Hiroto Suhara

Phylogenetical approach to isolation of white-rot fungi capable of degrading polychlorinated dibenzo- p -dioxin

A degradation experiment on PCDDs and phylogenetical analyses were carried out on newly isolated 2,7-dichlorodibenzo-p-dioxin (2,7-diCDD)-degrading white-rot fungi, strains BMC3014, BMC9152, and BMC9160. When these fungi were incubated with tri- or tetraCDDs, the substrates were degraded efficiently, and hydroxylated metabolites were detected. On the other hand, 1,3,6,8-tetrachlorodibenzo-p-dioxin was not decreased, and no metabolites were detected. Phylogenetic analysis of internal transcribed spacers (ITSs) containing rRNA gene sequence (ITS-rDNA) clarified that these strains belonged to the genus Phlebia and were closely related to the fungi Phlebia lindtneri, strains MZ-227 and MG-60, which had both been isolated as 2,7-diCDD-degrading fungi in our previous study. Based on this phylogenetical relationship, other Phlebia genera species were used for a degradation experiment on 2,7-diCDD and 1,3,6,8-tetraCDD. Phlebia acerina and Phlebia brevispora degraded 2,7-diCDD about 40 and 80%, respectively, over 14 days of incubation. It became clear that P. brevispora can degrade 1,3,6,8-tetraCDD and transform it to monohydroxy-tetraCDD, monomethoxy-tetraCDD, dimethoxy-tetraCDD, dimethoxy-triCDD, and 3,5-dichlorocatechol in the treatment cultures. In this paper, we could clearly prove for the first time by identifying the metabolites that white-rot fungus P. brevispora could degrade the recalcitrant dioxin, 1,3,6,8-tetraCDD.

Specific detection of a basidiomycete, Phlebia brevispora associated with butt rot of Chamaecyparis obtusa, by PCR-based analysis

In order to monitor the basidiomycetous fungus Phlebia brevispora isolated from butt rot of Chamaecyparis obtusa (Japanese cypress) in 1997 in Nagasaki Prefecture, a sensitive polymerase chain reaction (PCR)-based assay was developed to specifically detect the fungus on-site. A species-specific primer for P. brevispora was derived from the internal transcribed spacer (ITS) region (containing 5.8S ribosomal DNA, ITS1 and ITS2) sequences of the fungus. The PCR assay was able to detect down to 1 fg DNA (per 1 µl PCR reaction mixture) and down to 0.2 mg mycelium of P. brevispora (per 1 g of decayed wood). The samples for on-site monitoring were collected in 2002 from the decayed tree stump in which P. brevispora had first been isolated. From the decayed tree tissue, P. brevispora could be detected by PCR assay even when its mycelium could not isolated from the tree tissue by culturing. This indicates that the PCR amplification using the specific primer developed here is a useful method for monitoring P. brevispora on-site.

Corticioid fungi (Basidiomycota) in mangrove forests of the islands of Iriomote and Okinawa, Japan

Sixteen species of corticioid fungi (Basidiomycota) were collected from mangrove forests in the islands of Iriomote and Okinawa, Japan. All the species are new records for the Japanese mangrove forests. Of these species, 6 species are newly reported from Japan: Cerocorticium molle, Gloeocystidiellum moniliforme, G. wakullum, Hyphoderma ayresii, Phanerochaete tropica, and Phlebia acanthocystis. Their morphological descriptions, illustrations, and remarks based on the Japanese specimens are provided.

Indole-3-carbaldehyde: a tyrosinase inhibitor from fungus YL185

Indole-3-carbaldehyde (1) was isolated as a tyrosinase inhibitor from the ethyl acetate-soluble fraction of extracellular fluids of unknown fungus YL185. The partial sequencing data of 18S ribosomal DNA (18S rDNA) indicate that this isolate belongs to the family Polyporaceae or Corticiaceae sensu lato. Indole-3-carbaldehyde inhibited the oxidation of l-3,4-dihydroxyphenylalanine (l-DOPA) by mushroom tyrosinase with a 50% inhibitory concentration (IC50) of 1.3 mM and showed inhibitory activity on melanin production in B16 melanoma cells. The aldehyde group of 1 plays an important role in eliciting tyrosinase inhibitory activity.

Wild Mushrooms in Nepal: Some Potential Candidates as Antioxidant and ACE-Inhibition Sources

Twenty-nine mushrooms collected in the mountainous areas of Nepal were analyzed for antioxidant activity by different methods, including Folin-Ciocalteu, ORAC, ABTS, and DPPH assays. Intracellular H2O2-scavenging activity was also performed on HaCaT cells. The results showed that phenolic compounds are the main antioxidant of the mushrooms. Among studied samples, Inonotus andersonii, and Phellinus gilvus exhibited very high antioxidant activity with the phenolic contents up to 310.8 and 258.7 mg GAE/g extracts, respectively. The H2O2-scavenging assay on cells also revealed the potential of these mushrooms in the prevention of oxidative stress. In term of ACE-inhibition, results showed that Phlebia tremellosa would be a novel and promising candidate for antihypertensive studies. This mushroom exhibited even higher in vitro ACE-inhibition activity than Ganoderma lingzhi, with the IC50 values of the two mushrooms being 32 μg/mL and 2 μg/mL, respectively. This is the first time biological activities of mushrooms collected in Nepal were reported. Information from this study should be a valuable reference for future studies on antioxidant and ACE-inhibitory activities of mushrooms.

Changes in content of triterpenoids and polysaccharides in Ganoderma lingzhi at different growth stages

Ganoderma lingzhi is a traditional medicinal mushroom, and its extract contains many bioactive compounds. Triterpenoids and polysaccharides are the primary bioactive components that contribute to its medicinal properties. In this study, we quantified 18 triterpenoids, total triterpenoid content and total polysaccharide content in the ethanol and water extracts of G. lingzhi at different growth stages. Triterpenoids were quantified by liquid chromatograph–tandem mass spectrometry in the multiple-reaction-monitoring mode. Total triterpenoid and total polysaccharide content were determined by colorimetric analysis. The results indicated that the fruit bodies at an early growth stage had a higher content of ganoderic acid A, C2, I and LM2, as well as of ganoderenic acid C and D, than those at a later growth stage. In contrast, ganoderic acid K, TN and T–Q contents were higher in mature fruit bodies (maturation stage). The highest total triterpenoid and total polysaccharide contents were found in fruit bodies before maturity (stipe elongation stage or early stage of pileus formation). Our results provide information which will contribute to the establishment of an efficient cultivation system for G. lingzhi with a higher content of triterpenoids.