Mycosphere 191 Cladosporium sp. from phyloplane: a diversity evaluation on a Continental ecosystem Guimarães JB 1* , Chambel L 2 , Melzoch K 3 , Pereira P 4 and Tenreiro R 2 1 Molecular Biology Group, Genetic Resources, Ecophysiology and Plant Breeding Research Unit, L-INIA - Pólo do Lumiar, Instituto Nacional de Recursos Biológicos, I.P., Edifício .S Gab 2.061, Estrada do Paço do Lumiar, 22, 1649- 038, Lisboa, Portugal. 2 Universidade de Lisboa, Faculdade de Ciências, Centro de Genética e Biologia Molecular e Instituto de Ciência Aplicada e Tecnologia, Edifício ICAT, Campus da FCUL, Campo Grande, 1749-016 Lisboa, Portugal. 3 Department of Fermentation Chemistry and Bioengineering, ICT Prague, Technická 5, CZ-166 28 Prague 6, Czech Republic. 4 Unidade de Investigação de Tecnologia Alimentar, L-INIA - Pólo do Lumiar, Instituto Nacional de Recursos Biológicos, I.P., Edifício .S, Estrada do Paço do Lumiar, 22, 1649-038, Lisboa, Portugal. Guimarães JB, Chambel L, Melzoch K, Pereira P, Tenreiro R. 2011 – Cladosporium sp. from phyloplane: a diversity evaluation on a Continental ecosystem. Mycosphere 2(3), 191–201. The biodiversity of filamentous fungi on leaves of two genera of plants from a mixed Continental ecosystem (Český Kras, Czech Republic) was assessed. Quercus (four species) and Acer (three species) are frequent plant genera in the area. Fifteen fungal genera were identified by phenotypic methods (colony characterisation and morphology of reproductive structures). The study focused on Cladosporium, the dominant genus, but less common genera were found including Alternaria, Fusarium, Rhizopus, Penicillium, Aspergillus, Trichoderma, and Absidia. csM13 fingerprinting and ITS-ARDRA were used for genomic clustering of isolates of Cladosporium. Most Cladosporium isolates (99%) were identified as C. herbarum and C. macrocarpum. Diversity indexes and Chi- square analysis were applied to better analyse the fungal community. Continental environments are demonstrated to have some diversity of filamentous fungi, but not as large as in tropical, temperate or Mediterranean areas. Key words – Cladosporium – csM13 PCR “fingerprinting” – Diversity – Fungi – ITS – ARDRA Article Information Received 7 December 2010 Accepted 7 April 2011 Published online 9 June 2011 *Corresponding author: Joana Bagoin Guimarães – e-mail – joana.guimaraes@inrb.pt, bagoinj@ yahoo.com Introduction The phylloplane, or leaf surface, repre sents an important terrestrial habitat that harbours a wide range of microorganisms (Andrews 1991, Lindow & Brandl 2003). Fun- gi, encompassing both filamentous and yeast taxa, are a major component of the phylloplane microbiota (Dickinson 1976, Andrews 1991, Bills & Polishook 1994, De Jager et al. 2001, Gazis & Chaverri 2010). The leaf surface is a suitable environment for microbial growth because of a thin film of nutrients deposited on the leaf. The fungal communities are influen- ced by external and/or internal factors such as nutrient availability, humidity, temperature, leaf age and type, and presence of inhibitors (chemical compounds produced by the plant) (Andrews 1991, Fokkema 1991, Kinkel 1997, de Jager et al. 2001, Santamaria & Bayman 2005, Evueh & Ogbebor 2008). Filamentous fungi from the phylloplane may be either para- sites, saprophytes, endophytes or epiphytes.