Title Novel and Highly Diverse Fungal Endophytes in Soybean Revealed by the Consortium of Two Different Techniques
Author Tiago de Souza Leite1, Andréia Cnossen-Fassoni1, Olinto Liparini Pereira2, Eduardo Seiti Gomide Mizubuti2, Elza Fernandes de Araújo1, and Marisa Vieira de Queiroz1*
Address 1Laboratory of Microorganism Molecular Genetics, Department of Microbiology/Institute of Microbiology Applied to Agriculture and Livestock Raising (BIOAGRO), Federal University of Viçosa, Viçosa-MG, Brazil, 2Department of Phytopathology, Federal University of Viçosa, Viçosa-MG, Brazil
Bibliography Journal of Microbiology, 51(1),56-69, 2013,
DOI 10.1007/s12275-013-2356-x
Key Words basidiomycota diversity extinction culturing fragment plating rDNA ITS
Abstract Fungal endophytes were isolated from the leaves of soybean cultivars in Brazil using two different isolation techniques – fragment plating and the innovative dilution-to-extinction culturing – to increase the species richness, frequency of isolates and diversity. A total of 241 morphospecies were obtained corresponding to 62 taxa that were identified by analysis of the internal transcribed spacer (ITS) of the ribosomal DNA (rDNA). The Phylum Ascomycota predominated, representing 99% and 95.2% of isolates in the Monsoy and Conquista cultivars, respectively, whereas the Phylum Basidiomycota represented 1% and 4.8% of isolates, respectively. The genera Ampelomyces, Annulohypoxylon, Guignardia, Leptospora, Magnaporthe, Ophiognomonia, Paraconiothyrium, Phaeosphaeriopsis, Rhodotorula, Sporobolomyces, and Xylaria for the first time were isolated from soybean; this suggests that soybean harbours novel and highly diverse fungi. The yeasts genera Rhodotorula and Sporobolomyces (subphylum Pucciniomycotina) represent the Phylum Basidiomycota. The species richness was greater when both isolation techniques were used. The diversity of fungal endophytes was similar in both cultivars when the same isolation technique was used except for Hill’s index, N1. The use of ITS region sequences allowed the isolates to be grouped according to Order, Class and Phylum. Ampelomyces, Chaetomium, and Phoma glomerata are endophytic species that may play potential roles in the biological control of soybean pathogens. This study is one of the first to apply extinction-culturing to isolate fungal endophytes in plant leaves, thus contributing to the development and improvement of this technique for future studies.