scholarly journals First Report of Bipolaris oryzae Causing Leaf Spot of Switchgrass in Mississippi

Plant Disease ◽  
2010 ◽  
Vol 94 (5) ◽  
pp. 643-643 ◽  
Author(s):  
M. Tomaso-Peterson ◽  
C. J. Balbalian
Keyword(s):  
Leaf Spot ◽  
Panicum Virgatum ◽  
Morphological Characteristics ◽  
Seed Transmission ◽  
Forest Products ◽  
Its Region ◽  
Infested Soil ◽  
Bipolaris Oryzae ◽  
Diagnostic Laboratory ◽  
First Report

‘Alamo’ switchgrass (Panicum virgatum L.) seedlings growing in a soilless mix exhibiting dark brown, irregular-shaped foliar lesions with black borders were submitted to the Mississippi State University Plant Disease Diagnostic Laboratory in the summer of 2009 from a local forest products company. Symptomatic tissues were plated onto water agar (WA) and incubated for 21 days on a laboratory bench top with a 12-h photoperiod at 22°C. An asexual, sporulating, dematiaceous hyphomycete identified as Bipolaris oryzae (Breda de Haan) Shoemaker was observed. Conidiophores were single, mostly straight, multiseptate, brown, and ranging from 138 to 306 × 7.7 to 15.3 μm and averaged 221.6 × 10.7 μm. Conidia were golden brown, multiseptate, ranging from 3 to 10 septa, straight to slightly curved to fusoid, wider midway, and tapering toward the terminal cells. Conidia measured 40.8 to 109.7 × 10.2 to 20.4 μm and averaged 75.8 × 13.8 μm. Morphological characteristics of B. oryzae were similar to those described by Drechsler (1) and Sivanesan (3). The internal transcribed spacer (ITS) region of ribosomal DNA from four pure culture colonies derived from single conidia was amplified by PCR using ITS1 and ITS4 primers. The resultant 572 bp was sequenced for isolates 86 through 89 (GenBank Accession Nos. GU222690–93). The sequences were 99% similar to the sequence of B. oryzae strain ATCC-MYA 3330 (GenBank No. FJ746665) isolated from P. virgatum. Pathogenicity of isolates 86 and 88 was confirmed by inoculating sterile potting mix with a fungal slurry. Sterile Alamo switchgrass seeds were sown into the infested soil in Magenta boxes and incubated for 6 weeks in a growth chamber with a 14-h photoperiod at 30°C. Leaf lesions and leaf blight were observed in seedling stands growing in B. oryzae-infested soil. Lesions were excised and plated onto WA. Sporulation of B. oryzae was observed on symptomatic tissue. In the interim, 300 nonsterilized Alamo switchgrass seeds of the same seed lot as the original symptomatic seedlings and originating from Oklahoma were plated onto WA (10 seed per plate). The seeds were incubated on the bench top as previously described. The experiment was repeated and B. oryzae colonized 1.4% of the total switchgrass seeds evaluated, indicating seed transmission and subsequent seedling infection as previously observed in the original seedlings. Leaf spot, caused by B. oryzae, was first reported as a new disease of switchgrass in North Dakota (2). In the summer of 2009, the authors observed leaf spot in four cultivars of switchgrass, including Alamo, growing in research plots in Webster County, MS. Twenty-two isolates of B. oryzae were recovered from diseased leaves of these switchgrass cultivars. To our knowledge, this is the first report of B. oryzae causing leaf spot of switchgrass in Mississippi, which broadens the natural distribution of this disease. References: (1) C. Drechlser. J. Agric. Res. 24:641, 1923. (2) J. M. Krupinsky et al. Can. J. Plant Pathol. 26:371, 2004. (3) A. Sivanesan. Mycol. Pap. 158:201, 1987.

scholarly journals First Report of Leaf Spot caused by Bipolaris oryzae on Switchgrass in Tennessee

Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1654-1654 ◽  
Author(s):  
A. L. Vu ◽  
M. M. Dee ◽  
J. Zale ◽  
K. D. Gwinn ◽  
B. H. Ownley
Keyword(s):  
Leaf Spot ◽  
Panicum Virgatum ◽  
Morphological Characteristics ◽  
Its Region ◽  
Spore Production ◽  
Post Inoculation ◽  
Sterile Water ◽  
Bipolaris Oryzae ◽  
First Report ◽  
Symptomatic Leaf

Knowledge of pathogens in switchgrass, a potential biofuels crop, is limited. In December 2007, dark brown to black irregularly shaped foliar spots were observed on ‘Alamo’ switchgrass (Panicum virgatum L.) on the campus of the University of Tennessee. Symptomatic leaf samples were surface-sterilized (95% ethanol, 1 min; 20% commercial bleach, 3 min; 95% ethanol, 1 min), rinsed in sterile water, air-dried, and plated on 2% water agar amended with 3.45 mg fenpropathrin/liter (Danitol 2.4 EC, Valent Chemical, Walnut Creek, CA) and 10 mg/liter rifampicin (Sigma-Aldrich, St. Louis, MO). A sparsely sporulating, dematiaceous mitosporic fungus was observed. Fungal plugs were transferred to surface-sterilized detached ‘Alamo’ leaves on sterile filter paper in a moist chamber to increase spore production. Conidia were ovate, oblong, mostly straight to slightly curved, and light to olive-brown with 3 to 10 septa. Conidial dimensions were 12.5 to 17 × 27.5 to 95 (average 14.5 × 72) μm. Conidiophores were light brown, single, multiseptate, and geniculate. Conidial production was polytretic. Morphological characteristics and disease symptoms were similar to those described for Bipolaris oryzae (Breda de Haan) Shoemaker (2). Disease assays were done with 6-week-old ‘Alamo’ switchgrass grown from seed scarified with 60% sulfuric acid and surface-sterilized in 50% bleach. Nine 9 × 9-cm square pots with approximately 20 plants per pot were inoculated with a mycelial slurry (due to low spore production) prepared from cultures grown on potato dextrose agar for 7 days. Cultures were flooded with sterile water and rubbed gently to loosen mycelium. Two additional pots were inoculated with sterile water and subjected to the same conditions to serve as controls. Plants were exposed to high humidity by enclosure in a plastic bag for 72 h. Bags were removed, and plants were incubated at 25/20°C with 50 to 60% relative humidity. During the disease assay, plants were kept in a growth chamber with a 12-h photoperiod of fluorescent and incandescent lighting. Foliar leaf spot symptoms appeared 5 to 14 days post-inoculation for eight of nine replicates. Control plants had no symptoms. Symptomatic leaf tissue was processed and plated as described above. The original fungal isolate and the pathogen recovered in the disease assay were identified using internal transcribed spacer (ITS) region sequences. The ITS region of rDNA was amplified with PCR and primer pairs ITS4 and ITS5 (4). PCR amplicons of 553 bp were sequenced, and sequences from the original isolate and the reisolated pathogen were identical (GenBank Accession No. JQ237248). The sequence had 100% nucleotide identity to B. oryzae from switchgrass in Mississippi (GU222690, GU222691, GU222692, and GU222693) and New York (JF693908). Leaf spot caused by B. oryzae on switchgrass has also been described in North Dakota (1) and was seedborne in Mississippi (3). To our knowledge, this is the first report of B. oryzae from switchgrass in Tennessee. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/, 28 June 2012. (2) J. M. Krupinsky et al. Can. J. Plant Pathol. 26:371, 2004. (3) M. Tomaso-Peterson and C. J. Balbalian. Plant Dis. 94:643, 2010. (4) T. J. White et al. Pages 315-322 in: PCR Protocols: a Guide to Methods and Applications. M. A. Innis et al. (eds), Acad. Press, San Diego, 1990.

scholarly journals First Report of Leaf Spot of Sweet Basil Caused by Cercospora guatemalensis in Korea

Plant Disease ◽  
2012 ◽  
Vol 96 (10) ◽  
pp. 1580-1580
Author(s):  
J. H. Park ◽  
K. S. Han ◽  
J. Y. Kim ◽  
H. D. Shin
Keyword(s):  
Leaf Spot ◽  
Morphological Characteristics ◽  
Its Region ◽  
Culture Collection ◽  
Distilled Water ◽  
First Report ◽  
Sweet Basil ◽  
Organic Farm ◽  
Leaf Spots ◽  
Close Phylogenetic Relationship

Sweet basil, Ocimum basilicum L., is a fragrant herb belonging to the family Lamiaceae. Originated in India 5,000 years ago, sweet basil plays a significant role in diverse cuisines across the world, especially in Asian and Italian cooking. In October 2008, hundreds of plants showing symptoms of leaf spot with nearly 100% incidence were found in polyethylene tunnels at an organic farm in Icheon, Korea. Leaf spots were circular to subcircular, water-soaked, dark brown with grayish center, and reached 10 mm or more in diameter. Diseased leaves defoliated prematurely. The damage purportedly due to this disease has reappeared every year with confirmation of the causal agent made again in 2011. A cercosporoid fungus was consistently associated with disease symptoms. Stromata were brown, consisting of brown cells, and 10 to 40 μm in width. Conidiophores were fasciculate (n = 2 to 10), olivaceous brown, paler upwards, straight to mildly curved, not geniculate in shorter ones or one to two times geniculate in longer ones, 40 to 200 μm long, occasionally reaching up to 350 μm long, 3.5 to 6 μm wide, and two- to six-septate. Conidia were hyaline, acicular to cylindric, straight in shorter ones, flexuous to curved in longer ones, truncate to obconically truncate at the base, three- to 16-septate, and 50 to 300 × 3.5 to 4.5 μm. Morphological characteristics of the fungus were consistent with the previous reports of Cercospora guatemalensis A.S. Mull. & Chupp (1,3). Voucher specimens were housed at Korea University herbarium (KUS). An isolate from KUS-F23757 was deposited in the Korean Agricultural Culture Collection (Accession No. KACC43980). 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. The resulting sequence of 548 bp was deposited in GenBank (Accession No. JQ995781). This showed >99% similarity with sequences of many Cercospora species, indicating their close phylogenetic relationship. Isolate of KACC43980 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 6 days after inoculation, and were identical to the ones observed in the field. C. guatemalensis was reisolated from symptomatic leaf tissues, confirming Koch's postulates. No symptoms were observed on control plants. Previously, the disease was reported in Malawi, India, China, and Japan (2,3), but not in Korea. To our knowledge, this is the first report of C. guatemalensis on sweet basil in Korea. Since farming of sweet basil has recently started on a commercial scale in Korea, the disease poses a serious threat to safe production of this herb, especially in organic farming. References: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Ithaca, NY, 1953. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology & Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , May 5, 2012. (3) J. Nishikawa et al. J. Gen. Plant Pathol. 68:46, 2002.

scholarly journals First Report of Bipolaris oryzae Causing Leaf Spot on Cultivated Wild Rice (Oryza rufipogon) in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Yue Lian Liu ◽  
Jian Rong Tang ◽  
Ya Li ◽  
Hong Kai Zhou
Keyword(s):  
Leaf Spot ◽  
Wild Rice ◽  
Morphological Characteristics ◽  
Oryza Rufipogon ◽  
Likelihood Method ◽  
Leaf Spot Disease ◽  
Pathogenicity Test ◽  
Sterile Water ◽  
Bipolaris Oryzae ◽  
First Report

Wild rice (Oryza rufipogon) has been widely studied and cultivated in China in recent years due to its antioxidant activities and health-promoting effects. In December 2018, leaf spot disease on wild rice (O. rufipogon cv. Haihong-12) was observed in Zhanjiang (20.93 N, 109.79 E), China. The early symptom was small purple-brown lesions on the leaves. Then, the once-localized lesions coalesced into a larger lesion with a tan to brown necrotic center surrounded by a chlorotic halo. The diseased leaves eventually died. Disease incidence was higher than 30%. Twenty diseased leaves were collected from the fields. The margin of diseased tissues was cut into 2 × 2 mm2 pieces, surface-disinfected with 75% ethanol for 30 s and 2% sodium hypochlorite for 60 s, and then rinsed three times with sterile water before isolation. The tissues were plated on potato dextrose agar (PDA) medium and incubated at 28 °C in the dark for 4 days. Pure cultures were produced by transferring hyphal tips to new PDA plates. Fifteen isolates were obtained. Two isolates (OrL-1 and OrL-2) were subjected to further morphological and molecular studies. The colonies of OrL-1 and OrL-1 on PDA were initially light gray, but it became dark gray with age. Conidiophores were single, straight to flexuous, multiseptate, and brown. Conidia were oblong, slightly curved, and light brown with four to nine septa, and measured 35.2–120.3 µm × 10.3–22.5 µm (n = 30). The morphological characteristics of OrL-1 and OrL-2 were consistent with the description on Bipolaris oryzae (Breda de Haan) Shoemaker (Manamgoda et al. 2014). The ITS region, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and translation elongation factor (EF-1α) were amplified using primers ITS1/ITS4, GDF1gpp1/GDR1 gdp2 (Berbee et al. 1999), and EF-1α-F/EF-1α-R EF-1/EF-2 (O’Donnell 2000), respectively. Amplicons of OrL-1 and OrL-2 were sequenced and submitted to GenBank (accession nos. MN880261 and MN880262, MT027091 and MT027092, and MT027093 and MT027094). The sequences of the two isolates were 99.83%–100% identical to that of B. oryzae (accession nos. MF490854，MF490831，MF490810) in accordance with BLAST analysis. A phylogenetic tree was generated on the basis of concatenated data from the sequences of ITS, GAPDH, and EF-1α via Maximum Likelihood method, which clustered OrL-1 and OrL-2 with B. oryzae. The two isolates were determined as B. oryzae by combining morphological and molecular characteristics. Pathogenicity test was performed on OrL-1 in a greenhouse at 24 °C to 30 °C with 80% relative humidity. Rice (cv. Haihong-12) with 3 leaves was grown in 10 pots, with approximately 50 plants per pot. Five pots were inoculated by spraying a spore suspension (105 spores/mL) onto leaves until runoff occurred, and five pots were sprayed with sterile water and used as controls. The test was conducted three times. Disease symptoms were observed on leaves after 10 days, but the controls remained healthy. The morphological characteristics and ITS sequences of the fungal isolates re-isolated from the diseased leaves were identical to those of B. oryzae. B. oryzae has been confirmed to cause leaf spot on Oryza sativa (Barnwal et al. 2013), but as an endophyte has been reported in O. rufipogon (Wang et al. 2015).. Thus, this study is the first report of B. oryzae causing leaf spot in O. rufipogon in China. This disease has become a risk for cultivated wild rice with the expansion of cultivation areas. Thus, vigilance is required.

scholarly journals First Report of Septoria Leaf Spot of Lavandin Caused by Septoria lavandulae in Croatia

Plant Disease ◽  
2014 ◽  
Vol 98 (2) ◽  
pp. 282-282
Author(s):  
K. Vrandečić ◽  
J. Ćosić ◽  
D. Jurković ◽  
I. Stanković ◽  
A. Vučurović ◽  
...  
Keyword(s):  
Leaf Spot ◽  
Antibacterial Properties ◽  
Morphological Characteristics ◽  
Its Region ◽  
Morphological Identification ◽  
Conidial Suspension ◽  
Necrotic Tissue ◽  
First Report ◽  
Initial Symptoms ◽  
Septoria Leaf Spot

Lavandula × intermedia Emeric ex Loiseleur, commonly known as lavandin, is an aromatic and medicinal perennial shrub widely and traditionally grown in Croatia. The lavandin essential oil is primarily used in perfumery and cosmetic industries, but also possesses anti-inflammatory, sedative, and antibacterial properties. In June 2012, severe foliar and stem symptoms were observed on approximately 40% of plants growing in a commercial lavandin crop in the locality of Banovo Brdo, Republic of Croatia. Initial symptoms on lower leaves included numerous, small, oval to irregular, grayish brown lesions with a slightly darker brown margin of necrotic tissue. Further development of the disease resulted in yellowing and necrosis of the infected leaves followed by premature defoliation. Similar necrotic oval-shaped lesions were observed on stems as well. The lesions contained numerous, dark, sub-globose pycnidia that were immersed in the necrotic tissue or partly erumpent. Small pieces of infected internal tissues were superficially disinfected with 50% commercial bleach (4% NaOCl) and placed on potato dextrose agar (PDA). A total of 10 isolates from leaves and five from stems of lavandin formed a slow-growing, dark, circular colonies with raised center that produced pycnidia at 23°C, under 12 h of fluorescent light per day. All 15 recovered isolates formed uniform hyaline, elongate, straight or slightly curved conidia with 3 to 4 septa, with average dimensions of 17.5 to 35 × 1.5 to 2.5 μm. Based on the morphological characteristics, the pathogen was identified as Septoria lavandulae Desm., the causal agent of lavender leaf spot (1,2). Pathogenicity of one selected isolate (428-12) was tested by spraying 10 lavandin seedlings (8 weeks old) with a conidial suspension (106 conidia/ml) harvested from a 4-week-old monoconidial culture on PDA. Five lavandin seedlings, sprayed with sterile distilled water, were used as negative control. After 5 to 7 days, leaf spot symptoms identical to those observed on the source plants developed on all inoculated seedlings and the pathogen was successfully re-isolated. No symptoms were observed on any of the control plants. Morphological identification was confirmed by amplification and sequencing of the internal transcribed spacer (ITS) region of rDNA (3). Total DNA was extracted directly from fungal mycelium with a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and PCR amplification performed with primers ITS1F/ITS4. Sequence analysis of ITS region revealed at least 99% identity between the isolate 428-12 (GenBank Accession No. KF373078) and isolates of many Septoria species; however, no information was available for S. lavandulae. To our knowledge, this is the first report of Septoria leaf spot of lavandin caused by S. lavandulae in Croatia. Since the cultivation area of lavandin plants has been increasing in many continental parts of Croatia, especially in Slavonia and Baranja counties, the presence of a new and potentially harmful disease may represent a serious constraint for lavandin production and further monitoring is needed. References: (1) T. V. Andrianova and D. W. Minter. IMI Descriptions of Fungi and Bacteria, 142, Sheet 1416, 1999. (2) R. Bounaurio et al. Petria 6:183, 1996. (3) G. J. M. Verkley et al. Mycologia 96:558, 2004.

scholarly journals First Report of a Leaf Spot Caused by Bipolaris oryzae on Switchgrass in New York

Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1192-1192 ◽  
Author(s):  
K. D. Waxman ◽  
G. C. Bergstrom
Keyword(s):  
New York ◽  
North Dakota ◽  
Leaf Spot ◽  
Panicum Virgatum ◽  
Perennial Grass ◽  
Leaf Tissue ◽  
Nucleotide Identity ◽  
Bipolaris Oryzae ◽  
First Report ◽  
Mist Chamber

Switchgrass (Panicum virgatum L.) is a perennial grass with biofuel potential. From 2007 to 2010, foliar lesions were observed on first year and mature stands of switchgrass in various locations in New York. Foliar lesions were purple, elliptical (up to 1 cm) with either distinct or diffuse margins, and occasionally with yellow halos and/or white necrotic centers. After 2 to 5 days of moist chamber incubation, surface-sterilized, symptomatic leaf tissue produced conidia that when streaked onto potato dextrose agar containing 0.3 g of streptomycin per liter gave rise to cultures with gray-to-black mycelium that developed brown conidia. The fungus was identified as Bipolaris oryzae (Breda de Haan) Shoemaker on the basis of conidial morphology (1,2). Conidiophores were brown, straight, cylindrical, and multiseptate. Conidia were brown, curved, ellipsoidal tapering to rounded ends, with 3 to 14 septa. Conidia averaged 105 μm (54 to 160 μm) long and 16 μm (12 to 20 μm) wide. Sequences of the glyceraldehyde-3-phosphate dehydrogenase (GDP) gene of three isolates from Tompkins County (Cornell Accession and corresponding GenBank Nos.: Bo005NY07 [cv. Cave-in-Rock], JF521648; Bo006NY07 [cv. Kanlow], JF521649; and Bo038NY07 [cv. Shawnee], JF521650) exhibited 100% nucleotide identity to B. oryzae isolates (GenBank Nos. AY277282–AY277285) collected from switchgrass in North Dakota (1). Sequences of the rDNA internal transcribed spacer (ITS) regions of the isolates (Cornell Accession and corresponding GenBank Nos.: Bo005NY07, JF693908; Bo006NY07, JF693909; and Bo038NY07, JF693910) exhibited 100% nucleotide identity to B. oryzae isolates (GenBank Nos. GU222690–GU222693) collected from switchgrass in Mississippi (3). Pathogenicity of two of the sequenced isolates (Bo006NY07 and Bo038NY07) along with one other isolate (Bo116NY09 from ‘Cave-in-Rock’ in Cayuga County) was evaluated in the greenhouse. Six- to eight-week-old switchgrass plants were inoculated with conidial suspensions (40,000 conidia/ml) of B. oryzae. Inoculum or sterilized water was applied until runoff. There were three plants per treatment of each of ‘Blackwell’, ‘Carthage’, ‘Cave-in-Rock’, ‘Kanlow’, ‘Shawnee’, ‘Shelter’, and ‘Sunburst’. After inoculum had dried, plants were placed in a mist chamber for 24 h and then returned to the greenhouse. Symptoms developed 2 to 4 days after inoculation for all cultivars. No symptoms developed on the control plants. Foliar lesions closely resembled those observed in the field. B. oryzae was consistently reisolated from symptomatic tissue collected from greenhouse experiments. B. oryzae was first reported as a pathogen of switchgrass in North Dakota (1) and more recently in Mississippi (3). To our knowledge, this is the first report of B. oryzae causing a leaf spot on switchgrass in New York. Observation of severe leaf spot in several field plots suggests that switchgrass populations should be screened for their reaction to regional isolates of B. oryzae prior to expanded production of switchgrass as a biofuel crop. References: (1) J. M. Krupinsky et al. Can. J. Plant Pathol. 26:371 2004. (2) R. A. Shoemaker. Can. J. Bot. 37:883, 1959. (3) M. Tomaso-Peterson and C. J. Balbalian. Plant Dis. 94:643 2010.

scholarly journals First Report of Corynespora cassiicola Causing Leaf Spot of Cassava in China

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 916-916 ◽  
Author(s):  
X.-B. Liu ◽  
T. Shi ◽  
C.-P. Li ◽  
J.-M. Cai ◽  
G.-X. Huang
Keyword(s):  
Leaf Spot ◽  
Morphological Characteristics ◽  
Its Region ◽  
Pathogenic Fungi ◽  
Commonwealth Mycological Institute ◽  
Corynespora Cassiicola ◽  
Single Spore ◽  
First Report ◽  
Leaf Spots ◽  
Main Vein

Cassava (Manihot esculenta) is an important economic crop in the tropical area of China. During a survey of diseases in July and September of 2009, leaf spots were observed on cassava plants at three separate plantations in Guangxi (Yunfu and Wuming) and Hainan (Baisha) provinces. Circular or irregular-shaped leaf spots were present on more than one-third of the plants. Spots were dark brown or had white papery centers delimited by dark brown rims and surrounded by a yellow halo. Usually, the main vein or small veinlets adjacent to the spots were dark. Some defoliation of plants was evident at the Wuming location. A fungus was isolated from symptomatic leaves from each of the three locations and designated CCCGX01, CCCGX02, and CCCHN01. Single-spore cultures of these isolates were incubated on potato dextrose agar (PDA) for 7 days with a 12-h light/dark cycle at a temperature of 28 ± 1°C. Conidiophores were straight to slightly curved, unbranched, and pale to light brown. Conidia were formed singly or in chains, obclavate to cylindrical, straight or curved, subhyaline-to-pale olivaceous brown, 19.6 to 150.3 μm long and 5.5 to 10.7 μm wide at the base, with 4 to 13 pseudosepta. Morphological characteristics of the specimen and their conidia were similar to the descriptions for Corynespora cassiicola (2). The isolate CCCGX01 was selected as a representative for molecular identification. Genomic DNA was extracted by the cetyltrimethylammoniumbromide protocol (3) from mycelia and used as a template for amplification of the internal transcribed spacer (ITS) region of rDNA with primer pair ITS1/ITS4. The sequence (GenBank Accession No. GU138988) exactly matched several sequences (e.g., GenBank Accession Nos. FJ852715, EF198117, and AY238606) of C. cassiicola (1). Young, healthy, and fully expanded green leaves of cassava cv. SC205 were surface sterilized. Ten leaves were inoculated with 10-μl drops of 104 ml suspension of conidia and five leaves were inoculated with the same volume of sterile water to serve as controls. After inoculation, leaves were placed in a dew and dark chamber for 36 h at 25°C and subsequently transferred to the light for 5 days. All inoculated leaves with isolates showed symptoms similar to those observed in natural conditions, whereas the controls remained symptom free. The morphological characteristics of reisolated conidia that formed on the diseased parts were identical with the nature isolates. To our knowledge, this is the first report of leaf spot caused by C. cassiicola on cassava in China. References: (1) L. J. Dixon et al. Phytopathology 99:1015, 2009. (2) M. B. Ellis et al. Corynespora cassiicola. No. 303 in: CMI Description of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, UK 1971. (3) J. R. Xu et al. Genetics 143:175, 1996.

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

Plant Disease ◽  
2010 ◽  
Vol 94 (8) ◽  
pp. 1064-1064 ◽  
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.

scholarly journals First Report of a Leaf Spot on Hazel Leaves Caused by Phyllosticta coryli in Liaoning Province of China

Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1254-1254 ◽  
Author(s):  
J. Sun ◽  
D.-M. Wang ◽  
X.-Y. Huang ◽  
Z.-H. Liu
Keyword(s):  
Leaf Spot ◽  
Control Strategies ◽  
Morphological Characteristics ◽  
Leaf Tissue ◽  
Its Region ◽  
Liaoning Province ◽  
Single Spore ◽  
First Report ◽  
Leaf Spots ◽  
Microscopic Morphology

Hazel (Corylus heterophylla Fischl) is an important nut tree grown in China, especially in Liaoning Province, and is rich in nutritional and medicinal values. In August 2011, leaf spotting was observed on hybrid hazel (Dawei) leaves in Paotai Town, Wafangdian County of Liaoning Province. By August 2012, the disease had spread to Zhangdang Town, Fushun County. Symptoms initially appeared on both sides of leaves as pinpoint brown spots, which enlarged and developed into regular, dark brown lesions, 3 to 9 mm in diameter. The lesions were lighter in color in the center compared to the margin. To identify the pathogen, leaf pieces (3 to 5 mm) taken from the margins, including both symptomatic and healthy portions of leaf tissue, were surface-disinfected first in 75% ethanol for 5 s, next in 0.1% aqueous mercuric chloride for 50 s, and then rinsed with sterilized water three times. Leaf pieces were incubated on potato dextrose agar (PDA) at 25°C for 14 days in darkness. Single spore isolates were obtained from individual conidia. For studies of microscopic morphology, isolates were grown on synthetic nutrient agar (SNA) in slide cultures. Colonies grew up to 45 to 48 mm in diameter on PDA after 14 days. Pycnidia appeared on the colonies after 12 days. Conidiophores were short. Pycnidia were dark brown, subglobose, and 150 to 205 μm in diameter. Conidia were unicellular, colorless, ovoid to oval, and from 2.4 to 4.5 × 1.6 to 2.4 μm. On the basis of these morphological characteristics, the isolates were tentatively identified as Phyllosticta coryli Westend (2). The rDNA internal transcribed spacer (ITS) region was amplified using primers ITS1 and ITS4 and sequenced (GenBank Accession No. KC196068). The 490-bp amplicons had 100% identity to an undescribed Phyllosticta species isolated from Cornus macrophylla in Gansu, Tianshui, China (AB470897). On the basis of morphological characteristics and nucleotide homology, the isolate was tentatively identified as P. coryli. Koch's postulates were fulfilled in the growth chamber on hazelnut leaves inoculated with P. coryli conidial suspensions (107 conidia ml–1). Eight inoculated 1-year-old seedlings (Dawei) were incubated under moist conditions for 8 to 10 days at 25°C. All leaf spots that developed on inoculated leaves were similar in appearance to those observed on diseased hazel leaves in the field. P. coryli was recovered from lesions and its identity was confirmed by morphological characteristics. P. coryli was first reported as a pathogen of hazel leaves in Bull of Belgium (2). In China, P. coryli was first reported on Corylus heterophylla Fisch. in Jilin Province (1). To our knowledge, this is the first report of P. coryli causing leaf spot on hybrid hazel in Liaoning Province of China. The outbreak and spread of this disease may decrease the yield of hazelnut in northern regions of China. More studies are needed on control strategies, including the possible resistance of hazel cultivars to P. coryli. References: (1) Y. Li et al. J. Shenyang Agric. Univ. 25:153, 1994. (2) P. A. Saccardo. Sylloge Fungorum Vol. III, page 31, 1884.

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

Plant Disease ◽  
2012 ◽  
Vol 96 (6) ◽  
pp. 917-917 ◽  
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.

scholarly journals First Report of Alternaria alternata Causing Leaf Spot on Bruguiera gymnorrhiza in China

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 286-286 ◽  
Author(s):  
Q.-L. Lin ◽  
H.-R. Su ◽  
H. He
Keyword(s):  
Leaf Spot ◽  
Direct Sequencing ◽  
Morphological Characteristics ◽  
Its Region ◽  
Average Diameter ◽  
Morphological Observation ◽  
Leaf Spot Disease ◽  
Bruguiera Gymnorrhiza ◽  
First Report ◽  
Mangrove Tree

Bruguiera gymnorrhiza (L.) Savigny is an important mangrove tree species that grows in the intertidal regions of the tropical and subtropical coastlines. In a survey conducted in March 2014, a leaf spot disease on this plant was observed in Sea View Promenade in Zhanjiang, Guangdong Province, China. Symptoms on leaves initially appeared as small circular to irregular, dark brown, necrotic, sunken spots with an average diameter of 4 to 7 mm. The spots gradually enlarged in size, becoming irregular, or remained circular with concentric rings or zones. In the latter, the spots coalesced, and the leaves withered, dried, and fell from the plants. Leaf tissues (3 × 5 mm), cut from the margins of lesions, were surface-disinfected, placed on potato dextrose agar (PDA), and incubated at 28°C with a 12-h photoperiod. Five fungi (MLL1 to MLL5) with different morphological characteristics were obtained. To fulfill Koch's postulates, wounded and nonwounded leaves were inoculated. Fresh wounds were made with a sterile needle on 10 detached leaves and 10 leaves on five living plants for fungi MLL1 to MLL5 independently. Mycelial plugs of each fungus were applied to wounded and nonwounded leaves. For the control, 10 leaves on five living plants were inoculated with agar plugs in a similar manner, to both wounded and nonwounded leaves. All treatments were incubated in a humid chamber in the dark at 28°C. Leaf spots identical to those observed in the field were observed on the wounded leaves inoculated with fungus MLL3 after 3 to 4 days, while the other four fungi and the control remained symptomless. The 10 nonwounded leaves inoculated with fungus MLL3 were also infected after 5 days. The fungus, with the same colony and conidial morphology as MLL3, was re-isolated from the affected leaves. The pathogenic test was repeated three times under the same conditions. Hyphal tips of MLL3 were transferred to PDA for morphological observation. Colonies of white-to-dark-gray mycelia, black on the underside, formed on PDA. The colonies were further identified as Alternaria sp., 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 (3). Conidia varied from 22.5 × 40.26 to 3.95 × 5.79 μm and had three to eight transverse and zero to four longitudinal septa, with a beak length of 0 to 7.25 μm. For molecular identification, PCR was carried out using internal transcribed spacer (ITS) region primers ITS1/ITS4, partial sequences of the beta tubulin gene primers Bt1a-Bt1b (1), and A. alternata species-specific primers AAF2/AAR3 (2). The PCR products were subjected to direct sequencing. The resulting sequences were compared against the GenBank nucleotide database by using a BLAST alignment, which revealed that MLL3 had 99 to 100% identity with A. alternata for the ITS, Bt1a-Bt1b, and AAF2/AAR3 regions (GenBank Accession Nos. KF669893, GQ240308, and KJ716876, respectively). Sequences for MLL3 were deposited in GenBank under accession numbers KJ767515, KJ921779, and KJ921778. According to both morphological and sequence analyses, the pathogen of the leaf spot of B. gymnorrhiza was identified as A. alternata. To our knowledge, this is the first report of A. alternata on leaves of B. gymnorrhiza in China. This pathogen could cause serious foliar damage and threaten the survival, growth, and fitness of the local B. gymnorrhiza community. References: (1) N. L. Glass and G. C. Donaldson. Appl. Environ. Microbiol. 61:1323, 1995. (2) P. Konstantinova et al. Mycol. Res. 106:23, 2002. (3) E. G. Simmons. Alternaria: An identification Manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

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