scholarly journals Characterization of Leaf Spot Pathogens from Several Spinach Production Areas in the United States

Plant Disease ◽  
2020 ◽  
Vol 104 (7) ◽  
pp. 1994-2004
Author(s):  
Bo Liu ◽  
Larry Stein ◽  
Kimberly Cochran ◽  
Lindsey J. du Toit ◽  
Chunda Feng ◽  
...  

Leaf spot diseases have become a major concern in spinach production in the United States. Determining the causal agents of leaf spots on spinach, their prevalence and pathogenicity, and fungicide efficacy against these pathogens is vital for effective disease management. Spinach leaves with leaf spots were collected from Texas, California, Arizona, and South Carolina from 2016 to 2018, incubated in a moist chamber, and plated on potato dextrose and tryptic soy agar media. Fungal and bacterial colonies recovered were identified based on morphology and sequence analysis of the internal transcribed spacer rDNA and 16S rRNA, respectively. Two predominant genera were isolated: (i) Colletotrichum spp., which were identified to species based on sequences of both introns of the glutamate synthetase (GS-I) and glyceraldehyde-3-phosphate dehydrogenase (gapdh-I) genes; and (ii) Stemphylium spp., identified to species based on sequences of the gapdh and calmodulin (cmdA) genes. Anthracnose (Colletotrichum spinaciae) and Stemphylium leaf spot (Stemphylium vesicarium and S. beticola) were the predominant diseases. Additional fungi recovered at very limited frequencies that were also pathogenic to spinach included Colletotrichum coccodes, C. truncatum, Cercospora beticola, and Myrothecium verrucaria. All of the bacterial isolates were not pathogenic on spinach. Pathogenicity tests showed that C. spinaciae, S. vesicarium, and S. beticola caused significant leaf damage. The fungicides Bravo WeatherStik (chlorothalonil), Dithane F-45 (mancozeb), Cabrio (pyraclostrobin), and Merivon (fluxapyroxad and pyraclostrobin) were highly effective at reducing leaf spot severity caused by an isolate of each of C. spinaciae and S. vesicarium, when inoculated individually and in combination.

Plant Disease ◽  
2021 ◽  
pp. PDIS-04-20-0918
Author(s):  
Bo Liu ◽  
Larry Stein ◽  
Kimberly Cochran ◽  
Lindsey J. du Toit ◽  
Chunda Feng ◽  
...  

Leaf spot diseases of spinach, caused by Colletotrichum spinaciae, has become a major production constraint in several production areas, including Texas, in recent years. Leaf spot symptoms were observed in several fields in Texas in 2016 and 2017, with typical anthracnose-like symptoms and leaves with small, circular, and sunken lesions that appeared similar to injury from windblown sand. The lesions were plated on potato dextrose agar, from which fungal cultures were recovered. The fungi were identified based on morphology and sequence analysis of the introns of glutamate synthetase and glyceraldehyde-3-phosphate dehydrogenase (for isolates determined to be Colletotrichum spp.) and the internal transcribed spacer ribosomal DNA (for isolates determined to be Myrothecium spp.). Based on foliar symptoms, fungal colony and spore morphology, pathogenicity tests of fungal isolates on the spinach cultivar ‘Viroflay’, and DNA sequence analysis of the isolates, the symptoms on spinach leaves for two sets of samples were caused by Colletotrichum coccodes and Colletotrichum truncatum, and leaf spots resembling damage from windblown sand were caused by Myrothecium verrucaria. This is the first report of spinach leaf spot diseases caused by C. coccodes, C. truncatum, and M. verrucaria in the United States. C. coccodes and C. truncatum caused severe symptoms on the spinach cultivar ‘Viroflay’, whereas M. verrucaria caused symptoms of intermediate severity. Fungicide efficacy tests demonstrated that chlorothalonil, mancozeb, pyraclostrobin, fluxapyroxad + pyraclostrobin, and penthiopyrad were completely effective at preventing leaf spots caused by any of these pathogens when applied 24 h before inoculation of ‘Viroflay’ plants in greenhouse trials.


2021 ◽  
Vol 60 (2) ◽  
pp. 177-198
Author(s):  
Yamin DU ◽  
Xianhong WANG ◽  
Yashuang GUO ◽  
Feng XIAO ◽  
Yuhong PENG ◽  
...  

Diaporthe species are significant pathogens, saprobes, and endophytes, with comprehensive host association and geographic distribution. These fungi cause severe dieback, cankers, leaf spots, blights, and stem-end rot of fruits on different plant hosts. This study, explored the occurrence, diversity and pathogenicity of Diaporthe spp. associated with Actinidia chinensis and A. deliciosa in the main kiwifruit production areas of China. Diaporthe isolates (284) derived from 106 diseased leaf and branch samples were examined. Multi-locus phylogenetic analyses and morphology of 43 representative isolates revealed that seven Diaporthe species were obtained, including D. alangii, D. compactum, D. eres, D. hongkongensis, D. sojae, D. tectonae, and D. unshiuensis. Pathogenicity tests were performed on kiwifruit fruits, leaves and branches. Koch’s postulates confirmed all species were pathogenic. D. alangii and D. tectonae were the most aggressive species, followed by D. eres, D. sojae, D. hongkongensis, D. unshiuensis, and D. compactum. Host range evaluation showed that the seven Diaporthe species could also infect apricot, apple, peach, pear, and plum.  This is the first report of D. alangii, D. compactum, D. sojae, D. tectonae, and D. unshiuensis infecting kiwifruit in China, increasing understanding of the Diaporthe complex causing diseases of kiwifruit plants, to assist effective disease management.


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.


Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 875-875 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
D. Bertetti ◽  
R. Nicoletti ◽  
M. L. Gullino

Lantana camara is increasingly grown in northern Italy as a potted plant and contributes to the diversification of offerings in the ornamental market. During the spring of 2001, selections of L. camara cuttings growing at a commercial farm located at Albenga (Riviera coast) exhibited tan leaf spots of irregular size and shape. Spots were at first isolated, 4 to 8 mm in diameter, and later coalesced and affected the entire plant. Heavily infected leaves, stems, and branches became blighted and were killed. Infected rooted cuttings also eventually died. Diseased cuttings showed a progressive reduction (to less than 20%) in rooting ability. Isolations from infected leaves and stems on potato dextrose agar (PDA), supplemented with 100 mg/liter of streptomycin sulphate, consistently yielded a fungus with mycelial and cultural characteristics resembling Rhizoctonia solani. The fungal isolates were further characterized as R. solani Kühn AG-4 based on hyphal anastomoses with several AG-4 tester isolates. Pathogenicity tests were performed by placing 5-day-old-fungal mycelial plugs, grown on PDA, at the base of five healthy yellow-sage stems and holding plants in a dew chamber at 18 to 22°C. After 2 days, foliage blight appeared on leaves of inoculated plants, and after 3 days, stems also became infected and entire plants wilted. Five noninoculated plants remained healthy. The fungal pathogen was reisolated from all inoculated plants. R. solani has been observed on L. camara in the United States (1) and the Philippines (2). To our knowledge, this is the first report of R. solani on L. camara in Europe. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN, 1989. (2) F. T. Orillo and R. B. Valdez. Philipp. Agric. A. 42:292, 1958.


Plant Disease ◽  
2009 ◽  
Vol 93 (1) ◽  
pp. 108-108 ◽  
Author(s):  
A. J. Caesar ◽  
R. T. Lartey ◽  
D. K. Berner ◽  
T. Souissi

The herbaceous perennial Lepidium draba L. is an invasive weed of rangelands and riparian areas in North America and Australia. As of 2002, it had infested 40,500 ha of rangeland in Oregon and large areas in Wyoming and Utah. Little is known of plant pathogens occurring on L. draba, especially in the United States, that could be useful for biological control of the weed. Leaf spots were first noted on a stand of L. draba near Shepherd, MT in 1997. The spots were mostly circular but sometimes irregularly shaped and whitish to pale yellow. The pathogen was erroneously assumed to be Cercospora beticola since its morphological traits closely resembled that species and the area had large fields of sugar beet with heavy Cercospora leaf spot incidence. Diseased leaves of L. draba were collected in 1997 and 2007. Conidia, borne singly on dark gray, unbranched conidiophores produced on dark stromata late in the season, were elongate, hyaline, multiseptate, 38 to 120 × 2 to 6 μm (mostly 38 to 50 × 2 to 5 μm) and had bluntly rounded tips and wider, truncate bases. These characteristics were consistent with the description of C. bizzozeriana Saccardo & Berlese (2). To isolate the fungus, spores were picked from fascicles of conidiophores with a fine-tipped glass rod, suspended in sterile water, and spread on plates of water agar. Germinated spores were transferred to potato dextrose agar (PDA). The ITS1, 5.8S, and ITS2 sequences of this fungus (GenBank Accession No. EU887131) were identical to sequences of an isolate of C. bizzozeriana from Tunisia (GenBank Accession No. DQ370428). However, these sequences were also identical to those of a number of Cercospora spp. in GenBank, including C. beticola. We also compared the actin gene sequences of the Montana isolate of C. bizzozeriana (GenBank Accession No. FJ205397) and an isolate of C. beticola from Montana (GenBank Accession No. AF443281); the sequences were 94.6% similar, an appreciable difference. For pathogenicity tests, cultures were grown on carrot leaf decoction agar. Aqueous suspensions of 104 spores per ml from cultures were sprayed on 6-week-old L. draba plants. Plants were covered with plastic bags and placed on the greenhouse bench at 20 to 25°C for 96 h. Koch's postulates were completed by reisolating the fungus from the circular leaf spots that appeared within 10 days, usually on lower leaves. Spores of C. bizzozeriana were also sprayed on seedlings of sugar beet, collard, mustard, radish, cabbage, and kale under conditions identical to those above. No symptoms occurred. After the discovery of the disease in 1997, plants of L. draba in eastern Montana, Wyoming, and Utah were surveyed from 1998 to 2003 for similar symptoms and signs, but none were found. This, to our knowledge, is the first report of C. bizzozeriana in the United States. The initial report of the fungus in North America was from Manitoba in 1938 (1). It has recently been reported as occurring on L. draba in Tunisia (4) and Russia (3) and is reported as common in Europe (2). A voucher specimen has been deposited with the U.S. National Fungus Collections (BPI No. 878750A). References: (1) G. R. Bisby. The Fungi of Manitoba and Saskatchewan. Natl. Res. Council of Canada, Ottawa, 1938. (2) C. Chupp. A Monograph of the Fungus Genus Cercospora. C. Chupp, Ithaca, NY, 1953. (3) Z. Mukhina et al. Plant Dis. 92:316, 2008. (4) T. Souissi et al. Plant Dis. 89:206, 2005.


Plant Disease ◽  
2000 ◽  
Vol 84 (10) ◽  
pp. 1154-1154 ◽  
Author(s):  
G. E. Holcomb

Angular chlorotic spots were observed on adaxial leaf surfaces of Salvia splendens (scarlet sage cvs. Empire Purple, Empire White, Red Pillar, and Red Hot Sally) and S. coccinea (scarlet or Texas sage cv. Lady in Red) in early May in Baton Rouge area nurseries. Leaf spots sometimes became necrotic and resulted in leaf drop. Abaxial leaf surfaces contained scattered patches of white mycelia with brown spores. Microscopic examination of mycelia revealed irregular dichotomously branched conidiophores with pointed tips and brown oval conidia. Conidiophores averaged 485 × 9 µm and conidia averaged 21 × 18 µm (16 to 26 × 15 to 23 µm) in dimensions. The fungus was identified as Peronospora lamii A. Braun (= P. swinglei Ellis & Everh.) based on these characters and its known occurrence on Salvia spp. and five other genera in the family Lamiaceae (2). Pathogenicity tests were performed by washing conidia from infected leaves into distilled water and mistinoculating S. coccinea cv. Lady in Red and S. splendens cv. Empire Purple with 50,000 spores/ml. Plants were held in a dew chamber at 20°C for 3 days, then moved to a greenhouse where temperatures ranged from 18 to 32°C. Typical angular chlorotic leaf spots developed on inoculated plants within 6 to 8 days and noninoculated plants remained healthy. The fungus did not sporulate under these greenhouse temperatures, but infected leaves that were removed and placed in a moist chamber at 25°C produced conidiophores and brown conidia typical of P. lamii within 2 to 3 days. P. lamii has been reported previously on S. officinalis (3) and S. reflexa (1) in the United States. This is the first report of downy mildew on S. coccinea and S. splendens. Appearance of the disease in retail nurseries that obtained plants from out of state (Arkansas) suggests a widespread occurrence of the disease on these host plants. References: (1) D. F. Farr et al. 1989. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN. (2) S. M. Francis. 1981. Peronospora lamii. Descriptions of Pathogenic Fungi and Bacteria No. 688. Commonwealth Mycological Institute, Kew, England. (3) R. T. McMillan and W. R. Graves. Plant Dis. 78:317, 1994.


2018 ◽  
Vol 19 (2) ◽  
pp. 140-142 ◽  
Author(s):  
T. Garcia-Aroca ◽  
V. Doyle ◽  
R. Singh ◽  
T. Price ◽  
Keith Collins

During the summer of 2017, corn (Zea mays L.) in production areas throughout Louisiana exhibited symptoms similar to eyespot, caused by Kabatiella zeae (Narita & Y. Hirats). Symptoms included round to oval, light tan to light brown lesions (0.5 to 2.0-mm diameter) with reddish-brown margins often with chlorotic halos in the mid to upper canopy of corn at the brown silk stage. The disease was not severe enough to warrant management; however, it was a concern to corn producers. Symptomatic leaves were obtained from diseased corn, lesion margins were disinfested, and the suspected pathogen was isolated and tentatively identified as Curvularia lunata. Koch’s postulates were completed by inoculating V4 to V5 stage corn plants with a spore suspension and subjecting plants to a 16-h dew period at 25°C, observing symptomology, reisolating the pathogen, and identification via molecular analysis. To our knowledge this is the first report of the disease in Louisiana and the United States.


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.


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