• Title/Summary/Keyword: Growing substrate

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Effects of Harvest Time on Growth and Phytochemical Contents of Baby Leaf Vegetables in Multi-layer System (다단재배에서 수확시기가 어린잎 채소의 생육과 항산화물질 함량에 미치는 영향)

  • Kim, Jae Kyung;Kang, Ho Min;Kim, Il Seop;Choi, Eun Young;Choi, Ki Yong
    • Journal of Bio-Environment Control
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    • v.26 no.3
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    • pp.194-200
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    • 2017
  • This study aimed to determine the suitable of harvest time on the growth and quality of baby leafy vegetables (Agastsche rugosa O. Kuntze and Lepidium sativum L.) grown on rice seedling tray in a six-layered bench system at 30cm intervals in order to exploit the space during rice growing off-season. Seedlings were grown on the rice seedling tray for 10 days after sowing with coir substrate supplied with nutrient solution at EC $1.5dS{\cdot}m^{-1}$ every 2 days prior to placing the tray on the bench, which were at $1^{st}$ (Low) layer above 20cm and $6^{th}$ (High) layer above 170cm apart from the ground. Growth and phytochemical contents were measured at 7-day and 14-day harvest time. During the culture periods, daily average of integrated solar radiation and temperature were $9.3{\sim}9.6MJ{\cdot}m^{-2}$, $27.5^{\circ}C$ in the High layer and $5.1{\sim}6.2MJ{\cdot}m^{-2}$ in average, and $26.5{\sim}26.6^{\circ}C$ in the Low layer, respectively. For A. rugosa, the highest growth was observed in the Low layer bench at a 14-day harvest time, while their plant height in the High layer was shorter and the leaf number was lower. For L. sativum, the plant height, leaf length and width, leaf number and fresh weight were higher in the Low layer. For A. rugosa, a high yield was observed with the increase in integrated temperature and integrated solar radiation, while a higher yield of L. sativum was found with the increase in integrated temperature, but not with integrated solar radiation. For A. rugosa, both polyphenol and anthocyanin contents were higher in the High layer at a 14-day harvest time. For L. sativum, polyphenol contents were higher in the High layer, whereas no significant difference in anthocyanin and flavonoid contents was observed depending on the layer and harvest time. The highest chlorophyll content showed in Low layer at a 7-day harvest time in both A. rugose and L. sativum. All of the results suggest that in terms of growth and quality, it may be better growing in the high layer for 14 days after seedling in A. rugosa, and low layer for 7 days in L. sativum.

Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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Characterization of a new commercial strain "Noeul" by intra-specific hyphal anastomosis in Pleurotus salmoneostramineus (계통간 교잡에 의한 분홍느타리 품종 "노을"의 육성 및 그 특성)

  • Yoo, Young-Bok;Kong, Won-Sik;Jang, Kab-Yeul;Kim, In-Yeup;Oh, Se-Jong;Jhune, Chang-Sung
    • Journal of Mushroom
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    • v.5 no.1
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    • pp.7-12
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    • 2007
  • All intra-specific hybrids among ASI $2172{\times}2104$, $2172{\times}2307$, $2186{\times}2172$ and $2186{\times}2307$ in P. salmoneostramineus produced pink fruiting bodies as like wild parental types. However, three hybrids of them between ASI 2186 and 2104 were white fruiting bodies. A new commercial strain "Noeul" of Pink Oyster mushroom was developed by intra-specific hyphal anastomosis. It was improved with hybridization between monokaryotic strain derived from ASI 2172 and ASI 2104. The optimum temperature of mycelial growth and fruiting body development were $25{\sim}30^{\circ}C$ and $19{\sim}24^{\circ}C$, respectively. The pileus was bright reddish pink. Commercial strain "Noeul" was as prolific as the more commonly cultivated Pleurotus ostreatus in the conversion of substrate mass to mushrooms using bottle cultivation. Mushroom cultivator can save money for mushroom growing on summer in Korea. Mushrooms should be picked when moderately young, and handled carefully so as to not bruise the brilliantly colored gills. This pink color makes marketing an interesting challenge depending upon the market niche.

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Present status and prospect for development of mushrooms in Korea

  • Jang, Kab-Yeul;Oh, Youn-Lee;Oh, Minji;Im, Ji-Hoon;Lee, Seul-Ki;Kong, Won-Sik
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.27-27
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    • 2018
  • The production scale of mushroom cultivation in Korea is approximately 600 billion won, which is 1.6% of the Korean gross agricultural output. Annually, ca. 190,000 tons of mushrooms are harvested in Korea. Although the numbers of mushroom farms and cultivators are constantly decreasing, the total mushroom yields are increasing due to the large-scale cultivation facilities and automation. The recent expansion of the well-being trend causes increase in mushroom consumption in Korea: annual per capita consumption of mushroom was 3.9kg ('13) that is a little higher than European's average. Thus the exports of mushrooms, mainly Flammulina velutipes and Pleurotus ostreatus, have been increased since the middle of 2000s. Recently, however, it is slightly reduced. However, Vietnam, Hong Kong, the United States, the Netherlands and continued to export, and the country has increased recently been exported to Australia, Canada, Southeast Asia and so on. Canned foods of Agaricus bisporus was the first exports of the Korean mushroom industry. This business has reached the peak of the sale in 1977-1978. As Korea initiated trade with China in 1980, the international prices of mushrooms were sharply fall that led to shrink the domestic markets. According to the high demand to develop new items to substitute for A. bisporus, oyster mushroom (Pleurotus ostreatus) was received the attention since it seems to suit the taste of Korean consumers. Although log cultivation technique was developed in the early 1970s for oyster mushroom, this method requires a great deal of labor. Thus we developed shelf cultivation technique which is easier to manage and allows the mass production. In this technique, the growing shelf is manly made from fermented rice straw, that is the unique P. ostreatus medium in the world, was used only in South Korea. After then, the use of cotton wastes as an additional material of medium, the productivity. Currently it is developing a standard cultivation techniques and environmental control system that can stably produce mushrooms throughout the year. The increase of oyster mushroom production may activate the domestic market and contribute to the industrial development. In addition, oyster mushroom production technology has a role in forming the basis of the development of bottle cultivation. Developed mushroom cultivation technology using bottles made possible the mass production. In particular, bottle cultivation method using a liquid spawn can be an opportunity to export the F.velutipes and P.eryngii. In addition, the white varieties of F.velutipes were second developed in the world after Japan. We also developed the new A.bisporus cultivar "Sae-ah" that is easy to grown in Korea. To lead the mushroom industry, we will continue to develop the cultivars with an international competitive power and to improve the cultivation techniques. Mushroom research in Korea nowadays focuses on analysis of mushroom genetics in combination with development of new mushroom varieties, mushroom physiology and cultivation. Further studied are environmental factors for cultivation, disease control, development and utilization of mushroom substrate resources, post-harvest management and improvement of marketable traits. Finally, the RDA manages the collection, classification, identification and preservation of mushroom resources. To keep up with the increasing application of biotechnology in agricultural research the genome project of various mushrooms and the draft of the genetic map has just been completed. A broad range of future studies based on this project is anticipated. The mushroom industry in Korea continually grows and its productivity rapidly increases through the development of new mushrooms cultivars and automated plastic bottle cultivation. Consumption of medicinal mushrooms like Ganoderma lucidum and Phellinus linteus is also increasing strongly. Recently, business of edible and medicinal mushrooms was suffering under over-production and problems in distribution. Fortunately, expansion of the mushroom export helped ease the negative effects for the mushroom industry.

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Overexpression and Activity Analysis of Cystathionine γ-Lyase Responsible for the Biogenesis of H2S Neurotransmitter (새로운 신경전달물질 H2S 발생 효소, cystathionine γ-lyase의 대량발현 조건과 활성측정)

  • Kim, Kyoung-Ran;Byun, Hae-Jung;Cho, Hyun-Nam;Kim, Jung-Hyun;Yang, Seun-Ah;Jhee, Kwang-Hwan
    • Journal of Life Science
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    • v.21 no.1
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    • pp.119-126
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    • 2011
  • There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

Growth, Quality and Irrigation Requirements of Melon Cultivars in Hydroponic Cultivation Using Coir Substrate (코이어 배지를 이용한 멜론 수경재배 시 품종별 생육, 품질 및 급액 요구량)

  • Lim, Mi Young;Roh, Mi Young;Jeong, Ho Jeong;Choi, Gyeong Lee;Kim, So Hui;Choi, Su Hyun;Lee, Choung Keun
    • Journal of Bio-Environment Control
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    • v.30 no.3
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    • pp.188-195
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    • 2021
  • This study was conducted to investigate the growth and quality characteristics of melon (Cucumis melo L.) cultivars and the irrigation requirements for cultivars. In our previous study in 2019, twelve melon cultivars including 'Dalgona' were examined for their cultivar characteristics under the same irrigation condition for all cultivars, and sorted into several groups based on different growth condition; for the internode length (from 0 to 20th node), leaf area, and fruit weight, 'Kingstar' belonged to the largest group, 'Worldstar' the middle group, and 'Dalgona' the smallest group. After analyzing the results of the previous experiment, 'Dalgona', 'Worldstar', 'Kingstar', and 'Rubyball' were selected as test cultivars for the growth group in 2020, and irrigated according to different irrigation levels for each cultivar. The control of the irrigation volume for each melon cultivar by monitoring the drainage rate during the cultivation periods showed that all four cultivars required a similar amount of irrigation in the 'early growth' stage where crops grew at about the same rate. From 'flowering time', however, the change in irrigation requirements showed a similar tendency for 'Worldstar' and 'Kingstar' and for 'Rubyball' and 'Dalgona' respectively. A sudden change in each irrigation volume was observed from the fruit set; 'Dalgona' began first to decline and 'Rubyball' was second, followed by 'Worldstar' and 'Kingstar'. In conclusion, the irrigation volume was the largest in 'Kingstar', followed by 'Worldstar', 'Rubyball', and 'Dalgona' in the same order as the growing amount of plant length, leaf area, and fruit weight. Therefore, it is necessary to control exactly the irrigation volume by reflecting the unique growth characteristics of each cultivar for the production of high-quality fruit in melon hydroponics, and especially to use great care when different cultivars are cultivated together.

Nanoscale Pattern Formation of Li2CO3 for Lithium-Ion Battery Anode Material by Pattern Transfer Printing (패턴전사 프린팅을 활용한 리튬이온 배터리 양극 기초소재 Li2CO3의 나노스케일 패턴화 방법)

  • Kang, Young Lim;Park, Tae Wan;Park, Eun-Soo;Lee, Junghoon;Wang, Jei-Pil;Park, Woon Ik
    • Journal of the Microelectronics and Packaging Society
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    • v.27 no.4
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    • pp.83-89
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    • 2020
  • For the past few decades, as part of efforts to protect the environment where fossil fuels, which have been a key energy resource for mankind, are becoming increasingly depleted and pollution due to industrial development, ecofriendly secondary batteries, hydrogen generating energy devices, energy storage systems, and many other new energy technologies are being developed. Among them, the lithium-ion battery (LIB) is considered to be a next-generation energy device suitable for application as a large-capacity battery and capable of industrial application due to its high energy density and long lifespan. However, considering the growing battery market such as eco-friendly electric vehicles and drones, it is expected that a large amount of battery waste will spill out from some point due to the end of life. In order to prepare for this situation, development of a process for recovering lithium and various valuable metals from waste batteries is required, and at the same time, a plan to recycle them is socially required. In this study, we introduce a nanoscale pattern transfer printing (NTP) process of Li2CO3, a representative anode material for lithium ion batteries, one of the strategic materials for recycling waste batteries. First, Li2CO3 powder was formed by pressing in a vacuum, and a 3-inch sputter target for very pure Li2CO3 thin film deposition was successfully produced through high-temperature sintering. The target was mounted on a sputtering device, and a well-ordered Li2CO3 line pattern with a width of 250 nm was successfully obtained on the Si substrate using the NTP process. In addition, based on the nTP method, the periodic Li2CO3 line patterns were formed on the surfaces of metal, glass, flexible polymer substrates, and even curved goggles. These results are expected to be applied to the thin films of various functional materials used in battery devices in the future, and is also expected to be particularly helpful in improving the performance of lithium-ion battery devices on various substrates.

Assessment of Methane Production Rate Based on Factors of Contaminated Sediments (오염퇴적물의 주요 영향인자에 따른 메탄발생 생성률 평가)

  • Dong Hyun Kim;Hyung Jun Park;Young Jun Bang;Seung Oh Lee
    • Journal of Korean Society of Disaster and Security
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    • v.16 no.4
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    • pp.45-59
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    • 2023
  • The global focus on mitigating climate change has traditionally centered on carbon dioxide, but recent attention has shifted towards methane as a crucial factor in climate change adaptation. Natural settings, particularly aquatic environments such as wetlands, reservoirs, and lakes, play a significant role as sources of greenhouse gases. The accumulation of organic contaminants on the lake and reservoir beds can lead to the microbial decomposition of sedimentary material, generating greenhouse gases, notably methane, under anaerobic conditions. The escalation of methane emissions in freshwater is attributed to the growing impact of non-point sources, alterations in water bodies for diverse purposes, and the introduction of structures such as river crossings that disrupt natural flow patterns. Furthermore, the effects of climate change, including rising water temperatures and ensuing hydrological and water quality challenges, contribute to an acceleration in methane emissions into the atmosphere. Methane emissions occur through various pathways, with ebullition fluxes-where methane bubbles are formed and released from bed sediments-recognized as a major mechanism. This study employs Biochemical Methane Potential (BMP) tests to analyze and quantify the factors influencing methane gas emissions. Methane production rates are measured under diverse conditions, including temperature, substrate type (glucose), shear velocity, and sediment properties. Additionally, numerical simulations are conducted to analyze the relationship between fluid shear stress on the sand bed and methane ebullition rates. The findings reveal that biochemical factors significantly influence methane production, whereas shear velocity primarily affects methane ebullition. Sediment properties are identified as influential factors impacting both methane production and ebullition. Overall, this study establishes empirical relationships between bubble dynamics, the Weber number, and methane emissions, presenting a formula to estimate methane ebullition flux. Future research, incorporating specific conditions such as water depth, effective shear stress beneath the sediment's tensile strength, and organic matter, is expected to contribute to the development of biogeochemical and hydro-environmental impact assessment methods suitable for in-situ applications.

Studies on nutrient sources, fermentation and harmful organisms of the synthetic compost affecting yield of Agaricus bisporus (Lange) Sing (양송이 수량(收量)에 미치는 합성퇴비배지(合成堆肥培地)의 영양원(營養源), 발효(醱酵) 및 유해생물(有害生物)에 관((關)한 연구(硏究))

  • Shin, Gwan-Chull
    • 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 $60^{\circ}C$.

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