The depletion of stratospheric ozone has resulted in increased amount of ultraviolet-B radiation (UV-B: 280-320 nm) reaching the Earth’s surface and could cause significant biological effect in plants. In this study, putative quantitative trait loci (QTL), which is responsible to UV-B resistance in soybean, was identified using recently developed high-density 180K Axiom SoyaSNP genotyping array. A population of 115 recombinant inbred lines (RILs) derived from a cross between susceptible Keunolkong and resistant Iksan 10 was analyzed. A total 8,970 polymorphic SNP markers were used to construct linkage map. The both parents and RILs were grown with supplemental UV-B radiation in a greenhouse condition. Three categories of UV-B induced morphological damage, degree of leaf chlorosis, leaf shape change, and total plant damage were evaluated. Using composite interval mapping analysis, one major QTL associated with all of the phenotypic traits was detected on 7.7cM of soybean chromosome 7 with 22 of LOD score accounting for about 60% of phenotypic variance. Also, the allele from Iksan 10 were responsible for the UV-B resistance. Thus, the UV-B resistance QTL on chromosome 7 from Iksan 10 was designated to qUVBR1, corresponding to 30kb on the Williams 82 genome assembly (Glyma2.0) including 7 candidate genes. This result could be useful in breeding for new foxglove aphid resistant soybean cultivars. In addition, these results provided useful information not only for marker-assisted selection for UV-B resistance soybean, but also for the future identification of putative candidate genes, responsible for UV-B resistance in soybean.

Capsicum diversity is getting lower in modern crops because of the genetic erosion. In Capsicum, breeders have been mainly focused on agriculturally important traits such as disease resistances, high yield and pungency. This narrow breeding pool hampered to develop improved cultivars. It has become a hot issue to conservation of genetic diversity and exploitation of wild germplasm in Capsicum. However, although a large number of accessions are maintained in Capsicum germplasm collections, their use for crop improvement is limited by the scarcity of information on genetic diversity, population structure and proper phenotypic assessment. The identification of representative and manageable subset of accessions would facilitate access to the diversity available in large collections. A genome wide germplasm characterization using molecular markers can offer reliable tools for adjusting the quality and representativeness of core samples. We investigated patterns of molecular diversity at 48 single nucleotide polymorphisms (SNPs) in 4056 accessions from 11 Capsicum species from 89 different countries. Using these genetic variations and 32 different morphological traits, 250 core set was selected in whole Capsicum germplasm. The core collection could be a primary source for distributing germplasm to pepper breeders and other national programs as well as for evaluation

In plants, eukaryotic translation elongation factor 1B (eEF1B) is composed of three subunits, eEF1Bα, eEF1Bβ and eEF1B γ. Two subunits are nucleotide exchange subunits (eEF1Bα and eEF1Bβ) and one is a structural protein (eEF1Bγ). In the previous study, eEF1B was identified as a common host factor for several RNA viruses. To test which subunit of eEF1B is essential for Potato virus X (PVX) replication, the virus-induced gene silencing (VIGS) for eEF1Bα, β or γ was performed in Nicotiana benthamiana and green fluorescent protein (GFP)-tagged PVX was inoculated. PVX-GFP accumulation was decreased when eEF1Bβ or γ subunit was silenced, whereas eEF1Bα had no effect on PVX-GFP accumulation in inoculated leaves. Targeting induced local lesions in genome (TILLING) was performed using a Capsicum annuum EMS population to test whether mutations in eEF1Bβ subunit affect virus infection in pepper. We obtained 81 eEF1Bβ mutant lines consisted of 16,759 individuals. These mutant lines are being tested to validate the function of eEF1B β in PVX replication.

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.

sy-2 (Seychelles-2) is a temperature sensitive 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 lower than 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 of 1,010 F2 plants derived from crossing sy-2 and the wild type C. chinense ‘No.3341’. sy-2 gene is located on chromosome 1, 0.3 cM and 0.1cM away from cosII markers C2_At4g29120 and C2_At1g09070, respectively. The tomato genome sequence between those two markers was compared with pepper genome sequence. We found three of pepper scaffold sequences in this region. We developed seven ingle nucleotide polymorphism (SNP) markers on the pepper scaffold sequences, among which five SNP markers were co-segregated with sy-2. To fill the gap between the scaffolds which contains co-segregating markers, we screened a bacterial artificial chromosome (BAC) library, and end-sequences of total of 22 AC clones were i. We found that five clones were overlapped to cover the gap. We fully sequenced four AC clones and found that the physical distance between C2_At4g29120 and C2_At1g09070 is 343kb. This region contains 70 putative genes such as HSP90-like ATPase family proteins, lipid-transfer proteins, calmodulin-domain protein kinases, and zinc finger proteins (ZFPs). To identify the sy-2 gene, we performed RT-PCR and found that a ZFP-like gene is differentially expressed between WT and sy-2 leaves. This result suggests that the ZFP-like gene is a strong candidate for the sy-2 gene. We are currently characterizing this candidate gene.

Tomato yellow leaf curl disease is a devastating disease of tomato (Solanum lycopersicum), which is caused by begomoviruses generally referred to as tomato yellow leaf curl virus (TYLCV). The breeding for TYLCV resistance has been based on the introgression of the Ty-3 resistance locus. Knowledge about the exact location of the Ty-3 on tomato chromosome 6 is needed to understand the genomic organization of the Ty-3 locus. In this study, we conducted a genetic analysis using a segregating population derived from a cross between resistant accession S. lycopersicum “A45” and susceptible accession S. lycopersicum “A39”. The F1 plants showed resistance to TYLCV and F1 was self-pollinated to produce F2 progeny. To screen the TYLCV resistance in 145 F2 plants, a leaf agroinfiltration method was used. F2 plants showed a classical Mendelian seregation (106 resistance : 39 susceptibility) for resistance to TYLCV respectively. SCAR and CAPS markers linked to the Ty-3 were tested for genotyping F2 plants and .genotyping and agroinfiltration results were cosegregated in the newly developed F2 population.

This study was conducted to determine the optimal dose of gamma-ray for mutation breeding in sorghum (Sorghum bicolor M.). Gamma-rays irradiated to dry seeds with various doses (0 to 1000 Gy) at Korea Atomic Energy Research Institute. Lethal dosage (LD50) was approximately 256 Gy. Significant decreases in growth characteristics (plant height, tiller number and fresh weight) were observed by dose of increased over 300 Gy. Reduction doses (RD50) was approximately 363 Gy in plant height. We also conducted comet assay to observe nucleus DNA damage due to gamma irradiation. In comet assay, a clear difference was identified over 100 Gy treatments. With increasing doses of gamma-ray in the range of 100 to 400 Gy, the rate of head DNA was decreased significantly from 93.52% to 67.57%. The most of the Sorghum cells were severely damaged in the integrity of DNA by gamma-ray. These data provide valuable information when the optimal dose should be chosen for purpose of mutation breeding program of Sorghum.

This study was carried out to determine the effects of different harvesting stage on nutritive value and the quality of ensiled kenaf after fermentation among six kenaf cultivars. Six kenaf cultivars including two different maturity groups, mid-late maturing (Auxu, Jangdae and Jinju) and early-maturing (Baekma, Jeokbong and C14), were planted on May 14, 2013. Four harvesting times were made at intervals of 20 days from 15 July to 16 September, 2013. In all cultivars, the CP (crude protein) contents were decreased by a delayed harvest; the CP contents of kenaf silage were ranged from 92 to 184 g kg-1. Interestingly, there were no significant difference of NDF (neutral detergent fiber) and ADF (acid detergent fiber) content between the cultivars, however NDF and ADF content of kenaf silage were significantly increased by a late harvest. The silages of all cultivars displayed a low pH ranges less than 4.0, which is sufficient for stable storage. The lactic acid contents in Auxu were from 2.57 to 3.21%, which is higher than other cultivars. The harvesting stages did not affect to the concentrations of butyric acid and acetic acid in all cultivars. These results indicate that the harvesting stage is more important for the quality of kenaf silage than cultivar differences. And kenaf silage could be also used as fodder for ruminants.

Pepper mottle virus (PepMoV) is frequently occurring virus in pepper field. PepMoV infected plants show symptoms including mosaic leaf, distortion of foliage and fruit deformation. The dominant gene Pvr7 from Capsicum annuum ‘9093’ confers resistance to PepMoV. Previous research reveals that Pvr7 is located in 10 chromosome and linked to the dominant potyvirus resistance gene Pvr4 and Tomato spotted wilt virus (TSWV) resistance gene Tsw. To identify the Pvr7 gene, we constructed an intraspecific F2 mapping population from a cross between C. annuum ‘9093’ (PepMov resistant) and C. annuum ‘Jejujaerae’ (PepMoV susceptible). Resistance of F2 plants were screened with green flouorescent protein (GFP) tagged PepMoV. Genomic DNA was extracted from F2 individuals and markers were developed using C.annuum ‘CM334’ whole genome sequence (WGS). Several single nucleotide polymorphism (SNP) markers that were co-segregated with Pvr7 were developed. We are expecting that this Pvr7 SNP marker can be used to breeding PepMoV resistant cultivars and fine mapping of Pvr7.

Characteristics of agronomy, especially good yield productivity is very important in wheat (Triticum aestivum L.) as human food crop. Development of elite cultivar having good traits is required for food stability. In 2013, a new wheat line called Iksan370 has been developed by the National Institute of Crop Science, RDA. Iksan370 has hard red winter wheat characteristics and better features like resistance to biotic, abiotic stress and higher yield ability than Keumkang. Heading and maturing date were delayed about ten days and period to flowering date was shorter three or four days than Keumkang. Culm and spike length of Iksan370 were 95cm and 15.3cm, averagely. This means that culm and spike length were longer than Keumkang (79cm and 7cm, respectively). It showed higher milling rate (75.5%), protein content (13.5%) and SDS-sedimentation volume (77.5ml) than Keumkang. Genotypes of six genes, GBSSA, B, D, PPO18, Pina and Pinb, were represented that were similar to Keumkang, respectively. Average yield of Iksan370 in the preliminary trial test was 6.61 tons/ha in upland. We expect that yield productivity and farm income were increased by Iksan370.

Transforming growth factor (TGF) family is well known to induce the chondrogenic differentiation of mesenchymal stem cells (MSC). However, the precise signal transduction pathways and underlying factors are not well known. Thus the present study aims to evaluate the possible role of C2 domain in the chondrogenic differentiation of human mesenchymal stem cells. To this end, 145 C2 domains in the adenovirus were individually transfected to hMSC, and morphological changes were examined. Among 145 C2 domains, C2 domain of protein kinase C eta (PKCη) was selected as a possible chondrogenic differentiation factor for hMSC. To confirm this possibility, we treated TGFβ3, a well known chondrogenic differentiation factor of hMSC, and examined the increased-expression of glycosaminoglycan (GAG), collagen type II (COL II) as well as PKCη using PT-PCR, immunocytochemistry and Western blot analysis. To further evaluation of C2 domain of PKCη, we examined morphological changes, expressions of GAG and COL II after transfection of PKCη -C2 domain in hMSC. Overexpression of PKCη-C2 domain induced morphological change and increased GAG and COL II expressions. The present results demonstrate that PKCη involves in the TGF-β3-induced chondrogenic differentiation of hMSC, and C2 domain of PKCη has important role in this process.

Capsinoids which were found recently in non-pungent pepper show the same biological effects as capsaicinoid including anticancer and anti-obesity. A precursor of capsaicinoids, vanillyl alcohol, is known to be produced by mutations in the p-aminotransferase (pAMT) gene. In the previous study, we showed that capsinoid production is also controlled by the capsaicin synthase (CS) gene. However correlation between the CS gene expression and capsinoids contents has not been fully understood. This study was conducted to elucidate correlation between the expression level of CS gene and capsinoids contents. Through germplasm screening, we identified one C. chinese pepper cultivar, SNU11-001, which contained capsinoids as much as C. annuum ‘CH-19 Sweet’. SNU11-001 was crossed with five Capsicum cultivars (ECW, Takanotsume, Yuwolcho, Habanero and Jolokia) containing different levels of capsaicin, ‘ECW’ is non-pungent pepper line, and ‘Takanotsume’ and ‘Yuwolcho’ have mild pungency, and ‘Habanero’ and ‘Jolokia’ is known to be included in the most pungent pepper lines. When we analyzed the expression of CS and pAMT genes using the six Capsicum cultivars, the expression levels of CS were higher in pungent Capsicum cultivars. To test whether the expression levels of CS also control capsinoids contents, we will analyze several F2 populations derived from crosses between SNU11-001 and Capsicum cultivars containing different levels of capsaicin.

Capsicum diversity is getting lower in modern crops because of the genetic erosion. In Capsicum, breeders have been mainly focused on agriculturally important traits such as disease resistances, high yield and pungency. However, this narrow breeding pool hampered to develop improved cultivars. It has become a hot issue to conservation of genetic diversity and exploitation of wild germplasm in Capsicum. Analysis of genetic diversity and construction of core collection is the first step to make efficient use of germplasm. Although there have been several attempts to construct core collections in Capsicum, most of these works were limited due to handling small number of samples, relying mainly on the characterization of morphological traits or focusing only C. annuum species. To expand understanding of the structure and genetic diversity of germplasm in Capsicum, we need to have a highly efficient genotyping tool to handle large number of samples. Toward this end, we are analyzing 3,599 germplasm accessions including other cultivated species and wild species in Capsicum with 48 single nucleotide polymorphism (SNP) markers.

The precise, fast, and cost-effective identification of important fruit crop cultivars is essential for practical breeding and plant breeder’s rights. Traditional methods for identification of persimmon cultivars are based on the evaluation of sets of morphological characteristics. However, the identification using only morphological traits is difficult to distinguish among genetically closely related cultivars. This study was conducted to develop more reliable DNA markers for identification of the 32 persimmon cultivars in Korea and Japan. In total, 309 random amplified polymorphic DNA (RAPD) markers were identified using 40 different random primers. The 4 (OPP-08) to 14 (UBD159) polymorphic bands were detected with an average of 7.7. The resulting 57 RAPD fragments were selected, and their sequences were determined for developing sequence-characterized amplified region (SCAR) markers. As a result, 15 of 57 RAPD fragments were successfully converted to SCAR markers. A single polymorphic band of the same size as the RAPD fragments or smaller DNA fragments were amplified depending on primer combinations in the 15 SCAR markers. Among these markers, a combination of eight SCAR markers (PS225_200, PSN05_420, PSF13_523, PSN11_540, PS372_567, PS485_569, PSP08_635, and PS631_735) provided sufficient polymorphisms to identify 32 persimmon cultivars depending on number and size of amplicons. These newly developed markers will be useful as a fast and reliable tool to identify persimmon cultivars.

Mutant lines induced by ethyl methane sulfonate (EMS) have been used for crop improvement and functional genomics. Since pepper is very recalcitrant to be transformed, EMS mutagenesis could be an alternative method to generate useful mutant lines and to characterize the function of genes. We have developed mutant lines consisting of about 3,938 M2 mutant lines using Korea local landrace, C. annuum ‘Yuwolcho’. Yuwolcho has suitable traits for mutagenesis such as early flowering and maturation, large number of seeds per fruit, and susceptibility to various diseases. Up to now, 917 M2 mutant lines were evaluated to confirm the effect of EMS. M2 mutant lines have shown variations in plant stature (small size, dwarfism, and early death), leaf development (light color, variegation and morphological change) and flower (inflorescence, morphological change) and fruit (size and color). We observed the largest morphological variation in leaf development. Most of these mutant phenotypes were inherited recessively. In addition, we are applying cel1-based TILLING to identify useful mutant lines. We will apply cel1-based TILLING to identify useful mutant lines. We are expecting that these mutant lines will be very useful to study the function of genes in C. annuum.

This study was performed to investigate the expression of cathepsin B mRNA and protein in eutopic and ectopic endometrial tissues of patients with endometriosis and in normal endometrial tissues and to clarify the association between the cathepsin B expression and endometriosis. A total of 40 women with histologically confirmed endometriosis were recruited for study group. For controls, 20 women undergoing operative treatment for uterine myoma, cervical intraepithelial neoplasia (CIN) or benign gynecologic conditions other than endometriosis were recruited. Eutopic endometrial tissues of both groups and ectopic endometrial tissue of study group were collected during the operations. We employed real time reverse transcriptase - polymerase chain reaction (RT-PCR) to quantify mRNA levels of cathepsin B in these tissues. Then, we performed western blot analysis to measure the protein levels of cathepsin B. The expressions of cathepsin B mRNA and protein were significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls. These data suggest that the higher expression of cathepsin B in the endometrial tissues might be associated with the development of endometriosis. In addition, eutopic endometrium itself with higher expression cathepsin B may play a pivotal role in the histogenesis of endometriosis.