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Linkage grouping of Pleurotus eryngii by simple sequence repeats (SSR)
Chak Han Im,Kyung-Hee Kim,Asjad Ali,Min-Keun Kim,Wan-Kyu Joung,Yong Jo Choi,Jae-San Ryu 한국버섯학회 2015 버섯 Vol.19 No.1
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.]
Linkage grouping of Pleurotus eryngii by simple sequence repeats (SSR)
Chak Han Im,Kyung-Hee Kim,Asjad Ali,Min-Keun Kim,Wan-Kyu Joung,Yong Jo Choi,Jae-San Ryu 한국버섯학회 2015 버섯 Vol.19 No.2
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.]
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.
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.
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.
Asjad Ali,Chak Han Im,Bokyung Park,Min Keun Kim,Wan-Kyu Joung,Yong Jo Choi,Jae-San Ryu 한국버섯학회 2015 버섯 Vol.19 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 Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]
Asjad Ali,Chak Han Im,Bokyung Park,Min Keun Kim,Wan-Kyu Joung,Yong Jo Choi,Jae-San Ryu 한국버섯학회 2015 버섯 Vol.19 No.2
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 Gold Seed Project [Supported by a grant from the IPET (213003-04-3-WTI11), MIFAFF, Republic of Korea.]