Evaluation of foliar fungal endophyte diversity and colonization of medicinal plant Luehea divaricata (Martius et Zuccarini)

Abstract

Endophyte microorganisms are organisms that live inside plants without causing any apparent damage to their hosts. Since all plants exhibit endophyte microorganisms, it is believed that mutual association is of great importance in nature. Luehea divaricata (Martius & Zuccarini), known popularly in Brazil as açoita-cavalo, is a big-sized tree with a wide distribution in the country that possesses medicinal qualities for: dysentery, leucorrhea, rheumatism, blennorrhoea, tumors, bronchitis, and depuration. This research aims at isolating and molecularly characterizing fungi isolates from L. divaricata by sequence analysis of ITS1-5.8S-ITS2 rDNA. Further, the colonization of endophyte in the host plant by Light and Scanning Electron Microscopy will also be investigated. Whereas, genera Alternaria, Cochliobolus, Diaporthe, Epicoccum, Guignardia, Phoma, and Phomopsis, were identified; rDNA sequence analysis revealed intra-species variability among endophyte isolates of the genus Phomopsis sp. Light and Scanning Electron Microscopy techniques showed the presence of endophyte fungi inside L. divaricata leaves, inhabiting inter-and intra-cellular spaces. These types of extensive colonization and dissemination were reported throughout all the leaf parts in palisade parenchyma, esclerenchyma, spongy parenchyma, adaxial epidermis, and vascular bundle indicating colonization of endophytes in multiple structural sub-niches in the host plant.

Phylogenetic diversity of endophytic leaf fungus isolates from the medicinal tree Trichilia elegans (Meliaceae)

“…In relation to the percentage of the isolates, the data are consonant with other findings showing that Phomopsis is quite frequently found as an endophyte in several plants (Chareprasert et al, 2006;Bernardi-Wenzel et al, 2010).…”

“…Bernardi-Wenzel et al (2010), with Luhea divaricata isolates and phylogenetic analysis by rDNA sequencing, also observed the presence of intraspecific variability of the genus Phomopsis. The endophytes of T. elegans could be grouped into 2 class clades (Figure 1).…”

“…The superficial disinfection of the collected leaves was carried out using sodium hypochlorite (NaOCl), according to the protocols described by Bernardi-Wenzel et al (2010), which were slightly modified. The NaOCl concentration used was 5%, and the time was 5 min.…”

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

“…According to Kuldau and Bacon (2008), the intracellular spaces actually have many organic and inorganic nutrients, able to support the concentration of endophytic fungi observed in gramineae. Bernardi-Wenzel et al (2010), using light microscopy and SEM, visualized endophytic fungi that inhabit inter and intracellular spaces in leaves of Luhea divaricata. It was possible to observe extensive and disseminated colonization along all leaf parts, in the palisade parenchyma, sclerenchyma, adaxial epidermis and conductive vessels, indicating close interaction among the endophytes in multiple structural and trophic subniches in the host.…”

“…2b). We stained transversal cuts of S. saponaria leaves with and without preincubation (incubation of leaves in BDA for 24 h) and observed strong staining after incubation, indicating the presence of endophytes, as found by Bernardi-Wenzel et al (2010). We mainly considered observations of individual hyphae in the results and discussion sections.…”