First Report of Glomerella cingulata (Colletotrichum gloeosporioides) Causing Anthracnose and Tip Dieback of Lygodium microphyllum and L. japonicum in Australia.

The Old World climbing fern, Lygodium microphyllum (Cav.) R. Br., and Japanese climbing fern, L. japonicum (Thunb.) Sw., are invasive noxious weeds in Florida (1). Exploratory surveys for classical biological control agents of L. microphyllum in the fern's native range of Australia and Asia have focused on aboveground herbivores (1). From February to August 2006, fungi were isolated from symptomatic foliage, including lesions associated with leaf curls caused by the mite Flocarus perrepae Knihinicki & Boczek., obtained from L. microphyllum at sites across southeast Queensland, Australia and from both fern species grown at the CSIRO Long Pocket Laboratories in Brisbane, Australia. Anthracnose symptoms with chlorotic margins, initiating at the tip or base of the individual pinnules, were observed on fronds. Dieback symptoms affected growing tips, with sunken lesions and a gradual necrotic wilt as far as the next growth junction of pinnae. Sections from diseased margins were surface sterilized, placed onto water agar, and incubated at 23°C with a 16-h photoperiod. Variable colonies of white-to-gray mycelia, felted or tufted with complete margins, grew well on oatmeal agar and potato dextrose agar. Conidia were hyaline to light salmon, aseptate, straight, and cylindrical (10.4 to 18.2 × 2.6 to 5.2 μm), borne in salmon-to-bright orange masses at 25°C, and consistent with previous descriptions of Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (3), anamorph of Glomerella cingulata (2). Asci that formed after 3 to 4 weeks in culture were eight-spored, clavate to cylindrical (46.8 to 62.4 × 9.1 to 11.7 μm), and thickened at the apex, and ascospores were cylindrical (11.7 to 18.2 × 3.9 to 5.2 μm), slightly curved, unicellular and hyaline, which is consistent with descriptions of G. cingulata (2). No fruiting bodies were observed in planta; acervuli, setae, and perethecia were not observed. Identification was further confirmed by molecular analysis using the primer pair ITS1/ITS4 (4) (GenBank Accession No. EU697014), indicating 100% similarity to isolates of G. cingulata. To confirm pathogenicity, Koch's postulates were performed on three plants of L. japonicum and 12 plants of L. microphyllum, with an equal number of controls. Conidial suspensions were made to 1.7 × 10 conidia ml. During the experiments in the glasshouse, temperatures ranged from 12.6 to 40°C and relative humidity from 39 to 85%. Tips and fronds were collected after 2 to 8 weeks and isolation and identification performed. G. cingulata was consistently reisolated from diseased tissue. No symptoms appeared on controls and isolations did not yield the pathogen. To our knowledge, this is the first report of G. cingulata infecting L. microphyllum and L. japonicum in Australia. Its potential as a biological control agent in the ferns' introduced range remains to be tested. References: (1) J. A. Goolsby et al. Biol. Control. 28:33, 2003. (2) J. E. M. Mordue. Glomerella cingulata. No. 315 in: CMI Descriptions of Pathogenic Fungi and Bacteria. CAB, Kew, UK, 1971. (3) B. C. Sutton. The Genus Glomerella and its Anamorph Colletotrichum. In: Colletotrichum: Biology, Pathology and Control. J. A. Bailey and M. J. Jeger, eds. CAB International, Wallingford, UK, 1992. (4) T. M. White et al. Amplification and Direct Sequencing of Fungal Ribosomal RNA for Phylogenetics. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.