This study was aimed to improve the productivity and income of mushroom farming by developing a new casing material as a substitute for clay loam casing soil, which is becoming more difficult to acquire. When the new casing materials were used for the stable production of button mushroom (Agaricus bisporus), a 1:1 mixture of clay loam and button mushroom media obtained after harvest supported 13% greater mycelial growth (32.0 kg/3.3 m²). This material was better than clay loam soil in preventing contamination with environmental compounds and pests. The use of an inexpensive 1:1 mixture of peat moss and coco peat resulted superior mycelial growth with 4% better yield (32.9 kg/3.3 m²) compared with conventional clay loam soil. Advantages of these casing materials included ready availability and improved productivity. Mixtures of peat moss + coco peat + zeolite (50%:30%:20%) and coco peat + coal ash (75%:25%) could substitute for conventional casing soil. Additionally, the novel mixtures containing material obtained after cultivation might be used to produce organic fertilizer.
Good Agricultural Practices (GAP) is the world's leading quality certification for food safety. Since its introduction in Korea in 2006, its importance has been increasing every year. In particular, food safety issues are becoming increasingly important in society, and food safety is directly linked to health. The core of GAP certification is the traceability of the production, distribution, and consumption of hazardous materials, including pesticide residues, heavy metals, and microbes. In the present study, pesticides and heavy metals in button mushroom (Agaricus bisporus) and associated cultivation materials were analyzed. Tricyclozole (0.0144 ppm), flubendiamide (0.147 ppm), and trifloxystrobin (0.0340 ppm) were detected in rice straw and wheat straw, and carbendazim (0.0142 ppm) was detected in mixed wheat straw and rice straw medium. Lead and cadmium were detected at levels higher than the standard level in rice straw and mixed medium. However, lead and cadmium were not detected in mushrooms, and levels of arsenic and mercury were below the safety limit. Therefore, it was confirmed that the residual pesticides and heavy metals are safely managed in the investigated mushroom species. The results of the present study suggest that if these materials are adequately managed in the surroundings during cultivation, all hazardous materials can be managed during mushroom production.
A entomopathogenic ascomycetous fungus, Cordyceps militaris is distributed world wide and cultivated as a medicinal mushroom having many valuable biological and pharmaceutical activities. In the breeding of new C. militaris mushroom, single ascospores were isolated and tested their characteristics such as mycelial growth, mycelial density, fruiting body formation ability, the production of perithecia. Also two opposite mating types were assayed using two sets of primers specific for C. militaris, which were amplified a 191-bp fragment for MAT1-1 and 233-bp fragment for MAT1-1. F1 hybrids resulted in well-developed perithecial fruiting bodies and their crossings were confirmed by the PCR assays. New hybrids were produced showing high quality fruiting bodies in artificial media. Mycelial growth of new strain 'Dowonhongcho 2’ was better on SDAY and at 15-25°C when it was compared with that of 'Yedang 3'. The stromata of new strain were club-shaped and bright orange-red color. Its length was 7.1 cm and the cordycepin content was 0.33% on average. In comparison with 'Yedang 3', the new strain had a yield that was 7% higher and it produced fruiting bodies which were firmer. Days of fruiting body were similar with 50 days from inoculation. This cultivar may serve as a valuable one for artificial cultivation and industrial-scale production of C. militaris.
For heaths and vitality Cordyceps sinensis has been widely used as an important medicinal mushroom. C. militaris is popular for its substitute and can be cultivated artificially. Its new strains have been improved by crossing single spore strains. A bipolar heterothallic fungus, C. militaris has two strains of compatible mating types and they can be differentiated by crossing, fruiting body formation ability and the production of perithecia. Each isolate was determined to be monokaryotic or dikaryotic on the basis of perithecial formation by fruiting tests. This process was also carried out by molecular markers for the detection of two opposite mating types or F1 hybrids using two sets of primers specific for C. militaris amplifying a 191-bp fragment for MAT1-2 and 233-bp fragment for MAT1-1. Therefore the crossing of two compatible mating types resulted in well-developed perithecial fruiting bodies and their crossings were confirmed by the PCR assays for the rapid and specific detection of both MAT1-1 and MAT1-2. This results showed that molecular markers provide an efficient alternative for identification of hybridal status or mating types in the breeding program for artificial cultivation and industrial-scale production of C. militaris.
Button mushroom is one of the popular agricultural products and it is cultivated in 70 countries. It has been cultivated more than 50 years in Korea because it is applied many Korean dishes as nutritional foods. The screening criterion for qualities of button mushroom is very strict and difficult for passing evaluation in Korea; therefore, the mushroom growers are burdened with difficulty in production and consumption in price as well. In this study, the contents of crude fat, fiber, ash, and protein were analyzed in three different qualities of button mushroom as normal, immature, and developed button mushrooms. The contents of crude fat, fiber, ash, and protein were 0.21% 8.68% 0.79% and 2.22% normal product, 0.23% 8.95% 0.86% and 2.41% in immature product, and 0.22% 7.79% 0.82% and 2.22% in developed product, respectively. There was no significant difference in basic nutritional values among the products. Therefore, it is expected that will be loosened the screen criterion of button mushroom, and the producers of button mushroom will be more economically benefits for production as well as the mushroom consumers are more economically consumption for button mushroom by reasonable price.
Button mushroom(Agaricus bisporus) are cultivated on wheat straw or rice straw based compost. In this study different compost formular, sawdust as carbon source and chicken manure as nitrogen source were used for button mushroom cultivation. Two steps of sufficient composting process was required to be successful in the cultivation. 1) the substrates are needed to be mixed periodically under the aerobic condition in the phase I process. 2) the temperature is need to be maintained between 55 and 65? for more than three days in the phase II process. Results showed that button mushroom was possible to grow normally with sawdust based compost. However the mushroom yield was less than that of the conventional compost formular(rice straw or wheat straw). The new compost formular developed from this study may be useful but further research may need to improve button mushroom cultivation more efficiently.
Lactic acid bacteria (LAB) producing phenyllactic acid (PLA), which is known as antimicrobial compound, was isolated from button mushroom bed and the isolated LAB was identified to Lactobacillus casei by 16 rRNA gene sequence analysis. Cell-free supernatant (CFS) from L. casei was assessed for both the capability to produce the antimicrobial compound PLA and the antifungal activity against three fungal pathogens (Rhizoctonia solani, Botrytis cinerea, and Collectotricum aculatum). PLA concentration was investigated to be 3.23 mM in CFS when L. casei was grown in MRS broth containing 5 mM phenylpyruvic acid as precursor for 16 h. Antifungal activity demonstrated that all fungal pathogens were sensitive to 5% CFS (v/v) of L. casei with average growth inhibitions ranging from 34.58% to 65.15% (p < 0.005), in which R. solani was the most sensitive to 65.15% and followed by C. aculatum, and B. cinerea. The minimum inhibitory concentration (MIC) for commercial PLA was also investigated to show the same trend in the range of 0.35 mg mL-1 (2.11 mM) to 0.7 mg mL-1 (4.21 mM) at pH 4.0. The inhibition ability of CFS against the pathogens were not affected by the heating or protease treatment. However, pH modification in CFS to 6.5 resulted in an extreme reduction in their antifungal activity. These results may indicate that antifungal activities in CFS was caused by acidic compounds like PLA or organic acids rather than protein or peptide molecules.
Cordyceps species are important mushrooms traditionally used for heaths and vitality. C. sinensis has been widely used as a medicinal mushroom and C. militaris is popular for its substitute. C. militaris can be cultivated artificially and new strains has been improved by crossing single spore strains. As a bipolar heterothallic fungus C. militaris has two strains of compatible mating types and they can be differentiated by crossing, fruiting body formation ability and the production of perithecia. However this process is very laborious and time consuming to carry out. In this study, molecular markers were designed and used for the detection of two opposite mating types. Two mating types were assayed using two sets of primers specific for C. militaris, which were amplified a 191-bp fragment for MAT1-2 and 233-bp fragment for MAT1-1. After crossing of two compatible mating types F1 hybrids resulted in well-developed perithecial fruiting bodies and their crossings were confirmed by the multiplex PCR assays for the rapid and specific detection of both MAT1-1 and MAT1-2. This results may serve as a efficient process in the breeding program for artificial cultivation and industrial-scale production of C. militaris.
Mushroom flies are one of the most serious threats to mushroom production and cultivation in Korea, and they cause significant losses in yield and quality. In this study, we investigated the occurrence and control of mushroom flies in farms where Agaricus bisporus was cultivated in Chungnam. The cultivation period was more than 7 years in the 32 mushroom farms examined; about 28% were environment-friendly cultivation farms, and 72% were conventional cultivation farms. Mushroom flies were a major pest in about 84% of the examined farms. Most of the mushroom flies were sciarids and phorids (71% and 24%, respectively). The adult flies were found throughout the year, but their occurrence was the highest in September when the temperature increased. In general, the occurrence of mushroom flies in the examined farms was high; however, insect nets and post-crop steaming programs used as basic control methods were not implemented properly. Our findings show that integrated pest management of mushroom flies is urgently required for A. bisporus cultivation in Korea.
Mushroom flies are the most serious insect pest problems affecting the cultivation of button mushroom, Agaricus bisporus Lange, and among them sciarid flies are the most common. This study was set out to determine the occurrence of mushroom flies at the mushroom farming areas in Chungnam. The occurrence of mushroom flies were highest in September as temperature increases causing yield loss of up to 27%. Adult flies invade mushroom farms and oviposit in freshly spawned compost. They spoil the appearance of the fruiting bodies, spread various fungal and bacterial disease, and transmit mites etc. Damage is mainly caused by the larvae, which feed on growing mycelium and developing stipes and caps of mushrooms. Out of 32 examined farms most of cultivation years were more than 7 years and environmental friendly cultivating farms were about 28% and conventional cultivating farms were 72%. About 84% of the examined farms selected mushroom flies as a major pests. It was found that sciarid were 71%, phorid were 24% as main mushroom flies. The occurrence density of mushroom flies were highest in September as temperature increases. The occurrence of mushroom flies at the examined farms were serious in general. However insect nets and the hygiene program as basic control were not effective. These results offer good potential for control and could implement for the integrated pest management.
Single ascospores were isolated and tested their characteristics such as mycelial growth, mycelial density, fruiting body formation ability, the production of perithecia in the breeding of new Cordyceps militaris mushroom. Among them selected isolates were crossed and hybrids were produced showing high quality fruiting bodies in artificial media. New strain 'Dowonhongcho’ was better on SDAY and at 10~25°C when it was compared with that of 'Yedang 3' in mycelial growth. The stromata of new strain were club-shaped and bright orange-red color. Its length was 6.1 cm and the cordycepin content was 0.34% on average. In comparison with 'Yedang 3', the new strain had a yield that was 9% higher and it produced fruiting bodies which were firmer. The optimum temperature for mycelial growth was 22~25°C and the optimum temperature for stroma development was 18~22°C. Days of fruiting body were similar with 45 days from inoculation. This cultivar may serve as a valuable one for artificial cultivation and industrial-scale production of C. militaris.
Spawn is the vegetative growth of the mycelium and serves as the inoculum or seed for cultivating mushrooms. Various cereal grains, such as wheat, millet, barley, sorghum, brown rice, rye, and oat were assessed to compare their characteristics and suitability for spawn production of button mushroom (Agaricus bisporus). The mycelial growth rates, density, and the number of completely colonized grains were measured from the twentieth day of inoculation. Wheat grains showed fastest mycelial growth with 8.4 cm followed by rye, oat, barley with 8.2, 7.5 and 7.3 cm, respectively. In the mycelial density, foxtail millet, barley, and sorghum were best compared with that of wheat grains. Especially, the number of grains which were completely colonized by mycelia were greatest in foxtail millet with 5,123 grains followed by proso millet, and wheat with 3,052 and 914, respectively. Based on the results obtained, barley, foxtail millet, and sorghum grains would be appropriate substituting for wheat grain in spawn production of button mushroom.
The effects of environmental and nutritional requirement such as temperature, pH, different media, carbon, nitrogen, and carbon-to-nitrogen ratio on the mycelial growth of Cordyceps militaris strain ‘Yedang 3’ were studied. The optimum temperature and pH for the growth of mycelium were 20-25oC and pH 6-7, respectively. Out of ten media tested, mushroom complete media (MCM) was the best medium for fast mycelial growth, and Sabouraud’s dextrose agar yeast extract (SDAY), malt extract yeast extract agar (YMA) also were favored. The color and shape of colonies varied in different media. The best carbon sources for mycelial growth were fructose, mannitol, and sucrose, whereas the best nitrogen sources were tryptone and peptone. However, mycelia grew slowly in inorganic nitrogen compounds such as NH4Cl, (NH4)2SO4, NH4NO3, and NaNO3. The optimum C:N ratio observed on the culture media was 30-40 range. These results provided basic information on cultural characteristics of vegetative growth and might be useful for spawn production in Cordyceps militaris.
The button mushroom (Agaricus bisporus) is one of the most widely cultivated important edible mushroom species. In the breeding of new button mushroom, ‘Seolgang’ was developed by crossing two monokaryons ‘CM020913-27’ and ‘SSU423-31’. Because of the secondarily homothallism, only a small percentage of the basidia produce 3 or 4 spores, which are mostly haploid (n) and do not fruit. Single spore cultures derived from these types of spores produce a vegetative mycelium that also contain a variable number of genetically identical nuclei per cell called monokaryon. The lack of clamp connections between monokaryon and dikaryon required a series of mycelial culture and fruiting test. After crossing, hybrids were cultivated on a small scale and on a commercial scale at a farm. For this, the spawn was made by a commercial spawn producer and the spawned compost by a commercial compost producer. Mycelial growth of ‘Seolgang’ on CDA was better at 20℃ and 25℃ when it was compared with that of ‘505 Ho’. The mature cap shape of new strain ‘Seolgang’ is oblate spheroid and the immature cap shape is round to oblate spheroid. The cap diameter was 41.2 mm on average. In comparison with white strain ‘505 Ho’, the strain had a yield that was 9% higher. It produced fruiting bodies which had a higher weight on average per fruiting body and were 19% firmer with a good shelf life. Days of fruiting body were 3-4 days later than those of ‘505 Ho’. The physical characteristics such as elasticity, chewiness, adhesiveness were better than that of ‘505 Ho’. Genetic analysis of the new strain ‘Seolgang’ showed different profiles compared to ‘505 Ho’, CM02913-27, SSU413-31, when RAPD primers A02 and O04 were used.
Pleurotus ostreatus ‘Miso’ is a mutant strain showing white color in pileus from the known parent strain ‘Wonhyeong 1’. Shape and several other characters also vary with culture conditions. Mating experiments were performed to understand interstrain mating relationship using monokaryons of the parent and the mutant strains. All monokaryons were grown from single spores isolated from freshly collected fruit bodies. Pairings were performed in 90 mm petri dishes on PDA. They were allowed to grow at 25 until two fronts of the advancing mycelia met and developed a conspicuous contact zone. The contact zone and the outer edges of paired colonies on each plate were examined for clamp connections. The parent and the mutant resulted in tetrapolar incompatibility in intrastrain crosses. In interstrain crosses, each monokaryotic tester strain of the parent strain was out-crossed to monokaryotic tester strains of the mutant. As a result of these crosses it was found that both strains share the same A and B incompatibility factors yielding 25% compatibility.
두 종류의 목질진흙버섯인 P. linteus 와 P. baumii 균주를 가지고 뽕나무, 참나무, 느릅나무 등 3가지 재배원목을 이용하여 자실체를 발생시킨 후 무기물의 함유량을 ICP 법으로 정량하였다. Ca은 2종의 균주 모두 뽕나무에서 발생한 자실체의 함유량이 300 ㎎/100 g 이상으로 참나무나 느릅나무에서 발생한 자실체보다 2~4배 높았으며, K은 2종의 균주 모두 참나무에서 발생한 자실체의 함유량이 600 ㎎/100 g 이상으로 전체 무기물 중 가장 많은 함유량을 보이며 참나무 및 느릅나무에서 발생된 자실체보다 2배 이상 높았다. 특히 Na은 2종 균주 모두 참나무에서 발생된 자실체가 10배 수준 이상 월등히 높게 함유되어 있어서 Ca, K, Na의 함유량 분석이 재배 원목을 구분하는데 매우 유용한 것으로 기대한다. 또한 P. linteus와 P. baumii의 구분에는 linteus는 Fe의 함유량이 참나무 재배 자실체에서 유의성게 높았으며, baumii는 P의 함유량이 참나무 재배 자실체에서 200 ㎎/㎏ 으로 2배 이상 높게 검출되어 Fe와 P의 분석만으로 균주를 구분할 수 있을 것으로 기대한다. P은 2종의 균주, 3종의 재배 원목에서 모두 80 ㎎/㎏ 이상 함유되었으나 다른 버섯류의 무기성분 연구에서는 분석되지 않은 무기물로 목질진흙버섯에만 함유되어 있는지는 앞으로 더 연구가 필요하다고 판단한다. 그 밖의 Cr, Mn, Cu, Zn의 분석으로는 극미량이 함유되어 있어 이들 성분으로는 균주 및 재배원목의 구분이 현실적으로 어려울 것으로 판단되었다. 이상의 결과로 볼 때 ICP를 이용한 무기물 함유량 분석이 P. linteus와 P. baumii 2가지 균주를 가지고 발생된 뽕나무, 참나무 그리고 느릅나무 자실체의 사용균주와 재배원목을 구분하는데 매우 유용한 자료가 될 수 있을 것으로 기대한다.
We performed the present study to investigate whether Rehmannia glutinosa Libosch (RG) extracts (RGX) and Eleutherococcus senticosus Max (ES) extracts (ESX) play any roles in bone metabolism. We examined cellular activities of bone cells by measurement of osteoblastic cell viability, osteoprotegerin (OPG) secretion from osteoblasts, osteoclastogenesis, and osteoclastic activity. There is no cytotoxicity from osteoblasts after treatment with RGX and ESX. The secretion of OPG from the osteoblasts showed marked increases after treatment with RGX and ESX. In addition, RGX and ESX treatment decreased the number of tartrate-resistant acid phosphatase-positive multinucleated cells and the resorption areas. RGX and ESX, when mixed at optimal ratios, induced synergic effects, in vitro. OPB, which showed synergic effects, is the extract of natural ingredients RG and ES mixed at a raw material weight ratio of 4 : 1. It can be suspected that extracts of RG and ES mixtures contains active ingredients involved in bone tissue metabolism and may be effective in improving osteoporosis.