• 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 CROP SCIENCE
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• v.14
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• pp.1-40
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• 1973

There is a general tendency to increase nitrogen level in rice production to insure an increased yield. On the other hand, percentage of ripened grains is getting decreased with such an increased fertilizer level. Decreasing of the percentage is one of the important yield limiting factors. Especially the newly developed rice variety, 'Tongil' is characterized by a relatively low percentage of ripened grains as compared with the other leading varieties. Therefore, these studies were aimed to finding out of some measures for the improvement of ripening in rice. The studies had been carried out in the field and in the phytotron during the period of three years from 1970 to 1972 at the Crop Experiment Station in Suwon. The results obtained from the experiments could be summarized as follows: 1. The spikelet of Tongil was longer in length, more narrow in width, thinner in thickness, smaller in the volume of grains and lighter in grain weight than those of Jinheung. The specific gravity of grain was closely correlated with grain weight and the relationship with thickness, width and length was getting smaller in Jinheung. On the other hand, Tongil showed a different pattern from Jinheung. The relationship of the specific gravity with grain weight was the greatest and followed by that with the width, thickness and length, in order. 2. The distribution of grain weight selected by specific gravity was different from one variety to another. Most of grains of Jinheung were distributed over the specific gravity of 1.12 with its peak at 1.18, but many of grains of Tongil were distributed below 1.12 with its peak at 1.16. The brown/rough rice ratio was sharply declined below the specific gravity of 1.06 in Jinheung, but that of Tongil was not declined from the 1.20 to the 0.96. Accordingly, it seemed to be unfair to make the specific gravity criterion for ripened grains at 1.06 in the Tongil variety. 3. The increasing tendency of grain weight after flowering was different depending on varieties. Generally speaking, rice varieties originated from cold area showed a slow grain weight increase while Tongil was rapid except at lower temperature in late ripening stage. 4. In the late-tillered culms or weak culms, the number of spikelets was small and the percentage of ripened grains was low. Tongil produced more late-tillered culms and had a longer flowering duration especially at lower temperature, resulting in a lower percentage of ripened grains. 5. The leaf blade of Tongil was short, broad and errect, having light receiving status for photosynthesis was better. The photosynthetic activity of Tongil per unit leaf area was higher than that of Jinheung at higher temperature, but lower at lower temperature. 6. Tongil was highly resistant to lodging because of short culm length, and thick lower-internodes. Before flowering, Tongil had a relatively higher amount of sugars, phosphate, silicate, calcium, manganese and magnesium. 7. The number of spikelets of Tongil was much more than that of Jinheung. The negative correlation was observed between the number of spikelets and percentage of ripened grains in Jinheung, but no correlation was found in Tongil grown at higher temperature. Therefore, grain yield was increased with increased number of spikelets in Tongil. Anthesis was not occurred below 21$^{\circ}C$ in Tongil, so sterile spikelets were increased at lower temperature during flowering stage. 8. The root distribution of Jinheung was deeper than that of Tongil. The root activity of Tongil evaluated by $\alpha$-naphthylamine oxidation method, was higher than that of Jinheung at higher temperature, but lower at lower temperature. It is seemed to be related with discoloration of leaf blades. 9. Tongil had a better light receiving status for photosynthesis and a better productive structure with balance between photosynthesis and respiration, so it is seemed that tongil has more ideal plant type for getting of a higher grain yield as compared with Jinheung. 10. Solar radiation during the 10 days before to 30 days after flowering seemed enough for ripening in suwon, but the air temperature dropped down below 22$^{\circ}C$ beyond August 25. Therefore, it was believed that air temperature is one of ripening limiting factors in this case. 11. The optimum temperature for ripening in Jinheung was relatively lower than that of Tongil requriing more than $25^{\circ}C$. Air temperature below 21$^{\circ}C$ was one of limiting factors for ripening in Tongil. 12. It seemed that Jinheung has relatively high photosensitivity and moderate thermosensitivity, while Tongil has a low photosensitivity, high thermosensitivity and longer basic vegetative phase. 13. Under a condition of higher nitrogen application at late growing stage, the grain yield of Jinheung was increased with improvement of percentage of ripened grains, while grain yield of Tongil decreased due to decreasing the number of spikelets although photosynthetic activity after flowering was. increased. 14. The grain yield of Jinheung was decreased slightly in the late transplanting culture since its photosynthetic activity was relatively high at lower temperature, but that of Tonil was decreased due to its inactive photosynthetic activity at lower temperature. The highest yield of Tongil was obtained in the early transplanting culture. 15. Tongil was adapted to a higher fertilizer and dense transplanting, and the percentage of ripened grains was improved by shortening of the flowering duration with increased number of seedlings per hill. 16. The percentage of vigorous tillers was increased with a denser transplanting and increasing in number of seedlings per hill. 17. The possibility to improve percentage of ripened grains was shown with phosphate application at lower temperature. The above mentioned results are again summarized below. The Japonica type leading varieties should be flowered before August 20 to insure a satisfactory ripening of grains. Nitrogen applied should not be more than 7.5kg/10a as the basal-dressing and the remained nitrogen should be applied at the later growing stage to increase their photosynthetic activity. The morphological and physiological characteristics of Tongil, a semi-dwarf, Indica $\times$ Japonica hybrid variety, are very different from those of other leading rice varieties, requring changes in seed selection by specific gravity method, in milling and in the cultural practices. Considering the peculiar distribution of grains selected by the method and the brown/rough rice ratio, the specific gravity criterion for seed selection should be changed from the currently employed 1.06 to about 0.96 for Tongil. In milling process, it would be advisable to bear in mind the specific traits of Tongil grain appearance. Tongil is a variety with many weak tillers and under lower temperature condition flowering is delayed. Such characteristics result in inactivation of roots and leaf blades which affects substantially lowering of the percentage of ripened grains due to increased unfertilized spikelets. In addition, Tongil is adapted well to higher nitrogen application. Therefore, it would be recommended to transplant Tongil variety earlier in season under the condition of higer nitrogen, phosphate and silicate. A dense planting-space with three vigorous seedlings per hill should be practiced in this case. In order to manifest fully the capability of Tongil, several aspects such as the varietal improvement, culural practices and milling process should be more intensively considered in the future.he future.

• Korean Journal of Agricultural Science
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• v.4 no.2
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• pp.283-316
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• 1977

In an attempt to obtain a basic information to develop an effective integrated system of controlling sheath blight of rice in Korea, the transition of this disease, the variation of cultural characters and pathogenicity of the pathogen, environmental conditions affecting the disease outbreak and varietal resistance have been investigated. 1. Rice sheath blight which has been minor disease in the past was widely spread, especially since 1971. This disease has promptly spread all over the country and infected 65.2% of total rice growing area in 1976. Various factors are considered to be related to such transition of this disease. Above all, increace of application of nitrogenous fertilizer, early season and earlier cultivation of rice, introduction of more susceptible "Tongil" varieties etc. must be important factors influencing the outbreak of this disease. 2. Great variations in cultural characteristics-such as mycelial growth rate, color of the medium, amount of the aerial mycelium, shape and color of the sclerotia- and in the pathogenicity of isolates of the pathogen, Thanatephorus cucumeris Dank were observed. The optimum temperature for mycelial growth also varied with isolates, from $25^{\circ}C$ to $30^{\circ}C$. There were not necessarily any correlation between curtural characteristics and pathogenicity of isolates of Thanatephorus cucumens. 3. Mycelial grow th of isolates of Thanatephorus cucumens on the PDA medium were correlated with the air temperatures of the region where the isolates were collected. The isolates from the regions with high temperature grew well on PDA medium at $35^{\circ}C$ than those from the region with low temperature, on the other hand, the isolates from the regions with the low temperature grew well on the same medium at $12^{\circ}C$ than those from the regions with high temperature. 4. Pectin polygalacturonase (PG) and cellulase (Cx) were most active on the 3rd day after inoculation on the leaves of rice plant with Thanatephorus cucumeris, whereas pectin methylestrase (PE) was most active on the 4th day after inoculation. Relationship between the activities of PE of isolates and the strength of pathogenicity of isolates was obtained, but PG and cellulase activities were not correlated with pathogenicity of isolates. 5. The tolerence of sclerotia from in-vitro culture to low temperature varied with their water content, the dried cultural sclerotia were more tolerent than wet ones, Dried cultural sclerotia maintained almost 100% germinability for 45 days at $-20^{\circ}C$, whereas wet sclerotia lost viability at $-5^{\circ}C$. The germination ratio of the sclerotia after overwintering changed from 18% to 70% according to the water content of the test paddy fields and the ratio was low in wet paddy condition. 6. To investigate the host range of this fungi in and near paddy field, 17 weeds were inoculated with fungi. The lesions of sheath blight disease was obserbed on Sagittaria trifolia L., Echinochloa crusgalli P. Beauv., Monochoria vaginal is Presl, Polygonum Hydropiper L., Eclipta prostrata L., Digitaria sanguinalis Scapoli. 7. When the level of nitrogen applied was doubled over standard level, total nitrogen content in rice sheath increased, ami when silicate was applied, starch content in rice sheath decreased, inducing the rice plants more susceptible to sheath blight disease. Increased dressing of potash ferilizer reduced the incidence of sheat blight disease. 8. The percentage of infected stems in the early period increased more in the narrow hill plot than in the wide hill plot, but in the late period this tendency was inversed; the percentage of infected stems as well as severity in the wide hill plot increased more compared to the narrow hill plot, and the disease severity in the one plant per hill plot was also low. The number of stems in the wide hill plot was more than the number of stems in the narrow hill plot. This indicates that the microclimate, such as the relative humidity, in the narrow hill plot was more favorable for the development of this disease. 9. There was a high negative correlation between the disease severity of varieties to the sheath blight and the maturity of the varieties, that is, the early varieties were more susceptible than the late ones, and much-tillering varieties usually showed more infection than less tillering varieties. 10. No relationship was obtained between the percentage of infected stems in the early period and the severity after heading, whereas a distinct relationship was obtained between former and latter after Aug. 10.

• 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.

• Applied Biological Chemistry
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• v.17 no.3
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• pp.193-218
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• 1974

These studies were carried out for the elucidation of liming effect on the growth of rice seedlings and the chemical characteristics of an acid sulphate paddy that shows not only extremely high acidity of soil but also poor growth of rice plants, consequently low yield. Thus the liming effect on the changes of acidity, oxidation-reduction potential, and the contents of iron, aluminium, sulphate, and phosphorus fractions in the soil was investigated under the waterlogging and drying condition. The reclaimable or inhibitory effect of phosphorus, iron and aluminium on the growth of rice seedlings was also investigated under liming. The results are summarized as follows: 1. After liming, the pH of the acid sulphate subsoil decreased again on drying. 2. The oxidation-reduction potential reached a minimum after 5 days of flooding and greatly decreased on liming but increased after drying. 3. The contents of ferrous iron soluble in water-and Morgan's solution reached a maximum after 15 days of flooding and only the content of water soluble ferrous iron was greatly decreased. 4. The content of aluminium soluble in water-and Morgan's solution decreased by flooding and liming, and showed a tendency to increase on drying. 5. In the limed acid sulphate soil, the content of water soluble calcium showed a highly significant negative correlation with the content of sulphate and liming decreased sulphate content in the soil. 6. The contents of total phosphorus was 496.3 ppm in the acid sulphate topsoil and 387.5 ppm in the subsoil. The content of each phosphorus fraction was in the order of Fe-P>Occ. Fe-P>Ca-P>Occ. Al-P>Al-P and Fe-P content in the soil was the highest fraction among them. 7. Lime application increased greatly Ca-P and Al-P, and Occ. Fe-P and Occ. Al-P only slightly, but decreased Fe-P differently in each soil. 8. Effect of phosphorus on the dry matter yield of rice seedlings was great. The optimum amount of phosphorus to produce maximum dry matter yield of rice seedlings appeared to be 6.8% of maximum absorption (absorption coefficient) without liming and 10.0% with liming. 9. In rice seedlings liming increased the content and uptake of calcium and silica but decreased those of iron and aluminium. Phosphorus application increased the content and uptake of phosphorus and decreased iron while the application of iron and aluminium increased their contents and uptake but decreased those of phosphorus. 10. Liming greatly alleviated such toxicity of iron and aluminium. 11. When phosphorus was applied, the dry matter yield of rice seedlings showed highly significant positive correlations with uptake of phosphorus, calcium and silica each. When iron and aluminium were applied, dry matter yields indicated significant positive correlations with the contents or uptake of calcium and silica each, but significant negative correlations with the content or uptake of iron and aluminium. 12. Under the application of phosphorus and lime, dry matter yields showed significant positive correlations with pH and Morgan's extractable calcium each of the soil samples after harvest. Under the application of lime, iron and aluminium, dry matter yields showed significant positive correlations with pH, calcium and silica each, but negative correlations with iron and aluminium contents each of the soil samples after harvest.

• 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.