The mannanase, cellulase and xylanase-producing thermophilic bacteria, designated EJ3, was isolated from fresh spent mushroom (Pleurotus eryngii) substrates taken from the DOJUN farm located in Keyongnam, Korea. The isolate EJ3 was facultatively anaerobic and grew at temperature ranging from 20℃ to 60℃ with an optimal temperature of 37℃. The DNA G+C content of the isolate EJ3 was 45 mlo%. The major fatty acids were anteiso-15:0 (38.9%), 17:0 (7.4%), and iso-15:0 (38.2%). The 16S rRNA gene sequence similarity between the isolate EJ3 and other Bacillus strains varied from 97% to 99%. In the phylogenetic analysis based on these sequences, the isolate EJ3 and Bacillus subtilis clustered within a group and separated from other species of Bacillus. Based on the physiological and molecular properties, the isolate EJ3 was classified within the genus Bacillus as Bacillus subtilis EJ3. The optimal growth condition of B. subtilis EJ3 were pH 6.0 and 55℃, respectively.

This study was conducted to evaluate the feeding value of the spent mushroom (Pleurotus eryngii) substrates (SMS) in laying hens (Hy-Line Brown). The fresh spent mushroom (Pleurotus eryngii) substrates collected from the DOJUN farm were fermented with Bacillus subtilis EJ3 for 2 weeks. A total of twenty-four laying hens were fed corn-soy based experimental diets containing 0% (control), 5% (T1), 10% (T2) and 15%(T3) fermented SMS for 7 weeks. There were no significant differences among the treatments in egg production, egg weight, egg mass, feed conversion and viability during the experimental period. Feed intake was significantly lowered in control (118.3 g) than T1 (121.9 g), T2 (120.3 g) and T3 (122.4 g). There were no significant differences among the treatments eggshell breaking strength, thickness and haugh unit, whereas the yolk color of T1, T2 and T3 were significantly heavy than T0. The palatability of boiled meat was significantly better in the T3 laying hens than in the T0 laying hens. In conclusion, fermented SMS can be used as resource of feed in laying hen feed at 5.0-15% level without effect on performance and egg qualify.

The price of mushrooms harvested by bottle cultivation is rapidly dropping and the income of farm households is also rapidly decreasing due to the increase of production cost. Although some of these mushroom farms have employed the systems of mass production, they are in financial difficulty because of the investment they need to do for facilities such as cultivation room and automated systems. In the other hands, some retailers want to buy a small volume of mushrooms of many different mushrooms produced by the small farms and these small farms was required to investigate the common cultural condition for mushroom production of many different mushrooms. In this study, we investigated the common cultural conditions for production of many different mushrooms (e.g., Pleurotus eryngii and Agrocybe cylindracea) at the bottle cultivation farms. The cultural period of Pleurotus eryngii and Agrocybe cylindracea was 30~35 days. The optimum temperature of the mycelial growth was 25~28℃ with the growth room being maintained about 20~23℃ with the consideration of the respiration heat of the mycelium. The temperature of mushroom growth room was 16~18℃ for all growth periods. In our results, Pleurotus eryngii and Agrocybe cylindracea produced the highest yields at the substrate formulation of sawdust 75%, rice brain 20%, soybean cake wastes 5%, water contents 70% and 850 ml P.P. In the long run, our results will result in the development of new automatic cultivation model and increase the income of small production farms.

Microsatellite loci are increasingly used as markers in the human, animal and plant genomes. Being highly mutable, microsatellite regions are able to differentiate between related taxa, even at the level of individual isolates in a single species. Studies on mushroom population structure, gene flow and dispersal between natural and cultivated species have become central in breeding programmes and the knowledge of new polymorphic, codominant markers will be a promising avenue to exploit wild genetic resources. The molecular phylogeny in 50 different commercial cultivated strains of Pleurotus eryngii using PCR amplification with URP primers and mitochondrial microsatellite primer was studed. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD analysis techniques were able to detect genetic variation among the tested strains. With these isolated PCR amplification with URP primers we intend to analyse the population structure of the P. eryngii species complex and investigate the structure of the basidiomycete genome which deserves. A few single-locus microsatellite markers have been isolated in Pleurotus eryngii and Pleurotus ferulae. This technique is useful in those species where microsatellite loci are rare in the mitochondria.

The mating-type genes control formation of the dikaryon from two haploid strains. These genes are now used in mating-type-assisted breeding programs for economically important mushrooms, especially the oyster mushroom, Pleurotus ostreatus, aiming at high-yield and high-quality standard mushroom production. However, it improves the breeding program when the breeder is able to quickly identify compatible strains in a given set of progeny. The two mating factors with their mating-type loci are used as markers for breeding and have been incorporated in a chromosome mapping investigation. The linkage maps include not only genetic markers such as the mating types that can be cored, but also molecular markers such as PCR-assisted approaches, e.g. RAPD analyses, or RFLP markers. Once mating-type genes within progeny may be more easily identified by the use of PCR-directed cloning of partial mating-type genes. We analyzed homeodomain (HD1 and HD2) and pheromone receptor(rcb1, 2 and 3) genes as molecular markers for breeding using mating type A and B of Pleurotus eryngii and Pleurotus ferulae by direct PCR.

This study was carried to investigate the suitable conditions of Pleurotus eryngii through precooling for the more long-term freshness. The time to be same temperatures between P. eryngii and storage room through precooling at 0℃ and 4℃ were showed 2 hours and 5.5 hours, respectively. P. eryngii was cooled within 6 hours and 18 hours at 0℃ and 4℃ with two type, forced air cooling and pressure cooling. After precooling, P. eryngii was packed 400±10g with anti-fog film, and then stored at 4±1℃. In all treatment of precooling, the weight loss was peaked at 25 days and hardness of P. eryngii were decreased at 10 days during storage. There were no significant freshness differences between precooling type and conditions. It was found that the optimum stored period of P. eryngii at 4℃ after precooling was estimated to be 30~35days. Consequently, it is necessary to elucidate efficient and economic precooling conditions.

A new commercial strain "Dan Bi" of Pleurotus eryngii was developed by intra-specific crossing between monokaryotic spores derived from KNR2312 and KNR2596, respectively. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 -16℃. The primordia formation of "Dan Bi" was one day later than that of control strain KNR2312. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has a scale like as cobweb. The yield was 93±4.8g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial strain "Dan Bi" showed a different profile as that of the control strain KNR2312 when RAPD primer 8001, 8002, 8003, 8004, and 8005 were used. This new variety of King Oyster mushroom is characterized by a few primordia formation after scratching. It would be of much help to mushroom cultivators in a process of culling of P. eryngii.

최근 국제원자재가 상승에 따라 버섯 배지 가격이 50% 이상 상승하여 톱밥 대체 배지의 개발이 절실한 실정으로 충남도내에서 생산되는 은행잎부산물을 이용하여 버섯 배지로 개발하여 생산비 절감 및 경제적 이익 창출을 도모하기 위해 수행한 결과를 요약하면 다음과 같다. 은행잎박을 10~30% 첨가량에 따른 균사생장은 대조구 102 ㎜/28일에 비해 은행잎박 10~30% 첨가시 86~101㎜/28일로 균사생장이 느린 경향이었으며, 균배양일수는 은행잎박 10~20% 첨가시 29일로 대조구와 같았다. 초발이 소요일수는 은행잎박 10~20% 첨가시 7일, 생육 일수는 11일로 대조구와 같았지만, 은행잎박 30% 첨가시 초발이소요일수는 1일정도 늦었다. 갓의 크기는 은행잎박 첨가시 대조구에 비해 작은 경향이 있었고, 대의 길이는 은행잎박 10% 첨가시 98㎜로 대조구 91㎜에 비해 길었다. 대의 두께는 대조구 45㎜에 비해 은행잎박 10~30% 첨가시 45~48㎜로 두꺼운 경향이었다. 대의 경도는 대조구 32,156(g/㎠)에 비해 은행잎박 10~30% 첨가시 35,062g/㎠~41,065g/㎠로 높아 저장 및 유통에 유리하였고, 병당 수량은 대조구 85.8g/병에 비해 은행잎박 10~30% 첨가시 88.7~95.6g/병으로 3~11% 증수되었다. 따라서 큰느타리 병재배시 은행잎박 10%를 첨가하는 것이 수량 증수 효과가 있는 것으로 사료된다.

Human GH (hGH) has been available over 40 years for the treatment of children with GH deficiency. Human growth hormone (hGH) is mainly produced in the somatotrophic cells of the pituitary in brain and is the product of the GH-N gene. Among the edible mushrooms, the king oyster mushroom (Pleurotus eryngii ) is one of the most popular mushrooms in Asia, Europe and North America. The increasing popularity of P. eryngii among consumers is due to its flavor, texture and shelf life. We report a modified Agrobacteriummediated method for the efficient transformation of hGH2 in Pleurotus eryngii . The binary vector pCAMBIA1304 was used for the initial transformation and detected by GUS and GFP. Infiltrated samples transformed with pCAMBIA1304 showed a wider GUS response than the co-cultivated in the 50㎍/㎖ hygromycin selection medium. This transformation technique offers new prospects for the production of useful protein in the genetically improved mushroom.

Human recombinant IL-32 induces the production of large amounts of several proinflammatory cytokines and chemokines, even in macrophage cell lines by activation of NF-κB and MAPK. The ability of IL-32 to potentiate inflammation is not the sole action of this cytokine. IL- 32 stimulates prostaglandin (PG)E2 in human PBMCs, which are pivotal in inflammation. The development of recombinant protein consisting of IL-32 is of great medical and industrial importance. Agrobacterium strains GV3101 harboring pCAMBIA1304 include IL- 32 vectors were used to infect the mushroom fruiting body pieces. For transformation experiments, fruiting body gill tissue pieces were vacuum infiltrated with the suspension of induced bacteria until the air had been completely purged. Mushroom mycelium appeared at the margins of the tissue pieces after 9 to 14 days on selection medium with hygromycin at 50㎍/㎖. Transformants PCR analyses confirmed that the IL-32 gene was into the genome of Pleurotus eryngii. The present results demonstrating the usefulness of the transgenic technique s in the genetic manipulation and improvement of mushroom.

This study was conducted to research the effect of Cryptomeria japonica sawdust and branch on the Production of Fruiting body of Pleurotus eryngii. Quantity of fruit body was lower in 5.2things/ bottle but yield of fruit body was higher in 3.5things/ bottle about big fruit body than Douglas-fir sawdust medium on C. japonica sawdust medium, so yield of fruit body was more 45g than Duglas-fir sawdust medium. But in the C. japonica branch sawdust medium, both quantity of fruit body and yield of fruit body was remarkably lower 13.3things/bottle, 42.8g/bottle respectively than Duglas-fir sawdust medium.

큰느타리버섯 재배에 있어 원균의 보존 및 증식기술은 아주 핵심적인 부분으로 취급되고 있다. 이를 위해 대부분 의 농가에서는 상업용 합성배지를 구입하여 사용하고 있 으나 과도한 비용부담은 균주 관리 소홀로 이어져 종균제 조에 있어 많은 피해를 야기 시키고 있다. 본 연구에서는 버섯재배에 이용되는 배지재료를 이용하여 합성배지를 대체할 수 있는 원균증식용 천연배지 제조 가능성에 대해 조사하였다. 각각의 배지 재료를 이용하여 조합한 뒤 열수 추출된 것을 원균증식을 위한 고체배지로 이용한 결과 포 플러:밀기울:미강 (50:20:30, v/v) 또는 포플러:밀기울:미 강 (50:30:20, v/v) 조건에서 합성배지 (36.3mm/7일) 보다 우수한 균사생장능력을 보여준다는 사실을 확인하였다. 이들 천연배지에서 유래된 큰느타리버섯 원균을 이 용하여 버섯재배에 이용한 결과 에서도 균사배양 기간이 23.6일 정도로 합성배지를 이용한 배지 (24.7일) 보다 약 1.0 일정도 단축되었으며, 버섯의 정상적인 생육 및 수확 도 가능 하였다.