. 1503-1503 ◽

Cited By ~ 4

Author(s):

T. Thomidis ◽

E. Exadaktylou

Keyword(s):

Random Amplified Polymorphic Dna ◽

Morphological Characteristics ◽

Fungal Mycelium ◽

Adhesive Tape ◽

Petroleum Jelly ◽

Koch’S Postulates ◽

First Report ◽

Agar Plug ◽

Dna Primers ◽

Koch's Postulates

In the spring of 2010, in commercial orchards located in the Prefecture of Pieria in northern Greece, wilted shoots of kiwifruit cv. Hayward were observed. Blighted shoots took on a distinct dark color. Isolations from the lower margins of the cankers were made by plating sodium-hypochlorite-treated shoot tissue sections of approximately 3 mm on acidified (2.5 ml of 85% lactic acid per liter of nutrient medium to create a pH = 3.5 after autoclaving) potato dextrose agar. Plates were incubated at 23°C for 5 days, and a fast-growing, mouse-gray colored fungus was consistently isolated from diseased stems. Identification of the pathogen was based on morphological characteristics and confirmed by using the four random amplified polymorphic DNA primers (K19 [CAC AGG CGG A], K20 [GTG TCG CGA G], R13 [GGA CGA CAA G], and R15 [GGA CAA CGA G], suggested by Ma et al. (2). This fungus formed darkly pigmented pycnidia (170 × 155 μm), while the conidia observed in these bodies were one-celled, hyaline, ellipsoidal to fusoid with distinctly truncate bases, and measured 10.9 to 21.55 × 3.25 to 10.10 μm. The pycnidia exuded conidia in white tendrils. Koch's postulates were completed in the laboratory by inoculating 20 segments (6 cm long and 1.5 to 2 cm in diameter) of 1-year-old woody shoots of kiwifruit cv. Hayward. Using a cork borer, a 7-mm-diameter wound was created in the middle of each shoot segment by removing the bark and a 6-mm-diameter agar plug bearing mycelia from a 15-day-old culture of B. dothidea was inserted into the wound. The wound was covered with petroleum jelly and wrapped with adhesive tape to prevent desiccation. Ten control segments were similarly wounded and inoculated with an agar disk without fungal mycelium. All inoculated and noninoculated shoot segments were incubated at 25°C in moist chambers, after which the resulting necrosis was recorded. Koch's postulates were satisfied after reisolating the fungus from inoculated shoots that developed symptoms similar to those observed on shoots collected from orchards. Although B. dothidea has been previously reported to cause dieback on kiwifruit in Japan (1), to our knowledge, this is the first report of the occurrence of B. dothidea on kiwifruit in Greece. This pathogen can cause a high level of shoot blights in diseased plants and presents a significant threat to the commercial kiwifruit production in Greece. References: (1) M. Kinugawa and T. Sato. Ann. Phytopathol. Soc. Jpn. 69:373, 2003. (2) Z. Ma et al. Phytopathology 91:665, 2001.

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First Report of Phomopsis amygdali Causing Fruit Rot on Peaches in Greece

Plant Disease ◽

10.1094/pd-90-1551c ◽

2006 ◽

Vol 90 (12) ◽

pp. 1551-1551 ◽

Cited By ~ 3

Author(s):

T. J. Michailides ◽

T. Thomidis

Keyword(s):

Prunus Persica ◽

Morphological Characteristics ◽

Fruit Rot ◽

Peach Fruit ◽

Koch’S Postulates ◽

Canker Disease ◽

First Report ◽

Immature Fruit ◽

Diseased Fruit ◽

Koch's Postulates

In the summer of 2005, the fungus Phomopsis amygdali (Del.) Tuset & Portilla was frequently isolated from decayed peaches (Prunus persica cv. Andross) grown in the province of Imathia, Greece. Fruit infected by P. amygdali developed gray-to-brown decay lesions with white mycelium forming on the surface of lesions. Identification of the pathogen was based on morphological characteristics. Dark-pigmented pycnidia (flask-shaped, conidia-bearing fruiting bodies) were produced over the surface of potato dextrose agar. The pycnidia exuded conidia in white tendrils 7 days later. Koch's postulates were completed in the laboratory by inoculating mature and immature cv. Andross peach fruits with an isolate of P. amygdali isolated from decayed cv. Andross peaches. Thirty peach fruit were surface sterilized by dipping them into 0.1% chlorine solution and allowing them to dry in a laminar flow hood. The peach fruit were wounded with a 2-mm diameter glass rod and a 40-μl drop of 5 × 105 conidia of P. amygdali per milliliter suspension was applied to the wound. Thirty control fruits were similarly wounded and inoculated with a 40-μl drop of sterile water. All inoculated and noninoculated fruit were incubated at 24 to 26°C for 7 days. Koch's postulates were satisfied when the same fungus was reisolated from 100% of inoculated mature and immature fruit that developed symptoms similar to diseased fruit collected from orchards. Although P. amygdali has been previously reported as a causal agent of canker disease (2) and fruit rots of peaches (1) in other countries, to our knowledge, this is the first report of the occurrence of P. amygdali causing a fruit rot of peaches in Greece. References: (1) Y. Ko and S. Sun. Plant Pathol. Bull. 12:212, 2003. (2) E. I. Zehr, Constriction canker. Page 31 in: Compendium of Stone Fruit Diseases. J. M. Ogawa et al., eds. The American Phytopathological Society, St. Paul, MN, 1995.

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First Occurrence of Anthracnose Caused by Colletotrichum gloeosporioides on Pitahaya

Plant Disease ◽

10.1094/pdis-91-5-0631a ◽

2007 ◽

Vol 91 (5) ◽

pp. 631-631 ◽

Cited By ~ 9

Author(s):

A. J. Palmateer ◽

R. C. Ploetz ◽

E. van Santen ◽

J. C. Correll

Keyword(s):

Colletotrichum Gloeosporioides ◽

Morphological Characteristics ◽

Dade County ◽

Koch’S Postulates ◽

Free Moisture ◽

First Occurrence ◽

Alternaria Sp ◽

Hylocereus Undatus ◽

Research And Education ◽

Koch's Postulates

Pitahaya, Hylocereus undatus Britt. & Rose, is a columnar, climbing cactus that produces a commercially important fruit. In December 2004, a new disease was found on the crop in Miami-Dade County, FL. Reddish brown lesions with conspicuous chlorotic haloes developed concentrically on the edges of vine ribs. Lesion centers became white and coalesced to rot much of the vine column, and in severe cases, only the vascular column in the vine center was not diseased. Salmon-colored spores and waxy, subepidermal acervuli, typically with setae and simple, short, erect conidiophores, were observed in lesion centers. Tissue from lesion margins was surface disinfested and plated on potato dextrose agar (PDA; Difco Laboratories, Detroit, MI). Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. was isolated from all samples. Colonies produced abundant conidia that were hyaline, one celled, straight, cylindrical, and averaged 14.7 × 5.0 μm with ranges of 12.5 to 17.5 × 3.8 to 7.5 μm (1). Cultural and morphological characteristics of isolates matched those for C. gloeosporioides except for appressoria and hyphopodia (1,2); pitahaya isolates had a spherical rather than lobed hyphopodia reported for C. gloeosporioides and averaged 10.9 (8.5 to 12.7) × 9.1 (7.1 to 10.3) μm. Internal transcribed spacer sequences for the pitahaya isolates were nearly identical (98% homology) to those for C. gloeosporioides isolates occurring on Euphatorium thymifolia in Thailand (GenBank Accession No. AY266393). Koch's postulates were examined in greenhouse trials at the Tropical Research and Education Center, Homestead, FL. Treatments consisted of a noninoculated control, four C. gloeosporioides isolates, and an Alternaria sp. All isolates came from symptomatic pitahaya tissue collected in Miami-Dade County. Fungi were grown on PDA for 7 days at 27°C. A sterile dissecting needle was used to gently pinprick the epidermis of the stem and 2-mm-diameter plugs of C. gloeosporioides, an Alternaria sp., or clean PDA were placed over wounds. Plants were placed in a plastic tent in a greenhouse where the temperature was held at 25°C, and free moisture was maintained on plant surfaces with a household humidifier for 48 h following inoculation. Two isolates of C. gloeosporioides were shown, in repeated greenhouse experiments, to cause reddish brown lesions with conspicuous chlorotic haloes that coalesced to rot much of the vine column, and Koch's postulates were completed with the reisolation of isolates that were used to inoculate plants. The age of vine segments had no significant effect on lesion development. To our knowledge, this is the first report of C. gloeosporioides as a pathogen of pitahaya. References: (1) J. A. Bailey and M. J. Jeger. Colletotrichum: Biology, Pathology and Control. CAB International, Wallingford, UK, 1992. (2) M. Du et al. Mycologia 97:641, 2005.

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First Report of Magnaporthe poae, Cause of Summer Patch Disease on Annual Bluegrass, in Canada

Plant Disease ◽

10.1094/pdis-06-12-0530-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1698-1698 ◽

Cited By ~ 3

Author(s):

M. M. I. Bassoriello ◽

K. S. Jordan

Keyword(s):

Morphological Characteristics ◽

Root Tissue ◽

Golf Course ◽

Koch’S Postulates ◽

First Report ◽

Annual Bluegrass ◽

Streptomycin Sulfate ◽

Magnaporthe Poae ◽

Patch Disease ◽

Koch's Postulates

The ectotrophic, root-infecting fungus Magnaporthe poae Landschoot & Jackson, the causal agent of summer patch disease in the U.S. (2), is implicated in the damage and loss of annual bluegrass (Poa annua L.) on golf course greens. This pathogenic fungus, one of the important root pathogens of turfgrass, attacks and colonizes susceptible turfgrass roots suffering from environmental or cultural stresses. Over 100 turf samples that exhibited symptoms (chlorotic circular or irregular patches of ≥15 cm in diameter with necrotic crowns and discolored roots) reminiscent of summer patch were collected from 77 southwestern Ontario golf courses from July to August of 2009 and 2010. Roots and crowns were often covered with dark, ectotrophic runner hyphae, lobed hyphopodia, and growth cessation structures, characteristic of M. poae. Sections of root tissue were surface sterilized in 0.6% sodium hypochlorite (NaOCl) for 5 min. Sterilized root tissue was plated on potato dextrose agar (PDA) containing 50 mg L–1 streptomycin sulfate and incubated at 28°C for 7 to 10 days. A fungus with morphological characteristics (hyaline mycelium that appears gray or olive-brown when mature) similar to those of M. poae (1) was consistently isolated (≥100 isolates were obtained) and used to identify M. poae through molecular techniques and Koch's postulates. DNA was extracted from the fungal mycelium of the collected isolates using the PowerPlant DNA isolation kit (MO BIO Laboratories, Inc., Carlsbad, CA). The rDNA internal transcribed spacer (ITS) regions of the isolates (≥100 isolates) were amplified by PCR using universal fungal rDNA primers ITS 4 (5′-TCCTCCGCTTATTGATATGC-3′) and ITS 5 (5′- GGAAGTAAAAGTCGTAACAAGG-3′) (3). The purified PCR products were sequenced (GenBank Accession No. JX134588 through JX134601) and a BLAST search exhibited seven isolates with 99% (MAG3, MAG6, MAG13, MAG16, and MAG17) and 100% (MAG1 and MAG14) similarity to M. poae in the GenBank database. Pathogenicity of four isolates (MAG1, MAG3, MAG6, and MAG14) was confirmed with Koch's postulates. Sixteen healthy P. annua core samples (four replicates of each treatment/isolate) collected from an Ontario golf course were inoculated with 25 mg M. poae-infested Kentucky bluegrass seed (Poa pratensis L.; 12.5 mg inoculum applied at the surface of the potting medium and 12.5 mg inoculum applied on the foliar surface) and were placed in a growth chamber with 12-h day/night cycles at 30/25°C and approximate relative humidity. After 2 to 3 weeks, inoculated plants exhibited chlorotic foliage and necrotic roots covered with dark ectotrophic runner hyphae and lobed hyphopodia. Infected root sections from each replication were surface sterilized and placed on PDA containing 50 mg L–1 streptomycin sulfate. The fungal cultures exhibited morphological characteristics consistent with M. poae (1). To our knowledge, this is the first report of summer patch caused by M. poae in Canada. References: (1) B. B. Clarke and A. B. Gould, eds. Turfgrass Patch Diseases Caused by Ectotrophic Root-Infecting Fungi. The American Phytopathological Society, St. Paul, MN, 1993. (2) P. J. Landschoot and N. Jackson. Mycol. Res. 93:59, 1989. (3) T. J. White et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Pages 315-322 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al. eds. Academic Press, San Diego, CA, 1990.

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