Novel and highly diverse fungal endophytes in soybean revealed by the consortium of two different techniques

• Published in: The journal of microbiology 2013, 51(1), , pp.56-69
• Main Authors: de Souza Leite, Tiago, Cnossen-Fassoni, Andréia, Pereira, Olinto Liparini, Mizubuti, Eduardo Seiti Gomide, de Araújo, Elza Fernandes, de Queiroz, Marisa Vieira
• Format: Article
• Language: English
• Subjects: Ampelomyces; Ascomycota; Biodiversity; Biological control; Biomedical and Life Sciences; Brazil; Chaetomium; Cluster Analysis; cultivars; DNA; DNA, Fungal - chemistry; DNA, Fungal - genetics; DNA, Ribosomal Spacer - chemistry; DNA, Ribosomal Spacer - genetics; Endophytes; Endophytes - classification; Endophytes - cytology; Endophytes - genetics; Endophytes - isolation & purification; Fungi; Fungi - classification; Fungi - cytology; Fungi - genetics; Fungi - isolation & purification; Genomics and Molecular Biology; Glycine max - microbiology; Guignardia; internal transcribed spacers; isolation techniques; Leaves; Life Sciences; Magnaporthe; Microbial Genetics; Microbiology; Molecular Sequence Data; Pathogens; Phoma glomerata; Phylogeny; Plant Leaves - microbiology; Pucciniomycotina; Rhodotorula; ribosomal DNA; Sequence Analysis, DNA; Soybeans; Spacer; species diversity; Species richness; Sporobolomyces; Xylaria; yeasts; 생물학
• ISSN: 1225-8873; 1976-3794
• DOI: 10.1007/s12275-013-2356-x

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.