Radish is an important root vegetable in the world, and many cultivars have been developed with various molecular marker systems to identify these cultivars. Recently developed markers for radish cultivar identification require only 11 primer pairs, but they still use conventional PCR with different annealing temperatures and time-consuming gel electrophoresis. To improve the genotyping method, we applied touchdown PCR with 11 primers with M13 tails among 105 radish cultivars. Touchdown PCR successfully generated amplicons in all 11 M13-tailed primers with a condition of annealing temperature starting from 55℃, decreased by 1°C and 33 cycles at 53°C. The 11 M13-tailed primers followed by fragment analysis produced 71 amplicons, which produced more amplicons than gel electrophoresis that produced 23 amplicons. Especially, simple sequence repeats produced more amplicons, 12 on average, than the other marker types. The present study requires less effort and provides more accurate results compared to genotyping using gel electrophoresis. Besides, a database can be established using digitized genotyping results among radish cultivars.

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, GB19 의 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 hexanucleotide 와 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 (SSR), also referred to “microsatellites” consist of tandemly repeated short DNA sequence motifs and have been applied in various marker-based studies. SSRs were isolated and characterized from ‘Heuktari’ and ‘Miso’, which are major oyster mushroom cultivars in Korea, by genome sequencing and bioinformatic analysis. The genome sizes of ‘Heuktari’ and ‘Miso’ were estimated to be 40.8 and 40.3 Mb, respectively, which are larger than those of other P. ostreatus species (PC9 and PC10) and smaller than those of P. eryngii (KNR2312P5). In total, 949 and 968 SSRs were found in the ‘Heuktari’ and ‘Miso’ genomes, respectively. Comparative analysis of five mushrooms including P. ostreatus var. florida (PC9 and PC15) and P. eryngii revealed that the number of SSRs in ‘Heuktari’ and ‘Miso’ were the highest among them. All mushrooms studied showed similar SSR distribution patterns. Tri-, hexa-, and octanucleotide motifs accounted for the top three fractions of all SSRs.

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.]

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.]

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.]

한국꿩 (Korean ring-necked Pheasant, Phasianus colchicus karpowi)과 외국 아종의 유전적 유연관계를 파악하기 위해 야생 한국꿩, 사육 한국꿩, 사육 한국꿩과 외국꿩간의 잡종꿩, 외국꿩 4아종(중국 링넥, 흑 뮤탄트, 백 뮤탄트, 녹치)을 대상으로 ISSR 표지자 분석과 AMOVA 분석을 수행하였다. 야생 한국꿩의 전체 유전 다양성중 94.08%가 서식지 내 개체간 유전적 차이에 기인하고, 5.9% (Φ

This study was performed to analyze genetic relationship of the four major Cucurbitaceae crop. We used 120 Expressed Sequence Tag(EST)-Simple Sequence Repeat(SSR) primer sets of developed from watermelon and published in International Cucurbit Genomics Initiative (ICuGI) database. Among 120 EST-SSR primer, 51(49.17%) EST-SSR primer set successfully amplified and 49(40.8%) EST-SSR primer set showed polymorphisms among eight cultivars of Cucurbitaceae. In the first instance, amplified PCR products analysis was conducted by the agarose-gel electrophoresis then further analyzed by using Fragment Analyzer. A total 382 PCR band were producted by 49 EST-SSR primers in 24 plant panels, used the analysis of pairwise similarity and dendrogram construction. Assessment of the genetic relationships resulted in similarity index with range of 0.0103 to 0.8452. In dendrogram, 24 plant panels were formed three major groups (A, B, C) and 7 subgroups (A-1, A-2, B-1, B-2, B-3, C-1, C-2). Major group A was comprised of 2 subgroups, subgroup A-1 (6 watermelon cultivars, Citrullus lanatus var. vulgaris Schrad.) and subgroup A-2 (3 wild type watermelon, Citrullus lanatus var. citroides Mats. & Nakai). Major group B was comprised of 3 subgroups, subgroup B-1 (4 melon cultivars, Cucumis melo var. cantalupensis Naudin.), subgroup B-2 (2 oriental melon cultivars, Cucumis melo var. conomon Makino.) and subgroup B-3 (5 cucumber cultivars, Cucumis sativus L.). Major group C was comprised of 2 subgroups, subgroup C-1 [2 squash/ pumpkin cultivars, Cucurbita moschata (Duch. ex Lam.)/Duch. & Poir. and Cucurbita maxima Duch.] and subgroup C-2(2 squash/pumpkin cultivars, Cucurbita pepo L./Cucurbita ficifolia Bouche.)

Gastrodia elata, an achlorophyllous orchid plant, is rare medicinal plant. We investigated the genetic diversity in G. elata from 4 locations by using Inter-Simple Sequence Repeats (ISSR) markers. Shannon's information Index (S.I.) indicating genetic diversity ranged from 0.255 (Pocheon) to 0.322 (Muju) with the mean of 0.29. The level of genetic diversity was lower than other plant and most genetic diversity was allocated among individuals within populations (26.81%). The UPGMA dendrogram based on genetic distance failed in showing decisive geographic relationship. In the case of gastrodin (GA), the major components in G. elata, Sangju was highest. The ergothionine (ERG) was detected a lot of contents in Muju and Pocheon. In conclusion, our results is very important information for explaining relationship of genetic variation and functional substances without the effects of environment factors and developing genetic marker by ISSR in G. elata, which may be responsible for the development of breeds with a lot of functional substance in G. elata.

The genetic diversity and the genetic relationship among 30 genetic resources of T. officinale and T. coreanum collected from 20 regions in Korea were evaluated by using ISSR markers. Out of 127 loci detected overall, 122 were identified to be polymorphic with a rate of 96.0% at the 30 individuals. The intraspecific polymorphism between T. officinale and T. coreanum was 92.6% and 88.2%, respectively. The genetic similarity matrix (GSM) revealed a wide range of variablility among the 30 accessions, spanning from 0.179 to 922. According to the clustering analysis, different species T. officinale and T. coreanum, were divided into independent groups and all of the accessions could be classified into 7 categories. Especially, all of the mountain collected accessions belonged to independent groups. The study findings indicate that T. officinale and T. coreanum accessions have a high genetic diversity and accordingly carry a germ-plasm qualifying as good genetic resources for breeding.

The polymorphism and the genetic relationships among 32 genetic resources of genus Nelumbo from Korea, Japan, China, USA, India, Thailand and Gabong were thoroughly investigated and extensively examined using ISSR markers. Out of 103 loci detected overall, 94 were identified to be polymorphic with a rate of 91.2%. The genetic similarity matrix revealed a wide range of variability among the 32 accessions, spanning from 0.227 to 0.833. The study findings indicate that the Nelumbo accessions have a high genetic diversity, and accordingly carry a germplasm qualifying as good genetic resources for cross breeding. According to the clustering analysis, different subspecies, N. nucifera and N. lutea, were divided into independent groups and all of the N. nucifera accessions could be classified into five categories. Compared to RAPD analysis, ISSR method showed a clearer picture of polymorphism among the accessions and exhibited a definite distinction even among the subspecies. In this respect, ISSR analysis is considered to be more effective in differentiating the accessions and subspecies of the genus Nelumbo than RAPD test.

Cowpea might have been introduced from China to Korea and cultivated for several hundred years but it has never been a staple food crop in Korea. In this study, genetic diversity of 492 Korean cowpea landrace accessions that have passport information was estimated using six SSR markers. The mean of Weir's gene diversity was 0.665 from all accessions investigated in the study. Cowpea gene diversity of six local provinces in Korea was ranged from 0.370 in accessions of Gangwon to 0.680 in Jeonra provinces. Low gene diversity of the cowpea genepool of Gangwon province was probably derived from relatively few introductions. Especially SSR markers VM36 and VM39 seem to be good markers to distinguish the Gangwon accessions from others by occurring at a specific locus with higher than 78% of allele frequency. Except for the Gangwon province with the low genetic diversity, gene diversity of cowpea accessions from other provinces was ranged from 0.600 to 0.680 indicating no big differences among provinces. Distribution pattern of the allele frequencies was similar among the other provinces. This may reveal that Korean farmers might exchange cowpea seeds easily with even their neighbors with geographical barriers. A core collection, 100 landraces, ca. 20% of base collection, was developed at the 70% of a similarity coefficient level using random sampling approaches after stratification of the entire landrace collection based on the phenetic dendrogram. The variability of SSR in the base and core collections of Korean cowpea landrace was compared by calculating Weir's gene diversity. The mean of Weir's gene diversity of the core was 0.707 while that of the base collection was 0.665. The higher diversity index in the core collection indicates that it maintains the initial variability and well represents the base collection. The core collection included one of determinate accession (IT 216155) and two of no branching type accessions (IT 103959 and IT 161024). The core collection could be used to guide more efficient management and utilization of the entire collection. This core collection should be revised periodically as additional accessions are collected and further characterization is conducted.

In Korea, chilli pepper (Capsicum annum L.) is a major vegetable crop. The pepper seed market is about $35 million and the whole sale market including processed products is equivalent to $2 billion, representing the second highest market value among crops, next to rice in Korea. Since the development of elite pepper variety is so competitive, vegetable seed companies usually run two important programs to keep the credibility of seed quality. One program is to deliver F1 hybrid seeds with a high purity test to farmers. The purity control of parents and F1 hybrid to avoid any contamination is conducted by DNA markers because pepper seeds are obtained using MS line. The other program is to identify the F1 variety from other varieties by analyzing the polymorphism so that the company and/or breeder protects the intellectual property from copying by others or from non-intentional contamination. We have developed about 900 EST-SSR sets from pepper and used to both programs. A total of 66 markers were selected to identify 32 F1 varieties and their own parents. Using these markers, the purity control of F1 hybrid rose up to the highest degree. We also found several SSR markers to distinguish F1 variety from other varieties and these markers could be useful to find the uniqueness of F1 cultivar.

Diversity and relatedness of 64 rice accessions were evaluated using 85 simple sequence repeats (SSRs) markers. These64 accessions represent a set of international and Korean blast diferential cultivars, a set of blast monogenic lines, donors of blastresi