First Report of Anthracnose of Alocasia macrorrhiza Caused by Colletotrichum karstii in Guangdong, China

Abstract

Alocasia macrorrhiza (L.) Schott. (Araceae), native to South America, is a common, herbaceous perennial ornamental plant in tropical and subtropical areas (1). A severe leaf spot disease was observed on this plant in several places on the campus of authors' university in Guangzhou, Guangdong Province, China, in April 2013. Initial symptoms were water-soaked, dark green leaf spots. These small spots gradually expanded to 6- to 11-mm circular lesions. They were grayish-white in color with a yellow halo and many small, black, concentric dots were observed on them. Microscopic examination revealed that these small dots were acervuli, which were 100 to 300 μm in diameter, developing beneath the epidermis and becoming erumpent with age. By using routine tissue-isolation method and single-spore purification technique, four single-conidial isolates were obtained from each of four diseased leaves. These isolates formed a grayish-white colony with numerous pink spore masses on PDA at 28°C. Their mycelial radial growth rate was about 4.5 mm per day. Conidia were single-celled, hyaline, and cylindrical with an obtuse apex and protruding base; they were 12.7 to 14.2 × 4.8 to 5.9 μm in size. Conidial appressoria were irregular in shape, sepia to dark brown, solitary, and 6.9 to 8.5 × 4.6 to 5.9 μm. These morphological characteristics were consistent with the description of Colletotrichum karstii (2). The sequences of beta-tubulin gene (TUB2) and partial actin gene (ACT) of a representative isolate CAM1 were obtained by PCR amplification with primers BT2a/BT2b and ACT512F/ACT783R, respectively. These sequences were deposited in GenBank under the accession numbers of KF444947 and KF460435. BLAST searches showed a 99% homology with the TUB2 and ACT sequences of C. karstii (JX625209, KC843559). Therefore, the fungus isolated from A. macrorrhiza was identified as C. karstii by morphological and molecular characteristics. Pathogenicity tests were performed on 30-day-old plants of A. macrorrhiza grown in plastic pots (0.8 L) by spraying 15 ml conidial suspension (1 × 106 conidia ml–1) of this fungus onto each plant. The control plants were sprayed only with sterile distilled water. These plants then were placed in an intelligent artificial climate incubator with 12-h photoperiod and 100% relative humidity at 24 ± 1°C. Three replicates, each with five plants, were included in a test, and the test was repeated twice. Seven days after inoculation, the inoculated plants showed necrotic lesions on leaves similar to those observed on the campus, but no symptoms were observed on any non-inoculated controls. The same fungus C. karstii was re-isolated from the infected leaves. Although C. karstii is a well-known anthracnose pathogen on some plants belonging to family Orchidaceae (2), this is the first report of the same pathogen causing anthracnose on A. macrorrhiza in Guangdong, China. References: (1) S. Li et al. PLoS ONE 8(6):e66016, 2013. (2) Y. Yang et al. Cryptogr. Mycol. 32:229, 2011.