Diversity of foliar endophytic fungi from the medicinal plant Sapindus saponaria L . and their localization by scanning electron microscopy

Abstract

Endophytic fungi inhabit vegetable tissues or organs, without causing them any harm. Endophytes can co-evolve with plant hosts and possess species-specifi c interactions. They can protect the plant from insect attacks and diseases, and are also able to produce substances of biotechnological interest. In folk medicine, the bark, roots and fruits of Sapindus saponaria is used to produce substances with anxiolytic, astringent, diuretic and expectorant properties, as well as tonics, blood depuratives and cough medicine. This study evaluated the diversity of endophytic fungi present in the leaves of S. saponaria L. and observed the colonization of host plants by endophytes, using light and scanning electron microscopy. We verifi ed that these fungi are found in intercellular and intracellular spaces. The genera of some isolates of S. saponaria were identifi ed mainly by sequencing of ITS region of rDNA and, when possible, also by their microscopic features, as follows: Cochliobolus, Alternaria, Curvularia, Phomopsis, Diaporthe and Phoma. Phylogenetic analysis showed the existence of genetic variability of the genera Phomopsis and Diaporthe and interspecifi c variation among the Curvularia, Alternaria and Phoma, belonging to family Pleosporaceae.

Differential methods of localisation of fungal endophytes in the seagrasses

“…Different staining procedure for detecting the fungi from different plants are reported earlier (Petrini and Fischer (1990); Kowalski and Kehr (1992); Baum et al (2003); Gomez-Vidal et al (2006); Oses et al (2008); Bernardi-Wenzel et al (2010); Garcia et al (2012)). However, there is a dearth of knowledge of microscopical studies of endophytic fungal assemblage within the tropical seagrass.…”

“…Light microscopy evidenced the presence of endophytic fungal hyphae within the cells of seagrass tissue and thus allowed to understand the colonisation pattern of endophytic fungi. Techniques of staining, light microscopy and scanning electron microscopy detect the presence of fungal structures in the hosts (Garcia et al 2012) clearly. The present study founds evidence for the endophytic colonisation within seagrass tissue using three different staining methods through the visualisation of the hyphae.…”

“…This gives a clear idea about the abiotic stress tolerance exhibited by the seagrass are accompanied by the role of endophytic fungi by trapping the free oxygen radicals. Light microscopy and trypan blue staining identified the presence of hyphae and mycelial mass in the cells, in intercellular spaces and crossing from one cell to another (Garcia et al 2012) of S. saponaria . The data presented here also portrays the inter and intracellular hyphae of different diameters in the parenchyma cells, showing an extensive spread of hyphae both transversely and longitudinally.…”

“…The data presented here also portrays the inter and intracellular hyphae of different diameters in the parenchyma cells, showing an extensive spread of hyphae both transversely and longitudinally. The importance of detecting inter or intracellular colonisation by fungal endophytes is to better understand endophyte/host interaction (Garcia et al 2012). …”

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

“…Furthermore, because endophytic fungi have been shown to demonstrate high levels of phenotypic plasticity (Saikkonen at al., 1999), the role that they play in an ecosystem can vary substantially depending on both biotic and abiotic factors. The use of molecular markers has been useful in studies of endophytic variability, because these markers are capable of detecting polymorphisms in several species (Waldschimidt et al, 2002;Bernardi-Wenzel et al, 2010;Garcia et al, 2012;Orlandelli et al, 2012;Rhoden et al, 2012). Currently, the best tools available to detect endophyte variation effectively are ITS1-5.…”