• Korean Journal of Environmental Agriculture
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• v.32 no.3
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• pp.214-223
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• 2013

BACKGROUND: This study focused on the development of an analytical method about dichlorprop (DCPP; 2-(2,4-dichlorophenoxy)propionic acid) which is a plant growth regulator, a synthetic auxin for agricultural commodities. DCPP prevents falling of fruits during their growth periods. However, the overdose of DCPP caused the unwanted maturing time and reduce the safe storage period. If we take fruits with exceeding maximum residue limits, it could be harmful. Therefore, this study presented the analytical method of DCPP in agricultural commodities for the nation-wide pesticide residues monitoring program of the Ministry of Food and Drug Safety. METHODS AND RESULTS: We adopted the analytical method for DCPP in agricultural commodities by gas chromatograph in cooperated with Electron Capture Detector(ECD). Sample extraction and purification by ion-associated partition method were applied, then quantitation was done by GC/ECD with DB-17, a moderate polarity column under the temperature-rising condition with nitrogen as a carrier gas and split-less mode. Standard calibration curve presented linearity with the correlation coefficient ($r^2$) > 0.9998, analysed from 0.1 to 2.0 mg/L concentration. Limit of quantitation in agricultural commodities represents 0.05 mg/kg, and average recoveries ranged from 78.8 to 102.2%. The repeatability of measurements expressed as coefficient of variation (CV %) was less than 9.5% in 0.05, 0.10, and 0.50 mg/kg. CONCLUSION(S): Our newly improved analytical method for DCPP residues in agricultural commodities was applicable to the nation-wide pesticide residues monitoring program with the acceptable level of sensitivity, repeatability and reproducibility.

• Magazine of the Korean Society of Agricultural Engineers
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• v.19 no.1
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• pp.4296-4311
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• 1977

This study was conducted to derive an Instantaneous Unit Hydrograph for the accurate and reliable unitgraph which can be used to the estimation and control of flood for the development of agricultural water resources and rational design of hydraulic structures. Eight small watersheds were selected as studying basins from Han, Geum, Nakdong, Yeongsan and Inchon River systems which may be considered as a main river systems in Korea. The area of small watersheds are within the range of 85 to 470$\textrm{km}^2$. It is to derive an accurate Instantaneous Unit Hydrograph under the condition of having a short duration of heavy rain and uniform rainfall intensity with the basic and reliable data of rainfall records, pluviographs, records of river stages and of the main river systems mentioned above. Investigation was carried out for the relations between measurable unitgraph and watershed characteristics such as watershed area, A, river length L, and centroid distance of the watershed area, Lca. Especially, this study laid emphasis on the derivation and application of Instantaneous Unit Hydrograph (IUH) by applying Nash's conceptual model and by using an electronic computer. I U H by Nash's conceptual model and I U H by flood routing which can be applied to the ungaged small watersheds were derived and compared with each other to the observed unitgraph. 1 U H for each small watersheds can be solved by using an electronic computer. The results summarized for these studies are as follows; 1. Distribution of uniform rainfall intensity appears in the analysis for the temporal rainfall pattern of selected heavy rainfall event. 2. Mean value of recession constants, Kl, is 0.931 in all watersheds observed. 3. Time to peak discharge, Tp, occurs at the position of 0.02 Tb, base length of hlrdrograph with an indication of lower value than that in larger watersheds. 4. Peak discharge, Qp, in relation to the watershed area, A, and effective rainfall, R, is found to be {{{{ { Q}_{ p} = { 0.895} over { { A}^{0.145 } } }}}} AR having high significance of correlation coefficient, 0.927, between peak discharge, Qp, and effective rainfall, R. Design chart for the peak discharge (refer to Fig. 15) with watershed area and effective rainfall was established by the author. 5. The mean slopes of main streams within the range of 1.46 meters per kilometer to 13.6 meter per kilometer. These indicate higher slopes in the small watersheds than those in larger watersheds. Lengths of main streams are within the range of 9.4 kilometer to 41.75 kilometer, which can be regarded as a short distance. It is remarkable thing that the time of flood concentration was more rapid in the small watersheds than that in the other larger watersheds. 6. Length of main stream, L, in relation to the watershed area, A, is found to be L=2.044A0.48 having a high significance of correlation coefficient, 0.968. 7. Watershed lag, Lg, in hrs in relation to the watershed area, A, and length of main stream, L, was derived as Lg=3.228 A0.904 L-1.293 with a high significance. On the other hand, It was found that watershed lag, Lg, could also be expressed as {{{{Lg=0.247 { ( { LLca} over { SQRT { S} } )}^{ 0.604} }}}} in connection with the product of main stream length and the centroid length of the basin of the watershed area, LLca which could be expressed as a measure of the shape and the size of the watershed with the slopes except watershed area, A. But the latter showed a lower correlation than that of the former in the significance test. Therefore, it can be concluded that watershed lag, Lg, is more closely related with the such watersheds characteristics as watershed area and length of main stream in the small watersheds. Empirical formula for the peak discharge per unit area, qp, ㎥/sec/$\textrm{km}^2$, was derived as qp=10-0.389-0.0424Lg with a high significance, r=0.91. This indicates that the peak discharge per unit area of the unitgraph is in inverse proportion to the watershed lag time. 8. The base length of the unitgraph, Tb, in connection with the watershed lag, Lg, was extra.essed as {{{{ { T}_{ b} =1.14+0.564( { Lg} over {24 } )}}}} which has defined with a high significance. 9. For the derivation of IUH by applying linear conceptual model, the storage constant, K, with the length of main stream, L, and slopes, S, was adopted as {{{{K=0.1197( {L } over { SQRT {S } } )}}}} with a highly significant correlation coefficient, 0.90. Gamma function argument, N, derived with such watershed characteristics as watershed area, A, river length, L, centroid distance of the basin of the watershed area, Lca, and slopes, S, was found to be N=49.2 A1.481L-2.202 Lca-1.297 S-0.112 with a high significance having the F value, 4.83, through analysis of variance. 10. According to the linear conceptual model, Formular established in relation to the time distribution, Peak discharge and time to peak discharge for instantaneous Unit Hydrograph when unit effective rainfall of unitgraph and dimension of watershed area are applied as 10mm, and $\textrm{km}^2$ respectively are as follows; Time distribution of IUH {{{{u(0, t)= { 2.78A} over {K GAMMA (N) } { e}^{-t/k } { (t.K)}^{N-1 } }}}} (㎥/sec) Peak discharge of IUH {{{{ {u(0, t) }_{max } = { 2.78A} over {K GAMMA (N) } { e}^{-(N-1) } { (N-1)}^{N-1 } }}}} (㎥/sec) Time to peak discharge of IUH tp=(N-1)K (hrs) 11. Through mathematical analysis in the recession curve of Hydrograph, It was confirmed that empirical formula of Gamma function argument, N, had connection with recession constant, Kl, peak discharge, QP, and time to peak discharge, tp, as {{{{{ K'} over { { t}_{ p} } = { 1} over {N-1 } - { ln { t} over { { t}_{p } } } over {ln { Q} over { { Q}_{p } } } }}}} where {{{{K'= { 1} over { { lnK}_{1 } } }}}} 12. Linking the two, empirical formulars for storage constant, K, and Gamma function argument, N, into closer relations with each other, derivation of unit hydrograph for the ungaged small watersheds can be established by having formulars for the time distribution and peak discharge of IUH as follows. Time distribution of IUH u(0, t)=23.2 A L-1S1/2 F(N, K, t) (㎥/sec) where {{{{F(N, K, t)= { { e}^{-t/k } { (t/K)}^{N-1 } } over { GAMMA (N) } }}}} Peak discharge of IUH) u(0, t)max=23.2 A L-1S1/2 F(N) (㎥/sec) where {{{{F(N)= { { e}^{-(N-1) } { (N-1)}^{N-1 } } over { GAMMA (N) } }}}} 13. The base length of the Time-Area Diagram for the IUH was given by {{{{C=0.778 { ( { LLca} over { SQRT { S} } )}^{0.423 } }}}} with correlation coefficient, 0.85, which has an indication of the relations to the length of main stream, L, centroid distance of the basin of the watershed area, Lca, and slopes, S. 14. Relative errors in the peak discharge of the IUH by using linear conceptual model and IUH by routing showed to be 2.5 and 16.9 percent respectively to the peak of observed unitgraph. Therefore, it confirmed that the accuracy of IUH using linear conceptual model was approaching more closely to the observed unitgraph than that of the flood routing in the small watersheds.

• 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.