• The Korean Journal of Mycology
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• v.32 no.2
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• pp.152-157
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• 2004

In this study the ITS1, ITS2 and 5.8S ribosomal DNA sequences from 29 strains of the Genus Inonotus and its related genera were compared with 31 strains obtained from GenBank database. Using the neighbor-joining (NJ) method and most parsimonious analysis the phylogenetic tree was constructed. The hymenochaetales formed no monophyletic group and several non-hymenochaetales appeared as intermingled with the Hymenochaetales. Strains 6, 46, 49, 50, 53, 55 showed no certain affinities within the Hymenochaetales, whereas Inonotus sp. (51) was closely related to Phellinus baumii, and Inonotus sp. (52), and Inonotus glomeratus (10) was related to Phellinus linteus, and Fomes fomentarius (30) was related to Ganoderma lucidum. Inonotus sp. and Phellinus sp. formed no monophyletic groups and a subdivision in the following genera is accepted: Inonotus sp. Phellinus baumii, Phellinus linteus, Phellinus igniarius, Phellinus pini, Hericium erinaceum, Ganoderma lucidum and Sparassis sp. were confirmed and separated genera. The taxonomic status of Inonotus remained uncertain. Eight new combinations are proposed.

• KSBB Journal
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• v.23 no.1
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• pp.23-30
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• 2008

An extracellular pigment production of three Phellinus sp. (Phellinus 421, P. linteus and P. hartigil) through submerged cultivation was investigated. The maximum brown pigment from culture broth was obtained from the precipitate by addition of 10% 1M HCI solution. This precipitate showed absorption characteristics with ${\lambda}_{max}$ of 360nm. The maximum production of extracellular pigment obtained at optimum medium and culture condition was 3.54 ($A_{360}$). The precipitate was fractionated by Sephadex G-75 gel chromatography, and the isolated brown pigment contained a large amount of polyphenol and the small amounts of sugar and protein. The brown pigment fraction was stable in temperature range of $30{\sim}60^{\circ}C$, pH range of $4{\sim}6$, sugar addition ranges of $1{\sim}5%$ and salt addition concentration of 3 molarity. Antioxidative activity of the brown pigment by TBA method was better than that of vitamin E (${\alpha}$-tocopherol).

• Mycobiology
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• v.39 no.4
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• pp.299-300
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• 2011

Two species of cultivated Phellinus sp. were identified as P. baumii by internal transcribed spacer (ITS) sequence analysis. The fruit bodies of the examined strains were similar to those of naturally occurring strains, having a bracket-like form, yellow-to-orange color, and poroid hymenial surfaces. The DNA sequences of ITS region of both strains showed a homology of 99% with ITS1 to ITS2 sequences of P. (Inonotus) baumii strain PB0806.

• Biomolecules & Therapeutics
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• v.9 no.3
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• pp.194-200
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• 2001

Phellinus linteus (PL)-hot water extract (PL-W) or- methanol extract (PL-M) was orally administered alone (single dose of 400, 800, 1600 mg/kg; 800 mg/kg/day for 5 days) or with cyclophosphamide (CY, 20 mg/kg, i.p.) to female ICR mice. Within PL alone-treated group, WBC and plaque forming cells (PFC) to SRBC were slightly and significantly enhanced when compared with control group. The relative thymus and spleen weights, WBC and PFC numbers were significantly decreased by the treatment of CY, whereas those values were markedly increased by the concomitant treatment of CY and PL when compared with CY administration alone. To assess the effects of PL and/or CY on the mitogen response of splenocytes io LPS, mouse splenocytes were stimulated with or without LPS in the presence of various concentration of PL and/or CY in vitro and splenocytes proliferation (SP) was measured by MTT assay. PL alone increased both SP and LPS- stimulated SP. Moreover, SP and LPS-induced SP suppressed by the treatment of CY alone were significantly restored by PL-treatment. These activities were higher by PL-M than by PL-W, These results indicated that PL was able to increase humoral immunity and to inhibit immunotoxicity induced by CY.

• Mycobiology
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• v.29 no.4
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• pp.190-193
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• 2001

Four strains of Phellinus spp. was identified based on internal transcribed spacer(ITS) region of rDNA sequence analysis and morphological characteristics. Basidiocarps of all strains were effused-reflexed and hymenial surface was poroid. Hyphal system was dimitic and basidiospore was globose to ellipsoid. The amplification of ITS regions produced a DNA fragment of 500 to 780 by in all strains examined. The determined sequences were analyzed for the reconstruction of phylogenetic tree. From these results, Phellinus sp. KM-1, KM-2, and KM-4 was identified as P. hartigii, P. baumii, and P. linteus, respectively.

• Journal of Life Science
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• v.11 no.2
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• pp.126-134
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• 2001

The phylogenetic tree displayed the presence of five groups in the Phellinus genus, which were distinguished based on their morphology. Most of the p. linteus appeared a cluster which was highly significant with the exception of P. linteus KACC 500122 and KACC 500411. They formed the sister taxa of P 1inteus where P. baumii, Phellinus sp. MPNU 7003, MPNU 7007, and MPNU 7010 had similar morphological characteristics. Also, P. nigricans IMSNU 32024 and P. pini var, carniformans IMSNU 32031 were grouped in the same cluster with P. igniarius KCTC 6227, KCTC 6228, and P. chrysoloma KCTC 6225 extracted from the Gen-Bank database. P. torulosus IMSNU 32028 and Phellinus sp. MPNU 7011 formed a closed group, however, these species had a distant taxa when compared with the other Phellinus species. The nucleotide sequences of the internal transcribed spacer (ITS) regions of ribosomal DNA (rDNA) including the 5.85 rDNA were determined from 24 strains of the Phellinus genus in order to analyze their phylogenetic relationship. These fungi were divided into two basic groups based on their ITS length, however, this grouping was different from that based on their morphological characteristics. Although various ITS sequences were ambiguously aligned, conserved sites were also identified. Accordingly, a neighbor-joining tree was constructed using the nucleotide sequence data of the conserved sites of the ITS regions and the 5.8S rDNA.

• The Korean Journal of Mycology
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• v.26 no.2 s.85
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• pp.265-274
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• 1998

This study was carried out to obtain the basic data for an artificial cultivation of Phellinus sp.. The optimum conditions for the mycelial growth on the different sawdusts (Quercus aliena, Morns alba and Alnus japonica) substrate of an isolated Phellinus sp. were optimized by response surface methodology. The ratio of rice bran addition to sawdust and the suitable moisture content for the mycelial growth in the all sawdust media were about 30% (w/w) and $65{\sim}70%$ (w/v), respectively. The initial pHs for the mycelial growth of Quercus aliena and Morns alba were in the range of $pH\;5{\sim}6$, whereas Alnus japonica was obtained at pH 6. The optimum temperature for the mycelial growth was about $25{\sim}30^{\circ}C$, depending on the different kinds of wood substrates. From the response surface analysis, the values of independent variables of Quercus aliena at stationary points were determined to be 31.01 % (w/w) of rice bran, pH of 5.31 and 69.03% (w/v) of moisture content, and the expected value of mycelial growth was about 8.32 cm. Both the ratio of rice bran addition to sawdust $(X_1)$ and moisture content $(X_3)$ were effective to the mycelial growth. In the case of Morns alba, the ratio of rice bran addition to sawdust, initial pH and moisture content at the stationary points were 28.77% (w/w), 5.28 and 69.8 (w/v),respectively, and the expected mycelial growth of 7.60 cm was obtained. Stationary points for the mycelial growth in the sawdust media of Alnus japonica were 28.74% (w/w) of rice bran, pH of 6. 04 and 66.96% (w/v) of moisture content, and the expected values of mycelial growth was about 5.38 cm. Based on the above results, there was correlations between the mycelial growth and independent variables, and the effect of rice bran $(X_1)$ and initial pH $(X_2)$ for the mycelial growth were higher than the moisture content $(X_3)$. The optimum species of sawdust media for the my celial growth of Phellinus sp. was in the order of Quercus aliena > Morns alba > Alnus japonica.

• The Korean Journal of Mycology
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• v.25 no.4 s.83
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• pp.257-267
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• 1997

Fruit bodies similar to the Phellinus sp. residing on the mulberry were collected at Yang-yang in Kang-won-do province and one strain of Phellinus sp. was isolated from the fruit bodies. For mass production of the isolated mycelia in a submerged culture, the culture conditions, medium composition, and the effect of various culture systems on the mycelial growth, were investigated. The morphological characteristics of the fruit body were as follows: covered with blackish to black and rough, lower surface with yellowish-brown to dull-brown and smooth, 5-7 cm thick and hard woody. Also, the pure cultured mycelia showed yellowish-brown color, capability of purplish-brown pigment production on the PDA plate media, no-formation of clamp-connection, much binding branch, and enzyme activities such as laccase, tyrosinase and peroxidase. Therefore, pure cultured strain was identified to be Phellinus sp. In the flask culture, the optimum culture conditions for the mycelial production were obtained after cultivation of 8 days at inoculum level of 5%(v/v), media volume of 70 mL, 150 rpm, initial pH 6, and temperature of $30^{\circ}C$. Optimum medium composition from the response surface analysis were determined to be glucose 12.12 g/L, sucrose 12.12 g/L, yeast extract 11.15 g/L, malt extract 11.15 g/L, $KH_2PO_4$ 0.855 g/L and $CaCl_2$ 0.855 g/L. The production of the mycelia after 4 and 8 days of cultivation was 1.95 and 9.89 g/L, respectively. The maximum specific growth rate and productivity were $0.020\;hr^{-1}$ and 1.25 g/L/day, respectively. Among the three different culture systems for the growth of mycelia, the maximum mycelial dry weight of 7.5 g/L was obtained after cultivation of 4 days in the air-lift fermentor under aeration rate of 2.5 vvm. The maximum specific growth rate and productivity were $0.033\;hr^{-1}$ and 1.9 g/L/day, respectively, which were about 1.7 and 4.2 times higher than those of flask culture.

• The Korean Journal of Mycology
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• v.27 no.2 s.89
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• pp.124-131
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• 1999

This study was carried out to identify the phylogenetic relationship among Phellinus species by comparing the DNA sequences of the 5.8S ribosomal DNA (rDNA) and the internal transcribed spacers (ITSs), ITS1 and ITS2 regions. Two primers from the 3' end of 18S rDNA and the 5' end of 28S rDNA sequences were chosen to amplify the specific ITS regions of Phellinus spp. Phellinus strains used in the study were divided into four clusters by the phylogenetic tree based on the amplified regions of ITS and 5.8S rDNA sequences. The first cluster consist of Phellinus hartigii IMSNU 32041 and Phellinus robustus IMSNU 32068, and the second cluster consists of Phellinus linteus strains and Phellinus weirianus IMSNU 32021. Phellinus laevigatus KCTC 6229, KCTC 6230 and Phellinus igniarius KCTC 6227, KCTC 6228 belong to the third cluster. Finally, Phellinus chrysoloma KCTC 6225 and Phellinus chrysoloma KCTC 6226 are the fourth cluster. In the second cluster the differentiation between Phellinus linteus strains and Phellinus weirianus species were not possible by the comparison of the ITS sequences. These results revealed that Phellinus linteus and Phellinus weirianus cannot be established the concept of species level only by the ITS sequences. Therefore, both physiological and molecular biological methods as well as the sequences of type strains are necessary to classify the strains of these two species accurately. The comparison of the ITS sequences of four Phellinus species indicated that the sequences of the ITS1 generally are more divergent than those of the ITS2. Although the ITS sequences are varied in some species, the conserved regions in both ITS1 and ITS2 are useful tool to differentiate the species. Phellinus linteus and related species have their specific sequences in the ITS1 compared to the other species.

• 한국약용작물학회:학술대회논문집
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• 2008.11a
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• pp.332-333
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• 2008