First Report of Corynespora cassiicola-Incited Target Spot on Cotton in Alabama.

Abstract

Target spot symptoms were first observed on dryland and irrigated cotton (Gossypium hirsutum L.) statewide in Alabama in 2011. Leaf spots first appeared in the lower canopy and spread upward through the canopy toward the shoot tips. Individual leaf spots were roughly circular, formed concentric rings of alternating light and dark brown bands, and were up to 10 mm in diameter. Leaves with multiple lesions senesced prematurely. In 2012, target spot symptoms were observed as early as 68 days after planting in Tallapoosa County, Alabama. The possible combination of early disease onset and frequent showers/irrigation triggered rapid premature defoliation in some fields in excess of 75% in susceptible cultivars (Phytogen 499). Estimated yield losses in select cultivars (Deltapine 1050 and Phytogen 499) exceeded 336 kg/ha seed cotton. In 2012, symptomatic leaves were obtained from two separate locations in Alabama (Baldwin and Tallapoosa counties). The fungus was isolated from lesions by single spores plated on antibiotic V8 agar (1) and incubated at 21°C for 2 weeks under 12-h light cycles. Conidiophores arising from the gray, flocculose colonies were simple, erect, cylindrical, brown or olivaceous, unbranched, with two to seven septa. Conidia were borne singly, ranging from subhyaline to olivaceous, obclavate to cylindrical, straight to slightly curved, contained 4 to 15 pseudosepta, and were 50 to 209 μm long and 7 to 15 μm wide. These characteristics were consistent with the original description of Corynespora cassiicola on cotton (2). The internal transcribed spacer region (ITS) of two isolates, one representing each location, was amplified using primers 2234c and 3126t targeting a 550-bp region of the ITS1, 5.8S rRNA gene, and ITS2 (3). Sequences revealed 99% similarity to C. cassiicola in NCBI (Accession Nos. AY238606 and JQ717069). In greenhouse pathogenicity tests, 10 cotton seedlings (Phytogen 499) were inoculated by spraying a fungal suspension (2 × 104 spores/ml) of each of the two isolates prepared from 2-week-old cultures until runoff. Controls were inoculated with sterile water. Cotton seedlings were incubated in a moist chamber at 21°C for 72 h. All plants inoculated with the fungus developed leaf spot symptoms in 6 days. The fungus was reisolated from five inoculated plants. DNA was extracted from each isolate, amplified using primer pair 2234c/3126t, and sequenced. Sequences (550-bp) from all isolates shared 99% similarity to other C. cassiicola sequences in GenBank (Accession Nos. AY238606 and JQ717069). Nucleotide sequence data reported are available in GenBank under Accession Nos. KC544017 to 23. This pathogen has been reported previously to be economically important on a number of other hosts. To our knowledge, this is the first report of C. cassiicola on cotton in Alabama. Given the increasing prevalence of this disease in Alabama, its confirmation is a significant step toward developing management recommendations for growers.