Leaf Spot on Lettuce (Lactuca sativa) Caused by Stemphylium solani, a New Disease in Malaysia

In June 2011, lettuce (Lactuca sativa) plants cultivated in major lettuce growing areas in Malaysia, including the Pahang and Johor states, had extensive leaf spots. In severe cases, disease incidence was recorded more than 80%. Symptoms on 50 observed plants initially were as water soaked spots (1 to 2 mm in diameter) on leaves, and then became circular spots spreading over much of the leaves. In this research, main lettuce growing areas infected by the pathogen in the mentioned states were investigated and the pathogen was isolated onto potato dextrose agar (PDA). Colonies observed were greyish green to light brown. Single conidia were formed at the terminal end of conidiophores that were 28.8 to 40.8 μm long and 11.0 to 19.2 μm wide, and 2 to 7 transverse and 1 to 4 longitudinal septa. To produce conidia, the fungus was grown on potato carrot agar (PCA) and V8 juice agar media under 8-h/16-h light/dark photoperiod. Fourteen isolates were identified Stemphylium solani based on morphological criteria described by Kim et al. (1). To confirm morphological characterization, DNA of the fungus was extracted from mycelium and PCR was done using universal primers ITS5 (5′-GGAAGTAAAAGTCGTAACAAGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′), which amplified the internal transcribed spacer (ITS) region of rDNA (2). The sequencing result was subjected to BLAST analysis which was 99% identical to the other published sequences in the GenBank database (GenBank Accession Nos. AF203451 and HQ840713). The nucleotide sequence was deposited in GenBank under Accession No. JQ736022. Pathogenicity testing of representative isolate was done using 20 μl of conidial suspension with a concentration of 1 × 105/ml in droplets (three drops on each leaf) on four detached 45-day-old lettuce leaves cv. BBS012 (3). Fully expended leaves were placed on moist filter paper in petri dishes and were incubated in humid chambers at 25°C. The leaves inoculated with sterile water served as control. After 7 days, disease symptoms were observed, which were similar to those symptoms collected in infected fields and the fungus was reisolated and confirmed as S. solani based on morphological criteria (1) and molecular characterization (2). Control leaves remained healthy. Pathogenicity testing was completed twice. To our knowledge, this is the first report of S. solani on lettuce in Malaysia and it may become a serious problem because of its broad host range, variability in pathogenic isolates, and prolonged active phase of the disease cycle. Previous research has shown that S. solani is a causal agent of gray leaf spot on lettuce in China (4). References: (1) B. S. Kim et al. Plant Pathol. J. 20:85, 2004. (2) Y. R. Mehta et al. Current Microbiol. 44:323, 2002. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) F. L. Tai. Sylloge Fungorum Sinicorum, Sci. Press, Acad. Sin., Peking, 1979.

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An Outbreak of Leaf Spot Caused by Stemphylium solani on Eggplant in Malaysia

Plant Disease ◽

10.1094/pdis-10-12-0901-pdn ◽

2013 ◽

Vol 97 (5) ◽

pp. 689-689

Author(s):

A. Nasehi ◽

J. B. Kadir ◽

M. Nasr Esfahani ◽

F. Mahmodi ◽

H. Ghadirian ◽

...

Keyword(s):

Leaf Spot ◽

Direct Sequencing ◽

Disease Incidence ◽

Solanum Melongena ◽

Its Region ◽

Gray Leaf Spot ◽

Distilled Water ◽

Conidial Suspension ◽

Morphological Criteria ◽

Pathogenicity Testing

In 2011, a severe gray leaf spot was observed on eggplant (Solanum melongena) in major eggplant growing areas in Malaysia, including the Pahang, Johor, and Selangor states. Disease incidence was >70% in severely infected areas of about 150 ha of eggplant greenhouses and fields examined. Symptoms initially appeared as small (1 to 5 mm diameter), brownish-black specks with concentric circles on the lower leaves. The specks then coalesced and developed into greyish-brown, necrotic lesions, which also appeared on the upper leaves. Eventually, the leaves senesced and were shed. Tissue cut from the edges of leaf spots were surface-sterilized in 1% NaOCl for 2 min, rinsed in sterilized water, dried, and incubated on potato dextrose agar (PDA). Fungal colonies were greyish green to light brown, and produced a yellow pigment. Single, muriform, brown, oblong conidia formed at the terminal end of each conidiophore, were each 21.6 to 45.6 μm long and 11.5 to 21.6 μm wide, and contained 2 to 7 transverse and 1 to 4 longitudinal septa. The conidiophores were tan to light brown and ≤220 μm long. Based on these morphological criteria, 25 isolates of the fungus were identified as Stemphylium solani (1). To produce conidia in culture, 7-day-old single-conidial cultures were established on potato carrot agar (PCA) and V8 juice agar media under an 8-h/16-h light/dark photoperiod at 25°C (4). Further confirmation of the identification was obtained by molecular characterization in which fungal DNA was extracted and the internal transcribed spacer (ITS) region of ribosomal DNA amplified using primers ITS5 and ITS4 (2), followed by direct sequencing. A BLAST search in the NCBI database revealed that the sequence was 99% identical with published ITS sequences for two isolates of S. solani (Accession Nos. AF203451 and HQ840713). The amplified ITS region was deposited in GenBank (JQ736023). Pathogenicity testing of a representative isolate was performed on detached, 45-day-old eggplant leaves of the cv. 125066-X under laboratory conditions. Four fully expanded leaves (one wounded and two non-wounded leaflets/leaf) were placed on moist filter paper in petri dishes, and each leaflet inoculated with a 20-μl drop of a conidial suspension containing 1 × 105 conidia/ml in sterilized, distilled water (3). The leaves were wounded by applying pressure to leaf blades with the serrated edge of forceps. Four control leaves were inoculated similarly with sterilized, distilled water. Inoculated leaves were incubated in humid chambers at 25°C with 95% RH and a 12-h photoperiod. After 7 days, symptoms similar to those observed in the original fields developed on both wounded and non-wounded inoculated leaves, but not on control leaves, and S. solani was reisolated consistently from the symptoms using the same method as the original isolations. Control leaves remained asymptomatic and the fungus was not isolated from these leaves. The pathogenicity testing was repeated with similar results. To our knowledge, this is the first report of S. solani on eggplant in Malaysia. References: (1) B. S. Kim et al. Plant Pathol. J. 20:85, 2004. (2) Y. R. Mehta et al. Curr. Microbiol. 44:323, 2002. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiv. Series 6:775, 2007.

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First Report of Gray Leaf Spot on Pepper Caused by Stemphylium solani in Malaysia

Plant Disease ◽

10.1094/pdis-03-12-0262-pdn ◽

2012 ◽

Vol 96 (8) ◽

pp. 1227-1227 ◽

Cited By ~ 4

Author(s):

A. Nasehi ◽

J. B. Kadir ◽

M. A. Zainal Abidin ◽

M. Y. Wong ◽

F. Abed Ashtiani

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Leaf Tissue ◽

Gray Leaf Spot ◽

Universal Primers ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Morphological Criteria ◽

Pepper Leaves

Symptoms of gray leaf spot were first observed in June 2011 on pepper (Capsicum annuum) plants cultivated in the Cameron Highlands and Johor State, the two main regions of pepper production in Malaysia (about 1,000 ha). Disease incidence exceeded 70% in severely infected fields and greenhouses. Symptoms initially appeared as tiny (average 1.3 mm in diameter), round, orange-brown spots on the leaves, with the center of each spot turning gray to white as the disease developed, and the margin of each spot remaining dark brown. A fungus was isolated consistently from the lesions using sections of symptomatic leaf tissue surface-sterilized in 1% NaOCl for 2 min, rinsed in sterile water, dried, and plated onto PDA and V8 agar media (3). After 7 days, the fungal colonies were gray, dematiaceous conidia had formed at the end of long conidiophores (19.2 to 33.6 × 12.0 to 21.6 μm), and the conidia typically had two to six transverse and one to four longitudinal septa. Fifteen isolates were identified as Stemphylium solani on the basis of morphological criteria described by Kim et al. (3). The universal primers ITS5 and ITS4 were used to amplify the internal transcribed spacer region (ITS1, 5.8, and ITS2) of ribosomal DNA (rDNA) of a representative isolate (2). A 570 bp fragment was amplified, purified, sequenced, and identified as S. solani using a BLAST search with 100% identity to the published ITS sequence of an S. solani isolate in GenBank (1). The sequence was deposited in GenBank (Accession No. JQ736024). Pathogenicity of the fungal isolate was tested by inoculating healthy pepper leaves of cv. 152177-A. A 20-μl drop of conidial suspension (105 spores/ml) was used to inoculate each of four detached, 45-day-old pepper leaves placed on moist filter papers in petri dishes (4). Four control leaves were inoculated similarly with sterilized, distilled water. The leaves were incubated at 25°C at 95% relative humidity for 7 days. Gray leaf spot symptoms similar to those observed on the original pepper plants began to develop on leaves inoculated with the fungus after 3 days, and S. solani was consistently reisolated from the leaves. Control leaves did not develop symptoms and the fungus was not reisolated from these leaves. Pathogenicity testing was repeated with the same results. To our knowledge, this is the first report of S. solani causing gray leaf spot on pepper in Malaysia. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) M. P. S. Camara et al. Mycologia 94:660, 2002. (3) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (4) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002.

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First Report of Tomato Gray Leaf Spot Disease Caused by Stemphylium solani in Malaysia

Plant Disease ◽

10.1094/pdis-03-12-0223-pdn ◽

2012 ◽

Vol 96 (8) ◽

pp. 1226-1226

Author(s):

A. Nasehi ◽

J. B. Kadir ◽

M. A. Zainal Abidin ◽

M. Y. Wong ◽

F. Mahmodi

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Gray Leaf Spot ◽

Leaf Spot Disease ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Spot Disease ◽

Pcr Product ◽

Morphological Criteria

In June 2011, tomatoes (Solanum lycopersicum) in major growing areas of the Cameron Highlands and the Johor state in Malaysia were affected by a leaf spot disease. Disease incidence exceeded 80% in some severely infected regions. Symptoms on 50 observed plants initially appeared on leaves as small, brownish black specks, which later became grayish brown, angular lesions surrounded by a yellow border. As the lesions matured, the affected leaves dried up and became brittle and later developed cracks in the center of the lesions. A survey was performed in these growing areas and 27 isolates of the pathogen were isolated from the tomato leaves on potato carrot agar (PCA). The isolates were purified by the single spore technique and were transferred onto PCA and V8 agar media for conidiophore and conidia production under alternating light (8 hours per day) and darkness (16 hours per day) (4). Colonies on PCA and V8 agar exhibited grey mycelium and numerous conidia were formed at the terminal end of conidiophores. The conidiophores were up to 240 μm long. Conidia were oblong with 2 to 11 transverse and 1 to 6 longitudinal septa and were 24 to 69.6 μm long × 9.6 to 14.4 μm wide. The pathogen was identified as Stemphylium solani on the basis of morphological criteria (2). In addition, DNA was extracted and the internal transcribed spacer region (ITS) was amplified by universal primers ITS5 and ITS4 (1). The PCR product was purified by the commercial PCR purification kit and the purified PCR product sequenced. The resulting sequences were 100% identical to published S. solani sequences (GenBank Accestion Nos. AF203451 and HQ840713). The amplified ITS region was deposited with NCBI GenBank under Accession No. JQ657726. A representative isolate of the pathogen was inoculated on detached 45-day-old tomato leaves of Malaysian cultivar 152177-A for pathogenicity testing. One wounded and two nonwounded leaflets per leaf were used in this experiment. The leaves were wounded by applying pressure to leaf blades with the serrated edge of a forceps. A 20-μl drop of conidial suspension containing 105 conidia/ml was used to inoculate these leaves (3). The inoculated leaves were placed on moist filter paper in petri dishes and incubated for 48 h at 25°C. Control leaves were inoculated with sterilized distilled water. After 7 days, typical symptoms for S. solani similar to those observed in the farmers' fields developed on both wounded and nonwounded inoculated leaves, but not on noninoculated controls, and S. solani was consistently reisolated. To our knowledge, this is the first report of S. solani causing gray leaf spot of tomato in Malaysia. References: (1) M. P. S. Camara et al. Mycologia 94:660, 2002. (2) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (3) B. M. Pryor and T. J. Michailides. Phytopathology 92:406, 2002. (4) E. G. Simmons. CBS Biodiversity Series 6:775, 2007.

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First Report of Alternaria Leaf Spot Caused by Alternaria sp. on Highbush Blueberry (Vaccinium corymbosum) in South Korea

Plant Disease ◽

10.1094/pdis-04-14-0344-pdn ◽

2014 ◽

Vol 98 (10) ◽

pp. 1434-1434

Author(s):

J.-H. Kwon ◽

D.-W. Kang ◽

M.-G. Cheon ◽

J. Kim

Keyword(s):

South Korea ◽

Leaf Spot ◽

Disease Incidence ◽

Vaccinium Corymbosum ◽

Its Rdna ◽

Universal Primers ◽

Conidial Suspension ◽

First Report ◽

Representative Isolate ◽

Alternaria Sp

In South Korea, the culture, production, and consumption of blueberry (Vaccinium corymbosum) have increased rapidly over the past 10 years. In June and July 2012, blueberry plants with leaf spots (~10% of disease incidence) were sampled from a blueberry orchard in Jinju, South Korea. Leaf symptoms included small (1 to 5 mm in diameter) brown spots that were circular to irregular in shape. The spots expanded and fused into irregularly shaped, large lesions with distinct dark, brownish-red borders. The leaves with severe infection dropped early. A fungus was recovered consistently from sections of surface-disinfested (1% NaOCl) symptomatic leaf tissue after transfer onto water agar and sub-culture on PDA at 25°C. Fungal colonies were dark olive and produced loose, aerial hyphae on the culture surfaces. Conidia, which had 3 to 6 transverse septa, 1 to 2 longitudinal septa, and sometimes also a few oblique septa, were pale brown to golden brown, ellipsoid to ovoid, obclavate to obpyriform, and 16 to 42 × 7 to 16 μm (n = 50). Conidiophores were pale to mid-brown, solitary or fasciculate, and 28 to 116 × 3 to 5 μm (n = 50). The species was placed in the Alternaria alternata group (1). To confirm the identity of the fungus, the complete internal transcribed spacer (ITS) rDNA region of a representative isolate, AAVC-01, was amplified using ITS1 and ITS4 primers (2). The DNA products were cloned into the pGEM-T Easy vector (Promega, Madison, WI) and the resulting pOR13 plasmid was sequenced using universal primers. The resulting 570-bp sequence was deposited in GenBank (Accession No. KJ636460). Comparison of ITS rDNA sequences with other Alternaria spp. using ClustalX showed ≥99% similarity with the sequences of A. alternata causing blight on Jatropha curcas (JQ660842) from Mexico and Cajannus cajan (JQ074093) from India, citrus black rot (AF404664) from South Africa, and other Alternaria species, including A. tenuissima (WAC13639) (3), A. lini (Y17071), and A. longipes (AF267137). Two base substitutions, C to T at positions 345 and 426, were found in the 570-bp amplicon. Phylogenetic analysis revealed that the present Alternaria sp. infecting blueberry grouped separately from A. tenuissima and A. alternata reported from other hosts. A representative isolate of the pathogen was used to inoculate V. corymbosum Northland leaves for pathogenicity testing. A conidial suspension (2 × 104 conidia/ml) from a single spore culture and 0.025% Tween was spot inoculated onto 30 leaves, ranging from recently emerged to oldest, of 2-year-old V. corymbosum Northland plants. Ten leaves were treated with sterilized distilled water and 0.025% Tween as a control. The plants were kept in a moist chamber with >90% relative humidity at 25°C for 48 h and then moved to a greenhouse. After 15 days, leaf spot symptoms similar to those observed in the field developed on the inoculated leaves, whereas the control plants remained asymptomatic. The causal fungus was re-isolated from the lesions of the inoculated plants to fulfill Koch's postulates. To our knowledge, this is the first report of Alternaria sp. on V. corymbosum in South Korea. References: (1) E. G. Simmons. Page 1797 in: Alternaria: An Identification Manual. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands, 2007. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (3) M. P. You et al. Plant Dis. 98:423, 2014.

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First report of banana (Musa acuminate L. AAA Cavendish, cv. Formosana) leaf spot disease caused by Curvularia geniculata in China

Plant Disease ◽

10.1094/pdis-09-21-2001-pdn ◽

2021 ◽

Author(s):

Yanxiang Qi ◽

Yanping Fu ◽

Jun Peng ◽

Fanyun Zeng ◽

Yanwei Wang ◽

...

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Morphological Characteristics ◽

Universal Primers ◽

Leaf Spot Disease ◽

Leaf Margin ◽

Conidial Suspension ◽

Tropical Fruit ◽

First Report ◽

Spot Disease

Banana (Musa acuminate L.) is an important tropical fruit in China. During 2019-2020, a new leaf spot disease was observed on banana (M. acuminate L. AAA Cavendish, cv. Formosana) at two orchards of Chengmai county (19°48ʹ41.79″ N, 109°58ʹ44.95″ E), Hainan province, China. In total, the disease incidence was about 5% of banana trees (6 000 trees). The leaf spots occurred sporadically and were mostly confined to the leaf margin, and the percentage of the leaf area covered by lesions was less than 1%. Symptoms on the leaves were initially reddish brown spots that gradually expanded to ovoid-shaped lesions and eventually become necrotic, dry, and gray with a yellow halo. The conidia obtained from leaf lesions were brown, erect or curved, fusiform or elliptical, 3 to 4 septa with dimensions of 13.75 to 31.39 µm × 5.91 to 13.35 µm (avg. 22.39 × 8.83 µm). The cells of both ends were small and hyaline while the middle cells were larger and darker (Zhang et al. 2010). Morphological characteristics of the conidia matched the description of Curvularia geniculata (Tracy & Earle) Boedijn. To acquire the pathogen, tissue pieces (15 mm2) of symptomatic leaves were surface disinfected in 70% ethanol (10 s) and 0.8% NaClO (2 min), rinsed in sterile water three times, and transferred to potato dextrose agar (PDA) for three days at 28°C. Grayish green fungal colonies appeared, and then turned fluffy with grey and white aerial mycelium with age. Two representative isolates (CATAS-CG01 and CATAS-CG92) of single-spore cultures were selected for molecular identification. Genomic DNA was extracted from the two isolates, the internal transcribed spacer (ITS), large subunit ribosomal DNA (LSU rDNA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), translation elongation factor 1-alpha (TEF1-α) and RNA polymerase II second largest subunit (RPB2) were amplified and sequenced with universal primers ITS1/ITS4, LROR/LR5, GPD1/GPD2, EF1-983F/EF1-2218R and 5F2/7cR, respectively (Huang et al. 2017; Raza et al. 2019). The sequences were deposited in GenBank (MW186196, MW186197, OK091651, OK721009 and OK491081 for CATAS-CG01; MZ734453, MZ734465, OK091652, OK721100 and OK642748 for CATAS-CG92, respectively). For phylogenetic analysis, MEGA7.0 (Kumar et al. 2016) was used to construct a Maximum Likelihood (ML) tree with 1 000 bootstrap replicates, based on a concatenation alignment of five gene sequences of the two isolates in this study as well as sequences of other Curvularia species obtained from GenBank. The cluster analysis revealed that isolates CATAS-CG01 and CATAS-CG92 were C. geniculata. Pathogenicity assays were conducted on 7-leaf-old banana seedlings. Two leaves from potted plants were stab inoculated by puncturing into 1-mm using a sterilized needle and placing 10 μl conidial suspension (2×106 conidia/ml) on the surface of wounded leaves and equal number of leaves were inoculated with sterile distilled water serving as control (three replicates). Inoculated plants were grown in the greenhouse (12 h/12 h light/dark, 28°C, 90% relative humidity). Necrotic lesions on inoculated leaves appeared seven days after inoculation, whereas control leaves remained healthy. The fungus was recovered from inoculated leaves, and its taxonomy was confirmed morphologically and molecularly, fulfilling Koch’s postulates. C. geniculata has been reported to cause leaf spot on banana in Jamaica (Meredith, 1963). To our knowledge, this is the first report of C. geniculata on banana in China.

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First Report of Gray Leaf Spot Caused by Alternaria brassicae on Canola in Argentina

Plant Disease ◽

10.1094/pd-89-0207a ◽

2005 ◽

Vol 89 (2) ◽

pp. 207-207 ◽

Cited By ~ 2

Author(s):

S. Gaetán ◽

M. Madia

Keyword(s):

Leaf Spot ◽

Buenos Aires ◽

Disease Incidence ◽

Leaf Tissue ◽

Climatic Conditions ◽

Gray Leaf Spot ◽

Conidial Suspension ◽

Alternaria Brassicae ◽

First Report ◽

And Control

Canola (Brassica napus) is a developing oleaginous crop grown commercially in Argentina. During 2003, typical symptoms of a foliar disease were observed on canola plants in experimental field plots in Buenos Aires. Average disease incidence across 14 6-month-old canola cultivars was 27% (range 12 to 42%). Climatic conditions in Buenos Aires during August 2003 included moderate temperatures and periods with high humidity, which were apparently favorable for disease development. Symptoms were observed on leaves, stems, and pods. Leaf symptoms were randomly distributed on the adaxial surfaces and consisted of zonate lesions of alternating light gray and dark brown areas that were 6 to 10 mm in diameter. Remaining leaf tissue was chlorotic and affected leaves abscised. Stem infections appeared as irregular and elongated black lesions, 0.7 to 1.2 cm long. Pods lesions were circular, 6 to 8 mm in diameter, gray in the center, and surrounded by a diffuse dark brown margin. The disease developed progressively from the lower leaves to the pods, resulting in premature senescence of the tissues, chlorosis, and defoliation. Conidiophores bearing conidia colonized the lesions as a dark gray growth of spore masses. Segments (0.5 cm long) taken from leaves, stems, and pods of diseased plants were dipped in 70% ethanol, surface sterilized with NaOCl (1%) for 2 min, and rinsed in sterile water. Each segment was blotted dry and placed on potato dextrose agar. Plates were incubated in the dark at 25°C for 2 to 3 days, followed by incubation under NUV light and a 12-h light/dark photoperiod for 6 to 8 days. Six fungal isolates were obtained. Fungal colonies were pale gray with dark concentric rings. Conidia were yellow to pale brown, ellipsoid to ovoid, produced singly or in short chains, with 8 to 10 transverse septa and 2 to 6 longitudinal septa. The spore body measured 13 to 22 × 68 to 135 µm with a beak cell 42 to 101 µm long. On the basis of conidial and cultural characteristics, the fungus was identified as Alternaria brassicae (Berk.) Sacc (1). Koch's postulates were completed for three isolates by spray-inoculating foliage of 6-week-old canola plants of cvs. Caviar, Dunkeld, Eclipse, Impulse, Mistral, and Sponsor with a conidial suspension (1 × 105 conidia per ml). The experiment, which included four inoculated plants and two noninoculated control plants for each cultivar per isolate, was conducted in the greenhouse at 22 to 24°C and maintained at 75% relative humidity with no supplemental light. Inoculated and control plants were covered with polyethylene bags for 48 h after inoculation. Within 12 days, inoculated plants developed small, brown lesions on leaves and stems for all three isolates; the pathogen was successfully reisolated in all instances. Control plants, inoculated only with sterile distilled water, remained symptomless. The experiment was repeated with similar results. The results suggest that A. brassicae may be a threat to the main cultivars being grown in Argentina. To our knowledge, this is the first report of A. brassicae causing gray leaf spot of canola in Argentina. Reference: (1) J. Joly. Le genre Alternaria. Recherches Physiologiques, Biologiques, et Systématiques. Paul Lechevalier, ed. Paris, France, 1964.

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First Report of Pestalotiopsis microspora Causing Leaf Spot on Moyeam in China

Plant Disease ◽

10.1094/pdis-04-21-0859-pdn ◽

2021 ◽

Author(s):

Si-Qi Yuan ◽

icai Wang ◽

Ling Lei ◽

Ju-Yun Hong ◽

Tuyong Yi ◽

...

Keyword(s):

Nucleotide Sequence ◽

Leaf Spot ◽

Disease Incidence ◽

Universal Primers ◽

Molecular Characteristics ◽

Conidial Suspension ◽

Tubulin Gene ◽

First Report ◽

Wide Range ◽

Β Tubulin

Ampelopsis grossedentata, commonly known as moyeam, has been widely used as health care herbal tea since it contains natural plant protein cream, 17 amino acids, 14 micronutrients and lots of functional flavonoid and provides a wide range of pharmaceutical functions such as antioxidant, anti-inflammatory, antitumor (Carneiro et al. 2021; Zhang et al. 2020). Moyeam is primarily produced in Zhangjiajie, stretching over the area from between 109’40 to 110’20E to between 28’52 to 29’48N, at 1300 to1890 meter above the sea level, with subtropical humid monsoon climate. Its economic value surpasses $1.25 billion in China (Liang et al. 2020). In July 2020, leaf spots were observed on some moyeam plants in Zhangjiajie. Initial spots were pinhead-sized with a yellow halo margin. The spots developed into light brown necrotic spots 6 to 8 mm in diameter, often with a dark brown margin. After 4 days of development, the spots enlarged and coalesced into irregular shape, frequently falling out and giving the leaves a tattered appearance. The infected plants eventually died with disease incidence ranging from 18 to 23%. This disease resulted in production losses of up to $1.7 million in 2020. One fungal isolate was isolated from the symptomatic leaves based on our previously published methods (Yi et al. 2019). Colonies on potato dextrose agar (PDA) were thick and villous with white at the front of the plate and yellowish at the back. After 1 week, the fungus produced conidia, which were spindle-shaped, straight or slightly curved, with 5 cells, 4-euseptates and 2-3 apical accessory filaments. Morphologically, the fungus was similar to Pestalotiopsis spp. Aerial hyphae with vigorous growth were collected for molecular identification. ITS nucleotide sequence of the rDNA and β-tubulin gene were amplified and sequenced with universal primers ITS1/ITS4 and self-designed primers based on β-tubulin gene conserved motif. BLAST searches against GenBank indicated that the ITS nucleotide sequence shared 99% similarity with that of P. microspora (MG808374.1) and the β-tubulin gene sequence shared 99% similarity with that of P. microspora (AF115396.1). Based on morphological and molecular characteristics, the fungus was identified as P. microspora. ITS and the β-tubulin nucleotide sequences were deposited in GenBank (accession no. MW350011 and MW816914). Pathogenicity tests were carried out with the following procedure. Three healthy moyeam seedlings were sprayed with a conidial suspension of 1 x106 conidia/ml while the other three seedlings were sprayed with distilled water as the controls. Plants were maintained in a greenhouse at 28±1°C. After one day of inoculation, symptoms identical to those in the field developed in the plants inoculated with the fungus. In contrast, no symptoms developed on the control plants. P. microspora has been reported to cause diseases in many crops in China. However, this is the first report of P. microspora causing leaf spot in moyeam in China. Identifying the pathogen causing the disease is important to the development of effective disease management strategies for control of this disease.

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A Temperature and Leaf Wetness Duration-Based Model for Prediction of Gray Leaf Spot of Perennial Ryegrass Turf

Phytopathology ◽

10.1094/phyto.2003.93.3.336 ◽

2003 ◽

Vol 93 (3) ◽

pp. 336-343 ◽

Cited By ~ 22

Author(s):

W. Uddin ◽

K. Serlemitsos ◽

G. Viji

Keyword(s):

Perennial Ryegrass ◽

Leaf Spot ◽

Disease Incidence ◽

Severity Index ◽

Gray Leaf Spot ◽

Leaf Wetness ◽

Conidial Suspension ◽

Linear Quadratic ◽

Leaf Wetness Duration ◽

Functional Relationships

Gray leaf spot is a serious disease of perennial ryegrass (Lolium perenne), causing severe epidemics in golf course fairways. The effects of temperature and leaf wetness duration on the development of gray leaf spot of perennial ryegrass turf were evaluated in controlled environment chambers. Six-week-old Legacy II ryegrass plants were inoculated with an aqueous conidial suspension of Pyricularia grisea (approximately 8 × 104 conidia per ml of water) and subjected to four different temperatures (20, 24, 28, and 32°C) and 12 leaf wetness durations (3 to 36 h at 3-h intervals). Three days after inoculation, gray leaf spot developed on plants at all temperatures and leaf wetness durations. Disease incidence (percent leaf blades symptomatic) and severity (index 0 to 10; 0 = leaf blades asymptomatic, 10 = >90% leaf area necrotic) were assessed 7 days after inoculation. There were significant effects ( α = 0.0001) of temperature and leaf wetness duration on disease incidence and severity, and there were significant interactions ( α = 0.0001) between them. Among the four temperatures tested, 28°C was most favorable to gray leaf spot development. Disease incidence and severity increased with increased leaf wetness duration at all temperatures. A shorter leaf wetness duration was required for disease development under warmer temperatures. Analysis of variance with orthogonal polynomial contrasts and regression analyses were used to determine the functional relationships among temperature and leaf wetness duration and gray leaf spot incidence and severity. Significant effects were included in a regression model that described the relationship. The polynomial model included linear, quadratic, and cubic terms for temperature and leaf wetness duration effects. The adjusted coefficients of determination for the fitted model for disease incidence and severity were 0.84 and 0.87, respectively. The predictive model may be used as part of an integrated gray leaf spot forecasting system for perennial ryegrass turf.

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First Report of a Leaf Spot Disease of Bells-of-Ireland (Moluccella laevis) Caused by Cercospora apii in California

Plant Disease ◽

10.1094/pdis.2003.87.2.203a ◽

2003 ◽

Vol 87 (2) ◽

pp. 203-203

Author(s):

S. T. Koike ◽

S. A. Tjosvold ◽

J. Z. Groenewald ◽

P. W. Crous

Keyword(s):

Leaf Spot ◽

Sequence Data ◽

Disease Incidence ◽

Leaf Spot Disease ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Spot Disease ◽

Leaf Spots ◽

Initial Symptoms

Bells-of-Ireland (Moluccella laevis) (Lamiaceae) is an annual plant that is field planted in coastal California (Santa Cruz County) for commercial cutflower production. In 2001, a new leaf spot disease was found in these commercially grown cutflowers. The disease was most serious in the winter-grown crops in 2001 and 2002, with a few plantings having as much as 100% disease incidence. All other plantings that were surveyed during this time had at least 50% disease. Initial symptoms consisted of gray-green leaf spots. Spots were generally oval in shape, often delimited by the major leaf veins, and later turned tan. Lesions were apparent on both adaxial and abaxial sides of the leaves. A cercosporoid fungus having fasciculate conidiophores, which formed primarily on the abaxial leaf surface, was consistently associated with the spots. Based on morphology and its host, this fungus was initially considered to be Cercospora molucellae Bremer & Petr., which was previously reported on leaves of M. laevis in Turkey (1). However, sequence data obtained from the internal transcribed spacer region (ITS1, ITS2) and the 5.8S gene (STE-U 5110, 5111; GenBank Accession Nos. AY156918 and AY156919) indicated there were no base pair differences between the bells-of-Ireland isolates from California, our own reference isolates of C. apii, as well as GenBank sequences deposited as C. apii. Based on these data, the fungus was subsequently identified as C. apii sensu lato. Pathogenicity was confirmed by spraying a conidial suspension (1.0 × 105 conidia/ml) on leaves of potted bells-of-Ireland plants, incubating the plants in a dew chamber for 24 h, and maintaining them in a greenhouse (23 to 25°C). After 2 weeks, all inoculated plants developed leaf spots that were identical to those observed in the field. C. apii was again associated with all leaf spots. Control plants, which were treated with water, did not develop any symptoms. The test was repeated and the results were similar. To our knowledge this is the first report of C. apii as a pathogen of bells-of-Ireland in California. Reference: (1) C. Chupp. A Monograph of the Fungus Genus Cercospora. Cornell University Press, Ithaca, New York, 1954.

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First Report of Gray Leaf Spot on St. Augustinegrass in Italy

Plant Disease ◽

10.1094/pdis.2003.87.12.1536a ◽

2003 ◽

Vol 87 (12) ◽

pp. 1536-1536 ◽

Cited By ~ 1

Author(s):

G. Polizzi ◽

I. Castello ◽

A. M. Picco ◽

D. Rodino

Keyword(s):

Leaf Spot ◽

Magnaporthe Grisea ◽

Fungal Spore ◽

Gray Leaf Spot ◽

Blast Disease ◽

Leaf Spot Disease ◽

High Humidity ◽

Conidial Suspension ◽

First Report ◽

Pathogenicity Tests

St. Augustinegrass (Stenotaphrum secundatum (Walt.) Kuntze) is used for lawns in southern Italy because it is much more resistant to biotic and abiotic adversities than other turfgrass species. Because few seeds are viable, this species is established by vegetative propagation. A new disease was noticed during the spring of 2002 and 2003 on cuttings of St. Augustinegrass growing in three greenhouses in eastern Sicily. The disease affected leaves and culms and caused a progressive drying of the plants. The infection was first seen on leaves as gray, necrotic spots that enlarged in high-humidity conditions to form oval, and later, spindle-shaped lesions. In association with the lesions, it was possible to observe fungal spore development and sunken areas with blue-gray centers and slightly irregular, brown margins with yellow halos. Spots were concentrated without specific arrangement along longitudinal veins and the midrib and at the base, tip, and margins of the leaf blade. Symptoms on the culms consisted of brown-to-black blotches that sometimes extended throughout the internodes. From these infected tissues, 20 explants taken from leaves and culms were cut, washed with sterile water, and placed on 1.5% water agar (WA). Later, conidia and conidiophores were obtained from colonies with a sterile glass needle and placed on 4% WA. From these plates, two monoconidial isolates were obtained and transferred to rice meal medium (1). The colonies were identified as Pyricularia grisea Cooke (Sacc.), anamorphic state of Magnaporthe grisea (Hebert) Yeagashi & Udagawa, the cause of rice blast disease and gray leaf spot disease of turfgrasses. The conidia were pyriform to obclavate, narrowed toward the tip, rounded at the base, 2-septate, 21 to 31 μm × 6 to 10 μm (average 25.7 ×8.2 μm). Pathogenicity tests were performed by inoculating leaves and culms of six St. Augustinegrass plants with a conidial suspension of the fungus (1.5 ×105 conidia per ml). The same number of noninoculated plants was used as controls. All plants were incubated in a moist chamber with high humidity at 25°C. After 6 days, all inoculated plants showed typical symptoms of the disease. Koch's postulates were fulfilled by isolating P. grisea from inoculated plants. Gray leaf spot caused by P. grisea has been a chronic problem on St. Augustinegrass since it was first reported in 1957 (2). To our knowledge, this is the first report of P. grisea on St. Augustinegrass in Italy. While it does not appear to be an important disease in the field at this time in Sicily, it could cause losses in greenhouses where vegetative material is propagated for field planting. A preliminary molecular analysis has shown a clear distinction between the tested strain and other strains isolated from rice seeds and plants in northern Italy. References: (1) E. Roumen et al. Eur. J. Plant Pathol. 103:363, 1997. (2) L. P. Tredway et al. Plant Dis. 87:435, 2003.

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