큰느타리버섯 주요 수출시장인 유럽과 북미시장의 소비자에게 선호도가 높은 갓이 큰 형태의 큰느타리버섯 품종을 육종하기 위하여 육종모본 KNR2555를 자식교배하여 소규모 시험재배하여 갓형(Convex)와 갓직경(60.7 mm), 품질(4.9)을 기준으로 2×12계통을 선발하였다. 선발된 계통을 갓애린이라고 명명하고 대량재배로 큰느타리2호와 생육특성을 비교하였다. 병당수량은 갓애린이가 71.7 g으로 대조품종 71.4 g과 통계적 유의성 없었다. 품질의 경우 갓애린이는 6.8, 큰느타리2호는 6.5로 나타났다. 생육소요일, 길이, 갓직경은 독립 t test로 분석한 결과 통계적으로 유의성을 보였다(각각 P < 0.001, P < 0.05, P < 0.001, P < 0.05). 고유성에 있어서는 URP1와 URP10에서 대조 품종과 신품종이 다형성을 보였고, 대치배양에서도 뚜렷한 대선이 관찰되었다.

생육이 빠르고 중형인 큰느타리버섯 품종을 육종하기 위하여 육종모본 24×46과 KNR2539를 단교배하여 소규모 시험재배하여 생육소요일수(15.4일)와 수량(81.5 g/850 cc), 품질(7.5)을 기준으로 17×15계통을 선발하였다. 선발된 계통을 ‘애린이6’라고 명명하고 대량재배로 큰느타리2호와 생육특성을 비교하였다. 병당수량은 ‘애린이6’가 76.0 g으로 대조품종 61.4 g의 113% 수준이었다. 품질의 경우 ‘애린이6’는 6.8, ‘큰느타리2호’는 5.7로 나타났다. 생육소요일, 수량, 품질은, 독립 t test로 분석한 결과 통계적으로 유의성을 보였다. 갓의 명도(L)에 있어서 ‘애린이6’는 61.7로 대조품종의 58.3보다 높아서 밝은 색을 띄었다. 고유성에 있어서는 URP2와 URP11에서 대조품종과 신품종이 다형성을 보였고, 대치배양에서도 뚜렷한 대선이 관찰되었다.

A new commercial cultivar ‘GARES-HM-17-1’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from Hae-Sal 3Ho. The parental strains are characterized by the property of high quality and a short period of time in cultivation. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15~16°C. The period of harvesting including primordia formation was 23.0 days shorter than that of control strain Mangadak No. 2. The color of pileus and stipe surface was neutral-white, respectively. The yield was 82.0±5.8g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘GARES-HM-17-1’ showed a different profile as that of the control strain, Mangadak No.2, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘GARES-HM-17-1’ of H. marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of H. marmoreus.

A new commercial cultivar ‘GARES-HM-16-1’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR8047 and KNR8080. The parental strains are characterized by the property of high quality and a short period of time in cultivation respectively. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15~16°C. The period of harvesting including primordia formation was 20.0 days shorter than that of control strain Mangadak No.2. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The yield was 123.7±14.8g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘GARES-HM-16-1’ showed a different profile as that of the control strain, Mangadak No.2, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘GARES-HM-16-1’ of Hypsizigus marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of Hypsizigus marmoreus.

To breed a new Korean Pleurotus eryngii cultivar with high quality and yield, single crosses between 24×46 and KNR2539 were performed, and a new cultivar, 6×13, was selected based on the days to harvest (14.6 days), quality (7.2), and yield (85.9 g/850 cc bottle). The strain was named Aeryni 5 and cultivated on a large scale at the mushroom farms to compare with Keuneutari 2ho. The yield of Aeryni 5 (82.2 g) was 122.7% of Keuneutari 2ho, and the quality of the new cultivar was 7.7 while reference cultivar was 6.3. The yield and quality of the two cultivars were statistically different. The lightness of the pileus of Aeryni 5 (61.7) was greater than that of Keuneutari 2ho by 3.4 points; thus, the pileus of Aeryni 5 looked brighter. PCR with URP2 was used to discriminate between Aeryni 5 and Kenneutari 2ho.

큰느타리버섯 품종 육성을 위해 품질이 우수한 특성을 지니는 큰느타리버섯 KNR2598과 경도가 우수한 특성을 지니는 큰느타리버섯 KNR2610 모본으로부터 단핵균주를 분리 한 뒤 단포자간 교잡을 통해 고품질의 저온 저장성이 우수한 신품종 “단비5호”를 육성하였다. 신품종의 균사 생 육 적정온도는 25 o C이며 자실체 발생 적정 온도는 15~ 16 o C 이었다. 솎음 재배에서 발이소요일수를 포함한 수확 소요일수는 대조품종인 큰느타리버섯 2호에 비해 0.5~1.7 일 정도 빠른 특성을 보였다. 갓 색깔은 중간수준의 갈색이 며 대 색깔은 흰색을 나타내었다. 갓모양은 우산형으로 850cc 플라스틱 병재배에서 한 개체의 평균무게는 90.4 ±16.9 g이었다. “단비5호”와 대조품종간의 RAPD 분석결과 서로 다른 DNA 밴드양상을 보여 주었으며 대치배양을 통 해 두품종간에 대치선을 확인 하였다. “단비5호”의 경우 4 o C에서의 저장성이 대조품종 대비 24.6~30.6일 이상 길게 유지되는 특성이 있어 수출적합형 품종으로 가치가 있을 것으로 기대된다.

Pleurotus eryngii, an edible white-rot fungus, is widespread in Eurasia and northern Africa. It has become a major cultivated mushroom in Asia, with a current global production rate of approximately 3 × 10 5 metrictons/yr. To improve the quality or productivity through breeding, a genetic linkage map is an important component. In this study, genetic linkage map of the P. eryngii was constructed using 98 monokaryotic progeny derived from dikaryon of parental KNR2312 strain derived from haploid meiotic spores. The whole genome sequence of P5 monokaryon from P. eryngii KNR2312 strain by Next Generation Sequencing (NGS) strategy was used to design the SSR markers. 484 primers pairs were identified by SSR Locator I and tested polymorphism via PCR. A total of 241 loci were mapped using Joinmap 4.0, comprising 222 SSR markers, 2 mating type factors, and the 13 INDEL markers. The map consisted of 14 linkage groups spanning 1003 cM at an average marker interval of 4.2 cM. The mating loci, A and B were mapped on linkage groups 4 and 11, respectively. The established linkage map and the genetic information based on NGS could be used for QTL mapping of agronomic traits, marker-assisted breeding that may ultimately lead to outstanding phenotypic characteristics. [Supported by a grant from the IPET (213003-04-3-SBY20), MIFAFF, Republic of Korea.]

A new commercial cultivar ‘Hae Sal’ of Hypsizigus marmoreus was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR8047 and KNR8080. The parental strains are characterized by the property of high quality and a short period of time in cultivation respectively. The optimum temperature of mycelial growth was 25°C and that of fruiting body development was 15～16°C. The period of harvesting including primordia formation was 7.0 days shorter than that of control strain ‘Mangadak No.2’. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The yield was 117.6±23.2g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial cultivar ‘Hae Sal’ showed a different profile as that of the control strain, ‘Mangadak No.2’, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new cultivar ‘Hae Sal’ of Hypsizigus marmoreus is characterized by the improved quality and a short period of time in cultivation after scratching. It would be of much help to expand the cultivation area of Hypsizigus marmoreus.

The gene encoding an esterase enzyme was cloned from a metagenomic library of cow rumen bacteria. The esterase gene (est2R) was 2,120 bp in length, encoding a protein of 516 amino acid residues with a calculated molecular weight of 57,286 Da. The molecular weight of the enzyme was estimated to be 57,000 Da by SDS-PAGE. Est2R shared 35.6% amino acid identity with esterase (CAH19079) of uncultured prokaryote. The Est2R was most active at 20-40°C, and showed optimum at 30°C and pH 8.0. The most activity of Est2R for the different chain length of p-nitrophenyl ester group as substrate was p-nitrophenyl acetate. Moreover, the enzyme was found to be most active without organic solvent, followed by 98% active with ethanol, and the enzyme activity was highly affected by the acetonitrile. The enzyme was significantly inhibited by Zn2+ but stimulated by Ca2+. So, novel esterase gene est2R is likely to obtain from cow rumen metagenome and supposed to use for industrial purpose.

A new commercial variety "KNR12-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2312, KNR2594, and KNR2596. The parental strains are characterized by the property of high quality and a small number of primordia, respectively. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15～16℃. The period of harvesting including primordia formation was 1.0 - 1.5 days later than that of control strain Knneutari No.3 in the culling cultivation. 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 101.7±3.5g/850cc plastic bottle. Analysis of the genetic characteristics of the new commercial variety "KNR12-1" showed a different profile as that of the control strain, Knneutari No.3, when RAPD (Random Amplified Polymorphic DNA) primer was used. This new variety "KNR12-1" of Pleurotus eryngii is characterized by the improved storability and a small number of primordia formation after scratching. It would be of much help to mushroom cultivators in a process of culling and storage of P. eryngii.

A carboxymethyl cellulase gene, cel5B, was cloned, sequenced, and expressed in Escherichia coli. pRCS20 in E. coli was identified from metagenomic cosmid library of cow rumen for cellulase activity on a carboxymethyl cellulose agar plates. Cosmid clone (RCS20) was partially digested with Sau3AI, ligated into BamHI site of pBluescript II SK+ vector, and transformed into E. coli DH5α. The insert DNA of 1.3 kb was obtained, designated cel5B, which has the activity of hydrolyzation of CMC. The cel5B gene had an open reading frame (ORF) of 1,059 bp encoding 352 amino acids with a signal peptide of 48 amino acids and the conserved region, VIYEIYNEPL, belongs to the glycosyl hydrolase family 5. The molecular mass of Cel5B protein expressed from E. coli DH5α exhibited to be about 34 kDa by CMC-SDS-PAGE. The optimal pH was 8.0, and the optimal temperature was about 50℃ for its enzymatic activity.

A new cultivar "KNR11-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2598 and KNR2610. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR11-1" was one day faster 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 the teeth of a comb. The yield was 90.4 g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. It is able to store up to 67.5 days in a cold room. Analysis of the genetic characteristics of the new commercial strain "KNR11-1" showed a different profile as that of the control strain KNR2312 when Random Amplified Polymorphic DNA primers were used. The new cultivar of Pleurotus eryngii is characterized by the improved storability after harvesting.

A new cultivar "KNR10-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2594 and KNR2610, respectively. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR10-1" was one day faster 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 the teeth of a comb. The yield was 90±12.9g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. Analysis of the genetic characteristics of the new commercial strain "KNR10-1" showed a different profile as that of the control strain KNR2312 when RAPD (Random Amplified Polymorphic DNA) primers were used. This new variety of King Oyster mushroom is characterized by the improved storability after harvesting.

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.

큰느타리버섯의 재배면적 및 생산량 증가에 따라 많은 양의 폐배지가 발생되고 있다는 사실을 염두에 두고 그 활용도를 높이고자 농가로부터 수거된 폐배지에 대해 성분분석 및 버섯균사에 대한 생육저해 여부를 확인한 뒤 버섯재배에 있어 폐배지의 적정첨가 비율을 결정하고 그에 따른 버섯의 배양특성 및 생육특성을 관찰하였다. 폐배지 성분분석 결과 pH는 새배지에 비해 낮아지는 경향을 보였고 총 질소 함량은 폐배지 첨가비율 많아질수록 높아지는 경향을 나타내었다. 버섯의 배양 및 생육과정 중 배지로 분비된 대사산물에 의해서 일어 날수 있는 버섯 균사생육저해 현상을 조사하기 위해 폐배지 열수추출물을 이용하여 만들어진 고체배지에 버섯 균사를 접종한 결과 폐배지에 의한 버섯균사 저해현상은 확인되지 않았다. 폐배지 첨가에 따른 배양특성을 조사한 결과 첨가량이 증가할수록 균사생장률은 약간씩 낮아지는 경향을 나타내었으나 50% 이내로 폐배지가 첨가된 조건에서는 일반적인 배양기간 (35일) 내에 배양이 완료된다는 사실을 확인 할 수 있었다. 버섯의 발이특성을 조사한 결과에 있어서도 50% 이내로 폐배지가 첨가된 경우 평균 10일 이내에 발이가 완료되었으나 폐배지 100%로 제조된 배지에서는 정상적으로 발이가 되지 못하였다. 생육특성의 경우 평균 18일 이내에 수확이 완료되었으나 폐배지 첨가량이 30% 이상 일 경우 자실체수량이 감소하였다. 이상의 결과를 요약하면 큰느타리버섯 재배농가에서 정상적으로 재배되고 발생되는 폐배지의 경우 버섯 잔재물이나 배지 덩어리 및 오염배지와 같은 이물질을 제거한 뒤 10-30% 수준으로 새배지에 첨가하여 재활용 할 경우 정상적인 버섯의 배양 및 생육이 가능하였으며, 이러한 사실은 버섯 재배농가의 재료비 절감에 도움을 줄 수 있을 것을 기대된다.