• Title/Summary/Keyword: Variation Point

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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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North Korea's Overseas Transfer Dance - Focusing on Japan and China - (북한춤의 해외전파 : 일본과 중국을 중심으로)

  • Kim, Chae-Won
    • (The) Research of the performance art and culture
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    • no.22
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    • pp.185-221
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    • 2011
  • This study overseas compatriots in the North during the propagation and development of dance patterns and was going to be based, people living outside of the dance culture as a group of overseas Koreans, especially dance culture of Koreans in Japan, China are interested in and thought about the necessity of the study. Issues discussed in the literature of research results, autonomous community of Koreans in Japan ethnic Koreans and Chinese dance culture dance Choi Seung-hee starting point common was, two ethnic groups, directly or indirectly from the Choi Seung-hee learn to dance or have received specialized training to work, compiled by Choi Seung-hee Korea on the basis of basic dance training was learning the dance. In addition, specialized training and dance training institutions in the North Koreans in Japan Social Dance Group for the system, such as dance training in a separate place where talented people through the exhaust, to act in a professional troupes have maintained a system. In contrast, Chinese ethnic Koreans in Yanbian Autonomous University and Central University for Nationalities in the dance departments are stationed there, the transfer from the Joseon dance dance by educating gifted talents have been dispose, South Korea and similar aspects of the dance education system can be seen. Dance work based training and the tendency of Koreans in Japan in terms of social practice and dance in the North of basic training as basic training and specialized training, and work to represent the North korea's famous dance folk dance performances have been transmitted intact. In China, however, ethnic Koreans Choi Seung-hee compiled by borough basis and the work of the North korean dance training or specialized training received directly from her, she founded the dance student of Choi Seung-hee developed basic techniques of Chinese ethnic dance and ethnic Koreans in China, while receiving only Sewonaga dance training system as a deal on exchanges with the North Korean dance dancing free dance culture for creation peppered ethnic Koreans in China was formed. When passed down to the time, Koreans in Japan since the 1960s, society began to visit Pyongyang in the 1970s, subjected to a direct transfer, and education and through the 1990s, the North Koreans in Japan by inviting dancers and dance directly to basic training by getting education bukhanchum As can be seen in the spectacular aspects will have to reproduce. However, ethnic Koreans in China in the 1950s in districts in Beijing, Pyongyang and received direct guidance from Choi Seung-hee, Dancers from the North after Pyongyang rather than direct guidance on the occasion of his visit to China Dance Troupe was affected. On the other hand Korean dance since the 1990s, starting with Ethnic Koreans in China only began to absorb a different dance culture has been created. The same nation, yet living in the region and to configure the ethnic groups, the configuration of the system and political system, according to the North Dancing transfer process and the development pattern similar, but each of the identities to ensure their own traits with a dance culture, the formation and develop the arrival of You can find out. In other words, Koreans in Japan and Federation of Koreans in Japan under the control of social forces of the dance culture by Acculturation variation of dance culture, dance culture of the borough ethnic Koreans in China Acculturation by the voluntary and free borrowers were able to gauge the changes in development.

Agronomical studies on the major environmental factors of rice culture in Korea (수도재배의 주요환경요인에 관한 해석적 조사연구)

  • Yung-Sup Kim
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.3
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    • pp.49-82
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    • 1965
  • For the stable and high yields of low-land rice in Korea, the characteristics of rice plant for the vegetative and physiological responses, plant type formation, and yield components have been studied in order to obtain the fundamental data for the improvement of cultural practices, especially for the ideal fertilizer application. Furthermore the environmental conditions in Korea including temperatures, light, precipitation, and soil conditions have been compared in the broad sense with those in Japan, and the application of nitrogen, phosphorus, potassium, silicate and other micro-nutrients were described in relation to the characteristics of environmental conditions for the improvement of fertilizer application. 1. The average yield of polished-rice per 10 are in Korea is about 204 kg and this values are much less than those in Japan and Taiwan where they produce 77% to 13% more than in Korea. The rate of yield increase a year in Korea is 4.2 kg, but in Japan and Taiwan the rates of yield increase a year are 81 % and 62%, respectively. It was also found that the coefficient of variation of yield is 7.7% in Korea, 6.7% in Japan and 2.5% in Taiwan. This means that the stability of producing rice in Korea is very low when compared with those in Japan and Taiwan. 2. It was learned from the results obtained from the 'annual yield estimation experiment' that there are big differences in the respect of plant type formations between rice crops grown in Japan and Korea. The important differences found were as follows: (1) The numbers of spikelets per 3.3 square meters are 891 in Korea and 1, 007 in Japan(13% more than in Korea). (2) The numbers of tillers per 3.3 square meters at the stage of maximum tillering are 1, 150 in Korea, but in Japan they showed 19% more than in Korea. (3) The ratio of effective tillers to total tillers is 77.5% in Korea and 74.7% in Japan, which seems to be higher in Korea than in Japan. But the ratio in Korea is very low when considered the numbers of total tillers in both countries. (4) The ratio of grain to straw is 85.4% in Korea and 96.3% in Japan. 3. The average temperatures during the growing season at the area of Suwon, Kwangjoo and Taegu are almost same as those in the district of Jookokoo(Fookoo yama) in Japan, i.e., the temperatures during the rice-growing season in Korea are similar to those in the southern-warm regions of Japan. 4. Considering the minimum temperatures at the stage of limiting transplanting, 13$^{\circ}C$, the time of transplanting might be 30 to 40 days earlier than presently practicing transplanting time, which comes around June 10. 5. The temperatures during the vegetative growth in Korea were higher than those temperatures that needed in the protein synthesis which ate the main metabolism during this stage. However, the temperatures at the time of reproductive growth was lower than the temperatures that needed in the sugar assimilation which is main metabolism in this stage. In this point of view, it might be considered that the proper time of growing rice plant in Korea would be rather earlier. 6. The temperatures and the day light conditions at the time of first tillering stage of rice plant, when planted as presenting transplanting practices, are very satisfactory, but the poor day light length, high temperatures and too wet conditions in the time of last-tillering stage(mid or last July) might cause the occurrence of disease such as blast. 7. The heading stage of rice plants at each region through nations when planted as presently practicing method comes when the day light length is short. 8. It was shown that the accumulated average air-temperature at the time of maturing stage was not enough and the heading time was too late, when considered the annual deviations of mean temperatures and low minimum temperatures. 9. The nitrogen content of each plant part at the each growing stage was very high at the stage of vegetative growth when compared with the nitrogen content at the stage of reproductive growth after heading. In this respect it was believed to be important to prevent the nutrient shortages at the reproductive stages, especially after the heading. 10. The area of unsatisfactory irrigation paddy fields and natural rain-fed paddy fields are getting reduced in Korea. The correlation between the rate of reducing unsatisfactory irrigation and natural rain-fed paddy fields and the rate of yield increase were computed. The correlation coefficients(r) between the area of unsatisfactory irrigation paddy fields and yield increase were +0.525, and between the natural rain-fed paddy fields and yield increase, +0.832 and between the unsatisfactory irrigation plus natural rain-fed paddy fields and yield increase, +0.84. And there were. highly significant positive correlations between natural rain-fed paddy fields and yield increases indicating that the less the area of natural rain-fed paddy fields, the greater the yields per unit area. 11. The results obtained from the fertilizer experiments (yield performance trials) conducted in both Korea and Japan showed that the yield of non-fertilized plots per 10 are was 231 kg in Korea and 360 kg in Japan. On the basis of this it might be concluded that the fertility of soil in Korea is lower than that in Japan. Furthermore it was. also found that the yields of non-nitrogen applied plots per 10 are were 236 kg in Korea and 383 kg in Japan. This also indicates that the yields of rice in Korea are largely depending on the nitrogen content in the soil. 12. The followings were obtained when the chemical natures of soils in both Korea and Japan were compared. (1) The content of organic matter, total nitrogen, exchangeable calcium, and magnesium in Korea were no more than the half those in Japan. (2) The content of N/2 chloride and soluble silicate in low-land soil were on the average lower in Korea. (3) The exchange capacity of bases in Korea was no more than half that in Japan. 13. It was also observed by comparing the soil nature of the soil with high yielding capacity with the soil with low yielding capacity that the exchange capacity of bases, exchangeable calcium and magnesium, potassium, phosphorus, manganese, silicate and iron were low in the soil with low yielding capacity. 14. The depth of furrow slice was always deeper in the soil with high yielding capacity, and the depth of furrow slice in Korea was also shallower than that in Japan. 15. Summarizing the various conditions mentioned previously and considering the effects of silicate and trace elements such as manganese and iron besides three elements on the physiological and plant type formation of rice crops, more realistic and more ideal fertilizing practices were proposed. proposed.

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