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Studies on Sclerotium rolfsii Sacc. isolated from Magnolia kobus DC. in Korea (목련(Magnolia kobus DC.)에서 분리한 흰비단병균(Sclerotium rolfsii Sacc.)에 관한 연구)

  • Kim Kichung
    • 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.

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A Study on Forest Insurance (산림보험(山林保險)에 관한 연구(硏究))

  • Park, Tai Sik
    • Journal of Korean Society of Forest Science
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    • v.15 no.1
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    • pp.1-38
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    • 1972
  • 1. Objective of the Study The objective of the study was to make fundamental suggestions for drawing a forest insurance system applicable in Korea by investigating forest insurance systems undertaken in foreign countries, analyzing the forest hazards occurred in entire forests of Korea in the past, and hearing the opinions of people engaged in forestry. 2. Methods of the Study First, reference studies on insurance at large as well as on forest insurance were intensively made to draw the characteristics of forest insurance practiced in main forestry countries, Second, the investigations of forest hazards in Korea for the past ten years were made with the help of the Office of Forestry. Third, the questionnaires concerning forest insurance were prepared and delivered at random to 533 personnel who are working at different administrative offices of forestry, forest stations, forest cooperatives, colleges and universities, research institutes, and fire insurance companies. Fourth, fifty three representative forest owners in the area of three forest types (coniferous, hardwood, and mixed forest), a representative region in Kyonggi Province out of fourteen collective forest development programs in Korea, were directly interviewed with the writer. 3. Results of the Study The rate of response to the questionnaire was 74.40% as shown in the table 3, and the results of the questionaire were as follows: (% in the parenthes shows the rates of response; shortages in amount to 100% were due to the facts of excluding the rates of response of minor respondents). 1) Necessity of forest insurance The respondents expressed their opinions that forest insurance must be undertaken to assure forest financing (5.65%); for receiving the reimbursement of replanting costs in case of damages done (35.87%); and to protect silvicultural investments (46.74%). 2) Law of forest insurance Few respondents showed their views in favor of applying the general insurance regulations to forest insurance practice (9.35%), but the majority of respondents were in favor of passing a special forest insurance law in the light of forest characteristics (88.26%). 3) Sorts of institutes to undertake forest insurance A few respondents believed that insurance companies at large could take care of forest insurance (17.42%); forest owner's mutual associations would manage the forest insurance more effectively (23.53%); but the more than half of the respondents were in favor of establishing public or national forest insurance institutes (56.18%). 4) Kinds of risks to be undertaken in forest insurance It would be desirable that the risks to be undertaken in forest insurance be limited: To forest fire hazards only (23.38%); to forest fire hazards plus damages made by weather (14.32%); to forest fire hazards, weather damages, and insect damages (60.68%). 5) Objectives to be insured It was responded that the objectives to be included in forest insurance should be limited: (1) To artificial coniferous forest only (13.47%); (2) to both coniferous and broad-leaved artificial forests (23.74%); (3) but the more than half of the respondents showed their desire that all the forests regardless of species and the methods of establishment should be insured (61.64%). 6) Range of risks in age of trees to be included in forest insurance The opinions of the respondents showed that it might be enough to insure the trees less than ten years of age (15.23%); but it would be more desirous of taking up forest trees under twenty years of age (32.95%); nevertheless, a large number of respondents were in favor of underwriting all the forest trees less than fourty years of age (46.37%). 7) Term of a forest insurance contract Quite a few respondents favored a contract made on one year basis (31.74%), but the more than half of the respondents favored the contract made on five year bases (58.68%). 8) Limitation in a forest insurance contract The respondents indicated that it would be desirable in a forest insurance contract to exclude forests less than five hectars (20.78%), but more than half of the respondents expressed their opinions that forests above a minimum volume or number of trees per unit area should be included in a forest insurance contract regardless of the area of forest lands (63.77%). 9) Methods of contract Some responded that it would be good to let the forest owners choose their forests in making a forest insurance contract (32.13%); others inclined to think that it would be desirable to include all the forests that owners hold whenerver they decide to make a forest insurance contract (33.48%); the rest responded in favor of forcing the owners to buy insurance policy if they own the forests that were established with subsidy or own highly vauable growing stock (31.92%) 10) Rate of premium The responses were divided into three categories: (1) The rate of primium is to be decided according to the regional degree of risks(27.72%); (2) to be decided by taking consideration both regional degree of risks and insurable values(31.59%); (3) and to be decided according to the rate of risks for the entire country and the insurable values (39.55%). 11) Payment of Premium Although a few respondents wished to make a payment of premium at once for a short term forest insurance contract, and an annual payment for a long term contract (13.80%); the majority of the respondents wished to pay the premium annually regardless of the term of contract, by employing a high rate of premium on a short term contract, but a low rate on a long term contract (83.71%). 12) Institutes in charge of forest insurance business A few respondents showed their desire that forest insurance be taken care of at the government forest administrative offices (18.75%); others at insurance companies (35.76%); but the rest, the largest number of the respondents, favored forest associations in the county. They also wanted to pay a certain rate of premium to the forest associations that issue the insurance (44.22%). 13) Limitation on indemnity for damages done In limitation on indemnity for damages done, the respondents showed a quite different views. Some desired compesation to cover replanting costs when young stands suffered damages and to be paid at the rate of eighty percent to the losses received when matured timber stands suffered damages(29.70%); others desired to receive compensation of the actual total loss valued at present market prices (31.07%); but the rest responded in favor of compensation at the present value figured out by applying a certain rate of prolongation factors to the establishment costs(36.99%). 14) Raising of funds for forest insurance A few respondents hoped to raise the fund for forest insurance by setting aside certain amount of money from the indemnity paid (15.65%); others wished to raise the fund by levying new forest land taxes(33.79%); but the rest expressed their hope to raise the fund by reserving certain amount of money from the surplus money that was saved due to the non-risks (44.81%). 15) Causes of fires The main causes of forest fires 6gured out by the respondents experience turned out to be (1) an accidental fire, (2) cigarettes, (3) shifting cultivation. The reponses were coincided with the forest fire analysis made by the Office of Forestry. 16) Fire prevention The respondents suggested that the most important and practical three kinds of forest fire prevention measures would be (1) providing a fire-break, (2) keeping passers-by out during the drought seasons, (3) enlightenment through mass communication systems. 4. Suggestions The writer wishes to present some suggestions that seemed helpful in drawing up a forest insurance system by reviewing the findings in the questionaire analysis and the results of investigations on forest insurance undertaken in foreign countries. 1) A forest insurance system designed to compensate the loss figured out on the basis of replanting cost when young forest stands suffered damages, and to strengthen credit rating by relieving of risks of damages, must be put in practice as soon as possible with the enactment of a specifically drawn forest insurance law. And the committee of forest insurance should be organized to make a full study of forest insurance system. 2) Two kinds of forest insurance organizations furnishing forest insurance, publicly-owned insurance organizations and privately-owned, are desirable in order to handle forest risks properly. The privately-owned forest insurance organizations should take up forest fire insurance only, and the publicly-owned ought to write insurance for forest fires and insect damages. 3) The privately-owned organizations furnishing forest insurance are desired to take up all the forest stands older than twenty years; whereas, the publicly-owned should sell forest insurance on artificially planted stands younger than twenty years with emphasis on compensating replanting costs of forest stands when they suffer damages. 4) Small forest stands, less than one hectare holding volume or stocked at smaller than standard per unit area are not to be included in a forest insurance writing, and the minimum term of insuring should not be longer than one year in the privately-owned forest insurance organizations although insuring period could be extended more than one year; whereas, consecutive five year term of insurance periods should be set as a mimimum period of insuring forest in the publicly-owned forest insurance organizations. 5) The forest owners should be free in selecting their forests in insuring; whereas, forest owners of the stands that were established with subsidy should be required to insure their forests at publicly-owned forest insurance organizations. 6) Annual insurance premiums for both publicly-owned and privately-owned forest insurance organizations ought to be figured out in proportion to the amount of insurance in accordance with the degree of risks which are grouped into three categories on the basis of the rate of risks throughout the country. 7) Annual premium should be paid at the beginning of forest insurance contract, but reduction must be made if the insuring periods extend longer than a minimum period of forest insurance set by the law. 8) The compensation for damages, the reimbursement, should be figured out on the basis of the ratio between the amount of insurance and insurable value. In the publicly-owned forest insurance system, the standard amount of insurance should be set on the basis of establishment costs in order to prevent over-compensation. 9) Forest insurance business is to be taken care of at the window of insurance com pnies when forest owners buy the privately-owned forest insurance, but the business of writing the publicly-owned forest insurance should be done through the forest cooperatives and certain portions of the premium be reimbursed to the forest cooperatives. 10) Forest insurance funds ought to be reserved by levying a property tax on forest lands. 11) In order to prevent forest damages, the forest owners should be required to report forest hazards immediately to the forest insurance organizations and the latter should bear the responsibility of taking preventive measures.

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Studies on the physio-chemical properties and the cultivation of oyster mushroom(Pleurotus ostreatus) (느타리버섯의 생리화학적성질(生理化學的性質) 및 재배(栽培)에 관(關)한 연구(硏究))

  • Hong, Jai-Sik
    • Applied Biological Chemistry
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    • v.21 no.3
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    • pp.150-184
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    • 1978
  • Nutritional characteristics and physio-chemical properties of mycelial growth and fruitbody formation of oyster mushroom(Pleurotus ostreatus)in synthetic media, the curtural condition for the commerical production in the rice straw and poplar sawdust media, and the changes of the chemical components of the media and mushroom during the cultivation were investigated. The results can be summarized as follows: 1. Among the carbon sources mannitol and sucrose gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while lactose and rhamnose gave no mycelial growth. Also, citric acid, succinic acid, ethyl alcohol and glycerol gave poor fruit-body formation, and acetic acid, formic acid, fumaric acid, n-butyl alcohol, n-propyl alcohol and iso-butyl alcohol inhibited mycelial growth. 2. Among the nitrogen sources peptone gave rapid mycelial growth and rapid formation of fruit-body with higher yield, while D,L-alanine, asparatic acid, glycine and serine gave very poor fruit-body formation, and nitrite nitrogens, L-tryptophan and L-tyrosine inhibited mycelial growth. Inorganic nitrogens and amino acids added to peptone were effective for fruit-body growth, and thus addition of ammonium sulfate, ammonium tartarate, D,L-alanine and L-leucine resulted in about 10% increase fruit-body yield. L-asparic acid about 15%, L-arginine about 20%, L-glutamic acid, and L-lysine about 25%. 3. At C/N ratio of 15.23 fruit-body formation was fast, but the yield decreased, and at C/N ratio of 11.42 fruit-body formation was slow, but the yield increased. Also, at the same C/N ratio the higher the concentration of mannitol and petone, the higher yield was produced. Thus, from the view point of both yield of fruit-body and time required for fruiting the optimum C/N ratio would be 30. 46. 4. Thiamine, potassium dihydrogen phosphate and magnecium sulfate at the concentration of $50{\mu}g%$. 0.2% and 0.02-0.03%, respectively, gave excellent mycelial and fruit-body growth. Among the micronutrients ferrous sulfate, zinc sulfate and manganese sulfate showed synergetic growth promoting effect but lack of manganese resulted in a little reduction in mycelial and fruit-body growth. The optimum concentrati on of each these nutrients was 0.02mg%. 5. Cytosine and indole acetic acid at 0.2-1mg% and 0.01mg%, respectively, increased amount of mycelia, but had no effect on yield of fruit-body. The other purine and pyrimidine bases and plant hormones also had no effect on mycelial and fruit-belly yield. 6. Illumination inhibited mycelial growth, but illumination during the latter part of vegetative growth induced primordia formation. The optimum light intensity and exposure time was 100 to 500 lux and 6-12 hours per day, respectively. Higher intensity of light was injurous, and in darkness only vegetative growth without primordia formation was continued. 7. The optimum temperature for mycelial growth was $25^{\circ}C$ and for fruit-body formation 10 to $15^{\circi}C$. The optimum pH range was from 5.0 to 6.5. The most excellent fry it-body formation were produced from the mycelium grown for 7 to 10 days. The lesser the volume of media, the more rapid the formation of fruit-body; and the lower the yield of fruit-body; and the more the volume of media, the slower the formation of fruit-body, and the higher the yield of fruit-body. The primordia formation was inhibited by $CO_2$. 8. The optimum moisture content for mycelial growth was over 70% in the bottle media of rice straw and poplar sawdust. 10% addition of rice bran to the media exhibited excellent mycelial growth and fruit-body formation, and the addition of calciumcarbonate alone was effective, but the addition of calcium carbonate was ineffective in the presence of rice bran. 9. In the cultivation experiments the total yield of mushroom from the rice straw media was $14.99kg/m^2$, and from the sawdust media $6.52kg/m^2$, 90% of which was produced from the first and second cropping period. The total yield from the rice straw media was about 2.3 times as high as that from the sawdust media. 10. Among the chemical components of the media little change was observed in the content of ash on the dry weight basis, and organic matter content decreased as the cultivation progressed. Moisture content, which was about 79% at the time of spawning, decreased a little during the period of mycelial propagation, after which no change was observed. 11. During the period from spawning to the fourth cropping about 16.7% of the dry matter, about 19.3% of organic matter, and about 40% of nitrogen were lost from the rice straw media; about 7.5% of dry mallet, about 7.6% of organic matter, and about 20% of nitrogen were lost from the sawdust media. For the production of 1kg of mushroom about 232g of organic matter and about 7.0g of nitrogen were consumed from the rice straw media; about 235g of organic matter and about 6.8g of nitrogen were consumed from the sawdust media, 1㎏ of mushroom from either of media contains 82.4 and 82.3g of organic matter and 5.6 and 5.4g of nitrogen, respectively. 12. Total nitrogen content of the two media decreased gradually as the cultivation progressed, and total loss of insoluble nitrogen was greater than that of soluble nitrogen. Content of amino nitrogen continued to increase up to the third cropping time, after which it decreased. 13. In the rice straw media 28.0 and 13.8% of the total pentosan and ${\alpha}$-cellulose, respectively, lost during the whole cultivation period was lost during the period of mycelial growth; in the sawdust media 24.1 and 11.9% of the total pentosan and ${\alpha}$-cellulose, respectively, was lost during the period of mycelial growth. Lignin content in the media began to decrease slightly from the second cropping time, while the content of reduced sugar, trehalose and mannitol continued to increase. C/N ratio of the rice straw media decreased from 33.2 at spawining to 30.0 at ending; that of the sawdust media decreased from 61.3 to 60.0. 14. In both media phosphorus, potassium, manganese and zinc decreased, at magnesium, calcium and copper showed irregular changes, and iron had a tendency to be increased. 15. Enzyme activities are much higher in the rice straw media than in the sawdust media. CMC saccharifying and liquefying activity gradually increased from after mycelial propagation to the second cropping, after which it decreased in both media. Xylanase activity rapidly and greatly increased during the second cropping period rather than the first period. At the start of the third cropping period the activity decreased rapidly in the rice straw media, which was not observed in the sawdust media. Protease activity was highest after mycelial propagation, after which it gradually decreased. The pH of the rice straw media decreased from 6.3 at spawning to 5.0 after fourth cropping; that of the sawdust media decreased from 5.7 to 4.9. 16. The contents of all the components except crude fibre of the mushroom from the rice straw media were higher than those from the sawdust media. Little change was observed in the content of the components of mushroom cropped from the first to the third period, but slight decrease was noticed at the fourth cropping.

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