We have used bulked segregant analysis to screen the strain-specific DNA marker associated psychrophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 490 bp was amplified with OP-L18 primer from the psychrophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-L18-1. The PCR analysis with the OP-L18-1 primer showed that this SCAR marker clearly distinguish the psychrophilic strains from the control strains.

We have used bulked segregant analysis to screen the strain-specific DNA marker associated thermophilic strain of Pleurotus eryngii. Bulked genomic DNAs of Pleurotus eryngii were amplified by randomly amplified polymorphic DNA (RAPD) using OP-A, OP-B, OP-L, OP-P, OP-R and OP-S primers to screen the strain-specific DNA marker. A unique DNA fragment of 500 bp was amplified with OP-A11 primer from the psychrophilic strain and sequenced. A sequence characterized amplified region (SCAR) marker was designed on the basis of the determined sequence and named as OP-A11-1. The PCR analysis with the OP-A11-1 primer showed that this SCAR marker clearly distinguish the psychrophilic strains from the control strains.

Spent mushroom substrate (SMS) is a by-product remaining after a crop of mushrooms. The fresh SMS was sample within 24 hours of removal from the production facility in DOJUN farm located in Jinju. About 11 bacterial species were isolated from fresh SMS on TSA medium. Among of them, one isolate, designated YJ02, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain YJ02 was produced cellulase, xylanase, mannanase as hydrolase. The strain YJ02 was identified as members of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain YJ02 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with 16S rDNA gene sequence similarity of 98.7%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain YJ02 was classified within the genus Bacillus, for which the name Bacillus amyloliquefaciens YJ02 is proposed.

The Hypsizigus marmoreus has been used as medicinal and food source in worldwide. Viridans Streptococci, which include Streptococcus sanguinis and Streptococcus gordonii, are pioneer oral bacteria that initiate dental plaque formation. This study was to investigate the antimicrobial activity of Hypsizigus marmoreus Extracts. Antimicrobial activity of extracts were tested against pathogenic bacteria (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus sanguinis and Streptococcus gordonii) by paper disc methods. The Hypsizigus marmoreus Extracts were extracted with 80% methanol. The 80% methanol extracts of Hypsizigus marmoreus Extracts showed antimicrobial activity against S. sanguinis and S. gordonii. The 80% methanol extract of Hypsizigus marmoreus Extracts were fractioned by hexane, chloroform, ethylacetate and buthanol. The ethylacetate fraction of Hypsizigus marmoreus Extracts showed strong antimicrobial activity against S. sanguinis and S. gordonii with minimum inhibitory concentration (MIC) range of 0.8-1 mg/ml. Especially, the 80% methanol extract and ethylacetate fraction of Hypsizigus marmoreus Extracts were inhibited the biofilm formation of S. sanguinis and S. gordonii at a concentration of 1.0 mg/ml. These results suggest that Hypsizigus marmoreus extracts can be developed as a potent antimicrobial agent.

The mushrooms have been used as traditional medicines and food resources in many countries. The objective of this study was to determine antioxidant compounds and to evoluate tyrosinase inhibitory activity of extracts from Hypsizigus marmoreus. The pileus and stipe of Hypsizigus marmoreus were extracted with methanol and water, separately. The methanol extract of Hypsizigus marmoreus were fractioned by hexane, chloroform, ethylacetate and buthanol. The concentrations of total polyphenolics and flavonoids in methanol extracts were investigated by colorimetric methods. The concentrations of total polyphenolics and flavonoids in methanol extract of the pileus was higher than methanol extract of the stipe. The DPPH redical scavenging activity of the pileus extract was also higher than stipe extract in methanol extract. The IC50 of DPPH redical scavenging activity of the pileus and stipe in methanol extract were 18 mg/ml and 1 10 mg/ml, respectively. The IC50 of tyrosinase inhibitory activity of the pileus and stipe in methanol extract were 500 mg/ml and 1,000 mg/ml in methanol extract. These results suggest that Hypsizigus marmoreus can be potentially used as a source of natural antioxidant agent in the cosmetic industry as well as the food, pharmaceutical and medicinal industry.

In order to isolate thermophilic compost-promoting bacteria with high activity of cellulase and xylanase, spent mushroom substrates with sawdust were collected from mushroom cultivation farm, Jinju, Gyeongnam in Korea. Among of the isolates, one strain, designated HJ01 was selected by agar diffusion method. The strain HJ01 was identified as members of the Bacillus subtilis by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain HJ01 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus subtilis with 16S rDNA gene sequence similarity of 99.3%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain HJ01 was classified within the genus Bacillus, for which the name Bacillus subtilis HJ0 is proposed. The cellulase and xylanase activity of B. subtilis HJ0 was slightly increased according to bacterial population from exponential phase to stationary phase in growth curve for B. subtilis HJ01.

Spent mushroom substrate(SMS) is a by-product remaining after a crop of mushrooms. About 9 thermophilic strains were isolated from SMS(Flammulina velvtipes). Among of them, one isolate, designated UJ09, showed the antifungal activity against Aspergillus flavus and Aspergillus ochraceous producing mycotoxin on PDA medium, potentially. The strain UJ09 was produced cellulase and xylanase as hydrolase. The strain UJ21 was identified as members of the genus Bacillus by biochemical characteristics using Bacillus ID kit and VITEK 2 system. Comparative 16S rDNA gene sequence analysis showed that strain UJ09 formed a distinct phylogenetic tree within the genus Bacillus and was most closely related to Bacillus amyloliquefaciens with 16S rDNA gene sequence similarity of 99.2%. On the basis of its physiological properties, biochemical characteristics and phylogenetic distinctiveness, strain UJ09 was classified within the genus Bacillus, for which the name Bacillus amyloliquefaciens UJ09 is proposed.

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 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.

배추좀나방과 담배거세미나방 유충에 대한 30종 한방 식물체 메탄올 조추출몰의 살충활성 및 섭식 저해활성을 잎침지법을 이용하여 5,000ppm으로 검정하였다. 황련의 추출물은배추좀나방에 대해 살충작용을 보였다. 길경, 사삼, 세신, 오배자 및 자초의 추출물은 배추좀나방 유충에 대해, 목통 및 속새의 추출물은 담배거세미나방 유충에 대한 강한 섭식저해활성을 보였으나, 오배자와 황련의 추출물은 이들 나비목 유충 모두에 강한 섭식저해활성을 나타내었다.