Lignin-degrading enzymes from a pathogenic canker-rot fungus Inonotus obliquus strain IO-B2.

Inonotus obliquus is a pathogenic fungus found in living trees and has been widely used as a traditional medicine for cancer therapy. Although lignocellulose-degrading enzymes are involved in the early stages of host infection, the parasitic life cycle of this fungus has not been fully understood. In this study, we aimed to investigate the activities of laccase (Lac), manganese peroxidase (MnP), and lignin peroxidase (LiP) from I. obliquus cultivated in Kirk's medium. The fungus was subjected to genome sequencing, and genes related to wood degradation were identified. The draft genome sequence of this fungus comprised 21,203 predicted protein-coding genes, of which 134 were estimated to be related to wood degradation. Among these, 47 genes associated with lignin degradation were found to have the highest number of mnp genes. Furthermore, we cloned the cDNA encoding a putative MnP, referred to as IoMnP1, and characterized its molecular structure. The results show that IoMnP1 has catalytic properties analogous to MnP. Phylogenetic analysis also confirmed that IoMnP1 was closely related to the MnPs from Pyrrhoderma noxium, Fomitiporia mediterranea, and Sanghuangporus baumii, which belong to the same family of Hymenochaetaceae. From the above results, we suggest that IoMnP1 is a member of MnPs.

Geographic and longitudinal variations of anatomical characteristics and mechanical properties in three bamboo species naturally grown in Lombok Island, Indonesia.

The anatomical characteristics (fiber length and fiber area) and mechanical properties (modulus of elasticity, modulus of rupture, compressive strength, tensile Young's modulus, and tensile strength) of Bambusa vulgaris, Bambusa maculata, and Gigantochloa atter, naturally growing at four different sites in Lombok Island, Indonesia, were examined for evaluating geographic and longitudinal variations by mixed-effects modeling to effectively utilize bamboo culm resources for structural materials. We found geographic and longitudinal variations of bamboo culm properties in these three species. Based on the results, we concluded that, for utilization of bamboo culm as a structural material, variation of individual culm rather than site, and longitudinal variations should be considered for Bambusa species and G. atter, respectively.

Physical and mechanical properties of wood and their geographic variations in Larix sibirica trees naturally grown in Mongolia.

We examined the physical and mechanical properties of wood in Siberian larch (Larix sibirica) trees that grow naturally in five Mongolian provenances (Khentii, Arkhangai, Zavkhan, Khuvsgul, and Selenge) and the geographic variations between them. Five trees with stem diameters of 20 to 30 cm at 1.3 m above ground were collected from each provenance. The mean values of the modulus of elasticity (MOE), modulus of rupture (MOR), compressive strength parallel to grain (CS), and shearing strength (SS) ranged from 7.03 to 9.51 GPa, 79.8 to 103.9 MPa, 46.3 to 51.1 MPa, and 10.4 to 13.0 MPa, respectively. Significant differences were found in radial and tangential shrinkage, MOE, MOR, and SS in wood among the five provenances. In addition, juvenile wood had inferior physical and mechanical properties in comparison to mature wood within and among provenances. Furthermore, there were significant differences in all examined properties, except for CS, in mature wood among the five provenances. Higher correlation coefficients were also obtained in mature wood among all mechanical properties, except for SS.

The complete mitochondrial genome sequence of the medicinal fungus Inonotus obliquus (Hymenochaetaceae, Basidiomycota).

Inonotus obliquus is a medicinal fungus in the family Hymenochaetaceae and commonly known as chaga. The sclerotium of this fungus has been used as a traditional medicine for long time. In this study, we present the mitochondrial genome sequence of I. obliquus. The mitochondrial DNA (mtDNA) is 119,110 base pairs in length and contained genes for 58 Open reading frames, 2 ribosomal RNAs, and 30 transfer RNAs. Consequently performed phylogenetic analysis indicates that this fungus is closely related to Sanghuangporus sanghuang which belongs to the same family Hymenochaetaceae. We first reported about the complete mitochondrial genome of fungi belonging to the genus Inonotus.

Proteomic Analysis of Responsive Proteins Induced in Japanese Birch Plantlet Treated with Salicylic Acid.

The present study was performed to unravel the mechanisms of systemic acquired resistance (SAR) establishment and resistance signaling pathways against the canker-rot fungus (Inonotus obliquus strain IO-U1) infection in Japanese birch plantlet No.8. Modulation of protein-profile induced by salicylic acid (SA)-administration was analyzed, and SA-responsive proteins were identified. In total, 5 specifically expressed, 3 significantly increased, and 3 significantly decreased protein spots were identified using liquid chromatography/tandem mass spectrometry (LC/MS/MS) and the sequence tag method. These proteins were malate dehydrogenase, succinate dehydrogenase, phosphoglycerate kinase, diaminopimalate decarboxylase, arginase, chorismate mutase, cyclophilin, aminopeptidase, and unknown function proteins. These proteins are considered to be involved in SAR-establishment mechanisms in the Japanese birch plantlet No 8.

Oxidosqualene cyclases from cell suspension cultures of Betula platyphylla var. japonica: molecular evolution of oxidosqualene cyclases in higher plants.

Betula platyphylla var. japonica is a rich source of triterpenoid as it contains dammarane type triterpenes in the leaves, and lupane type and oleanane type triterpenes in the bark. Four oxidosqualene cyclase cDNAs (BPX, BPX2, BPW and BPY) were cloned by homology based PCR methods from cell suspension cultures of B. platyphylla var. japonica. Open reading frames consisting of 2274, 2304, 2268 and 2340 bp were ligated into yeast expression plasmid pYES2 under the control of GAL1 promoter and introduced into lanosterol synthase deficient (erg7) Saccharomyces cerevisiae strain GIL77. Analyses of in vitro enzyme activities and/or accumulated products in the transformants demonstrated that they encode cycloartenol synthase (BPX and BPX2), lupeol synthase (BPW) and beta-amyrin synthase (BPY) proteins. Phylogenetic tree was constructed for all the known oxidosqualene cyclases (OSCs) including the clones obtained in this study, revealing that OSCs having the same enzyme function form respective branches in the tree even though they derive from different plant species. Intriguing correlation was found between reaction mechanism and molecular evolution of OSCs in higher plants.