고온적응성 큰느타리(새송이)버섯 품종육성

임착한,김민근,김경희,조수정,이종진,정완규,이상대,최용조,알리아스자드,류재산 한국버섯학회 2014 한국버섯학회지 Vol.12 No.3

In order to breed a new P. eryngii cultivar with high temperature tolerance trait to cope with climate change, strains and cultivars were characterized at 20oC that is 5oC higher than normal condition followed by screening for the characteristics such as required days to harvest, quality and yield. Monokaryons from the selected strains were crossed. Da-32×KNR2322-15 derived from the crosses between KNR2322 having characteristics of short growing day and Da(Ga5Na5-4× KNR2312-7) having charateristics of high guality and yield at 20oC, showed 14.9 days for harvest, 120.6 g yield, and 7.0 quality in the first trial. The strains were named as Taeyangsongi and cultivated on a large scale to compare with Kenneutari No. 2 at a mushroom farm. Yield of Taeyangsongi (109 g) was significantly different(P=0.001) from Kenneutari No.2. Quality of the new (6.6) and the reference cultivar (3.5) was also statistically different (P=0.001) The brightness of pileus of Taeyangsongi (59.5) was 10 points less than the reference cultivar due to which it has an inability to bear high temperature stress. Thus, PCR reactions with URP2 discriminated between Taeyansongi and reference cultivars (Keneutari No. 2 and Aeryni).

큰느타리(Pleurotus eryngii) 품종 판별을 위한 초위성체유래 다중 표지 개발

임착한,김경희,제희정,알리 아스자드,김민근,정완규,이상대,신현열,류재산 한국균학회 2014 韓國菌學會誌 Vol.42 No.2

For development of a method for differentiation of Pleurotus eryngii cultivars, simple sequence repeats (SSR) fromwhole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSRprimer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypingresults. PCR using each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism informationcontent (PIC) values of the five markers were in the range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pairgroupmethod with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivarsinto two clusters. Cluster I and II were comprised of four and eight cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 andSSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivars and strains with 21 alleles and a PIC value of0.9090. These results might be useful in providing an efficient method for the identification of P. eryngii cultivars with separatePCR reactions. 큰느타리 품종구분을 위한 마커의 개발을 위하여 큰느타리 전체 유전자 염기서열을 바탕으로 제작한 484개의 SSR 마커를 사용하여 다형성 분석을 실시하였다. 그 결과 각275개의 primer에서 다형성이 관찰되었다. 이 중 품종간에다양한 패턴을 나타내는 5개의 마커를 최종 선발하였다. 이들 마커의 PIC 값은 0.6627에서 0.6848로 나타났고, 평균값은 0.6775였다. 이 결과를 밴드 이미지 인식 방법으로dendrogram을 작성하였다. UPGMA 집괴분석 결과, 큰느타리 품종은 크게 Cluster 1과 Cluster 2로 구분되었다. SSR primer를 이용한 PCR 결과 나타나는 품종별 고유의 DNA 밴드를 품종특이적 마커로 개발하기 위하여, 선발된 마커중에서 SSR312과 SSR366, SSR178과 SSR 277 마커를 조합하여 초위성체 유래 다중 표지 세트를 개발하였다. Multiplex- SSR 마커의 사용을 통해 두번의 PCR 반응만으로 본 연구에서 사용된 12개의 큰느타리 품종을 구분할 수 있었다.

큰느타리버섯(새송이)에서 역교배를 통한 속성형질의 도입

임착한,김민근,제희정,김경희,류재산 한국버섯학회 2012 한국버섯학회지 Vol.10 No.2

Two strains which have good traits in quality and speedy growing were selected based on previous report (Ryu, 2007) to breed a new strain carrying the traits in it. KNR2322 carrying high quality and KNR2503 carrying a speedy growing trait were cultivated to harvest monokaryons. The monokaryotic mycelia were characterized in growth rate, color, and stability. Selfing of monokaryons from KNR2322 and crossing between KNR2322- and KNR2503-derived monokaryons were performed. Two parental lines, KNR2322-4×37- 6×12(B) showing thremmatological values in weight and quality, 68.5g and 6.5 and KNR2322-30×KNR2503- 60(A) was harvested in 13.5 days after scraping old medium were selected. Crossing between A and B resulted in two high quality and speedy growing strain, A8B8 and A8B10. Their days for harvest after scraping, weight, and quality were 14.0 days, 88.8g and 92.7g, 7.0 and 7.1. Originality tests were conducted with genomic DNAs of A8B8 and Keuneutari 3ho(commercial strain) by DNA fingerprinting a confrontation culture. The results showed polymorphism and a inhibition line between them. It suggested that the new strain have originality from the commercial strain. A8B8 have been registered in Korea Seed and Variety Service with as “Saesongi 1ho”.

큰느타리(Pleurotus eryngii) 품종 판별을 위한 초위성체 유래 다중 표지 개발

임착한 ( Chak Han Im ),김경희 ( Kyung Hee Kim ),제희정 ( Hee Jeong Je ),알리아스자드 ( Asjad Ali ),김민근 ( Min Keun Kim ),정완규 ( Wan Kyu Joung ),이상대 ( Sang Dae Lee ),신현열 ( Hyunyeol Shin ),류재산 ( Jae San Ryu ) 한국균학회 2014 Mycobiology Vol.42 No.2

큰느타리 품종구분을 위한 마커의 개발을 위하여 큰느타리 전체 유전자 염기서열을 바탕으로 제작한 484개의 SSR마커를 사용하여 다형성 분석을 실시하였다. 그 결과 각 275개의 primer에서 다형성이 관찰되었다. 이 중 품종간에 다양한 패턴을 나타내는 5개의 마커를 최종 선발하였다. 이들 마커의 PIC 값은 0.6627에서 0.6848로 나타났고, 평균 값은 0.6775였다. 이 결과를 밴드 이미지 인식 방법으로 dendrogram을 작성하였다. UPGMA 집괴분석 결과, 큰느타리 품종은 크게 Cluster 1과 Cluster 2로 구분되었다. SSR primer를 이용한 PCR 결과 나타나는 품종별 고유의 DNA밴드를 품종특이적 마커로 개발하기 위하여, 선발된 마커중에서 SSR312과 SSR366, SSR178과 SSR 277 마커를 조합하여 초위성체 유래 다중 표지 세트를 개발하였다. Multi-plex-SSR 마커의 사용을 통해 두번의 PCR 반응만으로 본 연구에서 사용된 12개의 큰느타리 품종을 구분할 수 있었다. For development of a method for differentiation of Pleurotus eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotyping results. PCR using each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in the range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pairgroup method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into two clusters. Cluster I and II were comprised of four and eight cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivars and strains with 21 alleles and a PIC value of 0.9090. These results might be useful in providing an efficient method for the identification of P. eryngii cultivars with separate PCR reactions.

보문 : 큰느타리버섯 갓우수 품종 육종

임착한 ( Chak Han Im ),김민근 ( Min Keun Kim ),제희정 ( Hee Jung Je ),김경희 ( Kyung Hee Kim ),김선영 ( Sun Young Kim ),김계자 ( Kye Ja Kim ),박성자 ( Sung Ja Park ),하영아 ( Young A Ha ),김미진 ( Mi Jin Kim ),김설화 ( Sul Ha Kim 한국균학회 2012 韓國菌學會誌 Vol.40 No.3

갓모양이 우수한 계통과 대길이가 길고 속성형질을 가진 계통을 모본으로부터 교배육종으로 두 가지 형질을 모두 가진 우수품종을 육성하고자 하였다. 낮은 갓명도 형질의 동형접합체를 만들기 위하여 KNR2312 유래 단핵균사들로 자가교배를 실시하여 갓명도가 45.2, 품질이 6.5인 중간모본 KNR2312-2636-10 × 18을 선발하였고, 이를 A8B10(나) 유래 단핵균사와 이종교배 하였다. 이후 낮은 품질을 개선하기 위하여 KNR2312 유래 단핵균사와 여교배를 실시하여 갓명도가 49.5, 품질이 7.3 수확량이 95.1 g인 가5나5KNR2312-47KNR2312-12 × 38 계통을 선발하였다. 선발된 계통은 갓모양이 우수하고 특히 갓끝이 24mm로 다듬기나 유통중 파손이 적을 것으로 사료된다. Two strains Pleurotus eryngii KNR2312 and A8B10 (Na) which have good traits in cap quality and speedy growing were selected to breed a new strain carrying the two traits. KNR2312-2636-10 × 18 (Ga) with 45.2 cap lightness and 6.5 quality was breeded by a consecutive inbreeding between KNR2312-derived monokaryons. Ga5 × Na5 came from outcrossing between Ga and Na was backcrossed with KNR2312-derived monokaryons twice to improve quality in cap color and shape. Therethrough Ga5NaKNR2312-47KNR2312-12 × 38 carrying lightness of 49.5, quality of 7.3 and weight of 95.1 g was selected. The selected strain possesses good quality and dark color of cap. Especially its edge is 24 mm thick, therefore it is not likely damaged during processing and distribution.

[Erratum]큰느타리(Pleurotus eryngii) 품종 판별을 위한 초위성체 유래 다중 표지 개발

임착한 ( Chak Han Im ),김경희 ( Kyung Hee Kim ),제희정 ( Hee Jeong Je ),알리아스자드 ( Asjad Ali ),김민근 ( Min Keun Kim ),정완규 ( Wan Kyu Joung ),이상대 ( Sang Dae Lee ),신현열 ( Hyunyeol Shin ),류재산 ( Jae San Ryu ) 한국균학회 2014 韓國菌學會誌 Vol.42 No.3

큰느타리(새송이)버섯 다수확 속성 품종 육종 및 관능평가

임착한 ( Chak Han Im ),김민근 ( Min Keun Kim ),김경희 ( Kyung Hee Kim ),김선영 ( Sun Young Kim ),이성태 ( Seong Tae Lee ),허재영 ( Jae Young Heo ),권진혁 ( Jin Hyeuk Kwon ),김동성 ( Dong Sung Kim ),류재산 ( Jae San Ryu ) 한국균학회 2013 韓國菌學會誌 Vol.41 No.2

Two strains Pleurotus eryngii ``Aeryni`` and ``Na`` carrying superior traits of a pileus and a earliness of harvest were selected to improve previously bred strains by single crosses. New hybrid, Aeryni 3 (Aeryni10 × Na5) showed superiority to other hybrids in yield, fruit body shape and days for harvest. The new strain, Aeryni 3 was harvested earlier than Keunneutari No. 2 by 2~3 days, and yielded 110.5 g/bottle (850 mL) which was 108% of that of Keunneutari No. 2. The ratio of diameter of pileus and stipe was 1.8 indicating that new stain will be likely low damage rate of fruit body during a distribution, and that was better than 2.1 of Keunneutari No. 2. A sensory test of taste of the new strain showed that 84.7% of evaluation panels selected very good while that of Keunneutari No. 2 was 55.5%. In purchasing intent test, 86.9% of panels will buy the new stain whereas 46.8% will buy Keunneutari No. 2 implicating that the new strain will likely be more marketability than previously bred strain.

고온적응성 큰느타리(새송이)버섯 품종육성

임착한,김민근,김경희,조수정,이종진,정완규,이상대,최용조,알리아스자드,류재산 한국버섯학회 2014 한국버섯학회지 Vol.12 No.3

기후변화에 대비한 고온성 품종을 육성하기 위하여 유전자원을 수집하고 표준재배온도보다 5˚C 높은 20˚C에서의 자실체 특성을 평가하고 생육소요일, 품질, 수량을 기준으로 육종모본을 선발하고 단핵균사를 채취하여 단교배하였다. 생육소요일을 기준으로 선발한 KNR2523과 품질 수량으로 선발한 “다“계통간의 단교배에 의해 다×KNR2322-32×15가 생육소요일수(14.9일)과 수량(120.6 g), 품질(7.0)로 선발되었다. 선발된 계통을 태양송이라고 명명하고 대량재배로 큰느타리2호와 생육특성을 비교하였다. 수량은 태양송이가 109.0 g으로 대조품종의 70.6 g의 154% 수준이며, 통계적으로 분산분석결과 0.001% 수준에서 고도의 유의성을 보였다. 품질은 태양송이는 6.6, 큰느타리2호는 3.5로 나타나서 우수하였다. 갓의 명도에 있어서 태양송이는 고온임에도 불구하고 59.5의 수치를 보여서 대조구보다 10정도 낮았다. 고유성에 있어서는 URP2프라리머에서 큰느타리2호와 근친품종 애린이3과 다형성을 보였다. In order to breed a new P. eryngii cultivar with high temperature tolerance trait to cope with climate change, strains and cultivars were characterized at 20˚C that is 5˚C higher than normal condition followed by screening for the characteristics such as required days to harvest, quality and yield. Monokaryons from the selected strains were crossed. Da-32×KNR2322-15 derived from the crosses between KNR2322 having characteristics of short growing day and Da(Ga5Na5-4×KNR2312-7) having charateristics of high guality and yield at 20˚C, showed 14.9 days for harvest, 120.6 g yield, and 7.0 quality in the first trial. The strains were named as Taeyangsongi and cultivated on a large scale to compare with Kenneutari No. 2 at a mushroom farm. Yield of Taeyangsongi (109 g) was significantly different(P=0.001) from Kenneutari No.2. Quality of the new (6.6) and the reference cultivar (3.5) was also statistically different (P=0.001) The brightness of pileus of Taeyangsongi (59.5) was 10 points less than the reference cultivar due to which it has an inability to bear high temperature stress. Thus, PCR reactions with URP2 discriminated between Taeyansongi and reference cultivars (Keneutari No. 2 and Aeryni).

Development of molecular markers to differentiate Pleurotus eryngii cultivars using simple sequence repeats

박보경,임착한,김민근,김희대,최용조,류재산 한국버섯학회 2016 버섯 Vol.20 No.1

Differentiation of Pleurotus eryngii is laborious and time-consuming tasks especially in mycelial status. For development of a method for differentiation of P. eryngii cultivars, simple sequence repeats (SSR) from whole genomic DNA sequence analysis was used for genotyping and two multiplex-SSR primer sets were developed. These SSR primer sets were employed to distinguish 12 cultivars and strains. Five polymorphic markers were selected based on the genotypes. PCR with the each primer produced one to four distinct bands ranging in size from 200 to 300 bp. Polymorphism information content (PIC) values of the five markers were in range of 0.6627 to 0.6848 with an average of 0.6775. Unweighted pair-group method with arithmetic mean clustering analysis based on genetic distances using five SSR markers classified 12 cultivars into 2 clusters. Cluster I and II comprised of 4 and 8 cultivars, respectively. Two multiplex sets, Multi-1 (SSR312 and SSR366) and Multi-2 (SSR178 and SSR277) completely discriminated 12 cultivar and strains with 21 allele with a PIC value of 0.9090. These results might be useful to provide an efficient method for the identification of P. eryngii cultivars with separate PCR reactions. (This work was supported by a grant from the Golded Seed Project (213003-04-3-SBY20), MIFAFF, Republic of Korea.]

큰느타리(새송이)버섯 최적 생육온도 조건

김선영,김민근,임착한,김경희,박기관,송원두,류재산 한국버섯학회 2012 한국버섯학회지 Vol.10 No.4

The effects of fixed and variable temperatures on fruiting body formation and characters of mushrooms were investigated with normal and thinning treatment plots. In fixed temperature, as temperature was lower, period of harvest was longer as well as days for pinheading in the both of normal and thinning plots. Harvest rate were 91.0, 90.3, and 95.8% at 13℃, 17℃, and 15℃. In the thinning plot, qualities of mushrooms were 7.5, 8.1, and 7.7, and yields were 86.0, 105.9, 104.0g at 13, 15, and 17℃ respectively. In variable temperature with a variable temperature plot, condition Ⅱ(17℃ for pin- heading(before reverting, 7-8 days)→ 16℃ for extending primodia(before thinning, 4-5 days)→15℃ for extending (after thinning)) was the best for yield which was better than other conditions by 17.4% and 4.0% and quality of mushroom was highest 8.1 among all conditions.