First Report of Myrothecium roridum Causing Leaf Spot of Anubias barteri var. nana in Mainland China

Anubias barteri var. nana (belonging to family Araceae) is one of the most popular ornamental plants for aquaria. It has slow growth and small attractive ornamental leaves. It originated from West Africa, was first described in 1860 by Heinrich Wilhelm Schott (1), and is now commercially planted widely in southern China. A new disease of A. barteri var. nana was observed on leaves. Approximately 30% of the plants in an ornamental nursery in Baiyun district, Gaungzhou, China (113°22′45.15″ E, 23°23′41.15″ N) in July 2013 were found to be infected. The pathogen caused necrotic spots that progressed to form holes on leaves of the infected plants, negatively affecting their ornamental market value. Eventually the diseased plant died. White fungal fruiting bodies and black sporodochia were observed on the surfaces of the symptomatic leaves. A single-spore isolate (myr5) was obtained from the concentric lesions and cultured on potato dextrose agar (PDA). The floccose fungal colonies were white to buff, and black conidiomata were often visible on the surface after 25 days of incubation. The conidia had rounded ends and the average size of conidia was 5.98 ± 0.15 × 2.24 ± 0.08 μm. The rDNA internal transcribed spacer (ITS) region of isolate myr5 was amplified by PCR using ITS1 and ITS4 primer pairs. The amplified product was sequenced and deposited in GenBank (Accession No. KJ572115) and showed 99% identity to Myrothecium roridum isolates BBA 71015 (AJ302001) and BBA 67679 (AJ301995) (unpublished), and 100% identity to the isolate myr2-2 from Dieffenbachia picta ‘Camilla’ in Taiwan (2). On the basis of morphological and molecular characterization, the fungus causing leaf spot on A. barteri var. nana plants was identified as M. roridum Tode ex Fr (2,3). To confirm the etiology of the disease, Koch's postulates was performed in a greenhouse (28 ± 2°C) using 3-day-old cultures of isolate myr5 and fungal spore suspensions of 1 × 105 conidia ml−1 containing 0.05% of Tween 20. Sixteen healthy leaves, two each from eight plants, were infiltrated on two different parts close to the midribs with 100 μl of the fungal spore using needleless syringes. Water infiltration was used as the control treatment. Water-soaked brown lesions appeared on leaves 7 days after inoculation, followed by the development of dark concentric rings within the necrotic areas on the surface of the inoculated leaves after 15 days of incubation. These symptoms were similar to those in the naturally infected aquarium plants. No symptoms were observed on any of the water micro-infiltrated plants. The fungus was re-isolated from the inoculated plants, but not from control plants. To our knowledge, this is the first report of Myrothecium leaf spot caused by M. roridum on A. barteri var. nana in mainland China. References: (1) W. Crusio. Primitiae Africanae XII 79 (14):1-48, 1979. (2) C. F. Hong et al. Plant Dis. 97:1253, 2013. (3) M. Tulloch. Mycol. Pap. 130:1-42, 1972.