• Journal of Biosystems Engineering
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• v.1 no.1
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• pp.17-48
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• 1976

In analyzing the operational characteristics of a rice whitening machine, the internal radial pressure of the machine was measured using strain gage equipment. Changes in cylinder and feed screw configurations, screen type, cylinder speed and counter-pressure levels were examined to determine their impact on the quality and quantity of milled rice and the performance of the machine. The results are summarized as follows: 1. The internal radial pressure in the whitening chamber varied with the surface condition of the grain being processed. During the first or second pass through the machine, pressure was relatively low, reached a maximum after two to three passes with combinations I and II, three to six with combination III and then began to fall. 2. The pitch of the feed screw and the size of the feed gate opening which determine the rate of entry of grain into the whitening chamber, appeared to be the most important factor aff-::cting the degree of radial pressure, quality and quantity of milled rice and the efficiency of the machine. Using a feed screw with a wide pitch (4.8cm), radial pressure was relatively high and head rice recovery ratio \vere quite low. In this case capacity and machine effic\ulcorneriency were much higher than obtained when using a feed screw with a narrow pitch (2.3cm). Very significant responses in radial pressure, head rice recovery rates and machine capacity were observed with changes in cylinder speed and counter-pressure levels when using the wide pitch feed screw. 3. The characteristics of the screen which surrounds the whitening chamber had an important effect on whitening efficiency. The existence of small protuberances on the original screen resulted in significant increases in both machine capacity and efficiency but without a significant decrease in head rice recovery or development of excessive radial pressure. Further work is required to determine the effects of screen surface conditions and the shape of the cylinderical steel roller on the rate of bran removal, machine efficiency and recovery rates. The size of the slotted perforations 0:1 the screen affects total milled rice recovery. The opening size on the original screen was fabricated to accommodate the round shape of Japonica rice varieties but was not suitable for the more slender Indica type. Milling Indica varieties with this screen resulted in a reduction in total milled rice recovery. 4. An increase in cylinder speed from 380 to 820 rpm produced a positive effect on head rice recovery for all machine combinations at every level of counter-pressure used in the tests. Head rice recovery was considerably lower at 380rpm using a wide screw pitch when compared to the results obtained at speeds from 600 to 820 r.p.m. The effects of cylinder speed On radial pressure, capacity and machine efficiency showed contrasting results, depending on the width of the feed screw pitch. With a narrow feed screw pitch (2.3cm), a direct proportional relationship was observed bet\ulcornerween cylinder speed and both radial pressure and machine efficiency. In contrast, using a 4.8 centimeter pitch feed roller produced a series of inverse relationships between the above variables. Based on the results of this study it is recommended when milling Indica type long grain rice varieties that the cylinder speed of the original machine be increased from 500-600 rmp up to a minimum of 800 rpm to obtain a greater abrasive effect between the grain and the screen. The pitch of the feed screw should be also reduced to decr\ulcornerease the level of internal radial pressure and to obtain higher machine efficiency and increased quality of milled rice with increased cylinder speeds. Further study on the interaction between cylinder speed and feed screw pitch is recommended. 5. An increase in the counter pressure level produced a negative effect On the head rice recovery with an increase in radial pressure, capacity, and machine efficiency over all combinations and at every level of cylinder speed. 6. Head rice recovery rates were conditioned primarily by the pressure inside the whitening chamber. According to the empirical cha racteristics curve developed in this study, the relationships of head rice recovery ($Y_h$) and machine capacity ($Y_c$/TEX>) to internal radial pressure ($X_p$) followed an inverse quadratic function and a linear function respectively: $$Y_h^\Delta=\frac{1}{{1.4383-0.2951X_p^\ast+0.1425X_p^{\ast\ast}}^2} , (R^2=0.98)$$ $$Y_c^\Delta=-305.83+374.37X_p^{\ast\ast}, (R^2=0.88)$$ The correlation between capacity and power consumption per unit of brown rice expressed in the following exponential function: $$Y_c^\Delta=1.63Y_c^{-0.7786^\{\ast\ast}, (R^2=0.94)$$ These relationships indicate that when radial pressure increases above a certain range (1. 6 to 2.0 kg/$cm^2$ based On the results of the experiment) head ricerecovery decrea\ulcornerses in a quadratic relation with a inear increase in capacity but without any decrease in power consump tion per unit of brown rice. On the other hand, if radial pressure is below the range shown above, power consumption increases dramatically with a lin\ulcornerear decrease in capacity but without significant increases in head rice recovery. During the operation of a given whitening machine, the optimum radial pressure range or the correct capacity range should be selected by controlling the feed rate and/or counter-pressure keeping in mind the condition of the grain, particulary the hardness. It was observed that the total number of passes is related to radial pessure level, feed rate and counter-pressure level. The higher theradial pressure the fewer num\ulcornerber of pass required but with decreased head rice recovery. In particular, when using high feed rates, the total number of passes should be increased to more than three by reducing the counter-pressure level to avoid decreaseases in head rice recovery (less than 65 percent head rice recovery on the basis of brown rice) at every cylinder speed. 7. A rapid rise in grain temperature seemed to have a close relationship with the pressure generated inside the whitening chamber and, subsequently with head rice reco\ulcornervery rates. The higher the rate of increase, the lower were the resulting head rice recoveries.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.13 no.2
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• pp.65-80
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• 1980

Processing conditions of the krill products such as paste food, krill protein concentrate, seasoned dried krill, powdered seasoning, meat ball, and snack have been examined and the quality was evaluated chemically and organoleptically. In the processing of paste food, krill juice was yielded $71\%$ and krill scrap $29\%$. The yields of paste and broth from the krill juice showed $53\%$ and $43\%$, respectively. In amino acid composition of the krill paste, proline, glutamic acid, aspartic acid, lysine, and leucine were abundant, while histidine, methionine, tyrosine, serine and threonine were poor. The optimum condition for solvent extraction in the processing of krill protein concentrate was the 5 times repetitive extraction using isopropyl alcohol at $80^{\circ}C$ for 5 mins. The yield of krill protein concentrate when used fresh frozen materials was $10.2\%$ in isopropyl alcohol solvent and $8.8\% in ethyl alcohol, and when used preboiled frozen materials, the yield was $13.0\%$ in isopropyl alcohol and $11.8\%$ in ethyl alcohol. Amino acid composition of krill protein concentrate showed a resemblance to that of fresh frozen krill meat. In quality comparison of the seasoned dried krill, hot air dried krill was excellent as raw materials and sun dried krill was slightly inferior to hot air dried krill, but preboiled frozen krill showed the poorest quality. The result of quality evaluation for seasoning made by combination of dried powdered krill, parched powdered sesame, salt, powdered beef extract, monosodium glutamate, powdered red pepper and ground pepper showed that the hot air dried krill was good in color and sundried krill was favorable in flavor. When krill meat ball was prepared using wheat flour, monosodium glutamate and salt as side materials, the quality of the products added up to $52\%$ of krill meat was good and the difference in quality upon the results of the organoleptic test for raw materials was not recognizable between fresh frozen and preboiled frozen krill. In the experiment for determining the proper amount of materials such as dried Powdered krill, $\alpha-starch$, sweet potato starch, sugar, salt, monosodium glutamate, glycine, potassium tartarate, ammonium bicarbonate, and sodium bicarbonate in processing krill snack, sample B(containing $7.7\%$ of dried powdered krill) and sampleC (containing $10.8\%$ of dried powdered krill) showed the most palatable taste from the view point of organoleptic test. Sweet potato starch in testing side materials was good in the comparison of suitability for processing krill snack. Corn starch and kudzu starch were slightly inferior to sweet potato starch, while wheat flour was not proper for processing the snack. In the experiment on frying method, oil frying showed better effect than salt frying and the suitable range of frying temperature was $210-215^{\circ}C$.

• 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.

• Korean Journal of Soil Science and Fertilizer
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• v.10 no.3
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• pp.103-134
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• 1977

The physiological and biochemical role of potassium for upland crops according to recent research reports and the nutritional status of potassium in Korea were reviewed. Since physical and chemical characteristics of potassium ion are different from those of sodium, potassium can not completely be replaced by sodium and replacement must be limited to minimum possible functional area. Specific roles of potassium seem to keep fine structure of biological membranes such as thylacoid membrane of chloroplast in the most efficient form and to be allosteric effector and conformation controller of various enzymes principally in carbohydrate and protein metabolism. Potassium is essential to improve the efficiency of phoro- and oxidative- phosphorylation and involve deeply in all energy required metabolisms especially synthesis of organic matter and their translocation. Potassium has many important, physiological functions such as maintenance of osmotic pressure and optimum hydration of cell colloids, consequently uptake and translocation of water resulting in higher water use efficiency and of better subcellular environment for various physiological and biochemical activities. Potassium affects uptake and translocation of mineral nutrients and quality of products. potassium itself in products may become a quality criteria due to potassium essentiality for human beings. Potassium uptake is greatly decreased by low temperature and controlled by unknown feed back mechanism of potassium in plants. Thus the luxury absorption should be reconsidered. Total potassium content of upland soil in Korea is about 3% but the exchangeable one is about 0.3 me/100g soil. All upland crops require much potassium probably due to freezing and cold weather and also due to wet damage and drought caused by uneven rainfall pattern. In barley, potassium should be high at just before freezing and just after thawing and move into grain from heading for higher yield. Use efficiency of potassium was 27% for barley and 58% in old uplands, 46% in newly opened hilly lands for soybean. Soybean plant showed potassium deficiency symptom in various fields especially in newly opened hilly lands. Potassium criteria for normal growth appear 2% $K_2O$ and 1.0 K/(Ca+Mg) (content ratio) at flower bud initiation stage for soybean. Potassium requirement in plant was high in carrot, egg plant, chinese cabbage, red pepper, raddish and tomato. Potassium content in leaves was significantly correlated with yield in chinese cabbage. Sweet potato. greatly absorbed potassium subsequently affected potassium nutrition of the following crop. In the case of potassium deficiency, root showed the greatest difference in potassium content from that of normal indicating that deficiency damages root first. Potatoes and corn showed much higher potassium content in comparison with calcium and magnesium. Forage crops from ranges showed relatively high potassium content which was significantly and positively correlated with nitrogen, phosphorus and calcium content. Percentage of orchards (apple, pear, peach, grape, and orange) insufficient in potassium ranged from 16 to 25. The leaves and soils from the good apple and pear orchards showed higher potassium content than those from the poor ones. Critical ratio of $K_2O/(CaO+MgO)$ in mulberry leaves to escape from winter death of branch tip was 0.95. In the multiple croping system, exchangeable potassium in soils after one crop was affected by the previous crops and potassium uptake seemed to be related with soil organic matter providing soil moisture and aeration. Thus, the long term and quantitative investigation of various forms of potassium including total one are needed in relation to soil, weather and croping system. Potassium uptake and efficiency may be increased by topdressing, deep placement, slow-releasing or granular fertilizer application with the consideration of rainfall pattern. In all researches for nutritional explanation including potassium of crop yield reasonable and practicable nutritional indices will most easily be obtained through multifactor analysis.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.47
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• pp.15-32
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• 2002

The endeavors enhancing the grain quality of high-yielding japonica rice were steadily continued during 1980s-1990s along with the self-sufficiency of rice production and the increasing demands of high-quality rices. During this time, considerably great progress and success was obtained in development of high-quality japonica cultivars and quality evaluation techniques including the elucidation of interrelationship between the physicochemical properties of rice grain and the physical or palatability components of cooked rice. In 1990s, some high-quality japonica rice cultivars and special rices adaptable for food processing such as large kernel, chalky endosperm, aromatic and colored rices were developed and its objective preference and utility was also examined by a palatability meter, rapid-visco analyzer and texture analyzer, Recently, new special rices such as extremely low-amylose dull or opaque non-glutinous endosperm mutants were developed. Also, a high-lysine rice variety was developed for higher nutritional utility. The water uptake rate and the maximum water absorption ratio showed significantly negative correlations with the K/Mg ratio and alkali digestion value(ADV) of milled rice. The rice materials showing the higher amount of hot water absorption exhibited the larger volume expansion of cooked rice. The harder rices with lower moisture content revealed the higher rate of water uptake at twenty minutes after soaking and the higher ratio of maximum water uptake under the room temperature condition. These water uptake characteristics were not associated with the protein and amylose contents of milled rice and the palatability of cooked rice. The water/rice ratio (in w/w basis) for optimum cooking was averaged to 1.52 in dry milled rices (12% wet basis) with varietal range from 1.45 to 1.61 and the expansion ratio of milled rice after proper boiling was average to 2.63(in v/v basis). The major physicochemical components of rice grain associated with the palatability of cooked rice were examined using japonica rice materials showing narrow varietal variation in grain size and shape, alkali digestibility, gel consistency, amylose and protein contents, but considerable difference in appearance and texture of cooked rice. The glossiness or gross palatability score of cooked rice were closely associated with the peak, hot paste and consistency viscosities of viscosities with year difference. The high-quality rice variety "IIpumbyeo" showed less portion of amylose on the outer layer of milled rice grain and less and slower change in iodine blue value of extracted paste during twenty minutes of boiling. This highly palatable rice also exhibited very fine net structure in outer layer and fine-spongy and well-swollen shape of gelatinized starch granules in inner layer and core of cooked rice kernel compared with the poor palatable rice through image of scanning electronic microscope. Gross sensory score of cooked rice could be estimated by multiple linear regression formula, deduced from relationship between rice quality components mentioned above and eating quality of cooked rice, with high probability of determination. The $\alpha$-amylose-iodine method was adopted for checking the varietal difference in retrogradation of cooked rice. The rice cultivars revealing the relatively slow retrogradation in aged cooked rice were IIpumbyeo, Chucheongyeo, Sasanishiki, Jinbubyeo and Koshihikari. A Tonsil-type rice, Taebaegbyeo, and a japonica cultivar, Seomjinbyeo, showed the relatively fast deterioration of cooked rice. Generally, the better rice cultivars in eating quality of cooked rice showed less retrogradation and much sponginess in cooled cooked rice. Also, the rice varieties exhibiting less retrogradation in cooled cooked rice revealed higher hot viscosity and lower cool viscosity of rice flour in amylogram. The sponginess of cooled cooked rice was closely associated with magnesium content and volume expansion of cooked rice. The hardness-changed ratio of cooked rice by cooling was negatively correlated with solids amount extracted during boiling and volume expansion of cooked rice. The major physicochemical properties of rice grain closely related to the palatability of cooked rice may be directly or indirectly associated with the retrogradation characteristics of cooked rice. The softer gel consistency and lower amylose content in milled rice revealed the higher ratio of popped rice and larger bulk density of popping. The stronger hardness of rice grain showed relatively higher ratio of popping and the more chalky or less translucent rice exhibited the lower ratio of intact popped brown rice. The potassium and magnesium contents of milled rice were negatively associated with gross score of noodle making mixed with wheat flour in half and the better rice for noodle making revealed relatively less amount of solid extraction during boiling. The more volume expansion of batters for making brown rice bread resulted the better loaf formation and more springiness in rice breed. The higher protein rices produced relatively the more moist white rice bread. The springiness of rice bread was also significantly correlated with high amylose content and hard gel consistency. The completely chalky and large grain rices showed better suitability far fermentation and brewing. The glutinous rice were classified into nine different varietal groups based on various physicochemical and structural characteristics of endosperm. There was some close associations among these grain properties and large varietal difference in suitability to various traditional food processing. Our breeding efforts on improvement of rice quality for high palatability and processing utility or value-adding products in the future should focus on not only continuous enhancement of marketing and eating qualities but also the diversification in morphological, physicochemical and nutritional characteristics of rice grain suitable for processing various value-added rice foods.ice foods.

• Korean Journal of Agricultural Science
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• v.25 no.2
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• pp.181-198
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• 1998

In order to determine the optimum pasteurization conditions by microwave heating(MWH) at $50^{\circ}C{\sim}70^{\circ}C$ for 30 minute compared with water bath heating(WBH) at $65^{\circ}C$ for 30minute during storage at $5^{\circ}C$, the chemical composition, microbiological changes and keeping quality were examined and the results were as follows: 1. The fat protein lactose, total solid contents of raw milk, at $50{\sim}70^{\circ}C$ for 30 min. in MWH and at 65 for $30^{\circ}C$ min. in WBH did not changed significantly during the storage at $5^{\circ}C$. 2. The pH and acidity for the raw milk untreated were 6.75 and 0.16%, and those of MWH heated and WBH milk wee 6.75~6.50 and 0.16%~0.19%, phosphatase test were negative at $61^{\circ}C$ for 20 min. at $62^{\circ}C$ for 15 min. at $63^{\circ}C$ for 10 min. at $64^{\circ}C$ for 5 min. at $65^{\circ}C$ for 5 min. in MWH and at $65^{\circ}C$ for 30 min. in WBH. 3. Whey protein content was $18.53mg/m{\ell}$ in raw milk untreated, however, those were decreased as the heating temperature increased. The proteolytic activity of treated milk by WBH(44%) was lower than that by MWH(94%). 4. Total bacteria counts were $2.8{\times}10^5CFU/m{\ell}$ in raw milk untreated, $2.8{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $2.4{\times}10^3CFU/m{\ell}$ at $70^{\circ}C$ for 30 min. in MWH and $3.0{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. in WBH. Because total bacteria count did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30 min. and $65^{\circ}C$ for 30 min. in WBH during the 10 days storaging, Also, total bacteria counts for treated milk were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 5. Coliform bacteria counts were $2.6{\times}10^3CFU/m{\ell}$ in raw milk untreated. There were not detected at $55^{\circ}C{\sim}70^{\circ}C$ for 30 min. in MWH and at $65^{\circ}C$ for 30 min. in WBH. Coliform bacteria counts were not detected after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 6. Thermoduric bacteria counts were $5.2{\times}10^4CFU/m{\ell}$ in raw milk untreated, $2.0{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $1.9{\times}10^3CFU/m{\ell}$ at $70^{\circ}C$ for 30min. in MWH and $2.2{\times}10^3CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. in WBH. Because thermoduric bacteria counts did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30 min. and $65^{\circ}C$ for 30 min. in WBH during the 10days storaging. Also, thermoduric bacteria counts were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH. 7. Psychrotrophic bacteria counts were $2.8{\times}10^5CFU/m{\ell}$ in raw milk untreated, $2.0{\times}10^1CFU/m{\ell}$ at $65^{\circ}C$ for 30 min. $2.0{\times}10^1CFU/m{\ell}$ at $70^{\circ}C$ for 30 min. in MWH and $3.0{\times}10^1CFU/m{\ell}$ at $65^{\circ}C$for 30 min. in WBH. Because psychrotrophic bacteria counts did not increased in MWH at $65^{\circ}C$, $70^{\circ}C$ for 30min. and $65^{\circ}C$ for 30 min. in WBH during the 10 days storaging. Also, psychrotrophic bacteria counts were a most drastic decrease after $61^{\circ}C$, $62^{\circ}C$, $63^{\circ}C$, $64^{\circ}C$, $65^{\circ}C$ for 5 min. in MWH.

• Korean journal of applied entomology
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• v.13 no.3 s.20
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• pp.105-133
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• 1974

The present study is an attempt to solve the basic problems involved in the control of the Sclerotium disease. The biologic stranis of Sclerotium rolfsii Sacc., pathogen of Sclerotium disease of Magnolia kobus, were differentiated, and the effects of vitamins, various nitrogen and carbon sources on its mycelial growth and sclerotial production have been investigated. In addition the relationship between the cultural filtrate of Penicillium sp. and the growth of Sclerotium rolfsii, the tolerance of its mycelia or sclerotia to moist heat or drought and to Benlate (methyl-(butylcarbamoy 1)-2-benzimidazole carbamate), Tachigaren (3-hydroxy-5-methylisoxazole) and other chemicals were also clarified. The results are summarizee as follows: 1. There were two biologic strains, Type-l and Type-2 among isolates. They differed from each other in the mode of growth and colonial appearance on the media, aversion phenomenon and in their pathogenicity. These two types had similar pathogenicity to the Magnolia kobus and Robinia pseudoacasia, but behaved somewhat differently to the soybaen and cucumber, the Type-l being more virulent. 2. Except potassium nitrite, sodium nitrite and glycine, all of the 12 nitrogen sources tested were utilized for the mycelial growth and sclerotial production of this fungus when 10r/l of thiamine hydrochloride was added in the culture solution. Considering the forms of nitrogen, ammonium nitrogen was more available than nitrate nitrogen for the growth of mycelia, but nitrate nitrogen was better for sclerotia formation. Organic nitrogen showed different availabilities according to compounds used. While nitrite nitrogen was unavailable for both mycelial growth and sclerotial formation whether thiamine hydrochlioride was added or not. 3. Seven kinds of carbon sources examined were not effective in general, as long as thiamine hydrochloride was not added. When thiamine hydrochloride was added, glucose and saccharose exhibited mycelial growth, while rnaltose and soluble starch gave lesser, and xylose, lactose, and glycine showed no effect at all,. In the sclerotial production, all the tested carbon sources, except lactose, were effective, and glucose, maltose, saccharose, and soluble starch gave better results. 4. At the same level of nitrogen, the amount of mycelial growth increased as more carbon Sources were applied but decreased with the increase of nitrogen above 0.5g/1. The amount of sclerotial production decreased wi th the increase of carbon sources. 5. Sclerotium rolfsii was thiamine-defficient and required thiamine 20r/l for maximun growth of mycelia. At a higher concentration of more than 20r/l, however, mycelial growth decreased as the concentration increased, and was inhibited at l50r/l to such a degree of thiamine-free. 6. The effect of the nitrogen sources on the mycelial growth under the presence of thiamine were recognized in the decreasing order of $NH_4NO_3,\;(NH_4)_2SO_4,\;asparagine,\;KNO_3$, and their effects on the sclerotial production in the order of $KNO_3,\;NH_4NO_3,\;asparagine,\;(NH_4)_2SO_4$. The optimum concentration of thiamine was about 12r/l in $KNO_3$ and about 16r/l in asparagine for the growth of mycelia; about 8r/l in $KNO_3$ and $NH_4NO_3$, and 16r/l in asparagine for the production of sclerotia. 7. After the fungus started to grow, the pH value of cultural filtrate rapidly dropped to about 3.5. Hereafter, its rate slowed down as the growth amount increased and did not depreciated below pH2.2. 8. The role of thiamine in the growth of the organism was vital. If thiamine was not added, the combination of biotin, pyridoxine, and inositol did not show any effects on the growth of the organism at all. Equivalent or better mycelial growth was recognized in the combination of thiamine+pyridoxine, thiamine+inositol, thiamine+biotin+pyridoxine, and thiamine+biotin+pyridoxine+inositol, as compared with thiamine alone. In the combinations of thiamine+biotin and thiamine+biotin+inositol, mycelial growth was inhibited. Sclerotial production in dry weight increased more in these combinations than in the medium of thiamine alone. 9. The stimulating effects of the Penicillium cultural filtrate on the mycelial growth was noticed. It increased linearly with the increase of filtrate concentration up to 6-15 ml/50ml basal medium solution. 10. $NH_4NO_3$. as a nitrogen source for mycelial growth was more effective than asparasine regardless of the concentration of cultural filtrate. 11. In the series of fractionations of the cultural filtrate, mycelial growth occured in unvolatile, ether insoluble cation-adsorbed or anion-unadsorbed substance fractions among the fractions of volatile, unvolatile acids, ether soluble organic acids, ether insoluble, cation-adsorbed, cation-unadsorbed, anion-adsorbed and anion-unadsorbed. and anion-un-adsorbed substance tested. Sclerotia were produced only in cation-adsorbed fraction. 12. According to the above results, it was assumed that substances for the mycelial growth and sclerotial formation and inhibitor of sclerotial formation were include::! in cultural filtrate and they were quite different from each other. I was further assumed that the former two substances are un volatile, ether insotuble, and adsorbed to cation-exchange resin, but not adsorbed to anion, whereas the latter is unvolatile, ether insoluble, and not adsorbed to cation or anion-exchange resin. 13. Seven amino acids-aspartic acid, cystine, glysine, histidine, Iycine, tyrosine and dinitroaniline-were detected in the fractions adsorbed to cation-exchange resin by applying the paper chromatography improved with DNP-amino acids. 14. Mycelial growth or sclerotial production was not stimulated significantly by separate or combined application of glutamic acid, aspartic acid, cystine, histidine, and glysine. Tyrosine gave the stimulating effect when applied .alone and when combined with other amino acids in some cases. 15. The tolerance of sclerotia to moist heat varied according to their water content, that was, the dried sclerotia are more tolerant than wet ones. The sclerotia harvested directly from the media, both Type-1 and Type-2, lost viability within 5 minutes at $52^{\circ}C$. Sclerotia dried for 155 days at$26^{\circ}C$ had more tolerance: sclerotia of Type-l were killed in 15 mins. at $52^{\circ}C$ and in 5 mins. at $57^{\circ}C$, and sclerotia of Type-2 were killed in 10 mins. both at $52^{\circ}C$ or $57^{\circ}C$. 16. Cultural sclerotia of both strains maintained good germinability for 132 days at$26^{\circ}C$. Natural sclerotia of them stored for 283 days under air dry condition still had good germinability, even for 443 days: type-l and type-2 maintained $20\%$ and $26.9\%$ germinability, respectively. 17. The tolerance to low temperature increased in the order of mycelia, felts and sclerotia. Mycelia completely lost the ability to grow within 1 week at $7-8^{\circ}C$> below zero, while mycelial felts still maintained the viability after .3 weeks at $7-20^{\circ}C$ below zero, and sclerotia were even more tolerant. 18. Sclerotia of type-l and type-2 were killed when dipped into the $0.05\%$ solution of mercury chloride for 180 mins. and 240 mins. respectively: and in the $0.1\%$ solution, Type-l for 60 mins. and Type-2 for 30 mins. In the $0.125\%$ uspulun solution, Type-l sclerotia were killed in 180 mins., and those of Type-2 were killed for 90 mins. in the$0.125\%$solution. Dipping into the $5\%$ copper sulphate solution or $0.2\%$ solution of Ceresan lime or Mercron for 240 mins. failed to kill sclerotia of either Type-l or Type-2. 19. Inhibitory effect on mycelial growth of Benlate or Tachi-garen in the liquid culture increased as the concentration increased. 6 days after application, obvious inhibitory effects were found in all treatments except Benlate 0.5ppm; but after 12 days, distingushed diflerences were shown among the different concentrations. As compared with the control, mycelial growth was inhibited by $66\%$ at 0.5ppm and by $92\%$ at 2.0ppm of Benlate, and by$54\%$ at 1ppm and about $77\%$ at 1.5ppm or 2.0ppm of Tachigaren. The mycelial growth was inhibited completely at 500ppm of both fungicides, and the formation of sclerotia was checked at 1,000ppm of Benlate ant at 500ppm or 1,000ppm of Tachigaren. 20. Consumptions of glucose or ammonium nitrogen in the culture solution usually increased with the increment of mycelial growth, but when Benlate or Tachigaren were applied, consumptions of glucose or ammonium nitrogen were inhibited with the increment of concentration of the fungicides. At the low concentrations of Benlate (0.5ppm or 1ppm), however, ammonium nitrogen consumption was higher than that of the ontrol. 21. The amount of mycelia produced by consuming 1mg of glucose or ammonium nitrogen in the culture solution was lowered markedly by Benlate or Tachigaren. Such effects were the severest on the third day after their treatment in all concentrations, and then gradually recovered with the progress of time. 22. In the sand culture, mycelial growth was not inhibited. It was indirectly estimated by the amount of $CO_2$ evolved at any concentrations, except in the Tachigaren 100mg/g sand in which mycelial growth was inhibited significantly. Sclerotial production was completely depressed in the 10mg/g sand of Benlate or Tachigaren. 23. There was no visible inhibitory effect on the germination of sclerotia when the sclerotia were dipped in the solution 0.1, 1.0, 100, 1.000ppm of Benlate or Tachigaren for 10 minutes or even 20 minutes.