Molecular characterization of endophytes isolated from Saccharum spp based on esterase and ribosomal DNA (ITS1-5.8S-ITS2) analyses

Abstract

ABSTRACT. This study used esterases and ribosomal DNA (rDNA) markers to determine endophytic variability in order to better understand endophyte-host interactions. Polyacrylamide gel electrophoresis and esterase isoenzymes (EST; EC 3.1.1.3), with a-naphthyl acetate and β-naphthyl acetate as substrates, were used to assess relationships among endophytes. ITS1-5.8S-ITS2 sequencing data were used as rDNA markers. Thirty-two esterases were obtained from 37 isolates of Saccharum spp, which clustered into five endophyte groups. Esterase EST-06 was observed with the highest frequency, being present in 22 of the 37 isolates analyzed, followed by esterase EST-11, which was present in 20 isolates. The esterases EST-10 and EST-14 were present in 19 isolates and EST-09 was present in 18 isolates. The esterase EST-01 was unique to isolate 33 and can, therefore, be used as a marker for this isolate. None of the esterases identified were common to all isolates tested. Similarly, phylogenetic analysis, based on rDNA sequence data, classified the isolates into 5 genus groups: 1) Curvularia with a 100% bootstrap value (BP), 2) Alternaria with 100% BP, 3) Epicoccum with 60% BP, 4) Phoma with 89% BP, and 5) Saccharicola with 100% BP. This polyphyletic analysis based on several markers, therefore, proved to be a valuable approach in determining the relationship between variation in endophytes and their associated host plants. Furthermore, both the esterase and rDNA analyses obtained similar results and were equally effective in resolving relationships.

Extracellular enzymatic profiles and taxonomic identification of endophytic fungi isolated from four plant species

“…Several studies have reported the isolation of large numbers of endophytes from plants of medicinal and great economic importance. As in the current analysis, fungi of the genus Diaporthe were identified in investigations by Bernardi-Wenzel et al 2010, Rhoden et al (2012), García et al (2012), and Leme et al (2013). Rhoden et al (2012) found members of the genus Diaporthe (Phomopsis) to be the most frequently observed endophytic species.…”

“…The plants Luehea divaricata, Trichilia elegans, Sapindus saponaria, and Saccharum spp are used in traditional culture for the treatment of several diseases, and have also attracted commercial interest. Bernardi-Wenzel et al 2010, Rhoden et al (2012), García et al (2012), and Leme et al (2013) demonstrated the presence of endophytes in each of these plants, respectively. Although some endophytic fungi associated with these species have already been studied by our group, this represents only a fraction of the total number of fungal and bacterial isolates obtained from these plants belonging to our microbial laboratory collection, since endophytes were initially grouped by morphological characteristics, with representatives of each morphogroup being examined.…”

“…The endophytic fungi used in the current research were as follows: 8 isolates from L. divaricata Martius et Zuccarini (Bernardi-Wenzel et al, 2010); 10 isolates from T. elegans A. Juss (Rhoden et al, 2012); 11 isolates from S. saponaria L. (García et al, 2012); and 13 isolates from Saccharum spp (Leme et al, 2013), belonging to the collection of the Laboratory of Microbial Biotechnology of Universidade Estadual de Maringá, Maringá, PR, Brazil. The fungi were selected from the same morphogroup, considering colony morphology and coloration, mycelial form, and pigment production of each isolate.…”

Phylogenetic analysis of endophytic bacterial isolates from leaves of the medicinal plant Trichilia elegans A. Juss. (Meliaceae)

“…Endophytic microorganisms live inside plants without causing them any damage (Bernardi-Wenzel et al, 2010;Garcia et al, 2012;Orlandelli et al, 2012;Rhoden et al, 2012a;Leme et al, 2013). They can be found in the aerial parts of the plant, primarily in the leaves, and occupy intra-and intercellular spaces (Bernardi-Wenzel et al, 2010;Garcia et al, 2012), characterizing a symbiotic relationship.…”

In silico analysis of the 16S rRNA gene of endophytic bacteria, isolated from the aerial parts and seeds of important agricultural crops

“…This group of microorganisms can be found in different parts of the plant, such as leaves Rhoden et al, 2012;Leme et al, 2013), flowers, seeds, stems, and roots (Vega et al, 2005;Azevedo and Araújo, 2007;Compant et al, 2011). Furthermore, endophytes may also be used as an alternative to fertilizers and pesticides because these microorganisms have been shown to be a sustainable alternative, enhancing the growth of plants and biologically controlling insects (Sessitsch et al, 2004;Compant et al, 2005).…”