Malaysian Journal of Microbiology, Vol 11(4) 2015,pp. 324-340 Malaysian Journal of Microbiology Published by Malaysian Society for Microbiology (In since 2011) 324 ISSN (print): 1823-8262, ISSN (online): 2231-7538 Identification of phylloplane yeasts from paper mulberry (Broussonetia papyrifera (L.) L'Hér. ex Vent.) in Java, Indonesia Dalia Sukmawati 1 , Ariyanti Oetari 1,2 ,Dian Hendrayanti 1 , Mega Atria 1 , Wellyzar Sjamsuridzal 1,2 1 Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Kampus UI, Depok, 16424, Indonesia. 2 Center of Excellence for Indigenous Biological Resources-Genome Studies, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Kampus UI, Depok, 16424, Indonesia. Email:sjwelly@hotmail.com Received 12 November 2014; Received in revised form 23 April 2015; Accepted 24 April 2015 ABSTRACT Aims: Broussonetia papyrifera (Saeh plant) has many qualities, the inner bark was the material for ‘dluwang’ papers on which Indonesian historical manuscripts were written, and the leaves have bioactive constituents of medicinal value, and antifungal activities. We investigated the diversity of yeast species associated with leaves from 6 months and 1.5 year- old plants, which is prerequisite to understand the phylloplane yeasts and plant interaction. Methodology and results: The yeasts were isolated from fresh leaves by washing and membrane filtration methods. A total of 16 leaf samples of 6 months and 1.5 year-old plants were collected from four locations in Java, Indonesia, and 2,543 yeast isolates were obtained. Based on similarity of colony morphology, 82 representative yeast isolates were selected and identified based on the sequence analyses of internal transcribed spacer (ITS) regions of rDNA. The identification result showed that they consisted of 17 genera and 32 species. Thirty six of representative yeast isolates belong to 11 genera (18 species) of the phylum Ascomycota and forty six isolates belong to 6 genera (14 species) of the phylum Basidiomycota. Phylogenetic trees showed that the yeast isolates are phylogenetically diverse and distributed in the phyla of Ascomycota (classes Saccharomycetales and Dothideomycetes) and Basidiomycota (classes Microbotryomycetes, Tremellomycetes, and Ustilaginomycetes). Conclusions, significance and impact of study: The phylloplane yeasts of B. papyrifera (Saeh plant) were taxonomically heterogeneous. This is the first report of the isolation and identification of phylloplane yeasts from B. papyrifera. Phylloplane yeasts may possess antagonistic activity to fungal plant pathogens in their natural habitats. Keywords: Broussonetia papyrifera, ITS regions of rDNA, phylloplane yeast INTRODUCTION Broussonetia papyrifera (L.) Vent. (Moraceae) (paper mulberry) is a plant which grows naturally in Asian and Pacific countries (Thailand, China, Myanmar, Laos, Japan, Korea, and Indonesia) (Whistler and Elevitch, 2006). In Indonesia this plant was commonly found in Java, Sulawesi, Kalimantan, Bali, New Guinea, and Maluku islands (Permadi, 2010). This plant is known by several vernacular names, e.g. ‘saeh’ (Sunda), ‘galugu’ or ‘glugu’ (Java), ‘dhalubang or dhulubang’ (Madura), ‘sepukau’ (Basemah, Sumatera), ‘kembala, ‘rowa’ (Sumba), and ‘linggowas’ (Banggai) (Permadi, 1998). Broussonetia papyrifera has been widely exploited by humans (Whistler and Elevitch, 2006). The most significant part of this plantis its strong fibrous bark which was used as a raw material for paper-making and textiles for centuries throughout East and Southeast Asia, and Polynesia (Teijgeler, 1995; Permadi, 2005). According to Teijgeler(1995), in the past the inner bark of B. papyrifera was used by the Indonesians for making high-quality paper known as ‘daluang’ (‘dluwang’). Islamic manuscripts are written especially on ‘daluang’ . At the turn of last century the only production centres left were Tunggilis Village, Garut (West Java) and Tegalsari, Ponorogo (East Java). After World War II only one family in Tunggilis, near Garut, was still engaged in making ‘daluang’. Teijgeler (2000) reported that ‘daluang’ was used for clothing in Java. Old Javanese literature mentioned that in the pre- Islamic era, roughly before A.D. 1550, clothes made from ‘daluang’ were worn by clergymen, especially ascetics. Oetari et al. (2010) stated that nowadays, the plant is in scarcity and the traditional knowledge for ‘daluang’ paper making is not known anymore. All parts of this plant (root, leaves, bark, and fruit) are used in traditional Chinese medicines (Sun et al., 2012). *Corresponding author