Abstract
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.
합성배지(合成培地)에서 느타리 버섯균(菌)의 균사생육(菌絲生育)과 자실체형성(子實體形成)에 대한 영양적(營養的) 특성(特性)과 생리화학적(生理化學的) 제성질(諸性質)을 구명(究明)하고 볏짚과 톱밥 양(兩) 배지(培地)에서 느타리 버섯의 대량(大量) 생산(生産)을 위한 배양조건(培養條件)을 밝히고, 느타리 버섯 재배기간(栽培期間) 중 배지(培地)와 버섯중의 각종(各種) 성분(成分)의 추이(推移)를 알고자 실험을 수행하여 다음과 같은 결과를 얻었다. 1. 탄소원(炭素源) 중 mannitol과 서은 균사생육(菌絲生育)과 자실체(子實體) 형성(形成)이 빠르고 자실체(子實體)의 수량(收量)이 많았으나 lactose와 rhamnose는 균사(菌絲) 조차도 생육하지 못하였다. 또한 구연산, 호박산, ethyl alcohol 및 glycerol에서는 자실체(子實體) 형성(形成)이 매우 빈약(貧弱)하였고, 식초산, 개미산, 푸마르산, n-butyl alcohol, iso-butyl alcohol 및 n-propyl alcohol은 균사생육(菌絲生育)을 저해(阻害)하였다. 2. 질소원(窒素源)중 peptone은 균사생육(菌絲生育)과 자실체(子實體) 형성(形成)이 빠르고 자실체(子實體)의 수량(收量)이 많았으나 DL-alanine, asparagine, L-aspartic acid, glycine및 serine은 자실체형성(子實體形成)이 매우 빈약(貧弱)하였으며 아질산태질소(亞窒酸態窒素), L-tryptophan 및 L-tyrosine은 균(菌)의 생육을 저해(沮害)하였다. 또한 peptone에 무기태질소(無機態窒素)와 아미노산(酸)을 혼용(混用)한 결과 $(NH_4)_2SO_4$, $NH_4$-tartarate, DL-alanine및 L-leucine에서는 자실체(子實體)의 수량(收量)이 약 10% 증가되었고, L-aspartic acid는 약 15%. L-arginine은 약20%, L-glutamic acid와 L-lysine은 약 25%증가 되었다. 3. C/N율(率) 15.23에서 자실체(子實體) 형성(形成)은 빠르나 자실체(子實體)의 수량(收量)은 감소(減少)되었으며, C/N율(率) 11.42에서는 자실체형성(子實體形成)은 늦으나 자실체(子實體)의 수량(收量)은 증가되는 경향이 있었다. 또한 동일 C/N율(率)에서도 mannitol과 peptone의 농도(濃度)가 높은 편이 수량(收量)이 증가되었다. 그러므로 자실체(子實體)의 수량(收量)과 자실체형성(子實體形成) 소요일(所要日)의 관점(觀點)에서 보면 C/N율(率) 30.46이 어느정도 적당(適當)한 것 같다. 4. Thiamine $50{\mu}g%,\;KH_2PO_4$ 0.2%, $MgSO_4{\cdot}7H_2O$는 $0.02{\sim}0.03%$일때 균사(菌絲)와 자실체(子實體) 생육(生育)이 우수(優秀)하였으며 미량원소(微量元素)로서는 $FeSO_4{\cdot}7H_2O$,\;ZnSO_4{\cdot}7H_2O$ 및 $MnSO_4{\cdot}5H_2O$가 공존(共存)하면 생육촉진(生育促進)의 상승효과(相乘效果)가 인정되었으나 3이원소(元素)중 Mn이 결핍(缺乏)하면 균사(菌絲)와 자실체(子實體)의 생육(生育)이 다소 저하되었다. 이들 염류(鹽類)의 최적농도(最適濃度)는 각각 0.02mg%이었다. 5. Cytosine $0.2{\sim}1mg%$와 indole acetic acid 0.01mg%에서 균사량(菌絲量)은 증가되었으나 자실체(子實體)의 수량(收量)에는 효과 없었으며 그밖의 purine염기(鹽基), pyrimidine염기(鹽基) 및 식물(植物) hormone은 영향이 없었다. 6. 광조사(光照射영)에 의해서 균사생육(菌絲生育)은 저해(沮害)되었으며 영양생장(營養生長)의 후기에 광(光)을 조사(照射)하면 원기형성(原基形成)이 유도(誘導)되었다. 광(光)의 최적조도(最適照度)는 $100{\sim}500lux$, 조사시간(照射時間)은 매일 $6{\sim}12$시간이었고, 이 이상(以上)의 조도(照度)에서는 오히려 저해(沮害)되었으며, 암소(暗所)에서는 원기(原基)가 형성(形成)되지 않고 영양생장(營養生長)만 계속되었다. 7. 균사생육(菌絲生育)과 자실체(子實體) 형성(形成)의 최적온도(最適溫度)는 각각 $25^{\circ}C,\;10{\sim}15^{\circ}C$이었고 최적(最適)의 pH범위(範圍)는 $5.0{\sim}6.5$이었으며 균사(菌絲)는 $7{s\im}10$일간 배양(培養)했을 때가 자실체(子實體) 형성(形成)이 제일 우수(優秀)하였다. 또한 배지량(培地量)이 적을수록 자실체(子實體) 형성(形成)은 빠르나 자실체(子實體)의 수량(收量)은 감소(減少)되었고 배지량(培地量)이 많을수록 자실체(子實體) 형성(形成)은 늦은반면에 그 수량(收量)은 증가 되었으며, 원기형성(原基形成)은 $CO_2$에 의하여 저해(沮害)되었다. 8. 볏짚과 톱밥 병 배지(培地)에서 균사생육(菌絲生育)의 최적(最適) 수분량(水分量)은 70%이상 이었으며 미강(米糠) 10%를 배지(培地)에 첨가(添加)했을 때는 균사생육(菌絲生育)과 자실체형성(子實體形成)이 우수(優秀)하였다. 그리고 양배지(兩培地)에 $CaCo_3$를 단독(單獨)으로 첨가했을 때는 유효(有？)하였으나 미강(米糠)과 함께 첨가했을때는 효과(？果)를 볼 수 없었다. 9. 재배(栽培) 실험(實驗)에서 느타리 버섯의 전체(全體) 수량(收量)은 볏짚배지(培地)에서 $14.99kg/m^2$, 톱밥배지(培地)에서 $6.52kg/m^2$이었고 양배지(兩培地) 모두 90%이상이 1,2주기(週期)에서 얻어졌으며 볏짚배지(培地)(dry matter $20.96kg/m^2$)의 전수율(全收率)을 톱밥배지(培地)(dry matter $20.83kg/m^2$)의 약 2.3배(倍)이었다. 10. 재배기간(栽培期間)중 양(兩) 배지(培地)의 일반 성분을 고형물(固形物) 기준(基準)으로 볼때 회분(灰分)의 변화는 적었으나 유기물(有機物)은 감소(減少)되었으며, 수분(水分)은 종균접종시(種菌接種時) 약 79%이던것이 균사번식기간(菌絲繁殖期間)중에 다소 감소(減少)되었고 그 이후부터는 큰 변화가 없었다. 11, 종균접종시(種菌接種時) 부터 4주기(週期) 수확(收穫) 후까지 배지(培地) 성분(成分)의 소실(消失)을 보면 볏짚배지(培地)는 고형물(固形物) 약 19.7%, 유기물(有機物) 약 19.3%, 질소(窒素) 약 40%가 소실(消失)되었으며, 톱밥 배지(培地)에서는 고형물(固形物) 약 7.5%, 유기물(有機物) 약 7.6%, 질소(窒素) 약 20%가 소실(消失)되었다. 버섯 1kg을 생산(生産)하기 위하여 볏짚 배지(培地)에서는 유기물(有機物) 약 232g, 질소(窒素) 약 7.0g이 소실(消失)되었고, 톱밥 배지(培地)에서는 유기물(有機物) 약 235g, 질소(窒素) 약 6.8g이 소실(消失)되었으며, 버섯 1kg당(當) 함유된 유기물(有機物)은 각각 82.4g, 82.3g, 질소(窒素)는 각각 5.6g, 5.4g이었다. 12. 양배지(兩培地)의 전질소(全窒素)는 점차적으로 감소(減少)되었고 불용성질소(不溶性窒素)의 절대감소량(絶對減少量)은 수용성질소(水溶性窒素)보다 컸으며 아미노태(態) 질소(窒素)는 3주기(週期)까지는 계속 증가 되었으나 그 이후부터는 감소(減少)되었다. 13. 볏짚 배지(培地)에서는 재배기간(栽培其間)에 소실(消失)된 전(全) pentosan의 28%, ${\alpha}$-cellulose는 13.8%가 균사생육(菌絲生育)중에 소실(消失)되었고 톱밥배지(培地)에서는 전(全) pentosan의 24.1%, ${\alpha}$-cellulose는 11.9%가 소실(消失)되었으며 lignin은 양(兩) 배지(培地)의 2주기(週期) 수확(收穫)부터 다소 감소(減少)되었다. 환원당(還元糖), trehalose 및 mannitol은 계속 증가의 추세를 보였으며 C/N율(率)은 볏짚 배지(培地)에서 종균(種菌) 접종시(接種時) 33.2이었던 것이 폐상시(廢床時)에는 30.3이었고, 톱밥 배지(培地)는 61.3이었던 것이 60.0 이었다. 14. 양(兩) 배지(培地)에서 P, K, Mn, Zn은 감소(減少)되었고, Mg, Ca, Cu는 불규칙하게 변화되었으며, Fe는 증가되는 경향이었다. 15. 재배기간(栽培期間)중 각종효소(各種酵素)의 활성(活性)은 톱밥배지(培地)보다 볏짚배지(培地)가 월등히 높았다. 즉 CMC 당화활성(糖化活性)과 CMC액화활성(液化活性)은 균사번식(菌絲繁殖)후부터 2주기수확(週期收穫)까지는 양배지(兩培地)에서 점차적으로 증가 되었으나 그 이후부터는 감소(減少)되었다. xylanase활성(活性)은 1주기(週期)보다 2주기(週期)에서 급격히 상승되었고 3주기(週期)가 되면서 볏짚 배지(培地)에서는 신속히 감소(減少)되었으나 톱밥 배지(培地)에서는 이와같은 감소(減少)를 볼 수 없었다. protease 활성(活性)은 균사번식(菌絲繁殖)후 최고의 활성도(活性度)를 보였다가 점차로 감소(減少)하였다. 또한 볏짚 배지(培地)의 pH는 종균접종시(種菌接種時) 6.3이던 것이 4주기(週期)후는 5.0이었고 톱밥배지(培地)의 pH는 5.7에서 4.9로 떨어졌다. 16. 볏짚 배지(培地)에서 생육한 버섯은 섬유소(纖維素)를 제외한 모든 성분량(成分量)이 톱밥배지(培地)에서 생육한 버섯보다 높은 경향이있었으며 양배지(兩培地)에서 버섯의 각주기별(各週期別) 성분(成分) 변화는 $1{\sim}3$주기(週期)까지는 거의 비슷하였으나 4주기(週期)에서는 다소 감소(減少)의 추세를 보였다.