scholarly journals First Report of Alternaria alternata Causing Leaf Spot on Aloe vera in Louisiana

Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1379-1379 ◽  
Author(s):  
W. L. da Silva ◽  
R. Singh

Aloe vera (L.) Burm. f. is a perennial succulent plant that is grown worldwide mainly for medicinal and cosmetic uses. In the USA, it is mainly cultivated in some southern states to produce aloe gel for the cosmetic industry (3), and in Louisiana it is also sold commercially as an ornamental. During the summer of 2011, several A. vera plants infected with leaf spots were observed on the campus of Louisiana State University, Baton Rouge. Large, necrotic, sunken, circular to oval, dark brown spots were present on both surfaces of the leaves. Infected leaf tissue pieces were surface disinfested with 1% NaOCl solution for 1 min and plated on potato dextrose agar (PDA). Plates were incubated at 28°C in the dark for 4 days. A dark olivaceous fungus with profuse golden brown, branched, and septate hyphae was consistently isolated from the infected tissue on PDA. The fungus produced conidia with longitudinal and transverse septa, and was morphologically identified as an Alternaria sp. (4). Conidia were produced in long chains, pale to light brown, obpyriform, with a beak (6.0 μm long), one to seven transverse and up to three longitudinal septa, and measured 10 to 45 μm long × 7 to 18 μm wide. Conidiophores were straight, septate, light to olive golden brown with conidial scar, and measured 35 to 100 μm long × 2 to 5 μm wide. Genomic DNA from a single-spored isolate was extracted and the internal transcribed spacer (ITS1-5.8s-ITS2) regions were amplified and sequenced using primers ITS1 and ITS4. BLASTn analysis of a 486-bp sequence (GenBank Accession No. JQ409455) resulted in 100% homology with A. alternata strain DHMJ16 (GenBank Accession No. JN986768) from China and several other Alternaria spp. The fungus was identified as A. alternata based on mycelial and conidia characters after being grown under standard, previously described conditions (4). Pathogenicity tests were carried out by inoculating six potted aloe plants with 0.5-cm diameter discs taken from a 6-day-old culture grown on PDA. Four discs were placed on the upper surface of each of the bottom leaves of every plant. Inoculated plants were individually covered with a plastic bag and maintained in a greenhouse for 1 week at 25 ± 2°C. Six control plants received only agar plugs. Seven days after inoculation, necrotic leaf spots were observed on the inoculated plants and A. alternata was reisolated from these spots. No leaf spots were observed on control plants. To the best of our knowledge, this is the first report of leaf spot caused by A. alternata on A. vera in Louisiana. Several outbreaks of the disease have been reported in Pakistan and India as damaging aloe gel production in those countries (1,2). An outbreak of this disease in Louisiana could represent a serious issue for the state's A. vera ornamental commerce. References: (1) R. Bajwa et al. Can. J. Plant Pathol. 32:490, 2010. (2) A. Kamalakannan et al. Australas. Plant Dis. Notes 3:110, 2008. (3) T. Reynolds. Aloes: The Genus Aloe. CRC Press, Boca Raton, FL, 2004. (4) E. G. Simmons. Alternaria: An Identification Manual: Fully Illustrated and with Catalogue Raisonné 1796-2007. CBS Fungal Biodiversity Centre, Utrecht, The Netherlands, 2007.

Plant Disease ◽  
2021 ◽  
Author(s):  
LiLi Zhang ◽  
Yan Zhong Li

A disease was observed on alfalfa cultivar WL168 characterized by white to brown leaf spots of regular to round shapes, in Aluhorqin County, Inner Mongolia, China (120°13′23″ to 120°29′14″ E, 43°27′52″to 43°35′16″ N, 281.71m to360.13 m Altitude) during 2019 to 2020. The disease mainly presented in spring one month after re-greening and the incidence was 78.30% in this field. Twenty alfalfa plants with severe symptoms were used for pathogen isolation. The infected tissue was cut into 2 × 2 mm pieces, surface-sterilized (in 75% ethanol and 5% commercial bleach (NaClO) for 30 s and 2 min, respectively), rinsed five times with sterilized distilled water, and dried between sterile filter paper (Wang et al. 2019). The diseased tissue from each plant sample were cultured on potato dextrose agar (PDA) and incubated at 25 °C with 12 h light/day for ten days. A fungus was isolated from the diseased leaves at a 100% frequency. Fungal growth on PDA was round with a black surface, radial edge, and a dirty white center. The ascocarps were moved to a clean microscope slide to release asci and ascospores. Ascocarps were spheroidal, subglobose brown, 120 to 160 µm × 160 to 180 µm, which contain several ascus. The size of ascus were 31.0 to 41.6 μm × 75.0 to 87.5 μm and each asci having eight ascospores. Ascospores were ellipsoid to oblong with a gelatinous sheath, brown, 8.8 to 15.0 µm × 29.9 to 43.0 µm with 2 to 3 horizontal septums, and 0 to 2 vertical septums. A phylogenetic tree was constructed after DNA extraction, PCR with primers to amplify the ITS (VG9: 5′- TTACGTCCCTGCCCTTTGTA-3′ and ITS4: 5′-TCCTCCGCTTATTGATATGC-3′) and LSU (LR7: 5′-TACTACCACCAAGATCT-3′ and LROR: 5′- GTACCCGCTGA ACTTAAGC -3′) regions. The LSU (SUB8273071) and ITS (SUB8218291) amplicons showed 99% similarity with L. australis (EU754166.1) in the GenBank. To verify the pathogenicity, fungs plugs were inverted on three compound leaves of 20 alfalfa WL168 for two days. Agar plugs (PDA) were inverted on another 20 alfalfa WL168 three compound leaves which were control. All plants were maintained at 22 °C and 44% relative humidity in a growth chamber. Similar disease symptoms were observed on infected leaves ten days after inoculation, while control plants showed no symptoms. The same fungus was re-isolated from the lesions, and further morphological characterization and molecular assays, as described above. L. australis has been reported on various plants, including Prunusarmeniaca, Dolichos, Poa, Lolium, and Vitis in Australia (Graham and Luttrell., 1961), and also from Korean soil in 2018 (Weilan et al., 2018). Additionally, L. briosiana, which is common in the USA, China, and other countries, causes Leptosphaerulina leaf spot (Samacet al., 2015). L. trifolii is newly reported to occur in China (Liu et al., 2019). To the best of our knowledge, this is the first report of L. australis infecting alfalfa in China. Considering the large planting area in Inner Mongolia, this pathogen may losses to alfalfa cultivation. Hence, future studies should explore aspects of effective management of this disease.


Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1257-1257 ◽  
Author(s):  
A. D. A. Silva ◽  
D. B. Pinho ◽  
B. T. Hora Junior ◽  
O. L. Pereira

Yucca filamentosa L. (Agavaceae), commonly known as Adam's needle, is known in Brazil as “agulha-de-adão.” It is an ornamental garden plant with medicinal properties (4). In 2010, 100% of Y. filamentosa seedlings and plants were observed with a severe leaf spot disease in two ornamental nurseries located in the municipality of Viçosa, Minas Gerais, Brazil. Initially, lesions were dark brown, elliptical, and scattered, and later became grayish at the center with a reddish brown margin, irregular and coalescent. Infected leaf samples were deposited in the herbarium at the Universidade Federal de Viçosa (Accession Nos. VIC32054 and VIC32055). A fungus was isolated from the leaf spots and single-spore pure cultures were obtained on potato dextrose agar (PDA). The sporulating single-spore cultures were deposited at the Coleção de Culturas de Fungos Fitopatogênicos “Prof. Maria Menezes” (CMM 1843 and CMM 1844). On the leaf, the fungus produced pycnidial conidiomata that were scattered or gregarious, usually epiphyllous, immersed, dark brown, unilocular, subglobose, and 95 to 158 × 108 to 175 μm, with a minute, subcircular ostiole. Conidiogenous cells were blastic, hyaline, conoidal, or short cylindrical. Conidia were aseptate, hyaline, smooth walled, coarsely granular, broadly ellipsoidal to subglobose or obovate, usually broadly rounded at both ends, occasionally truncate at the base or indented slightly at the apex, and 7.5 to 13.5 × 6 to 10 μm. Conidia were also surrounded by a slime layer, usually with a hyaline, flexuous, narrowly conoidal or cylindrical, mucilaginous apical appendage that was 10 to 16 μm long. Spermatia were hyaline, dumbbell shaped to cylindrical, both ends bluntly rounded, and 3 to 5 × 1 to 1.5 μm. These characteristics matched well with the description of Phyllosticta yuccae Bissett (1). To confirm this identification, DNA was extracted using a Wizard Genomic DNA Purification Kit and amplified using primers ITS1 and ITS4 (2) for the ITS region (GenBank Accession Nos. JX227945 and JX227946) and EF1-F and EF2-R (3) for the TEF-1α (JX227947 and JX227948). The sequencing was performed by Macrogen, South Korea. The ITS sequence matched sequence No. JN692541, P. yuccae, with 100% identity. To confirm Koch's postulates, four leaves of Y. filamentosa (five plants) were inoculated with 6-mm-diameter plugs from a 7-day-old culture growing on PDA. The leaves were covered with plastic sack and plants were maintained at 25°C. In a similar manner, fungus-free PDA plugs were placed on five control plants. Symptoms were consistently similar to those initially observed in the nurseries and all plants developed leaf spots by 15 days after inoculation. P. yuccae was successfully reisolated from the symptomatic tissue and control plants remained symptomless. P. yuccae has been previously reported in Canada, the Dominican Republic, Guatemala, Iran, and the United States of America. To our knowledge, this is the first report of P. yuccae causing disease in Y. filamentosa in Brazil and it may become a serious problem for the nurseries, due to the severity of the disease and the lack of chemical products to control this pathogen. References: (1) J. Bissett. Can. J. Bot. 64:1720, 1986. (2) M. A. Innis et al. PCR Protocols: A guide to methods and applications. Academic Press, 1990. (3) Jacobs et al. Mycol. Res. 108:411, 2004. (4) H. Lorenzi and H. M. Souza. Plantas Ornamentais no Brasil. Instituto Plantarum, 2001.


Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1254-1254 ◽  
Author(s):  
J. Sun ◽  
D.-M. Wang ◽  
X.-Y. Huang ◽  
Z.-H. Liu

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.


Plant Disease ◽  
2008 ◽  
Vol 92 (3) ◽  
pp. 486-486
Author(s):  
M. Zhang ◽  
H. L. Li ◽  
A. L. Zhao ◽  
J. X. Zhang

Tree peony (Paeonia suffruticosa) is known as “the king of flowers” for its beautiful and showy flowers. It is regarded as the symbol flower of China and is cultivated throughout the country. During the summer of 2006, a leaf spot was observed on tree peony cultivated in the Zhengzhou area of Henan Province, and in 2007, the leaf spot was observed in the Luoyang area. In some gardens, the leaf spot affected more than 50% of the plants. Early symptoms appeared as small, round, water-soaked lesions on the leaves. Lesions expanded into 5 to 35-mm-diameter spots that were circular or irregular, brown to dark brown, with pale brown margins. Later, the center of some lesions dropped out. Signs of the suspected pathogen were usually seen on the leaf spots after an abundant rainfall. Lesions contained numerous, pale brown, cupulate conidiomata with salmon-colored spore masses. Conidiophores (70 × 1 to 2 μm) were hyaline, branched, septate, and filiform. Conidia (5.5 to 7.5 × 1.5 to 2 μm) were hyaline, aseptate, and cymbiform to allantoid. The pathogen was identified as Hainesia lythri on the basis of the morphology. This fungus infects a wide variety of hosts including P. suffruticosa, Acer pseudoplatanus, Calluna sp., Dissotis paucistellata, Epilobium angustifolium, and Eucalyptus saligna (3). The fungus was isolated on potato dextrose agar (PDA) medium using conidia from conidiomata found on symptomatic leaf tissue; the fungus produced gray-to-brown colonies. Pathogenicity was tested by inoculating 10 leaves on one 5-year-old tree with a mycelia plug from the colony (0.5 cm in diameter); leaves inoculated with plugs of PDA medium served as controls. Inoculated leaves were covered with plastic for 24 h to maintain high relative humidity and incubated at 25 to 28°C. After 5 days, 100% of the inoculated leaves showed symptoms identical to those observed on leaves from P. suffruticosa infected in the field while controls remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was H. lythri. Thus, we concluded that H. lythri is the causal agent of leaf spots of P. suffruticosa. To our knowledge, this is the first report of H. lythri infecting P. suffruticosa in China. H. lythri has been previously reported on Paeonia in Japan and Korea (1,2). References: (1) W. D. Cho and H. D. Shin, eds. List of Plant Diseases in Korea. 4th ed. Korean Society of Plant Pathology, 2004. (2) M. E. Palm. Mycologia 83:787, 1991. (3) B. C. Sutton. The Coelomycetes. CAB International Publishing, New York, 1980.


Plant Disease ◽  
2013 ◽  
Vol 97 (9) ◽  
pp. 1256-1256 ◽  
Author(s):  
L. F. Zhai ◽  
J. Liu ◽  
M. X. Zhang ◽  
N. Hong ◽  
G. P. Wang ◽  
...  

Aloe vera L. var Chinese (Haw) Berg is a popular ornamental plant cultivated worldwide, whose extracts are used in cosmetics and medicine. Aloe plants are commonly affected by leaf spot disease caused by Alternaria alternata in Pakistan, India, and the United States (1). An outbreak of Alternaria leaf spot recently threatened aloe gel production and the value of ornamental commerce in Louisiana (1). During the summer of 2011, leaf spot symptoms were observed on A. vera plants growing in several greenhouses and ornamental gardens in Wuhan, Hubei Province, China. In two of the greenhouses, disease incidence reached 50 to 60%. The initial symptoms included chlorotic and brown spots that expanded to 2 to 4 mm in diameter and became darker with age. Lesions also developed on the tips of 30 to 50% of the leaves per plant. In severe infections, the lesions coalesced causing the entire leaf to become blighted and die. In September of 2012 and February of 2013, 10 symptomatic A. vera leaves were collected randomly from two greenhouses and gardens in Wuhan. A fungus was consistently recovered from approximately 80% of the tissue samples using conventional sterile protocols, and cultured on potato dextrose agar (PDA). The colonies were initially white, becoming grey to black, wool-like, and growing aerial mycelium covering the entire petri dish (9 cm in diameter) plate within 5 days when maintained in the dark at 25°C. The conidia were brown or black, spherical to subspherical, single celled (9 to 13 μm long × 11 to 15 μm wide), borne on hyaline vesicles at the tip of conidiophores. The conidiophores were short and rarely branched. These colonies were identified as Nigrospora oryzae based on the described morphological characteristics of N. oryzae (2). Genomic DNA was extracted from a representative isolate, LH-1, and the internal transcribed spacer region was amplified using primer pair ITS1/ITS4 (3). A 553-bp amplicon was obtained and sequenced. The resulting nucleotide sequence (GenBank Accession No. KC519728) had a high similarity of 99% to that of strain AHC-1 of N. oryzae (JQ864579). Pathogenicity tests for strain LH-1 were conducted in triplicate by placing agar pieces (5 mm in diameter) containing 5-day-old cultures on A. vera leaves. Four discs were placed on each punctured surface of each leaf. Noncolonized PDA agar pieces were inoculated as controls. Leaves were placed in moist chambers at 25°C with a 12-h photoperiod. After 3 days, the inoculated leaves showed symptoms similar to those observed in the greenhouses. N. oryzae was reisolated from these spots on the inoculated leaves. No visible symptoms developed on the control leaves. The pathogenicity tests were performed twice with the same results. Based on the results, N. oryzae was determined as a pathogen responsible for the leaf spots disease on A. vera. N. oryzae has been described as a leaf pathogen on fig (Ficus religiosa), cotton (Gossypium hirsutum) and Kentucky bluegrass (Poa pratensis) (4), and to our knowledge, this is the first report of N. oryae causing leaf spot disease on A. vera worldwide. References: (1) W. L. da Silva and R. Singh. Plant Dis. 86:1379, 2012. (2) M. B. Ellis. Dematiaceous Hyphomycetes, CAB, Kew, Surrey, England, 1971. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990. (4) L. X. Zhang et al. Plant Dis. 96:1379, 2012.


Plant Disease ◽  
2009 ◽  
Vol 93 (4) ◽  
pp. 425-425 ◽  
Author(s):  
M. Zhang ◽  
T. Tsukiboshi ◽  
I. Okabe

European columbine, Aquilegia vulgaris L., Ranunculaceae, is an herbaceous flower widely used in gardens, parterres, and courtyards and is a traditional herbal plant. During the summer of 2008, leaf spots were observed on a plant cultivated along a roadside area in Nasushiobara, Tochigi, Japan. In some courtyards, the leaf spot affected more than 60% of the plants. Early symptoms appeared as small, round or elliptic, brown lesions on the leaves. Lesions expanded to 5 to 15 × 4 to 10 mm, irregular spots that were dark brown to black in the middle, with pale yellow-brown or purple-brown margins. In continuously wet or humid conditions, thick, gray mycelium and conidia appeared on the surface of leaf spots. Conidiophores were melanized at the base and hyaline near the apex, branched, and septated (approximately 3 mm × 16 to 18 μm). Conidia were hyaline, aseptate, ellipsoidal to obovoid, with a slightly protuberant hilum, and ranged from 9 to 14.5 × 5.5 to 6.5 μm. The pathogen was identified as Botrytis cinerea Pers.:Fr on the basis of morphology and sequence of ITS1-5.8s-ITS2 region of rDNA. The sequence (GenBank Accession No. FJ424510) exactly matched the sequences of two Botryotinia fuckeliana (anamorph Botrytis cinerea), (e.g., GenBank Accession Nos. AY686865 and FJ169666) (2). The fungus was isolated on potato dextrose agar (PDA) from a single conidium found on the symptomatic leaf tissue. Colonies of B. cinerea were first hyaline and later turned gray to black when the spores differentiated. Koch's postulates were performed with three whole plants of potted aquilegia. Leaves were inoculated with mycelia plugs harvested from the periphery of a 7-day-old colony; an equal number of plants were inoculated with the plugs of PDA medium only and served as controls. All plants were covered with plastic bags for 24 h to maintain high relative humidity and incubated at 25°C. After 8 days, all mycelium-inoculated plants showed symptoms identical to those observed on leaves from A. vulgaris infected in the field, whereas controls remained symptom free. Reisolation of the fungus from lesions on inoculated leaves confirmed that the causal agent was B. cinerea. B. cinerea has been previously reported on A. vulgaris in the United States and China (1,3). To our knowledge, this is the first report of leaf spots caused by B. cinerea on A. vulgaris in Japan. References: (1) Anonymous. Index of Plant Diseases in the United States. USDA Agric. Handb. No 165, 1960. (2) M. B. Ellis. Dematiaceous Hyphomycetes. Commonwealth Mycological Institute, Kew, England, 1971. (3) Z. Y. Zhang. Flora Fungorum Sinicorum. Vol. 26. Botrytis, Ramularia. Science Press, Beijing, 2006.


Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 998-998 ◽  
Author(s):  
R. Carrieri ◽  
G. Pizzolongo ◽  
G. Carotenuto ◽  
P. Tarantino ◽  
E. Lahoz

During spring 2013, lamb's lettuce plants (Valerianella locusta) cv. Calarasi (Rijk Zwaan) in a commercial greenhouse in Sele Valley (Salerno Province, southern Italy) exhibited small, black-brown, irregular spots (1 mm2) that became necrotic, enlarged, and coalesced. The spots were mostly on the tips of leaves, and were surrounded by a yellow halo. The disease was severe under greenhouse conditions of 60 to 90% RH and maximum air temperature of 26°C, and affected up to 70% of the plants. The greenhouses covered an area of ~3,000 ha where many salad species are grown. Tissue fragments were excised from symptomatic leaves, sterilized by sequential dipping in 70% ethanol for 30 s and in 1% NaOCl for 30 s, rinsed in sterilized distilled water, and placed in 9-cm-diameter petri dishes containing potato dextrose agar (PDA) medium amended with streptomycin sulfate (0.1 g/liter). The plates were incubated at 24°C in the dark. A fungus was isolated consistently from infected leaf tissue after 4 days. Each colony was whitish to orange. Mycelium was hyaline, branched, septate, 3 to 4 μm wide, with numerous anastomosis-forming hyphal coils. Conidiophores were solitary, hyaline, smooth, thin-walled, unbranched or rarely irregularly branched. Conidiogenous cells were phialidic, determinate, discrete, smooth, solitary, and formed on hyphal coils. Phialides were aseptate or occasionally 1-septate near the base. Conidia (n = 100) were ellipsoidal, hyaline, smooth, septate or aseptate, and 6.6 ± 0.9 × 2.8 ± 0.4 μm. On the basis of morphological criteria (3), the fungus was ascribed to Plectosphaerella cucumerina (L.) Laterr. (anamorph Plectosporium tabacinum). An aliquot (50 ng) of genomic DNA extracted from mycelium of five cultures obtained by monosporic isolation on PDA was used as template for a PCR reaction with primers ITS5/ITS4, specific for the ITS 5.8S rDNA region of fungi (3). The 500-bp sequences amplified from the five isolates were identical, and the sequence of isolate Val-2 was submitted to GenBank (KF753234). Sequence analysis with BLASTn showed 100% identity of this sequence to the ITS-5.8S rDNA sequences of 11 isolates of P. cucumerina in GenBank. Three isolates were selected for pathogenicity tests on the lamb's lettuce cv. Calarasi. Before planting, seeds were surface-disinfected in 1% NaOCl and rinsed with sterilized distilled water. Plants (35 days old, 30 plants tested/isolate) were grown in 0.7-liter pots filled with a sterilized (autoclaved at 112°C for 1 h on each of two consecutive days) mixture of soil:sand:perlite (2:1:1), and inoculated by spraying the leaves with a spore suspension (106 CFU/ml, ~3 ml applied/plant) of each isolate prepared from 7-day-old cultures on PDA. As a control, five plants were sprayed with sterilized water. All plants were incubated in a growth chamber at 90% RH with a 12-h photoperiod at 24°C. Leaf spots typical of those on the original symptomatic plants appeared 7 to 10 days after inoculation on all inoculated plants. No symptoms were observed on control plants. P. cucumerina was re-isolated only from symptomatic leaves, as described for the original isolations. P. cucumerina has been associated with root and collar roots of some horticultural crops (1), and a leaf spot on Diplotaxis tenuifolia (2), often grown in rotation with lamb's lettuce in southern Italy. To our knowledge, this is the first report of P. cucumerina as a pathogen of V. locusta in Italy or elsewhere. The disease caused economic loss to lamb's lettuce, primarily used in Italy in fresh-cut, mixed salads. References: (1) A. Carlucci et al. Persoonia 28:34, 2012. (2) A. Garibaldi et al. Plant Dis. 96:1825, 2012. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.


Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 763-763 ◽  
Author(s):  
A. L. Vu ◽  
M. M. Dee ◽  
T. Russell ◽  
J. Zale ◽  
K. D. Gwinn ◽  
...  

Field-grown seedlings of ‘Alamo’ switchgrass (Panicum virgatum L.) from Vonore, TN exhibited light brown-to-dark brown leaf spots and general chlorosis in June 2009. Symptomatic leaf tissue was surface sterilized (95% ethanol for 1 min, 20% commercial bleach for 3 min, and 95% ethanol for 1 min), air dried on sterile filter paper, and plated on 2% water agar amended with 10 mg/liter rifampicin (Sigma-Aldrich, St. Louis, MO) and 5 μl/liter miticide (2.4 EC Danitol, Valent Chemical, Walnut Creek, CA). Plates were incubated at 26°C for 4 days in darkness. An asexual, dematiaceous mitosporic fungus was isolated and transferred to potato dextrose agar. Cultures were transferred to Alternaria sporulation medium (3) to induce conidial production. Club-shaped conidia were produced in chains with branching of chains present. Conidia were 27 to 50 × 10 to 15 μm, with an average of 42.5 × 12.5 μm. Morphological features and growth on dichloran rose bengal yeast extract sucrose agar were consistent with characteristics described previously for Alternaria alternata (1). Pathogenicity studies were conducted with 5-week-old ‘Alamo’ switchgrass plants grown from surface-sterilized seed. Nine pots with approximately 20 plants each were prepared. Plants were wounded by trimming the tops. Eight replicate pots were sprayed with a conidial spore suspension of 5.0 × 106 spores/ml sterile water and subjected to high humidity by enclosure in a plastic bag for 7 days. One pot was sprayed with sterile water and subjected to the same conditions to serve as a control. Plants were maintained in a growth chamber at 25/20°C with a 12-h photoperiod. Foliar leaf spot symptoms appeared 5 to 10 days postinoculation for all replicate pots inoculated with A. alternata. Symptoms of A. alternata infection were not observed on the control. Lesions were excised, surface sterilized, plated on water agar, and identified in the same manner as previously described. The internal transcribed spacer (ITS) region of ribosomal DNA and the mitochondrial small sub-unit region (SSU) from the original isolate and the reisolate recovered from the pathogenicity assay were amplified with PCR, with primer pairs ITS4 and ITS5 and NMS1 and NMS2, respectively. Resultant DNA fragments were sequenced and submitted to GenBank (Accession Nos. HQ130485.1 and HQ130486.1). A BLAST search (BLASTn, NCBI) was run against GenBank isolates. The ITS region sequences were 537 bp and matched 100% max identity with eight A. alternata isolates, including GenBank Accession No. AB470838. The SSU sequences were 551 bp and matched 100% max identity with seven A. alternata isolates, including GenBank Accession No. AF229648. A. alternata has been reported from switchgrass in Iowa and Oklahoma (2); however, this is the first report of A. alternata causing leaf spot on switchgrass in Tennessee. Switchgrass is being studied in several countries as a potentially important biofuel source, but understanding of the scope of its key diseases is limited. References: (1) B. Andersen et al. Mycol. Res. 105:291, 2001. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , September 22, 2011. (3) E. A. Shahin and J. F. Shepard. Phytopathology 69:618, 1979.


Plant Disease ◽  
2021 ◽  
Author(s):  
Viet-Cuong Han ◽  
Nan Hee Yu ◽  
Hyeokjun Yoon ◽  
Youn Kyoung Son ◽  
Buoung Hee Lee ◽  
...  

Flowering cherry (FC, Prunus x yedoensis Matsumura; Somei-yoshino cherry) is an ornamental tree, planted across South Korea and producing stunning flowers in spring. The seasonal blooms are annually celebrated during cherry blossom festivals in many locations across the country. The leaf spot disease is among the most common and important diseases affecting FC trees every year, resulting in premature defoliation and reduced flowering of cherry blossoms in the following year. In May 2018, brown spots (2 to 5 mm), circular to irregular and with dark borders were observed on FC leaves in Hadong, Gyeongsangnamdo, South Korea (35°07'48.9"N, 127°46'53.8"E), with a disease incidence of 55%. Single lesions often coalesced and were sometimes perforated, leaving shot holes. Sampled leaves were surface sterilized with 1% NaOCl for 1 min and 70% ethanol for 30 s, and then rinsed twice with sterile distilled water. About 2-mm-long infected leaf pieces from the margins of lesions were put onto water agar (WA, 1.5% agar) plates and incubated at 25oC for 72 h. Mycelia grown from symptomatic tissue were transferred to PDA plates, and five similar fungal isolates were obtained from hyphal tips. They produced a strong reddish-orange diffusible pigment on PDA after 5 d and exudates after 8 d. Conidia were globular to pear-shaped, dark, verrucose, multicellular, and 14.8 to 23.5 μm in diameter (av. = 18.7 μm, n = 30) for isolate JCK-CSHF10. These morphological characteristics were consistent with the Epicoccum genus. Three loci, ITS, tub2, and rpb2, from three isolates JCK-CSHF8, JCK-CSHF9, and JCK-CSHF10 were amplified using the primer pairs ITS1F/LR5 (Gardes and Bruns 1993; Vilgalys and Hester 1990), Btub2Fd/Btub4Rd (Woudenberg et al. 2009), and RPB2-5F2/RPB2-7cR (Liu et al. 1999; Sung et al. 2007), respectively. The ITS, tub2, and rpb2 sequences of the three isolates were deposited in Genbank (MW368668-MW368670, MW392083-MW392085, and MW392086-MW392088, respectively), showing 99.6 to 100% identity to E. layuense (E33), a later synonym for E. tobaicum (Hou et al. 2020). The phylogenetic tree using concatenated sequences of the three loci placed the three isolates in a cluster of E. tobaicum (CBS 232.59, CGMCC 3.18362, and CBS 384.36; Hou et al. 2020). Taken together, the three isolates were identified as E. tobaicum. The pathogenicity of JCK-CSHF10 was tested on 15 healthy leaves on three FC trees (cv. Somei-yoshino, 1.2 m in height) kept in a greenhouse. Five-mm-diameter plugs from 7-d-old fungal cultures grown on PDA or mycelia-free PDA plugs as controls were placed on the abaxial side of a leaf at three points, previously wounded by a sterile needle (Zlatković et al. 2016). Inoculation sites were covered with moist cotton plugs. Trees were then covered with a clear plastic bag and maintained in high humidity at 25oC in darkness for 24 h, followed by a 12-h photoperiod. Brown spots appeared on inoculated leaves after 7 d, identical to those observed in the field, while control leaves remained symptomless. This experiment was repeated three times. A fungus with the same morphology as JCK-CSHF10 was recovered from lesions, thus confirming Koch’s postulates. E. layuense (syn. E. tobaicum) has been reported as a leaf spot-causing agent on Perilla sp. (Chen et al. 2017) and Camellia sinensis (Chen et al. 2020). To date, there is no report on the occurrence of E. tobaicum from leaf spots on FC. To our knowledge, this is the first report of E. tobaicum causing leaf spot on FC in South Korea.


Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1116-1116 ◽  
Author(s):  
V. Parkunan ◽  
S. Li ◽  
E. G. Fonsah ◽  
P. Ji

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


Sign in / Sign up

Export Citation Format

Share Document