Genotyping-by-sequencing (GBS) is a robust and rapid tool to develop SNP markers. Reduced sequencing complexity and multiplex sequencing of GBS has reduced genotyping cost for complex genome. However multiplex sequencing brings low sequencing depth which can lead to reduced number of markers. Therefore to find the appropriate condition for GBS is needed. In this research we demonstrated the use of ApeKI and selective primers for GBS of pepper (Capsicum spp.). Selective primers which amplify the GBS library with one or two flanking sequences to ApeKI site were used to increase sequencing depth. By in silico digestion, we developed six different selective primers amplify 4,000-400,000 regions. We made the GBS library with eight pepper accessions in four species using six selective primers and sequenced. Proper selective primers and pooling rate for each species will be determined. This approach will be useful for genotyping Capsicum breeding lines or populations by developing high quality SNP markers.

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

Radiation technologies have been widely used in mutation breeding of crops. Although gamma-radiation has been predominantly applied in radiation breeding, ion beam irradiation is recently emerging as a methodology highly effective in induction of mutations that are useful for plant breeding. Therefore, we investigated the biological effects caused by irradiation of two types of ion beams, which were proton and ion beams, respectively, in pepper. In the evaluation of survival rate, LD50 values were between 300 and 350 Gy, 30 and 40 Gy in irradiation of proton and carbon beams, respectively, while LD50 for gamma-ray was determined to be between 50 and 100 Gy. Growth traits including shoot length, root length, and root width were also examined in pepper seedlings according to does of proton and carbon ion beams to estimate biological effectiveness of each radiation. The result was applied in the construction of pepper mutant population which will be used to develop pepper breeding materials containing novel characteristics.

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 p-aminotransferase (p-AMT) gene. In the previous study, SNU11-001 (C. chinense) containing high levels of capsinoids was found in germplasm collections of Seoul National University. We found that this collection has a unique mutation in the p-AMT gene. In order to develop a cultivar containing high capsinoids contents, marker-assisted foreground and background selection were performed in this study. Backcrossing is an effective breeding method for introducing useful traits to an elite cultivar. Compared to conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. To obtain background selection markers, a total of 412 single nucleotide polymorphism (SNP) markers was screened to obtain polymorphic SNP markers between ‘Takanotsume (C. annuum)’ and ‘SNU11-001’. Of the 412 SNP markers, 96 polymorphic SNP markers evenly distributed in pepper genome were finally selected. Plants carrying the pAmt/pamt genotype were selected from a BC1F1 population using SCAR markers derived from the unique p-AMT mutation of SNU11-001. BC1F1 plants carrying the pAmt/pamt genotype were subjected to background selection. Multiple genotype analysis was done using Fluidigm platform (BioMark). Once we obtain plants carrying most similar genetic background to recurrent parent, capsinoids contents will be measured and another round of MABC be done to obtain plants containing high levels of capsinoids.

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.

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 p-aminotransferase (p-AMT) gene. In the previous study, SNU11-001 (C. chinense) containing high levels of capsinoids was found in germplasm collections of Seoul National University. We found that this collection has a unique mutation in the p-AMT gene. In order to develop a cultivar containing high capsinoids contents, marker-assisted foreground and background selection were performed in this study. Backcrossing is an effective breeding method for introducing useful traits to an elite cultivar. Compared to conventional backcrossing, marker-assisted backcrossing (MABC) is extremely useful for recovery of a recurrent parent’s genetic background. To obtain background selection markers, a total of 412 single nucleotide polymorphism (SNP) markers was screened to obtain polymorphic SNP markers between ‘Takanotsume (C. annuum)’ and ‘SNU11-001’. Of the 412 SNP markers, 96 polymorphic SNP markers evenly distributed in pepper genome were finally selected. Plants carrying the pAmt/pamt genotype were selected from a BC1F1 population using SCAR markers derived from the unique p-AMT mutation of SNU11-001. BC1F1 plants carrying the pAmt/pamt genotype were subjected to background selection. Multiple genotype analysis was done using Fluidigm platform (BioMark). Once we obtain plants carrying most similar genetic background to recurrent parent, capsinoids contents will be measured and another round of MABC be done to obtain plants containing high levels of capsinoids

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.

In Africa, rice is the fastest demanding staple food with a high growth rate of consumption at 4.5% per annum. Currently Africa occupies 32% of world rice imports, and pressure on rice is expected to rise in the near future. Faced with this deficit, Korea has launched the rice breeding project through Korea-Africa Food & Agriculture Cooperation Initiative (KAFACI) of which goal is to increase rice productivity through accelerated development of improved germplasm and varieties in Africa. As of 2013, the ten African member countries joined the rice breeding projects of KAFACI program; they are Cameroon, DR Congo, Ghana, Kenya, Malawi, Mali, Nigeria, Sudan, Tanzania, and Uganda. We adopted the breeding strategy of intervarietal cross and anther culture to speed up high-yielding secondary populations using the Korean and African germplasm. Korean germplasm is composed of 1) temperate japonicas adaptable to tropical conditions of short-day length and high temperature 2) wide-cross derivatives from African wild species, O.glaberrima and O,longistaminata, and 3) tongil-type varieties. These germplasm is evaluated for adaptability in Africa and African partners make crosses with local germplasm with the best selections. Korea produce double-haploid populations from these crosses for another cycle of selection for high-yielding lines in Africa. Inter-varietal crosses and double-haploid technology could accelerate the development of high-yielding germplasm and new rice varieties in Africa within short project period.

본 연구는 한국교육개발원에서 제공하는 「한국교육종단연구」의 자료를 활용하여 교사들의 소명감이 교사의 직업만족도에 주는 영향이 어떠한지 위계적 선형 모형(HLM)을 통해 분석해 보았다. 본 연구의 주요 연구결과는 다음과 같다. 첫째, 모든 학년에서 교사의 교직경력, 교장의 지도성 유형, 동료교사와의 대화와 갈등이 교사의 직업만족도에 유의미한 영향을 미치는 것으로 나타났다. 둘째, 교사의 소명감은 인화중심 교장지도성 다음으로 교사의 직업만족도에 유의미한 영향력을 가진 변인으로 나타났다. 다시 말해, 소명감을 가진 교사들이 그렇지 않은 교사들보다 자신들의 직업에 더욱 만족하고 있었다. 이러한 연구결과는 교사의 소명감이 교사의 만족도에 미치는 영향력을 밝혔다는 점에서 의미를 가진다. 그리고 예비교사 선별과정과 교사 양성과정에서 교사의 역할과 직무에 대한 의미와 목적으로서의 소명감이 더욱 강조되고 강화할 수 있는 접근이 필요하다는 것을 시사한다.

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.

The term euphemism is not polysemous. However it is frequently employed in a 2-way form; sometimes as a channel to convey Speaker's innocuous indirect message (euphemism) and in other times as a device to hide Speaker's malicious intention (doublespeak). The former is intended to promote politeness from Speaker to Hearer and the other is a manipulated form to secure Speaker‘s unduly profit at the cost of Hearer. This paper discusses what distinguishes benign euphemism from malicious doublespeak. This writer probes into the similarities and dissimilarities between euphemism and doublespeak by comparing Allen & Burridge (1991) and Lutz (1989) and proposes Leech's (1987) cost-benefit module as a possible criterion for telling benign euphemism from malicious doublespeak. Overall this research tries to shed light on how language can be manipulated to Speaker's purposes.