고추의 적색소는 고추의 상품성을 가늠하는 중요한 척도이면서 식재료 뿐 아니라 상업적으로도 다양하게 활용되고 있다. 본 연구는 적색소 성분의 함량과 관계하는 QTL 마커를 개발하기 위하여 적색소 성분 분석을 위한 mapping 집단을 육성하였고, 적색소 성분에 대한 QTL mapping을 수행하였다. 적색소 분석을 위한 mapping 집단인 ‘만다린’과 ‘블랙클러스터’를 양친으로 하는 F7 RIL 집단에서의 색도(ASTA value) 분포는 1.64에서 117.26의 범주에 있으며 그 분포 양상은 정규분포를 보여 QTL분석에 적합한 것으로 확인되었다.Mapping 집단의 양친들에 대해서 454 GS-FLX pyrosequencing을 이용한 NGS를 수행하였고, 그 결과 ‘만다린’과 ‘블랙클러스터’각각 120.44Mb와 142.54Mb의 염기서열 데이터를 확보할 수 있었으며, ‘만다린’에서 1,025개, ‘블랙클러스터’에서 1,059개의 SNP들을 확보하게 되었다. 이 SNP들을 HRM 분석에 용이하도록 프라이머를 제작하여 유전자 지도 작성을 수행한 결과 총 246개의 SNP 마커를 이용하여 약 512cM을 설명할 수 있는 21개 연관군의 유전자 지도가 작성되었다. 분석 집단 93계통들에서 측정된 ASTA 값을 이용하여 수행한 QTL 분석 결과 총 6개의 QTL 을 확인하였다. 이들 QTL과 근접한 마커들은 향후 고추의 적색소 함량 연구에 매우 유용한 정보로 활용될 것이며, 아직까지 개발된 바 없는 적색소 함량 연관 마커 개발에 가능성을 열어줄 것으로 기대한다.

Soybean [Glycine max (L.) Merr.] seeds are abundant in high-quality proteins and fats. In addition, soybean seeds are also rich in secondary metabolites, such as isoflavones, lecithin, and saponins. Triterpene saponins are major components of these physiologically active metabolites in soybean seeds. Soybean saponins are classified as group A and DDMP saponins. Among them group A saponins are undesirable component of food products due to bitterness and astringency and also cause foaming in tofu production. Whereas, DDMP saponins and their derivatives are less bitter and astringent and beneficial to human health when consumed as regular diet. Therefore, reducing the group A saponins or increasing the DDMP saponins are required to improve the food quality. The present study focused to identify and characterize the gene which is encoding a protein responsible for biosynthesis of DDMP saponins. EMS mutant lines (sg-7-1 & sg-7-2) which lack DDMP saponins were developed. The breeding cross has been made with these two mutants with two cultivars, Pungsannamul and Wooram to study the segregation and genetic linkage analysis, respectively. The segregation analysis showed that the mutant phenotype is controlled by single recessive gene. TLC analysis for phenotyping F2 population of Wooram X sg-7-1 showed mutant, wild and heterozygous types. To surprise two more patterns were detected and they were named as strange type1 (ST1) and strange type2 (ST2). Further, SSR marker analysis will be carried out to locate the gene which encoding a protein responsible for biosynthesis of DDMP saponins.

Soybean germplasm have diverse accessions with great variation in their ability to survive and reproduce under salt stress conditions. In general, cultivated soybeans are more sensitive to salt stress than their wild relatives, however exceptions are found in both the groups. These variations in response to salt stress makes soybean germplasm an interesting collection of genetic resources to be explored for the identification of salt-tolerance genes, and their mechanism of action. Here, in this report we presented a data showing differential response of selected accessions of both cultivated and wild soybeans to salt stress. Two modes of salt treatment; gradual salt stress (GS) as well as salt shock (SS) were used in this study. The GS was found more effective in finding the difference in response of soybean accessions to salt stress. Various genetic marker based methods are in use to identify and isolate the potential genes contributing to the salt tolerance in soybean. Even then there is a paucity of knowledge on the key genes contributing to the salt tolerance in soybean. We expect that a recently developed functional screen based method, like yeast based functional screen, using cDNA library generated from different salt tolerant accessions of soybean could lead to identification of novel genes responsible for salt tolerance in soybean. Also, we propose for the use of RNA isolated from different stages of GS and SS for making cDNA library to be used for functional screening.

Soybean [Glycine max (L.) Merr.] have a variety of flower colors which are controlled by six different genes (W1,W2,W3,W4,Wm, and Wp). Among these genes, mutation in W3 gene causes near white flowers in the background of w4 genotype whereas the genotype W3w4 does purple throat flowers. Earlier studies showed that dihydroflavonol 4-reductase1 (DFR1) gene was closely linked to the flower color variants for W3 locus. In order to find out the W3 gene responsible for w3 phenotype, we first, studied the candidate gene Glyma14g07940 (DFR1) which is having 100% similarity with DFR probe sequence. Sequence analysis of DFR1 between W3 and w3 soybeans showed one base substitution in exon 6 of w3 mutant soybean resulting in one amino acid change in the amino acid sequence. However, comparison of amino acid sequences of DFR proteins from various crop plants showed that there is no functional change in the protein. Besides, the promoter analysis showed that, 311 bp of indel was traced in 5’-upstream promoter region of DFR1 gene in the w3 mutant. Here, we show that the near white or purple throat phenotypes in G. max is associated with existence or nonexistence of indel at 5’- upstream promoter region and low or high expression of DFR1, respectively. These results suggest that w3 phenotype may be caused by certain regulator of DFR1 gene located near or distant from DFR1 in G. max. In further study, we need to check the correlation between promoter indel with W3 expression level through GUS analysis.

Grain color distinguishes between the pigmentation of the outer layer of the kernel. It is known that environmental factors affects the production of anthocyanins and abiotic stresses like high light intensity, low temperature, high salinity and/or drought stress, and others increase their amounts. After 7 days the germination rate between yellow and dark-purple seeds were almost the same with and without stress (100% yellow seeds under stress and without stress germinated, 93.3% under stress and 96.6% without stress of purple seeds germinated), even though at the final stage the germination was almost the same, we can conclude base on our observations that the germination takes place at a different rate. We think that this might be related to the seed color, since the germination of purple seeds under salt stress started earlier than the yellow ones, until both reached the same point. The antioxidant activity was higher in seedlings from dark-purple seeds than the yellow ones, and they were higher under salt stress than without it, supporting our hypothesis that the purple color in wheat seeds works as a protection under salt stress. Furthermore, the qRT-pCR showed that some genes related to the flavonoid pathway were expressed or had more expression in the seedlings from dark-purple seeds than yellow ones.

Capsinoids, low-pungent compounds, have the same biological effects as capsaicinoids such as anticancer and anti-obesity. A precursor of capsinoids, vanillyl alcohol, is known to be produced by mutations in the putative-aminotransferase (pAMT) gene. In the previous study, ‘SNU11-001’ (Capsicum chinense) containing high levels of capsinoids was identified in germplasm collections of Capsicum. This collection has a unique mutation in the pAMT gene that can cause dysfunction of this gene. In order to develop pepper varieties containing high capsinoids contents, marker-assisted foreground and background selections were performed during backcross breeding. Compared to the conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. For foreground selection, plants carrying the pAMT/pamt genotype were selected from a BC1F1 and BC2F1 populations using SCAR markers derived from the unique pAMT mutation of ‘SNU11-001’. To obtain background selection markers, a total of 412 single nucleotide polymorphism (SNP) markers was screened on ‘Shinghong’ parental lines and ‘SNU11-001’ to obtain polymorphic SNP markers. Of the 412 SNP markers, 144 and 204 polymorphic SNP markers evenly distributed in pepper genome were finally selected. BC1F1 and BC2F1 plants carrying the pAMT/pamt genotype were subjected to background selection using the selected marker sets. Multiple genotype analysis was done using a high-throughput genotyping system (EP1TM, Fluidigm®, USA). As a result, one BC1F1 plant 84% similar to the recurrent parent and several BC2F1 plants more than 96% recovery rate of the recurrent parent were selected. Genetic backgrounds of the selected BC2F1 plants were evaluated by the genotype-by-sequencing (GBS) method in order to confirm the background selection results using the SNP marker set. GBS results showed that recovery rate and positions of introgressed segments were well matched between two methods demonstrating MABC can be successfully done with a couple hundred SNP markers.

Establisment of rice library is an essential approach for rice functional genomics study. Utilizaing maize transposable element Ac/Ds is a promising method to construct insertional mutagenesis library of rice. Ac/Ds tagging system has received extensive application in rice during the past several years. The maize Ds element is one of the main tagging vehicles used in rice. Narrow leaf mutant have short height, narrow leaf width and large angle. To compare with wild type and narrow leaf mutant in detail, we observed the leaves under microscope. In specific portion(large and small vein), no significantly reduce cell size and number of cell. Knock-out of the OsNLR(narrow leaf ribokinase) gene inhibits internodes, panicles, angle(between leaf and stem), leaf, seed. OsNLR was shown to specifically expressed on leaf. In real time PCR analysis with mature leaf of wild type and mutant, there might be a functional association between OsAGO7, NRL1, NAL1 and NAL7 in regulating leaf development. We tested on the experimental field using wild type and mutant plants. In agricutural traits that contain leaf and seed related traits(except angle) significantly reduce in mutant plants. These results demonstrate that OsNLR gene may be associated with leaf development.

R2R3 MYB transcription factors play regulatory roles in plant responses to various environmental stresses and nutrient deficiency. In this study, we isolated MYB-like gene respond to phosphorus deprivation in rice and designated OsMYB4P, an R2R3 MYB transcription factor, from rice under low-phosphate conditions. OsMYB4P is 993bp long and encodes a 330 amino acid polypeptide. OsMYB4P was localized in the nucleus and acted as a transcriptional activator. Transcriptional levels of OsMYB4P in cell suspension, shoots, and roots of rice increased under low phosphate conditions. Shoots and roots of OsMYB4P overexpressing plants grew well in high and low phosphate conditions. In addition, root system architecture was altered considerably as a result of OsMYB4P overexpression. Under both phosphate sufficient and deficient conditions, more Pi accumulated in shoots and roots of OsMYB4P overexpressing plants than in the wild type. Overexpression of OsMYB4P led to greater expression of Pi transporter-family proteins OsPT1, OsPT2, OsPT4, OsPT7, and OsPT8 in shoots, and to decreased or unchanged expression of these proteins in roots, with the exception of OsPT8. These results demonstrate that OsMYB4P may be associated with efficient utilization of Pi in rice.

We used an efficient system to create rice mutant by Ac/Ds transposon insertion mutagenesis, such as selected homozygous mutant in dwarf phenotypes. We reported here the identification of function of dwarf OsGASD gene(Oryza sativa Gibberellin Acid Sensitive Dwarf). OsGASD gene encodes a 344 amino acid polypeptide and no homology proteins in Gene Bank. The osgasd mutnat was sensitive to exogenous gibberellic acid(GA) level. We performed experiment to controlled expression the OsGASD gene, its role in plant development, a quantitative analysis of endogenous GA content and sensitivity to GA. The osgasd mutant includes smaller amount of active GAs than wild-type. osgasd mutant plant of GA biosynthesis pathway causes GA deficiency and dwarf plants, and endogenous GA suppliance can restore the wild type phenotype in this mutant. There result indicated that OsGASD gene regulated the elongation of shoot, stem and plant height. The increased expression of OsGASD gene dramatically induces expression of the factors associated with GA biosynthesis, whereas osgasd mutant suppression of the factors associated with GA biosynthesis, loading to dwarf phenotypes. That applied GA3 at the plant development stage to survey the response of OsGASD gene to GA3. We suggest that OsGASD gene is related to factors of GA biosynthesis pathway regulating rice internodes development.

The ubiquitin-26S proteasome system is important in the quality control of intracellular proteins. The ubiquitin-26S proteasome system includes the E1 (ubiquitin activating), E2 (ubiquitin conjugating) and E3 (ubiquitin ligase) enzymes. U-box proteins are a derived version of RING-finger domains, which have E3 enzyme activity. Here, we present the isolation of a novel U-box protein, OsUPS, from rice (Oryza sativa).The cDNA encoding the O.sativa U-box protein(OsUPS) comprises 1338bp, with an open reading frame of 445 amino acids. The open reading frame of the OsUPS protein is comprised of notable domains: a single ~70-amino acid domain and a GKL domain that contains conserved glycine, lysine/ arginine residues and leucine-rich feature. We found that full-length expression of OsUPS was up-regulated in both rice plants and cell culture in the absence of inorganic phosphate (Pi). A self-ubiquitination assay indicated that the bacterially expressed OsUPS protein had E3 ligase activity, and subcellular localization results showed that OsUPS was located in the chloroplast. Suppression of OsUPS resulted in servre signs of toxicity caused by the over-accumulation of Pi. These results support the notion that OsUPS plays an important role in the Pi signaling pathway through the ubiquitin-26S proteasome system.

Fibroin silk proteins from spider or silkworm are attractive biomaterials that are of particular biotechnological interest for industrial and medical purposes because of their unique physical and mechanical properties. In this study, we generated and characterized the transgenic rice plant expressing a spider silk protein. Spider silks have great potential as biomaterials with extraordinary properties. Here, we report the cloning and characterization of the major ampullate silk protein gene from the spider Araneus ventricosus. A cDNA encoding the partial major ampullate silk protein (AvMaSp) was cloned from A. ventricosus. An analysis of the cDNA sequence shows that AvMaSp consists of a 240 amino acid repetitive region and a 99 amino acid C-terminal non-repetitive domain. The peptide motifs that were found in the spider major ampullate silk proteins, (A)n, (GA)n, and (GGX)n, were conserved in the repetitive region of AvMaSp. Phylogenetic analysis further confirmed that AvMaSp belongs to the spider major ampullate spidroin family of proteins. Recombinant AvMaSp-R was degraded abruptly by trypsin. However, AvMaSp-R was stable at 100 °C for at least 30 min. Additionally, the AvMaSp-R was stable at pH values from 2 to 12 for at least 1 h. Taken together, our findings describe the molecular structure and biochemical properties of the A. ventricosus major ampullate silk protein and demonstrate its potential as a biomaterial.

Space has many distinguishable characteristics from earth such as strong cosmic radiation, microgravity, supervaccum and weak magnetic field. For this reason, space environments can be used an efficient mutagen for plant breeding nowadays. To identify the affected genes by condition in space with outer space, Brachypodium seeds were placed in the Russia Segment (RS) Biorisk module of International Space Station (ISS). Brachypodium distachyon is a model system for temperature grass, because they represent the characteristics for annual winter grass. Seeds and organs of plants carried by satellite or spacecraft to space can be genetically mutated by exposing space environment. We performed a duplicated RNA sequencing to profile the differentially expressed genes. As a results, about 700 genes were upregulated and 250 genes were downregulated by cosmic environments, respectively. In the molecular function category, protein kinase and transcription activity related genes were upregulated. Among the many transcription factors (TFs), stress related TFs such as ERF, NAC and WRKY were differentially expressed in space exposed samples. In the future, their expression will be identified by using qRT_PCR.

sy-2 (Seychelles-2) is a temperature sensitive natural mutant of Capsicum chinense and native to Seychelles Island in Africa. Previously we showed that sy-2 leaves were irregularly shaped and defective in chlorophyll development at temperatures below 24℃. A segregation test revealed that the sy-2 gene is controlled by a single recessive gene. To identify the sy-2 gene, we performed a map-based cloning approach using a total 600 individual F2 plants derived from crossing sy-2 and the wild type C. chinense ‘No.3341’. Fine-mapping of the locus allowed us to position sy-2 to an approximately 170-kb region flanked by markers IN2-1-1 and SNP-3-7 on chromosome 1. Among the approximately 36 hypothetical genes in this region several candidate genes including: HSP90-like ATPase family proteins, lipid-transfer proteins, calmodulin-domain protein kinases, and zinc finger proteins (ZFPs) were identified. RT-PCR and sequencing of the hypothetical genes are under way to identify sy-2.

Rose (Rosa Hybrida Hort.) are of a high symbolic value and a great cultural importance in different societies. They are widely used as garden ornamental plants and as cut flowers. For the induction of mutation, gamma-rays are widely used as a mutagen. This study was carried out to establish a system for mutation breeding by irradiation of gamma-ray in rose. The rooted cuttings of five cultivar roses (Lovelydia, Vital, Aqua, Yellowbabe and Haetsal) are grown by in a greenhouse. They were two difference treatment (Before rooting gamma-ray irradiation, After rooting gamma-ray irradiation) were exposed to dose of 70 Gy using a 60Co gamma-irradiator (150 TBq of capacity ; ACEL, Canada) at the Korea Atomic Energy Research Institute. The irradiated plants were planted in a greenhouse, and investigated survival rate, mutation rate, flower buds number, and shoot length were planted after 80days. The two treatments of and growth characters was significantly reduced to 20% to 40% compared with the control. In addition, survival rate and mutation rate were ‘after rooting γ-ray irradiation (37.4~67.3% and 0.5~5.6%)’ higher than ‘before rooting γ-ray irradiation (18.3~50.8% and 0.3~3.4%)’. Mutation types were solid type, chimeric and mosaic petal mutants with various colors were induced from five rose. These results indicate that efficiency of mutation induction in rose by gamma-ray irradiation on petal colors and petal shapes in two difference treatment with rooted cutting system.

Assessing genetic diversity, population structure, and linkage disequilibrium is important in identifying potential parental lines for breeding programs. In this study, we assessed the genetic and phenotypic variation of 174 normal maize (Zea mays) inbred lines and made association analyses with respect to nine agronomical traits, using 150 simple sequence repeats (SSR). From population structure analysis, the lines were divided into three groups. Association analysis was done with a mixed linear model and a general linear model. Twenty one marker-trait associations involving 19 SSR markers were observed using the mixed model, with a significance level of P<0.01. All of these associations, as well as 120 additional marker-trait associations involving 77 SSR markers, were observed with the general model. Two significant marker-trait associations (SMTAs) were detected at P ≤ 0.0001. In the mixed linear model, one locus was associated with water content, two loci were associated with 100-kernel weight, setted ear length, ear thickness and stem thickness; three loci were associated with ear height, four loci were associated with total kernel weight and five loci were associated with plant height. These results should prove useful to breeders in the selection of parental lines and markers.

For understanding the genetic diversity and genetic relationship between cultivated and weedy types, we evaluated genetic variation of 80 accessions of rice (O. Sativa). This included 42 cultivated accessions and 38 weedy accessions with the help of AFLP and CACTA-TD. A total of 542 loci were analyzed (255 for AFLP and 287 for CACTA-TD) of which AFLP markers exhibited 75% of polymorphism and transposon based CACTA-TD markers exhibited 93% of polymorphism. The average genetic diversity value for all 80 accessions, using AFLP markers was 0.226 (Cultivated – 0.210; Weedy 0.241) and based on CACTA-TD markers was 0.281 (Cultivated – 0.294; Weedy 0.269). A UPGMA phylogenetic tree revealed three major groups for both the marker system. The average polymorphic content value obtained with AFLP and CACTA-TD markers were 0.21 and 0.232, Effective multiplex ratio (AFLP – 47.50; CACTA-TD – 66.75), Marker Index (AFLP – 9.94; CACTA-TD – 21.13) and Resolving power (AFLP – 19.53; CACTA-TD – 34.62) indicated that the CACTA-TD markers were relatively efficient than AFLP markers.

Anthocyanin has antioxidant and radical-scavenging effects which may protect cells from oxidative damage and reduce risk of cardiovascular diseases and cancer. A new peanut variety “Heuksaeng”(Arachis hypogaea ssp. hypogaea L.) with dark purple peanut skin was developed at the Department of Southern Area Crop Science, NICS, in Milyang in 2014. This variety was developed from the crossing line between cultivar “Iksan 31” with short stem and erect plant type and “Iksan35” with large grain and purple skin. “Heuksaeng” which is semi erect Virginia plant type has 32cm of main stem length and 25 branch number per plant. This also show more resistant to late leaf spot, web blotch and lodging, compared with check variety “Daekwang”. Each pod has two grains with ellipse shape of purple testa and its yield components is composed of 60 mature pods of per plant, 69g of 100-seed weight, 77% of pod shelling ratio in the regional yield trials(RYT). For 3 year regional yield trials the average kernel yield of “Heuksaeng” had 4.25 MT/ha similar to that of check variety. Its seed quality show 26.9% of crude protein and 46.0% of crude oil and 53.4% of oleate in fatty acid composition. Peanut skin of variety “Heuksang” consist of 2 kind of anthocyanin compounds such as 4.67mg/100g of delpinidine-3-glucoside (D3G) and 1.18mg/100g of cyanidine-3-glucoside(C3G). Peanut variety with high anthocyanin conent in skin will be useful to the recent preference of colorful food with healthful functional compounds.

Peanut is grown worldwide in the tropics and temperate zones primarily as an oilseed crop (38-54%) and protein source(25-30%). A new peanut variety “Daan”(Arachis hypogaea ssp. fastigiata L.) with the high yield potential was developed at the Department of Southern Area Crop Science, NICS, in Milyang in 2014. This was developed from the crossing line between cultivar “Sangpyeong” with short stem and high quality and “Dakwang” with large grain. “Daan” which is Shinpung plant type has 44cm of main stem length and 13 branch number per plant. Each pod has two grains with long ellipse shape of brown testa and yield components is composed of 34 mature pods of per plant, 127g of 100-seed weight, 75% of pod shelling ratio in the regional yield trials(RYT). Seed quality showed 47.8% of crude oil and 28.3% of protein content. This variety also showed more resistant to early leaf spot, late leaf spot, web blotch, stem rot and lodging compared with check variety “Daekwang”. In the regional yield trials “Daan” outyielded check variety by 16% with 5.00 MT/ha for kernel yield.

To define whole genome-level of structural variation by ionization energies and radiation doses in plant, the seeds of Ilpum rice cultivar were acutely irradiated with gamma rays (100Gy, 200Gy, and 400Gy) and ion-beams (20Gy, 40Gy, and 80Gy), respectively. Six M1 rice plants were re-sequenced by Hi-Seq2500 with Ilpum cultivar as control. The average sequencing coverage of the individuals was 10.6X, and the average mapping rate to the rice reference genome (IRGSP-1.0) sequence was 96.95%. The individual plants were irradiated with gamma-400Gy and ion-50Gy had highest variation of SNP with 471,837 and 469,147, respectively. The number of insertion/deletion was 77,500 and 77,106, the synonymous and frame-shift were 7,859 and 7,763 in above two individuals. Although high genome variation shown between Ilpum cultivar and irradiated individuals, there were non-correlation between number of variation and radiation doses. However, five individuals, except ion-20Gy, showed 33 common variant blocks (CVBs) spanning 6 Mb in whole rice genome (1.6%). The CVBs were distributed on 12 rice chromosomes, Chromosome 6 had biggest CVB (5 blocks, 1.3Mb), whereas chromosome 9 had smallest CVB (0.01Mb). Total five hundred fifty one genes were in CVBs which can regard radiation sensitive genes or may be regarded as radiation hot spots in rice genome. This study will contribute to the improvement of the radiation mutation breeding research in genetic and genomic aspect.

Clubroot is a devastating disease caused by Plasmodiophora brassicae and results in severe losses of yield and quality in Brassica crops including Brassica oleracea. Therefore, it is important to identify resistance gene for CR disease and apply it to breeding of Brassica crops. In this study, we applied genotyping-by-sequencing (GBS) technique to construct high resolution genetic map and mapping of clubroot resistance (CR) genes. A total of 18,187 GBS markers were identified between two parent lines resistant and susceptible to the disease, of which 4,103 markers were genotyped in all 78 F2 plants generated from crossing of both parent lines. The markers were clustered into nine linkage groups spanning 879.9 cM, generating high resolution genetic map enough to refine reported reference genome of cabbage. In addition, through QTL analysis using 78 F2:3 progenies and mapping based on the genetic map, two and single major QTLs were identified for resistance of race 2 and race 9 of P. brassicae, respectively. These QTLs did not show collinearity with CR loci found in Chinese cabbage (Brassica rapa) but roughly overlapped with CR loci identified in cabbage for resistance to race 4. Taken together, genetic map and QTLs obtained in this study will provide valuable information to improve reference genome and clubroot resistance in cabbage.