Reynoutria japonica and R. sachalinensis have been used as medicinal resources in Korea. However, it is difficult to identify and determine these medicinal herbs correctly because they are usually customized and purchased as the fragmented rhizomes types. To develop molecular markers for distinguishing two species, we analyzed and compared the chloroplast DNA sequences of seven loci (atpB, matK, ccD-psaI, atpF-H, trnL-trnF, psbK-I and rpl32-trnL). Among them, we found two effective SNPs in psbK-I region for R. japonica and atpF-H region for R. sachalinensis. Based on these SNP sites, we designed the new R. japonica- specific primer which is able to amplify 300 bp fragment in psbK-I region. A similar strategy was applied for the atpF-H region of R. sachalinensis. These molecular markers would be successfully applied to recognize R. japonica and R. sachalinensis.

Although it is well known that low-molecular-weight glutenin subunits (LMW-GS) affects bread and noodle processing quality, the function of specific LMW-GS proteins mostly remain unclear. It is important to find a corresponding gene for a specific LMW-GS protein in order to understand the function of the specific LMW-GS protein. The objective of this study was to identify LMW-GS genes and haplotypes using well known Glu-A3, Glu-B3 and Glu-D3 gene specific primers and to interlink their protein products by proteomic approaches in a wheat variety. A total of 36 LMW-GS genes and pseudo-genes were amplified including 11 Glu-3 gene haplotypes, designated as GluA3-13K and GluA3-22K (pseudogene) at Glu-A3 loci, GluB3-33K and GluB3-43K at Glu-B3 loci and GluD3-11K, GluD3-21K, GluD3-31K, GluD3-42K, GluD3-5K, GluD3-6K and GluD3-393K (pseudogene) at Glu-D3 loci. To determine the relationship between gene haplotypes and their protein products (to identify the corresponding LMW-GS proteins), we conducted N-terminal amino acid sequencing and tandem mass spectrometry (MS/MS) analysis of the 17 LMW-GS spots separated by 2-DGE. Successfully, LMW-GS proteins of the Glu-3 gene haplotypes except pseudo-genes mentioned above were identified. This is the first report on comprehensive characterization of LMW-GS genes and their corresponding proteins and establishment of specific correspondence between each other in a single wheat cultivar. Our approach will be useful to understand the molecular basis of the LMW-GS and to study their contribution to the end-use quality of flour.

Amplified fragment length polymorphism (AFLP) is one of molecular marker technique based on DNA and is extremely useful in detection of high polymorphism between closely related genotypes like Korean wheat cultivars. Six Korean wheat cultivar specific marker sets have been developed from inter simple sequence repeat (ISSR) analysis and we can identify the 13 Koran wheat cultivars form other cultivars using six that (Son et al., 2013). We used four combinations of primer sets in our AFLP analysis for developing additional cultivar specific markers in Korean wheat. Twenty-one of the AFLP bands were isolated from ACG/M-CAC primer combination and 19 bands were isolated from E-AGC/M-CTG primer combination, respectively. We used forty bands to design sequence characterized amplified region (SCAR) primer pairs for Korean wheat cultivar identification. Only one of 40 amplified primer pairs, C2, were able to use for wheat cultivar identification. The DNA band of 215bp length was amplified by C2 primer pairs in ten cultivars, Eunpa, Olgeuru, Gobun, Saeol, Milsung, Sinmichal, Jokyung, Sugang, Goso, and Joah. Then C2 primer was applied to these primer sets as newly SCAR marker, six cultivars are identifying from other cultivars, additionally. Finally, to use the C2 and six primer sets, 19 Korean wheat cultivars are identified.

To select genes associated with the high-temperature tolerance from Brassica, two transcriptomic analyses have been used: microarray and RNA Seq. Using two contrasting inbred lines of B. rapa, Chiifu and Kenshin, version 3 microarray (135 K microarray) was conducted to RNA samples extracted from series of 45℃-treated leaves and 29 genes were selected for genomic DNA cloning of cabbage. Of 29 genes, 8 genes contain 40 SNPs, 11 SSRs and 23 In-Del markers that distinguish high-temperature tolerant and susceptible cabbages, BN1 and BN2. These 8 genes include a unknown gene, AP2, SMP, FBD, SKP2B, IAA16, HSP21 and OLI2-2. We also selected 16 cabbage genes from RNA Seq analysis using two inbred lines, BN1 and BN2: 5 genes for BN1-high expression, 5 genes for BN1-specific expression, 5 genes for BN2-specific expression, and BoCaMB. Using RNA sequences, genomic DNAs corresponding to 16 genes have been clones and analyzed to find out molecular markers. Markers were further transformed into PCR-based marker and confirmed with additional cabbage genetic lines. We are currently transforming PCR-makers into SNP markers. To examine function of high-temperature tolerant genes, we also transformed 5 genes into Arabidopsis plants. We will describe detailed methods and results in a poster. [This work was supported by a grant from the Next-Generation BioGreen 21 Program (the Next-Generation Genomics Center No. PJ009085), Rural Development Administration, Republic of Korea]

This study describes the identification of Panax species using mitochondrial consensus primers. Initially, a total of thirty primers were tested in ten Korean ginseng cultivars and two foreign Panax species, P. quinquefolius and P. notoginseng. In the polymerase chain reaction (PCR) amplification results, three primers (cox1, nad1/2-3 and nad2/1-2) generated co-dominant polymorphic banding patterns discriminating Korean ginseng cultivars from P. quinquefolius and P. notoginseng. However, these primers could not generated polymorphisms among the Korean ginseng cultivars, and simply represented species-specific polymorphisms for P. quinquefolius and P. notoginseng. Primers PQ91 and PN418 were designed from the consensus sequence of nad1/2-3 region. Two banding patterns (A or B) were detected in PQ91. Korean ginseng cultivars and P. notoginseng shared the same banding pattern (A type) and P. quinquefolius was identified another banding pattern (B type). In the case of PN418, two banding patterns (A or B) were detected in the Korean ginseng cultivars and two foreign Panax species. Korean ginseng cultivars and P. quinquefolius shared the same banding pattern (A type) and P. notoginseng was identified another banding pattern (B type). The combination banding patterns of three Panax species, Korean ginseng cultivars (Panax ginseng C. A. Mey.), P. quinquefolius and P. notoginseng, was identified as 'AA', 'BA' and 'AB', respectively. Consequently, PQ91 and PN418 primer sets can be used to distinguish among Panax species.

This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.

In direct-seeding cultivation of rice, the emergence and establishment of seedlings are important for determining the actual yield. These traits depend principally upon elongation of both the mesocotyl and coleoptile. Mesocotyl elongation in rice is controlled by several genetic factors and is also affected by environmental factors. In this study, we mapped QTL for mesocotyl elongation using F8 lines from a cross between the cultivated rice, Ilpumbyeo and a weedy rice, PBR. One of the Korean weedy rice, PBR showed the long mesocotyl length than that of cultivars, Ilpumebyeo under soil and agar media conditions. This weedy rice showed long mesocotyl than the elite japonica cultivars. After a phenotyping of 150 F7 lines for mesocotyl length, a subset of 20 lines selected from the two extreme phenotypic tails was used for the bulked segregant analysis. Two QTL were identified on chromosomes 1 and 3. These two QTL were confirmed using 120 F8 lines. Two QTL, qMel-1 and qMel-3 on chromosomes 1 and 3 accounted for 37.3% and 6.5% of the phenotypic variance, respectively. The PBR alleles were associated with an increase in mesocotyl elongation at both loci. It is noteworthy that two QTL for mesocotyl elongation were colocalized with the QTL for mesocotyl length reported in the previous QTL reports. These QTLs can be introgressed into cultivar background using marker assisted backcrossing in an effort to enhance the level of mesocotyl elongation.

미성숙 종자로부터 추출된 전체 RNA를 이용하여 합성한 cDNA와 LMW-GS 특이 프라이머세트를 이용하여 43개의 LMW-GS 유전자를 분리하였다. 각각의 유추 아미노산은 상동성이 높은 20개의 시그널 펩타이드, N-말단 영역, 반복서열영역 그리고 C-말단 영역을 가지며 C-말단 영역에 분자내 혹은 분자간 이황화 결합을 형성하는 전형적인 8개의 시스테인을 가지고 있었다. 이들 시스테인의 위치는 첫번째, 일곱번째를 제외하고는 보존되어 있었다. Ikeda

The objective of this study was to map gene/QTL for photoblastism in a weedy rice (photoblastic rice: PBR) using DNA markers. Light-induced effect on germination of seeds was compared among three accessions (Oryza sativa L.), PBR, Milyang 23 and Ilpum. Results showed that PBR seeds started to show photoblastism during seed development, different from Ilpum and Milyang 23. Frequency distribution of germination in the F4 lines from crosses between Ilpum and PBR and, Milyang 23 and PBR revealed bimodal distributions suggesting that photoblastism was controlled by a few genes. Bulked segregant analysis using F4 populations derived from the above two crosses was conducted to identify gene/QTL for photoblastism. Two QTL were identified on chromosomes 1 and 12 explaining 11.2 and 12.8% of the phenotypic variance, respectively. Two QTL were further mapped between two SSR markers, RM8260 and RM246 on chromosome 1, and between RM270 and 1103 on chromosome 12. It is noteworthy that two QTL for photoblastism were colocalized with the QTL for seed dormancy reported in the previous QTL studies. The clustering of two genes for photoblastism and dormancy possibly indicates that these regions constitute rice phytochrome gene clusters related to germination. Because PBR has a low degree of dormancy, a pleiotropic effect of a single gene controlling dormancy and photoblastism can be ruled out. The linked markers will provide the foundation for positional cloning of the gene.

Lectin protein is a main antinutritional factor in mature soybean seed. The Le gene controls a lectin protein. Plant breeders can use molecular markers to select indirectly individuals in segregating populations that carry a gene for a favorable trait if a tight linkage exists between a marker locus and the genetic locus controlling that trait. The objective of this research was to identify RAPD markers linked to Le allele using bulked segregant analysis. Cultivar "Gaechuck#2" (LeLe) was crossed with PI548391(lele, absence of lectin protein) and F1 seeds were planted. The F1 plants were grown in the greenhouse to produce F2 seeds. Each F2 seed from F1 plants was analysed electrophoretically to determine the presence of the lectin protein band. F2 individual plants were grown in the greenhouse. Young leaf tissues from each F2 plant were collected. At maturity, single F2 plants were harvested. Random F3 seeds from individual F3 seeds harvested were selected and were used to confirm the presence of the lectin protein band. The dominant and recessive F2 plant leaf bulks consisted with ten F2 individual plants were made. 1,000 Operon random primers were used to screen polymorphic band between dominant and recessive bulk. The presence of lectin protein is dominant to the lack of a lectin protein and lectin protein was controlled by a single locus. A few primers that shows polymorphism in bulked samples were selected and were used to obtain segregating data in F2 individual plants.