• Title/Summary/Keyword: in vitro condition

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Screening for Effective Organic Farming Materials for the Control of Cucumber Scab Caused by Cladosporium cucumerinum (오이 검은별무늬병 방제에 효과적인 유기농업자재 선발)

  • Park, So-Hyang;Hong, Sung-Jun;Shim, Chang-Ki;Kim, Min-Jeong;Park, Jong-Ho;Han, Eun-Jung;Kim, Yong-Ki
    • Research in Plant Disease
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    • v.23 no.2
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    • pp.159-167
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    • 2017
  • This study was conducted to evaluate the control efficacy of the organic farming materials on cucumber scab caused by Cladosporium cucumerinum PT1 (KACC 48094). The antifungal activities in vitro as well as the suppressive effect of 43 organic farming materials on the spore germination and germ tube growth by inoculating spore suspension on cucumber seedlings in vivo were investigated. Thirteen organic farming materials inhibited the mycelial growth of C. cucumerinum and nine of these were microbial agents. In the screening using cucumber seedlings, six organic farming materials were very effective with control efficacy value of 90%. Among them, Bacillus amyloliquefaciens M27 provided suppressive effect on both mycelial growth and spore germination against cucumber scab. Finally, nine organic farming materials were selected to test the protective and curative effects, and all chosen organic farming materials significantly suppressed disease incidence when applied in the preventive action, in comparison with the curative action. Especially, Bordeaux mixture I and III gave excellent protective control efficacy with control values of 96.7% and 73.3%, respectively, whereas its curative control effect was significant low. Among these, only Thymus quinquecostatus+Sophora extract showed curative activity, although the control value was as low as 50%. This study suggests that cucumber scab can be controlled by some organic farming materials in the farmhouses under comparatively cold and wet condition and protective treatment is more important and efficient.

Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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In Vitro Differentiation of Human Amniotic Membrane-derived Stem Cells into Hepatocyte-like Cells (양막 유래 줄기세포의 간세포로의 분화 유도)

  • Kook, Min-Ji;Park, Soo-Yeon;Kang, Hyun-Mi;Kim, Hae-Kwon
    • Development and Reproduction
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    • v.10 no.1
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    • pp.63-73
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    • 2006
  • This study aimed to find out suitable culture conditions for the differentiation of human amniotic membrane-derived stem cells(HAM) into hepatocyte-like cells. Almost homogenous population of fibroblast-like cells was successfully isolated from the amniotic membrane. In comparison to the non-coated plates and in the absence of insulin/transferrin/selenium(ITS), HAM cultured on the fibronectin-coated plates and in the presence of ITS showed the more intense immunocytochemical staining against the albumin. Addition of both fibroblast growth factor(FGF)-1 and -2 to the differentiation medium gave stronger staining compared to the treatment with FGF-1 or -2 alone. Periodic acid Schiff's base staining of glycogen and morphological turnover of fibroblast-like appearance of HAM into round shape matched the results of immunocytochemical studies. When the efficiency of two-step culture method was examined on the differentiation of HAM into hepatocyte-like cells, all of the results of immunocytochemical staining, periodic acid Schiff's staining and morphological change exhibited effective hepatic differentiation of HAM compared to the continuous culture method. Immunoblot analyses of HAM- conditioned media against the albumin showed that the culture of HAM in the presence of both ITS and fibronectin always gave a stronger staining intensity than those in the absence of them, and that the addition of ether mixture of FGF-4 and either FGF-2 or transforming growth $factor(TGF)-{\alpha}$ to the culture medium significantly enhanced the albumin secretion by HAM. Based on these observations, it is suggested that HAM could differentiate into hepatocyte-like cells under a culture condition consisting of fibronectin and ITS, and addition of FGF-4 with either one of FGF-2 or $TGF-{\alpha}$ could enhance the hepatic differentiation of HAM.

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Effect of Electric fusion Methods on Cell Fusion Rate and Embryo Development by Somatic Cell Nuclear Transfer in Korean Native Cattle(KNC) (한우 체세포를 이용한 핵이식에서 전기융합 방법이 융합율 및 배발달율에 미치는 영향)

  • 김은국;김정욱
    • Journal of Embryo Transfer
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    • v.18 no.3
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    • pp.171-178
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    • 2003
  • This study was conducted to investigate the effect of electric fusion methods on cell fusion rate and embryo development by somatic cell nuclear transfer in Korean Native Cattle. The KNC ear cell was cultured in vitro for confluence in serum starvation condition(DMEM+0.05% FBS) for cell confluence. The zona pellucida of IVM oocytes were partially dissection using micro pipette. Ear cells were transferred into an enucleated oocyte. The reconstructed embryos were electrically fused with Zimmermann Cell Fusion Medium(ZCFM). Nuclear transfer embryos were activated with a combination of 10${\mu}{\textrm}{m}$ calcium ionophore(5 min) and 2.0mM 6-DMAP(3 hr). The activated embryos were cultured in CR1 -aa medium contains 0.3% BSA or 10% FBS at 37$^{\circ}C$, 90% $N_2$, and 5% $CO_2$in incubator for 6 days. The fusion rates were 51.6%(chamber) and 68.9%(needle), respectively and there were significantly difference between the fusion method(P<0.05). But, lysis rates were not significantly different(10.7%, 11.5%), respectively. The cleavage rates were significantly different between the chamber method(73.2%) and needle method(80.3%), respectively(P<0.05). The rates of early embryos(2∼4cells) and blastocysts of chamber and needle methods were 54.1%, 61.1% and 18.4%, 26.3% respectively, and needle method was significantly higher than chamber method(P<0.05). But, morulae formation rate were not significantly differences between the chamber(6.7%) and needle(6.2) method(P <0.05). These result suggest that electric fusion of needle method was to be profitable for nuclear transfer embryo fusion rate, blastocyst formation rate and reduce of oocyte lysis.

Suitable hormone-free medium for in vitro mass propagation via bioreactor culture of ever-bearing strawberry (Bioreactor를 이용한 사계성 딸기 조직배양묘 대량증식을 위한 적정 무호르몬 배지)

  • Kim, Hye-Jin;Lee, Jong-Nam;Kim, Ki-Deog;Im, Ju-Sung;Lim, Hak-Tae;Yeoung, Young-Rok
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.221-227
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    • 2011
  • This study was carried out to determine optimal medium conditions for mass propagation by bioreactor culture of ever-bearing strawberry (Fragaria ${\times}$ ananassa D.). Two different type of nitride were that mixed $NH_4NO_3$ and $KNO_3$ or added $KNO_3$ only. And nitride concentrations were at the 4 levels of $1/2{\times}$, $1{\times}$, $2{\times}$ and $3{\times}$ that was included $NH_4NO_3$ and $KNO_3$. Sucrose content ranged at 3 levels of $10g{\cdot}L^{-1}$, $20g{\cdot}L^{-1}$ and $30g{\cdot}L^{-1}$ and medium pH were at the 3 levels of 4.6, 5.6 and 6.6. In bioreactor culture, medium that are included $NH_4NO_3$ and $KNO_3$ together in MS medium was suitable for mass propagation. Medium EC rose rapidly when the nitride concentration was increased. For that reason, plantlet growth was inhibited. Shoots of nitride $1/2{\times}$ concentration was 10.8 ea and $1/2{\times}$ concentration was suitable for shoot propagation. Fresh weight of sucrose $30g{\cdot}L^{-1}$ was 3,101 mg which was heaviest and aerial and ground part were higher than the other concentration. Shoots were increased in proportion to the increasing concentration of sucrose. In the pH condition, from pH 5.6 to 6.8 were appropriate for the optimum growth of aerial and ground part of plant. From the results, in bioreactor culture for mass propagation, MS medium was suitable $1/2{\times}$ concentration that was included $NH_4NO_3$ and $KNO_3$ together, and added $30g{\cdot}L^{-1}$ of sucrose, and then adjusted pH between 5.6 and 6.6.

The effect of silane treatment timing and saliva contamination on shear bond strength of resin cement to porcelain (Silane의 처리시기와 타액오염이 도재-레진 시멘트의 전단 결합강도에 미치는 영향)

  • Ro, Young-Seon;Ryu, Jae-Jun;Suh, Kyu-Won
    • The Journal of Korean Academy of Prosthodontics
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    • v.47 no.1
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    • pp.61-69
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    • 2009
  • Statement of problem: Porcelain veneers have become a popular treatment modality for aesthetic anterior prosthesis. Fitting porcelain veneers in the mouth usually involve a try-in appointment, which frequently results in salivary contamination of fitting surfaces. Purpose: An in vitro study was carried out to investigate the effect of silane treatment timing and saliva contamination on the resin bond strength to porcelain veneer surface. Material and methods: Cylindrical test specimens (n=360) and rectangular test specimens (n=5) were prepared for shear bond test and contact angle analysis. Whole cylindrical specimens divided into 20 groups, each of which received a different surface treatment and/or storage condition. The composite resin cement stubs were light-polymerized onto porcelain adherends. The shear bond strengths of cemented stubs were measured after dry storage and thermocycling (3,000 cycles) between 5 and $55^{\circ}C$. The silane and their reactions were chemically monitored by using Fourier Transform Infrared Spectroscopy analysis (FTIR) and contact angle analysis. One-way analysis of variance (ANOVA) and Dunnett's multiple comparison were used to analyze the data. Results: FT-IR analysis showed that salivary contamination and silane treatment timing did not affect the surface interactions of silane. Observed water contact angles were lower on the saliva contaminated porcelain surface and the addition of 37% phosphoric acid for 20 seconds on saliva contaminated porcelain increased the degree of contact angle. Silane applied to the porcelain, a few days before cementation, resulted in increasing the bond strength after thermocycling. Conclusion: Within the limitation of this study, it can be concluded that it would be better to protect porcelain prosthesis before saliva contamination with silane treatment and to clean the contaminated surface by use of phosphoric acid.

ELASTIC CONSTANTS, SHEAR BOND STRENGTH OF TUNNEL RESTORATIVE MATERIALS AND MARGINAL RIDGE STRENGTH OF RESTORED TEETH (터널형 2급와동 충전재의 탄성계수와 전단결합강도 및 수복치의 변연융선 파절강도에 관한 연구)

  • Lee, Ka-Yean;Park, Yeong-Joon;Yang, Kyu-Ho
    • Journal of the korean academy of Pediatric Dentistry
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    • v.23 no.3
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    • pp.746-763
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    • 1996
  • An alternative design to conventional class II cavity preparation for proximal carious lesions is the tunnel preparation. It preserves the marginal ridge intact, thus making it possible to maintain the natural contact relationship with the adjacent tooth and minimize tooth reduction. This in vitro study was purposed to evaluate the effect of the materials' elastic constants and shear-bond strength on the marginal ridge fracture resistance of teeth restored by the tunnel technique, and to find the materials of choice for tunnel restorations. $Resinomer^{(R)}$, $Ketac-silver^{(R)}$, $Miracle-Mix^{(R)}$, and Tytin were used as restorative material. The elastic constants of each restorative material were evaluated by ultrasonic pulse measurement. Young's modulus and bulk modulus of the restorative materials were evaluated in three specimens for each material type. The shear-bond strength of the restorative materials to the dentin surface was measured after thermocycling 400 times between 6 and $60^{\circ}C$, using ten specimens for each material type. For measuring marginal ridge strength, 60 sound extracted molar teeth were distributed into six groups by size. Sound molar teeth were used as a Control group and unfilled prepared teeth were grouped as Unrestored. Another four groups were named Resinomer group, Ketac-Silver group, Miracle Mix group, and Tytin group by type of restorative material. Tunnel cavity preparation was done with ' 1/2, 2, and 4 round burs in sequence. Initial access to proximal surface was made through an occlusal access preparation started at least 2mm from the marginal ridge, and the proximal opening was formed about 2.5mm below the marginal ridge. After restoration and thermocycling, marginal ridge strength was measured using a universal testing machine. The results were as follows: 1. The Young's modulus of $Tytin^{(R)}$ was 63.95 GPa, followed by $Ketac-Silver^{(R)}$ 27.60 GPa, $Miracle-mix^{(R)}$ 18.48 GPa, and $Resinomer^{(R)}$ 10.74 GPa showing significant differences between the groups(P<0.05). The bulk modulus of the materials showed the same order as Young's modulus. The value of $Tytin^{(R)}$ showed 59.57 GPa indicating that it will deform less than other materials under the same stress. It was followed by $Ketac-Silver^{(R)}$ 23.57 GPa, Miracle $Mix^{(R)}$ 12.50 GPa, and $Resinomer^{(R)}$ 11.60 GPa. 2. The Resinomer group had a shear-bond strength of 7.41 MPa which was significantly higher than those of the Ketac-Silver group (1.80 MPa) and the Miracle Mix group (2.84 MPa) (P<0.01). All the specimens of Tytin group detatched from the dentin surface during thermocycling. 3. The mean marginal ridge strength of the Unrestored group(46.14 kgf) was significantly lower than that of the Control group (84.24 kgf) (P<0.01). The marginal ridge strength of teeth restored by the tunnel technique was, in order, Ketac-Silver group 74.06 kgf, Miracle Mix group 73.36 kgf, Resinomer group 63.47 kgf, and Tytin group 58.76 kgf. The Ketac-Silver, Miracle Mix, and Resinomer groups showed no significant difference with the Control group (P>0.05), but the Tytin group showed significantly lower strength compared to the Control group(P<0.05). The results showed that the marginal ridge strength of the teeth restored by the tunnel technique was not significantly lower than that of sound teeth. They also demonstrated that the bonding strength of the restorative material to the tooth surface should be high and the modulus of elasticity should not be lower than that of the tooth in order to restore the marginal ridge strength to its natural condition.

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Survival Rate of Lactic Acid Bacteria and the Change of ${\beta}-Galactosidase$ Activity in Commercial Yogurts Under the Acidic Conditions (산성조건하에서 시판요구르트의 유산균 생존률과 ${\beta}-galactosidase$의 활성도)

  • Shin, Yong-Seo;Sung, Hyun-Ju;Kim, Dong-Han;Lee, Kap-Sang
    • Applied Biological Chemistry
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    • v.37 no.3
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    • pp.143-147
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    • 1994
  • Four samples of commercially manufactured yogurts (plain, drinking type) were purchased and evaluated their physico-chemical properties, buffering capacity. And the survival rate of lactic acid bacteria and their ${\beta}-galactosidase$ activity under the acidic conditions (in vitro) were investigated. The values of pH, titratable acidity, viscosity and viable cell counts of yogurts were $3.71{\sim}4.08$, $0.990{\sim}1.045%$, $256{\sim}3164\;cps.$ and $10^8{\sim}10^9\;cfu/ml$, respectively. The volume of 1.0 M-HCl required to reduce the pH of yogurt (50 ml) to minus 2 value was $3.58{\sim}4.33\;ml$. When commercial yogurts were incubated at $37^{\circ}C$ for 120 minutes under the acidic conditions (pH 3.5, 2.5, 1.5), the survival rates of lactic acid bacteria in yogurt were $3.5{\times}10^{-2}{\sim}3.6{\times}10^{-1}%$ at pH 2.5, $8.3{\times}10^{-5}{\sim}4.2{\times}10^{-3}%$ at pH 1.5, respectively, but there was no significant difference at pH 3.5. The remaining activities of ${\beta}-galactosidase$ were $9.4{\sim}36.2%$ at pH 2.5, $4.2{\sim}19.0%$ at pH 1.5, respectively. These results suggested that a significant number of lactic acid bacteria in yogurt might be destroyed in the hostile environment of the stomach, but ${\beta}-galactosidase$ activity from yogurt might be somewhat maintained probably due to the protecting effect by its cell wall and membrane.

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Ameliorating effect of the ethyl acetate fraction of Pteridium aquilinum on glucose-induced neuronal apoptosis (포도당으로 유도된 신경세포 손상에 대한 고사리 아세트산에틸 분획물의 개선 효과)

  • Park, Seon Kyeong;Guo, Tian Jiao;Kim, Jong Min;Kang, Jin Yong;Park, Sang Hyun;Kang, Jeong Eun;Kwon, Bong Seok;Lee, Chang Jun;Lee, Uk;Heo, Ho Jin
    • Korean Journal of Food Science and Technology
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    • v.49 no.4
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    • pp.430-437
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    • 2017
  • The protective effect of Pteridium aquilinum on high glucose-induced cytotoxicity was examined in vitro to investigate the relationship between diabetic condition and neuronal dysfunction. The ethyl acetate fraction of P. aquilinum (EFPA), with total phenolic content of 265.08 mg gallic acid equivalent/g, showed higher 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)/2,2-diphenyl-1-picrylhydrazyl radical scavenging activities and lipid peroxidation inhibitory effect than any other fraction. In addition, EFPA showed a significant reduction in the inhibitory effect on ${\alpha}$-glucosidase activity ($IC_{50}$ value=$205.26{\mu}g/mL$) compared to the acarbose positive control. The anti-oxidative effect in PC12 cells, protective effects on high glucose-induced oxidative stress in neuronal cells, and neurotoxicity were measured using 2',7'-dichlorofluorescin diacetate, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazoliumbromide, and lactate dehydrogenase assays, respectively. EFPA showed conspicuous inhibitory effect on cellular reactive oxygen species production and neuronal cell apoptosis. Finally, kaempferol-3-glucoside was identified as the main phenolic compound of EFPA using high performance liquid chromatography.

Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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