Pleurotus속 균주들의 미토콘드리아 플라스미드 특성

김은경,구용범,차동렬,하영칠,노정혜 한국미생물학회 1993 미생물학회지 Vol.31 No.2

백색 부후균인 plaurotus ostreatus 의 4가지의 균주로부터 각각 10.2 kb 와 7.2 kb (NFFA 2), 두 종류의 10.2 kb (NFFA 4001) 11.2 kb (NFFA 4501), 10.2kb 와 11.2 kb(KFCC 11635) 크기의 미토콘드리아 플라스미드들을 분리해 내었다. NFFA 2 의 변종인 NFFA 2m1 과 NFFA 2m2 에서는 이들 플라스미드가 관찰되지 않았다. 분별 원심분리에 의해 얻은 미토콘드리아에서 핵산을 추출하여 agarose gel 에서 전기영동시키면 플라스미드가 관찰되지 않았으나 proteinase K 를 처리하고 핵산을 추출하여 전기영동한 결과 이들 플라스미드가 관찰되었는에, 이는 플라스미드상에 단백질인 결합되어 있음을 시사한다. Proteinase K 를 처리한 플라스미드 DNA 와 exonuclease 를 반응시킨 결과, 이들 플라스미드들은 5'말단에 단백질이 결합된 성형 이중가닥 DNA의 구조를 가진 것을 확인하였다. 각 플라스미드들의 상호관계를 조사하기 위하여 Southern hybridization 을 수행한 결과 최소 3가지 종류의 플라스미드들로 분류할 수 있었다. 이중 한 그룹 (group I) 은 모든 P. ostreatus 균주들에서 공통적으로 발견되었다. Pleurotus 속의 5가지 다른 종(P. cornucopiae, P. florida, P. pulmonarius, P. sajor-cuja, P. spodoleucus) 의 균주들로부터 미토콘드리아 플라스미드들을 분리하였다. 이들은 한균주당 1-4 개 까지의 플라스미드를 가지며, 플라스미드의 크기는 7.2 kb-14kb 범위에 있었다. P. ostreatus 의 NFFA 2 의 10.2 kb(group I) 플라스미드와 hybridization 을 수행한 결과 P. cornucopiae ASI 2011을 제외한 다른 모든 균주들이 유사한 염기서열의 플라스미드를 갖고 있음을 알았다. Plasmid DNAs were detected from the mitochondrial fraction of four strains of whiterot fungus, Pleurotus ostreatus. The size of the plasmids were 10.2 and 7.2 kb in strain NFFA 2, 10.2 kb in NFFA 4001, 11.2 kb in NFFA 4501, and 10.2 and 11.2 kb in KFCC 11635. The two strains,NFFA 2ml and NFFA 2m2, which are mutant derivatives of NFFA 2, did not contain any plasmids. The cleavage by proteinase K indicated that these plasmids have DNA ends associated with proteins. In digestion with proteinase K all the plasm ids remained resistant to lambda exonuclease which hydrolyzes DNA from 5' ends and were sensitive to exonuclease III which hydrolyzes DNA from 3' ends. This suggests that the plasmids are linear double-stranded DNA and the terminal proteins are covalently linked to 5' ends of plasm ids. In order to find relationship between these plasmids, hybridization of plasm ids by each separate plasmid DNA was done. The result indicated that the plasmids can be classified into at least 3 groups. Plasmids of group I were present in all the P ostreatus. More mitochondrial plasmids were detected in P cornucopiae. P ,florida, P pulmonarius, P sajor-caju, and P spodoleucus. The size of plasmids ranged between 7.2 kb and 14 kb. All the species except P cornucopiae contained plasmids of approximately 10 kb which hybridized with the 10.2 kb plasmid (group I) of P ostreatus NFFA 2.

산느타리(Pleurotus pulmonarius) 병 · 봉지재배에 적합한 배지조성 연구

이재홍,이남길,박영학,문윤기,정태성,권순배,김재록,김진원 한국버섯학회 2014 한국버섯학회지 Vol.12 No.4

Pleurotus pulmonarius usually grows on the dead wood of deciduous trees from mid summer to early fall. Cultivation of this mushroom is recently increasing due to its good flavour and fragrance. This study was conducted to find out the favorable substrate formulation in the bottle and bag cultivation of P. pulmonarius. The V formulation (10 : 50 : 20 : 20 mixing ratio of poplar sawdust, cottonseed hull, cottonseed meal and beet pulp) showed the fastest mycelial growth in the test tube column medium and highest yields in both bottle cultivation and bag cultivation. Those chemical conditions were pH 5.52, 29.07% organic matter contents, 1.12% total nitrogen, 25.95 OM/N ratio, 0.16% CaO, 0.51% K2O, 0.17 MgO and 0.26 P2O5.

산느타리버섯(Pleurotus pulmonarius) 수확후배지의 첨가량에 따른 수량특성

이남길,이재홍,문윤기,정태성,권순배 한국버섯학회 2015 한국버섯학회지 Vol.13 No.4

산느타리버섯 ‘호산’, ‘화산’ 품종을 대상으로 강원도농 업기술원에서 개발한 배지와 농가에서 사용하는 배지를 가지고 수확후배지 첨가율에 따른 생육 및 수량특성 조사 결과, 강원도에서 개발한 배지에서는 호산품종에서는 대조구에 비해 수량 차이가 거의 없었고, 화산에서는 20% 까지 수량이 증가하였다. 농가에서 사용하는 배지에서는 호산품종은 20%까지 수량 차이가 없었고 30%에서 다소 적어진 것으로 나타났다. 화산품종에서는 첨가율이 높을 수록 수량이 점차 떨어지는 경향이 나타났다. 이 시험연 구를 통해 산느타리버섯 수확후배지를 20%까지 재사용한 다면 생산비 절감 뿐만 아니라 폐배지 폐기처리 비용 절 감 등 농가소득에 기여할 것으로 여겨진다. This study was carried out to re-use the post-harvest substrate of Pleurotus pulmonarius. In this study, we used two mixing ratio. First, a mixing ratio is developed by Gangwon Province Agriculturl Research and Extension Services[Poplar sawdust(10)+Cottonseed hull(50)+Cottonseed mal(20)+Beet pulp(20)]. Second, mixing ratio is developed by farmers[Poplar sawdust(60)+Cottonseed hull(10)+Cottonseed mal(10)+Beet pulp(20)]. First mixing ratio research results, Hosan was no difference in the yield by 30%, Yield of the Hwasan has increased by 20%. Second mixing ratio research results, Hosan was no difference in the yield by 20%. However, Hwasan has decreased.

산느타리버섯(Pleurotus pulmonarius) 수확후배지의 첨가량에 따른 수량특성

이남길,이재홍,문윤기,정태성,권순배 한국버섯학회 2015 한국버섯학회지 Vol.13 No.4

This study was carried out to re-use the post-harvest substrate of Pleurotus pulmonarius. In this study, we used two mixing ratio. First, a mixing ratio is developed by Gangwon Province Agriculturl Research and Extension Services[Poplar sawdust(10)+Cottonseed hull(50)+Cottonseed mal(20)+Beet pulp(20)]. Second, mixing ratio is developed by farmers[Poplar sawdust(60)+Cottonseed hull(10)+Cottonseed mal(10)+Beet pulp(20)]. First mixing ratio research results, Hosan was no difference in the yield by 30%, Yield of the Hwasan has increased by 20%. Second mixing ratio research results, Hosan was no difference in the yield by 20%. However, Hwasan has decreased.

산느타리버섯 원목재배 시 수종별 생육 및 수량특성

한국버섯학회 한국버섯학회 2016 한국버섯학회지 Vol.14 No.3

This study was carried out to develop techniques for thelog cultivation of Pleurotus pulmonarius. Soil landfill cultivation of the in plastic container boxes containing yield per log thanbut there was no difference in the yield from both spawn Hyangsan” variety. In the caseof soil landfill cultivation in a shaded vinyl house, an Ailanthus tree gave a higher yield than that usingpoplar tree, and the yield of the Hosan” variety washigher than that of the Hyangsan” variety. With regard to proper tree species selection, willow and cherry trees were good for both the Hosan” and “Hyangsan” variet.

산느타리버섯 원목재배 시 수종별 생육 및 수량특성

이재홍,이남길,문윤기,정태성,권순배,박영학,김인종 한국버섯학회 2016 한국버섯학회지 Vol.14 No.3

This study was carried out to develop techniques for thelog cultivation of Pleurotus pulmonarius. Soil landfill cultivation of the in plastic container boxes containing yield per log thanbut there was no difference in the yield from both spawn Hyangsan” variety. In the caseof soil landfill cultivation in a shaded vinyl house, an Ailanthus tree gave a higher yield than that usingpoplar tree, and the yield of the Hosan” variety washigher than that of the Hyangsan” variety. With regard to proper tree species selection, willow and cherry trees were good for both the Hosan” and “Hyangsan” variet.

산느타리(Pleurotus pulmonarius) 품종의 초위성체(simple sequence repeats) 특성구명

최종인,나경숙,오민지,류재산,Choi, Jong In,Na, Kyeong Sook,Oh, Min-Ji,Ryu, Jae-San 한국버섯학회 2021 한국버섯학회지 Vol.19 No.4

국내의 주요 산느타리 품종인 호산47(일핵, 산타리 배우자), GB19(일핵, 산타리 배우자), 호산, 여름느타리1호, 삼복, 강산, 약산, 자산, 향산, 여름느타리2호의 유전체를 Hiseq을 이용하여 해독하였고 이 서열 정보에서 SSR을 분리하여 특성구명을 하였다. 일핵균사인 호산 47, GB19의 유전체의 크기는 각각 37.3와 37.2 Mbp이고, 이핵균사인 나머지 산느타리 품종의 유전체 크기는 47.1~61.1 Mbp인 것으로 밝혀졌다. 품종별 총 SSR의 수는 HS47이 711개로 가장 적고, 강산이(GS)이 1.5배 많은 1,106개로 최다를 기록하였다. SSR의 repeat motif 중에서 hexanucleotide와 octanucleotide가 가장 많은 빈도로 관찰되었고, 가장 많이 관찰되는 반복서열은 CGA/TCG, A/T, CTC/GAG이었다. SSR의 길이는 모든 품종에서 변이가 많아 유용성이 높은 20~30 nt가 가장 높은 비중인 70%를 차지하였다. Simple sequence repeats (SSRs) were isolated from major Pleurotus pulmonarius cultivars in Korea, namely 'HS47' (monokaryon, gamete of 'Santari'), 'GB19' (monokaryon, gamete of 'Santari'), 'Hosan,' 'Yeoleumneutali1,' 'Sambok,' 'Gangsan,' 'Yaksan,' 'Jasan,' 'Hyangsan,' and 'Yeoleumneutali2,' and characterized via HiSeq genome sequencing and bioinformatic analysis. The genome sizes of the monokaryons 'HS47' and 'GB19' were estimated to be 37.3 and 37.2 Mb, respectively, and those of the other dikaryotic cultivars ranged from 47.1 to 61.1 Mb. A total of 711 (smallest) and 1,106 (1.5 times the smallest) SSRs were found in the 'HS47' and 'Gangsan' genomes, respectively. Hexanucleotide and octanucleotide motifs accounted for the top two fractions of all SSRs. CGA/TCG, A/T, and CTC/GAG were the most frequently detected nucleotides in the SSRs. Most of the SSRs were 21~30 nucleotides long (hypervariable for application), accounting for 70% of all lengths of SSRs.

Evaluation of Antioxidant, Anti-cholinesterase, and Anti-inflammatory Effects of Culinary Mushroom Pleurotus pulmonarius

( Trung Kien Nguyen ),( Kyung Hoan Im ),( Jaehyuk Choi ),( Pyung Gyun Shin ),( Tae Soo Lee ) 한국균학회 2016 Mycobiology Vol.44 No.4

Culinary mushroom Pleurotus pulmonarius has been popular in Asian countries. In this study, the anti-oxidant, cholinesterase, and inflammation inhibitory activities of methanol extract (ME) of fruiting bodies of P. pulmonarius were evaluted. The 1,1-diphenyl-2-picryl-hydrazy free radical scavenging activity of ME at 2.0 mg/mL was comparable to that of butylated hydroxytoluene, the standard reference. The ME exhibited significantly higher hydroxyl radical scavenging activity than butylated hydroxytoluene. ME showed slightly lower but moderate inhibitory activity against acetylcholinesterase (AChE) and butyrylcholinesterase than galantamine, a standard AChE inhibitor. It also exhibited protective effect against cytotoxicity to PC-12 cells induced by glutamate (10~100 μg/mL), inhibitory effect on nitric oxide (NO) production and inducible nitric oxide synthase protein expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, and carrageenan-induced paw edema in a rat model. Highperformance liquid chromatography analysis revealed the ME of P. pulmonarius contained at least 10 phenolic compounds and some of them were identified by the comparison with known standard phenolics. Taken together, our results demonstrate that fruiting bodies of P. pulmonarius possess antioxidant, anti-cholinesterase, and inflammation inhibitory activities.

원형질체융합 기법을 이용한 산느타리 계통육성

권희민,이윤혜,김정한,백일선,강희완,최종인,Gwon, Hee-Min,Lee, Yun-Hae,Kim, Jeong-Han,Baek, Il-Sun,Kang, Hee-Wan,Choi, Jong-In 한국버섯학회 2021 한국버섯학회지 Vol.19 No.3

원형질체 융합 기술은 종·속간 유전적 한계를 넘어 육종과 그 소재로 활용하고데 목적이 있다. 본 연구에서는 '흑타리'(P. ostreatus)와 '호산'(P. pulmonarius)의 단핵균사를 이용하여 원형질체를 나출하고 나출된 원형질체를 융합하여 종간 교배 계통을 육성하였다. 육성계통의 균사생장속도는 '호산', '흑타리', PF160313, PF160306 계통 순으로 빠른 편이었다. 균사 밀도는 PF160306 계통이 가장 높았고, 나머지는 중간 수준의 밀도를 나타내었다. 원형질체 융합계통인 PF160306과 PF160313 계통은 '흑타리' 품종보다 배양 기간이 10일, '호산' 품종보다 2일 단축되었다. 자실체 생장 기간은 '흑타리'와 '호산에 비하여 각각 3일, 1일 단축되었다. PF160306 계통의 생산량은 135.9 g/병으로 '호산'에 비하여 높았으나 통계적으로 유의차가 없었다. 자실체 발생기간은 15℃에서 9일, 25℃에서 4.5일로 온도가 높아짐에 빨라졌다. 자실체의 갓색은 21℃ 노란색이 가장 선명하게 발현되었다. URP primer 7을 사용하여 PCR 밴드 패턴을 비교하였을 때, 전체적으로 '호산' 품종과 유사하였다. DPPH radical 소거능과 폴리페놀 함량에 있어 '순정'은 각각 62.5%, 43.5 mg/mL였으며, PF160313 계통은 각각 65.7%, 49.9 mg/mL를 나타내어 계통간 유의차가 있었다. ACE 활성은 '순정' 74%, PF160313 계통 75%로 유사한 수준이었다. In this study, monokaryons of "Heukari" (Pleurotus ostreatus) and "Hosan" (Pleurotus pulmonarius) were separated to remove the cell wall, and a cross-species protoplast fusion was developed through chemical treatment with polyethylene glycol. The protoplast-fused PF160306 and PF160313 strains have a culture period of 10 and 2 days shorter than that of the "Heuktari" and "Hosan" cultivars, respectively. Furthermore, the growth of the strains was faster than that of the existing cultivars. The yield was 135.9 g per bottle, which was approximately 8% higher than that of the commercially available "Hosan" cultivar; however, it was not statistically significant. A growth survey was conducted after treatment at five temperatures (15, 18, 21, 23, and 25℃). The growth of the strains accelerated with the increase in temperature. However, at 21℃, the yellow color of pileus was the brightest. Band pattern, assessed using URP Primer 7, was similar to that of the "Hosan" cultivar. The DPPH radical scavenging capacity and polyphenol content were 62.5% and 43.5 mg/mL, respectively, for "Sunjung" and 65.7% and 49.9 mg/mL, respectively, for PF160313. Furthermore, the antihypertensive activities of the "Sunjung" cultivar and PF160313 were similarly high at 74% and 75%, respectively. In conclusion, cross-species hybridization via the protoplast fusion technique can be used for obtaining primary data for mushroom breeding to develop new varieties. In addition, the protoplast fusion technique might aid in expanding the market for yellow mushrooms.

Upcycling the Spent Mushroom Substrate of the Grey Oyster Mushroom Pleurotus pulmonarius as a Source of Lignocellulolytic Enzymes for Palm Oil Mill Effluent Hydrolysis

( Nurul Anisa Mat Yunan ),( Tan Yee Shin ),( Vikineswary Sabaratnam ) 한국미생물생명공학회(구 한국산업미생물학회) 2021 Journal of microbiology and biotechnology Vol.31 No.6

Mushroom cultivation along with the palm oil industry in Malaysia have contributed to large volumes of accumulated lignocellulosic residues that cause serious environmental pollution when these agroresidues are burned. In this study, we illustrated the utilization of lignocellulolytic enzymes from the spent mushroom substrate of Pleurotus pulmonarius for the hydrolysis of palm oil mill effluent (POME). The hydrolysate was used for the production of biohydrogen gas and enzyme assays were carried out to determine the productivities/activities of lignin peroxidase, laccase, xylanase, endoglucanase and β-glucosidase in spent mushroom substrate. Further, the enzyme cocktails were concentrated for the hydrolysis of POME. Central composite design of response surface methodology was performed to examine the effects of enzyme loading, incubation time and pH on the reducing sugar yield. Productivities of the enzymes for xylanase, laccase, endoglucanase, lignin peroxidase and β-glucosidase were 2.3, 4.1, 14.6, 214.1, and 915.4 U g^-1, respectively. A maximum of 3.75 g/l of reducing sugar was obtained under optimized conditions of 15 h incubation time with 10% enzyme loading (v/v) at a pH of 4.8, which was consistent with the predicted reducing sugar concentration (3.76 g/l). The biohydrogen cumulative volume (302.78 ml H₂.L^-1 POME) and 83.52% biohydrogen gas were recorded using batch fermentation which indicated that the enzymes of spent mushroom substrate can be utilized for hydrolysis of POME.