A rapid and inexpensive method for estimating lipid content of yeasts is needed for screening large numbers of yeasts samples. Nile red is a fluorescent lipophilic dye used for detection and quantification of intracellular lipid droplets in various biological system including algae, yeasts and filamentous fungi. However, a published assay for yeast is affected by variable diffusion across the cell membrane, and variation in the time required to reach maximal fluorescence emission. In this study, parameters that may influence the emission were varied to determine optimal assay conditions. An improved assay with a high-throughput capability was developed that includes the addition of dimethyl sulfoxide (DMSO) solvent to improve cell permeability, elimination of the washing step, the reduction of Nile red concentration, kinetic readings rather than single time-point reading, and utilization of a black 96-well microplate. The improved method was validated by comparison to gravimetric determination of lipid content of a broad variety of ascomycete and basidiomycete yeast species.
Polyol esters of fatty acids (PEFA) are amphiphilic glycolipids produced by yeast that could play a role as natural, environmentally friendly biosurfactants. We recently reported discovery of a new PEFA-secreting yeast species, Rhodotorula babjevae, a basidiomycetous yeast to display this behavior, in addition to a few other Rhodotorula yeasts reported on the 1960s. Additional yeast species within the taxonomic order Sporidiobolales were screened for secreted glycolipid production. PEFA production equal or above 1 g L were detected in 19 out of 65 strains of yeast screened, belonging to 6 out of 30 yeast species tested. Four of these species were not previously known to secrete glycolipids. These results significantly increase the number of yeast species known to secrete PEFA, holding promise for expanding knowledge of PEFA synthesis and secretion mechanisms, as well as setting the groundwork towards commercialization.
Several species of Aquilaria and Gyrinops are native to Indonesia and well known as agarwood-producing trees with a high economic value. Their bioactive compounds have a wide spectrum of uses, such as in medicine and cosmetics. These genera have undergone extensive search for novel bioactive compounds. The purpose of this study was to isolate, identify, and characterize the endophytic fungi community associated with Aquilaria malaccensis, A. microcarpa, Gyrinops versteegii, and A. crassna trees and investigate their bioactive properties as antioxidant agents and antagonists. A total of 50 fungi were successfully isolated from different tissues of the four species of agarwood-producing trees. Two isolates exhibited strong antioxidant activity, namely, Apodus oryzae (R2MC3A, IC 50 60.92 mg/mL) and Diaporthe sp. (P1DS1[C], IC 50 76.65 mg/mL). Two isolates, Pestalotiopsis theae (P3BS3[B]) and Curvularia sp. (P2CD3A), showed >75% antifungal activity against pathogenic Fusarium solani. The results revealed that endophytic fungi associated with the studied agarwood-producing trees had potential antioxidant and antifungal activities for further applications in biotechnology.
Styrax benzoin is a native tree to Indonesia, particularly in North Sumatra. This plant species produces benzoin resin, which is beneficial for medicinal treatments, hence its commercial value. Endophytic fungi help produce bioactive metabolites and contribute to resin production. However, the diversity of endophytic fungal species from S. benzoin grown in North Sumatra remained largely unexplored. This study aims to determine the distribution and diversity of culturable endophytic fungi from two kemenyan-producing locations in North Sumatra, Simalungun and North Tapanuli, as well as their tree part origin. A total of 7 and 8 endophytic fungal species were obtained from barks, stems, and/or leaves of S. benzoin grown in Simalungun and North Tapanuli, respectively, and identified by internal transcribed spacer sequence analysis. Endophytic fungi from North Tapanuli showed higher diversity, with a Shannon-Wiener index of 2.31 than those from Simalungun (1.95). Morisita-Horn similarity indices for bark-stem, stem-leaf, and bark-leaf were 0.47, 0.08, and 0, respectively, hinting at organ-specificity colonization. This study offers insights into the diversity of endophytic fungi isolated from S. benzoin which may contribute to future improvement of benzoin resin production.
The restoration of peat swamp forests in Sumatra island has become Indonesian government’s priority to restore ecological functions and their utilization. Indigenous. However, little information is available on the status of arbuscular mycorrhizal (AM) fungi in Sumatera. The objective of this research was to know AM fungi colonization in indigenous of tropical peat swamp forests. Root samples of 28 tree species in 14 families grown in a peat swamp forest of Jambi, Riau, and South Sumatera. All soil and tree roots were grown in zeolite media and trapped in Pueraria javanica and Shorgum bicolor as host plants for four months in a green house. Roots were stained with 0.1% trypan blue and vesicles, arbuscles and internal hyphae of AM fungi observed under a compound microscope. The results have shown that 20 tree species (72%) were colonized by AM fungi, 4 tree species of the dipterocarps family (14%) were colonized by ectomycorrhizal fungi (ECM), and four other tree species (14%) did not find FMA or ECM colonization. It is suggested that utilization of mycorrhizas can increase early growth of some tree species grown in peat swamp forests and mycorrhizal application will be expected as a key technology to restore degraded peatlands.
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