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First Report of Curvularia Leaf Spot of Corn, Caused by Curvularia lunata, in the
United States

Article in Plant Health Progress · April 2018

DOI: 10.1094/PHP-02-18-0008-BR

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Plant Health Brief

First Report of Curvularia Leaf Spot of Corn, Caused

by Curvularia lunata, in the United States
T. Garcia-Aroca, V. Doyle, and R. Singh, Department of Plant Pathology and Crop Physiology, LSU AgCenter, Baton Rouge, LA 70803;
T. Price, † Macon Ridge Research Station, LSU AgCenter, Winnsboro, LA 71295; and Keith Collins, Richland Parish Extension Office,
Rayville, LA 71269

Accepted for publication 11 April 2018.

During the summer of 2017, corn (Zea mays L.) in production Curvularia and Bipolaris (Manamgoda et al. 2014) was used to de-
areas throughout Louisiana exhibited symptoms similar to eyespot, termine the corn isolates are nested within the clade that includes the
caused by Kabatiella zeae (Narita & Y. Hirats). Symptoms included ex-type of C. lunata (CBS 730.96) with maximum bootstrap
round to oval, light tan to light brown lesions (0.5 to 2.0-mm di- support. The shorter conidiophores and wider conidia distinguish
ameter) with reddish-brown margins often with chlorotic halos in our isolate from closely related species C. chiangmaiensis and
the mid to upper canopy of corn at the brown silk stage (Fig. 1A). C. dactyloctenicola, respectively (Marin-Felix et al. 2017). To our
The disease was not severe enough to warrant management; however, knowledge this is the first report of the disease in Louisiana and the
it was a concern to corn producers. Symptomatic leaves were ob- United States, because our isolate is the only one from the United
tained from diseased corn (hybrid DKC 66-19) near Start, Richland States conspecific with the ex-type of C. lunata (Fig. 3). Previous
Parish, Louisiana. Lesion margins were disinfested with a 10% reports of C. lunata in the United States on Poaceae with molecular
sodium hypochlorite solution for 45 to 75 s, rinsed in sterile distilled data do not appear to be accurate. For example, the isolate reported
water, and aseptically transferred to potato dextrose agar (PDA)
(Cole-Parmer) amended with chloramphenicol (C, 75 ppm) and
streptomycin sulfate (S, 125 ppm). Colonies that originated from
surface-sterilized diseased tissue were isolated via hyphal tip trans-
fer for subsequent analysis. An isolate was tentatively identified as
Curvularia lunata (Ellis 1971) based on the presence of smooth-
walled, curved, three-septate, melanized conidia lacking a distinctly
protuberant hilum and measuring 20 to 34 × 7 to 17 mm (Fig. 2) borne
on conidiophores ranging in length from 240 to 446 mm. This isolate
was grown on PDA-CS for 14 days at 25°C to prepare inoculum. A
spore suspension was prepared to a final concentration of 2 × 105
conidia/ml from five rinsed cultures of the isolate in 50 ml of water
plus 0.01% Silwet L-77 (Helena Agri-Enterprises, Collierville,
TN), an organosilicone surfactant. Ten corn plants at the V4 to V5
stage were inoculated with the suspension using a compressed-air-
powered atomizer (Crown SpraTool, Aervoe Industries, Gardnerville,
NV), and 10 others were sprayed with water plus Silwet L-77 (0.01%)
only. Plants were incubated in a dew chamber (Percival I-35D,
Boone, IA) at 100% relative humidity and 25 ± 2°C for 16 h.
Lesions resembling those found in the field were observed within
3 days of inoculation on all 10 inoculated plants (Fig. 1B and C).
Noninoculated plants remained symptomless. The entire process
was repeated twice. The causal agent was reisolated, and morpho-
logical characters of the reisolated fungus were similar to those pre-
viously described. DNA from original isolate DMCC2087 was extracted,
and the internal transcribed spacer region (nrITS, MG971304),
elongation factor 1-alpha (TEF1, MG979800), and glyceraldehyde FIGURE 1
3-phosphate dehydrogenase (GAPDH, MG979801) were amplified
Symptoms of Curvularia leaf spot on Zea mays. A, Light tan to light brown
and sequenced using universal primers. We also sequenced nrITS for lesions on a corn leaf caused by Curvularia lunata in the field. Picture was
the isolate recovered from leaves after confirming Koch’s postulates taken the same day that symptoms were first observed in the field and
(MG971305). Phylogenetic analysis of partial ITS, TEF1, and GAPDH brought to the lab for isolation (6/15/2017). B, Light tan to light brown
from representative and ex-type sequences for all known species of lesions on a corn leaf caused by C. lunata in growth chambers, 7 days after
reinoculation with conidia from DMCC2087 isolate, suspended in Silwet L-77
†
Corresponding author: T. Price; E-mail: pprice@agcenter.lsu.edu (0.01%) at a concentration of 20,000 cells/ml. C, Symptoms of Curvularia
observed under a stereoscope 10 days after inoculation. D, Closeup of
© 2018 The American Phytopathological Society a symptomatic leaf observed under the stereoscope.

PLANT HEALTH PROGRESS ¿ 2018, Vol. 19, No. 2 ¿ Page 140

FIGURE 2
Typical conidia of Curvularia from Zea mays observed at 60× under a compound microscope. A, Conidia of Curvularia lunata harvested from within lesions of
inoculated plants. Conidia were harvested 10 days after inoculation with the original Curvularia isolate DMCC2087. Scale bar = 50 mm. B, Conidia harvested from
original Curvularia isolate DMCC2087 from Louisiana, grown on potato dextrose agar (PDA) for 19 days. Scale bar = 25 mm. C, Conidia harvested from reisolated
Curvularia after infected plants showed symptoms in the growth chambers. Culture was grown on PDA for 25 days. Scale bar = 25 mm.

FIGURE 3
Maximum likelihood phylogeny of Curvularia inferred from sequences of the internal transcribed spacer region, elongation factor 1-alpha, and glyceraldehyde
3-phosphate dehydrogenase. Arrow indicates isolates from Louisiana causing leaf spot on Zea mays, which are conspecific with the ex-type sequence indicated
in bold. Terminals in red font are those sequences deposited in GenBank as C. lunata from the United States and other representative isolates that have been
published. The tree was rooted with Alternaria alternata, Dreschlera brizae, Johnalcornia aberrans, and 84 sequences representing Bipolaris. Outgroup taxa were
pruned from the tree for visualization purposes. Scale bar represents the estimated number of nucleotide substitutions per site. Bootstrap support values
greater than 50% are shown above the branches.

PLANT HEALTH PROGRESS ¿ 2018, Vol. 19, No. 2 ¿ Page 141

to be C. lunata var. aeria from Panicum in Tennessee (Fajolu et al. Fajolu, O. L., Dee, M. M., Zale, J., Gwinn, K. D., and Ownley, B. H.
2012) is conspecific with C. asiensis, and NRRL 6406 from corn seed 2012. First report of leaf spot and necrotic roots on switchgrass caused
by Curvularia lunata var. aeria in the United States. Plant Dis. 96:
(Wicklow and Poling 2009) is conspecific with C. pseudobrachyspora. 1372.
Curvularia leaf spot has been reported in hot, humid areas and is Manamgoda, D. S., Rossman, A. Y., Castlebury, L. A., Crous, P. W., Madrid, H.,
reported to cause significant damage in tropical areas (Munkvold and Chukeatirote, E., and Hyde, K. D. 2014. The genus Bipolaris. Stud. Mycol.
White 2016). It appears that the disease is not yield-limiting in Lou- 79:221-288.
isiana; however, it has prompted many concerns from stakeholders, and Marin-Felix, Y., Senwanna, C., Cheewangkoon, R., and Crous, P. W. 2017. New
species and records of Bipolaris and Curvularia from Thailand. Mycosphere
anecdotal evidence indicates that hybrids may vary in susceptibility. 8:1556-1574.
Munkvold, G. P., and White, D. G. 2016. Page 46 in: Compendium of
Corn Diseases, 4th Ed. American Phytopathological Society, St. Paul,
Literature Cited MN.
Wicklow, D. T., and Poling, S. M. 2009. Antimicrobial activity of pyrrocidines
Ellis, M. B. 1971. Dematiaceous Hyphomycetes. Commonwealth Mycological from Acremonium zeae against endophytes and pathogens of maize. Phy-
Institute, Kew, England. topathology 99:109-115.

PLANT HEALTH PROGRESS ¿ 2018, Vol. 19, No. 2 ¿ Page 142

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