• 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.

• Journal of The Korean Society of Grassland and Forage Science
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• v.32 no.1
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• pp.59-74
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• 2012

Buffer solubility and protein fractionation were evaluated from the hays (timothy, alfalfa and klein) and straws (tall fescue and rice), and $In$ $vitro$ trial was conducted to examine the effect of buffer extraction on fermentation characteristics, degradability and gas ($CO_2$ and $CH_4$) production. Buffer soluble protein (SP) content and A fraction in total protein were highest in alfalfa hay as 61% and 41.77%, respectively while lowest in rice straw (42.8% and 19.78%, respectively). No difference was observed in B1 fraction among forages but B2 fraction was slightly increased in klein hay (12.34%) and tall fescue straw (10.05%) compared with other forages (6.34~8.85%). B3 fraction of tall fescue was highest as 38.49% without difference among other forages while C fraction was highest in rice straw. pH in incubation solution was higher in all forages after extraction than before extraction at 3h (P<0.01) and 6h (P<0.05), and pH from hays of timothy and alfalfa was higher than the other forages at 6h (P<0.05) and 12h (P<0.001). Regardless of extraction, ammonia-N concentration from alfalfa hay was increased at all incubation times and extraction effect was appeared only at 3h incubation time (P<0.01). Total VFA concentration from alfalfa hay was highest up to 24h incubation while those from tall fescue straw and rice straw were lowest. Buffer extraction decreased (P<0.01~P<0.001) the total VFA concentration. Acetic acid proportion was increased (P<0.001) before extraction of forages but no difference was found between forages. Propionic acid($C_3$) proportion was also increased(P<0.001) before extraction in all forages than in straws at 3h, 24h and 48h incubations, and $C_3$ from hays were mostly higher (P<0.05) than from straws. Butyric acid proportion, however, was not affected by extraction at most incubation times. Parameter 'a' regarding to the dry matter (DM) degradation was increase (P<0.001) in all forages before extraction, and was decreased (P<0.05) in tall fescue straw and rice straw compared with hays. Parameter 'b' was also increased (P<0.001) before extraction but no difference was found between forages. Effective degradability of DM (EDDM) was higher (P<0.001) before extraction in most forages except for rice straw. Buffer extraction decreased (P<0.05) all parameters (a, b, and c) regrading to the crude protein (CP) degradation but no difference was found between forages. Effective degradation of CP (EDCP) was lower (P<0.05) in straws than in hays. Parameters 'a' and 'b' regarding to the NDF degradation (P<0.01) and effective degradability of NDF (EDNDF, P<0.001) were also higher in forages before extraction than after extraction but no difference was found between forages. Buffer extraction reduced (P<0.05~P<0.001) $CO_2$ production from all the forages uo to 24h incubation and its production was greater (P<0.05~P<0.01) from hays than straws. Methane ($CH_4$) production was also greater (P<0.01~P<0.001) in all forages at all incubation times, and its production was greater (P<0.05) from hays than from straws at most incubation times. Based on the results of the current study, it can be concluded that buffer solubility and CP fractionation might be closely related with $In$ $vitro$ VFA concentration, degradability and gas ($CO_2$ and $CH_4$) production. Thus, measurement of buffer solubility and protein fractionation of forages might be useful to improve TMR availability in the ruminants.

• The Korean Journal of Mycology
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• v.7 no.1
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• pp.13-73
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• 1979

These studies were conducted to investigate nutrient sources and supplementary materials of synthetic compost media for Agaricus bisporus culture. Investigation were carried out to establish the optimum composition for compost of Agaricus bisporus methods of out-door fermentation and peakheating with rice straw as the main substrate of the media. The incidence and flora of harmful organisms in rice straw compost and their control were also studied. 1. When rice straw was used as the main substrate in synthetic compost as a carbon source. yields were remarkably high. Fermentation was more rapid than that of barley straw or wheat straw, and the total nitrogen content was high in rice straw compost. 2. Since the morphological and physico-chemical nature of Japonica and Indica types of rice straw are greatly dissimilar. there were apparent differences in the process of compost fermentation. Fermentation of Indica type straw proceeded more rapidly with a shortening the compost period, reducing the water supply, and required adding of supplementary materials for producing stable physical conditions. 3. Use of barley straw compost resulted in a smaller crop compared with rice straw. but when a 50%, barley straw and 50% rice straw mixture was used, the yield was almost the same as that using only rice straw. 4. There were extremely high positive correlations between yield of Agaricus bisporus and the total nitrogen, organic nitrogen, amino acids, amides and amino sugar nitrogen content of compost. The mycerial growth and fruit body formation were severely inhibited by ammonium nitrogen. 5. When rice straw was used as the main substrate for compost media, urea was the most suitable source of nitrogen. Poor results were obtained with calcium cyanamide and ammonium sulfate. When urea was applied three separate times, nitrogen loss during composting was decreased and the total nitrogen content of compost was increased. 6. The supplementation of organic nutrient activated compost fermentation and increased yield of Agaricus bisporus. The best sources of organic nutrients were: perilla meal, sesame meal, wheat bran and poultry manure, etc. 7. Soybean meal, tobacco powder and glutamic acid fermentation by-products which were industrial wastes, could be substituted for perilla meal, sesame meal and wheat bran as organic nutrient sources for compost media. B. When gypsum and zeolite were added to rice straw. physical deterioration of compost due to excess moisture and caramelization was observed. The Indica type of straw was more remarkable in increase of yield of Agricus bisporus by addition of supplementing materials than Japonica straw. 9. For preparing rice straw compost, the best mixture was prepared by 10% poultry manure, 5% perilla meal, 1. 2 to 1. 5% urea and 1% gypsum. At spring cropping, it was good to add rice bran to accelerate heat generation of the compost heap. 10. There was significantly high positive correlation (r=0.97) between accumulated temperature and the decomposition degree of compost during outdoor composting. The yield was highest at accumulated temperatures between 900 and $1,000^{\circ}C$. 11. Prolonging the composting period brought about an increase in decomposition degree and total nitrogen content, but a decrease in ammonium nitrogen. In the spring the suitable period of composting was 20 to 25 days. and about 15 days in autumn. For those periods, the degree of decomposition was 19 to 24%. 12. Compactness of wet compost at filling caused an increase in the residual ammonium nitrogen. methane and organic acid during peak heating. There was negative correlation between methane content and yield (r=0.76)and the same was true between volatile organic acid and yield (r=0.73). 13. In compost with a moisture content range between 69 to 80% at filling. the higher the moisture content, the lower the yield (r=0.78). This result was attributed to a reduction in the porosity of compost at filling the beds. The optimum porosity for good fermentation was between 41 and 53%. 14. Peak heating of the compost was essential for the prevention of harmful microorganisms and insect pests. and for the removal of excess ammonia. It was necessary to continue fer mentatiion for four days after peak heating. 15. Ten species of fungi which are harmful or competitive to Agaricus bisporus were identified from the rice compost, including Diehliomyces microsporus, Trichoderma sp. and Stysanus stemoites. The frequency of occurrance was notably high with serious damage to Agaricus bisporus. 16. Diehliomyces microsporus could be controlled by temperature adjustment of the growing room and by fumigating the compost and the house with Basamid and Vapam. Trichoderma was prevented by the use of Bavistin and Benomyl. 17. Four species of nematodes and five species of mites occured in compost during out-door composting. These orgnanisms could be controlled through peakheating compost for 6 hours at $60^{\circ}C$.

• Journal of the Korean Wood Science and Technology
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• v.16 no.2
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• pp.1-41
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• 1988

For the purpose of utilizing the sawdust having poor combining properties as board raw material and resulting in dimensional instability of board, polypropylene chip (abbreviated below as PP chip) or oriented PP thread was combined with sawdust particle from white meranti(Shorea sp.). The PP chip was prepared from PP thread in length of 0.25, 0.5, 1.0 and 1.5 cm for conventional blending application. Thereafter, the PP chip cut as above was combined with the sawdust particle by 3, 6, 9, 12 and 15% on the weight basis of board. Oriented PP threads were aligned with spacing of 0.5, 1.0 and 1.5cm along transverse direction of board. The physical and mechanical properties on one, two and three layer boards manufactured with the above combining conditions were investigated. The conclusions obtained at this study were summarized as follows: 1. In thickness swelling, all one layer boards combined with PP chips showed lower values than control sawdustboard, and gradually clear decreasing tendendy with the increase of PP chip composition. Two layer board showed higher swelling value than one layer board, but the majority of boards lower values than control sawdustboard. All three layer boards showed lower swelling values than control sawdustboard. 2. In the PP chip and oriented thread combining board, the swelling values of boards combining 0.5cm spacing oriented thread with 1.0 or 1.5cm long PP chip in 12 and 15% by board weight were much lower than the lowest of one or three layer. 3. In specific gravity of 0.51, modulus of rupture of one layer board combined with 3% PP chip showed higher value than control sawdustboard. However, moduli of rupture of the boards with every PP chip composition did not exceed 80kgf/cm2, the low limit value of type 100 board, Korean Industrial Standard KS F 3104 Particleboards. Moduli of rupture of 6%, 1.5cm-long and 3% PP chip combined boards in specific gravity of 0.63 as well as PP chip combined board in specific gravity of 0.72 exceeded 80kgf/$cm^2$ on KS F 3104. Two layer boards combined with every PI' chip composition showed lower values than control sawdustboard and one layer board. Three layer boards combined with.1.5cm long PP chip in 3, 6 and 9% combination level showed higher values than control sawdustboard, and exceeded 80kgf/$cm^2$ on KS F 3104. 4. In modulus of rupture of PP thread oriented sawdustboard, 0.5cm spacing oriented board showed the highest value, and 1.0 and 1.5cm spacing oriented boards lower values than the 0.5cm. However, all PP thread oriented sawdustboards showed higher values than control saw-dustboard. 5. Moduli of rupture in the majority of PP chip and oriented thread combining boards were higher than 80kgf/$cm^2$ on KS F 3104. Moduli of rupture in the boards combining longer PP chip with narrower 0.5cm spacing oriented thread showed high values. In accordance with the spacing increase of oriented thread, moduli of rupture in the PP chip and oriented thread combining boards showed increasing tendency compared with oriented sawdustboard. 6. Moduli of elasticity in one, two and three layer boards were lower than those of control sawdustboard, however, moduli of elasticity of oriented sawdustboards with 0.5, 1.0 and 1.5cm spacing increased 20, 18 and 10% compared with control sawdustboard, respectively. 7. Moduli of elasticity in the majority of PP chip and oriented thread combining boards in 0.5, 1.0 and 1.5cm oriented spacing showed much higher values than control sawdustboard. On the whole, moduli of elasticity in the oriented boards combined with 9% or less combination level and 0.5cm or more length of PP chip showed higher values than oriented sawdustboard. The increasing effect on modulus of elasticity was shown by the PP chip composition in oriented board with narrow spacing. 8. Internal bond strengths of all one layer PP chip combined boards showed lower values than control sawdust board, however, the PP chip combined boards in specific gravity of 0.63 and 0.72 exceeded 1.5kgf/$cm^2$, the low limit value of type 100 board and 3kgf/$cm^2$, type 200 board on KS F 3104, respectively. And also most of all two, three layer-and oriented boards exceeded 3kgf/$cm^2$ on KS F. 9. In general, screw holding strength of one layer board combined with PP chip showed lower value than control sawdustboard, however, that of two or three layer board combined with PP chip did no decreased tendency, and even screw holding strength with the increase of PP chip composition. In the PP chip and oriented PP thread combining boards, most of the boards showed higher values than control sawdustboard in 9% or less PP chip composition.