First Report of Anthracnose on Giant Philodendron Caused by Colletotrichum siamense in Thailand.

Abstract

Giant philodendron (Philodendron giganteum Schott) is cultivated in Thailand and has become an important ornamental houseplant with great economic value. During the rainy season in July 2022, anthracnose disease on this plant was observed at a nursery in Saraphi District, Chiang Mai Province (18°40'18" N, 99°03'17" E), Thailand. The area investigated was approximately 800 m². The disease incidence was estimated at above 15% according to the total number of plants (220 plants). The disease severity of each plant was between 25 and 50% of the necrotic lesion on the leaf. Initially, symptoms with brown spots, appeared on leaves, gradually becoming enlarged, elongate, 1 to 11 cm long by 0.3 to 3.5 cm wide, irregular, sunken, dark brown, with a yellow halo surrounding each lesion. Then, the diseased leaves eventually withered and died. Leaf pieces (5 × 5 mm2) of the margins between lesions and the healthy tissue were surface sterilized in 1% NaClO for 1 min, 70% ethanol for 30 s, and rinsed three times with sterile distilled water. Tissues were placed on potato dextrose agar (PDA) and incubated at 25°C in darkness. After three days of incubation, pure fungal colonies were purified by a single hyphal tip method on PDA (Korhonen and Hintikka 1980). Two fungal isolates (SDBR-CMU471 and SDBR-CMU472) with similar morphology were obtained. Fungal colonies on PDA were white and 38 to 40 mm in diameter after 3 days of incubation at 25 °C, then grayish white with cottony mycelia, the reverse side pale yellow after one week of incubation. Both isolates produced asexual structures on PDA. Setae were brown with 1 to 3 septa, 50 to 110 × 2.4 to 4.0 µm, with a cylindrical base, and acuminate tip. Conidiophores were hyaline to pale brown, septate, and branched. Conidiogenous cells were hyaline to pale brown, cylindrical to ampulliform, 9.5 to 35 µm long (n = 50). Conidia were single-celled, straight, hyaline, smooth-walled, cylindrical, ends rounded, guttulate, 9.1 to 19.6 × 3.5 to 5.6 µm (n = 50). Appressoria were brown to dark brown, oval to irregular, smooth-walled, 5 to 10 × 5 to 7.5 µm (n = 50). Morphologically, both fungal isolates resembled members of the Colletotrichum gloeosporioides species complex (Weir et al. 2012; Jayawardena et al. 2021). The internal transcribed spacer (ITS) region of the ribosomal DNA, actin (act), β-tubulin (tub2), calmodulin (CAL), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were amplified using primer pairs ITS5/ITS4 (White et al. 1990), ACT-512F/ACT-783R (Carbone and Kohn 1999), T1/T22 (O'Donnell and Cigelnik 1997), CL1C/CL2C (Weir et al. 2012), and GDF1/GDR1 (Templeton et al. 1992), respectively. Sequences were deposited in GenBank (ITS: OQ699280, OQ699281; act: OQ727122, OQ727123; tub2: OQ727124, OQ727125; CAL: OQ727126, OQ727127; GAPDH: OQ727128, OQ727129). Multi-gene (combined data set of ITS, GAPDH, CAL, act, and tub2) maximum likelihood phylogenetic analyses demonstrated that both isolates were identified as C. siamense with 100% support. In a pathogenicity test, leaves of healthy plants were surface sterilized with a 0.1% NaClO solution for 3 min, rinsed three times with sterile distilled water. After being air-dried, a uniform wound (5 pores, 3 mm in width) was made at the equator of each leaf using aseptic needles. Conidial suspensions were collected from two-week-old cultures and suspended in sterile distilled water with 0.05% Tween-20. Fifteen microliters of the conidial suspension (1 × 106 conidia/ml) were placed on wounded attached leaves. As well, wounded control leaves were mock inoculated with sterile distilled water. Ten replications were conducted for each treatment and experiments were repeated twice. The inoculated plants were stored in a greenhouse at conditions of 25 to 30°C and 75 to 85% relative humidity. After 14 days, all the inoculated leaves showed disease symptoms, brown lesions with yellow halos, whereas control leaves remained asymptomatic. The pathogen C. siamense was consistently re-isolated on PDA from the inoculated tissues to complete Koch's postulates. Colletotrichum siamense has been reported as a causal agent on a wide range of host plants in Thailand and throughout the world (Farr and Rossman 2021; Jayawardena et al. 2021). Prior to this study, C. endophytica, C. karsti, C. orchidearum, C. philodendricola, and C. pseudoboninense were identified as causal agents of anthracnose on philodendrons (Xue et al. 2020; Zhang et al. 2023). However, anthracnose caused by Colletotrichum species on giant philodendron (P. giganteum) has not been previously reported. Thus, we propose C. siamense as a new causal agent of anthracnose disease on giant philodendron. This study provides information for further investigation into the epidemiology and management of this disease. Moreover, further investigations should be carried out in other philodendron growing areas of Thailand in order to specifically search for this pathogen.