RESUMO
The native 'ohi'a lehua (Metrosideros polymorpha) has cultural, biological and ecological significance to Hawai'i, but it is seriously threatened by a disease commonly referred to as rapid 'ohi'a death (ROD). Preliminary investigations showed that a Ceratocystis species similar to C. fimbriata s.lat. was the cause of the disease. In this study, we used a combination of the phylogenetic, morphological and biological species concepts, as well as pathogenicity tests and microsatellite analyses, to characterise isolates collected from diseased 'ohi'a trees across Hawai'i Island. Two distinct lineages, representing new species of Ceratocystis, were evident based on multigene phylogenetic analyses. These are described here as C. lukuohia and C. huliohia. Ceratocystis lukuohia forms part of the Latin American clade (LAC) and was most closely associated with isolates from Syngonium and Xanthosoma from the Caribbean and elsewhere, including Hawai'i, and C. platani, which is native to eastern USA. Ceratocystis huliohia resides in the Asian-Australian clade (AAC) and is most closely related to C. uchidae, C. changhui and C. cercfabiensis, which are thought to be native to Asia. Morphology and interfertility tests support the delineation of these two new species and pathogenicity tests show that both species are aggressive pathogens on seedlings of M. polymorpha. Characterisation of isolates using microsatellite markers suggest that both species are clonal and likely represent recently-introduced strains. Intensive research is underway to develop rapid screening protocols for early detection of the pathogens and management strategies in an attempt to prevent the spread of the pathogens to the other islands of Hawai'i, which are currently disease free.
RESUMO
Miconia calvescens (Melastomataceae), from the Neotropics, is a noxious forest weed in Hawaii. We evaluated an isolate of Colletotrichum gloeosporioides that causes leaf spots on Miconia spp. in Brazil for its potential in biological control. Hawaii has no native Melastomataceae genera but does have members of 12 introduced genera. Following Wapshere's centrifugal phylogenetic method (2), eight species of Melastomataceae genera in Hawaii were inoculated in addition to Miconia spp. Naturalized and native Hawaiian members of the order Myrtales also were inoculated to determine host specificity, including Terminalia catappa (Combretaceae); Cuphea hysopifolia and C. ignea (Lythraceae); Arthrostema ciliatum, Clidemia hirta, Dissotis rotundifolia, Heterocentron subtriplinervium, Medinilla scortechenii, Melastoma candidum, Pterolepsis glomerata, and Tibouchina herbaceae (Melastomataceae); Eucalyptus grandis, Eucalyptus microcorys, Eugenia reinwardtiana, Eugenia uniflora, Leptospermum laevigatum, Melaleuca quinquenervia, Metrosideros polymorpha, Psidium guajava, and Syzgium malaccanse (Myrtaceae); Fuchsia magellanica and Oenothera stricta (Onagraceae); and Wikstroemia oahuensis and W. uva-ursi (Thymelaeaceae). All M. calvescens plants were grown from seed collected in Hawaii. Other test plants were grown from seeds or cuttings in artificial potting medium in a greenhouse. Plants had 6 to 8 mature leaves when inoculated. C. gloeosporioides was cultured on 10% potato dextrose agar supplemented with plain agar (35 g/liter) and incubated under constant fluorescent illumination at 20°C. Conidia were harvested by flooding 10-to 14-day-old cultures with sterile tap water, followed by light scraping with a scalpel. Conidial suspensions were adjusted to 106 conidia per ml and applied to both leaf surfaces with a hand-held sprayer. Inoculated plants were kept at 100% relative humidity and 16 to 25°C for 48 h. Four replicate plants and one plant of M. calvescens per species were inoculated. Plants were observed for symptom development for up to 6 weeks. The entire test was repeated once. Lesions were visible after 7 to 10 days. Young lesions had chlorotic halos and expanded in a roughly circular pattern to diameters of 5 to 10 mm. Mature lesions developed necrotic centers, coalesced, and became dry and brittle with age, resulting in extensive leaf necrosis. Defoliation of moderately to severely infected leaves occurred ≈ 30 days after inoculation. With the exception of M. calvescens, C. gloeosporioides did not produce visible symptoms on test plants. The failure of Clidemia hirta, the taxonomic species most closely related to M. calvescens, to become symptomatic was particularly significant relative to the centrifugal phylogenetic concept. The results demonstrate that our pathogen (VIC 19306) is distinct from C. gloeosporioides f. sp. clidemiae (1), which did not infect M. calvescens. We designate our pathogen C. gloeosporioides f. sp. miconiae. Voucher specimens (VIC 19306, Sana, RJ, 24.II.1998, and R. W. Barreto) and cultures are maintained at the Departamento de Fitopatologia, Universidade Federal de Viçosa MG, Brazil. References: (1) E. E. Trujillo et al. Plant Dis. 70:974, 1986. (2) A. J. Wapshere. Ann. Appl. Biol. 77:201, 1974.
