CRISPR/Cas9-based genome editing technology fast replaces the previous methods that require protein engineering such as Zinc Finger Nucleases (ZFNs) and TALE nucleases (TALENs). Conventional genome editing of plant cells using CRISPR/Cas9 technology largely depends on Agrobacterium-mediated transformation of the plant cells and subsequent regeneration of whole plants from the edited cells. During this process, unwanted foreign DNAs including the antibiotics gene and fragments of the T-DNA can be introduced into plant genome. Insertion of these unwanted DNA causes lots of regulatory restrictions when commercializing the LMO products. To step aside these issues, we designed DNA-free ribonucleoprotein-based method and regenerated whole plants from the successfully engineered cells. We will share our discovery on the successful implement of this technology in lettuce protoplasts.

In order to breakthrough upcoming challenges for the food production, the efficient use of rice germplasm would be a indispensible. These rice germplasm, adapted from diverse eco-systems, are undiscovered treasures for rice breeders/researchers, potentially providing a broad array of useful alleles that enrich gene pools of current cultivated rice varieties. Although growing ex-situ conservation efforts are an important for preserving diverse rice genetic resources, the activity on finding the novel and favorable genetic variants from the vast genebank collection is greatly challenging, requiring extensive screening processes. Therefore, rice core collection is a powerful solution to accelerate utilizations of the exotic germplasm of the entire population. In addition, The application of whole genome re-sequencing technology would establish a potent platform for fast forward genetic study, such as genome wide association study (GWAS). The GWAS has been implemented to efficiently identify candidate genes related to various useful agricultural traits in many crop species including rice. Given the significant associations between genetic variations and phenotypic diversity does not require prior knowledge, GWAS using high genome coverage of SNP markers provides a genomics platform to dissect previously unknown adaptive or other useful genetic variation accumulated in plant germplasm resources over the times. Once pinpointing candidate genes, GWAS allows informed choice of parents for QTL analysis based on the haplotype information, along with suggesting targets for following mutagenesis and transgenics. Here, we are to report our current achievements and perspectives from GWAS and post-GWAS undertaken to dissect and exploit useful alleles underlying many agricultural traits from Rice core set, including PHS (Pre-Harvest Sprouting), salt tolerance and disease resistance and so forth. Also, we will introduce the integrated Omics based GWAS case study using transcriptomes, proteomes, metabolomes and ionomes of our rice core set.

Pepper (Capsicum spp.) germplasm shows diverse phenotypic variations including fruit size, color, pungency, and many other horticultural traits. Traditional markers including SSR, AFLP, and RFLP have been used to construct genetic maps using biparental populations. However to assess the genetic diversity of large number of germplasm, a robust and rapid marker development and genotyping approach is needed. We used six pepper accessions including C. annuum, C. chinense, C. baccatum and C. frutescens and performed genotyping-by-sequencing (GBS). To select the most appropriate condition, eight different 2 bp selective nucleotides were used to make GBS libraries. Selective nucleotide ‘OO’ showed the largest number of reads in all samples, and 11,026 to 47,957 high-quality SNPs were called in six accessions. When C. annuum ‘CM334’ genome sequence was used as a reference, C. annuum showed the smallest number of SNPs, while C. baccatum which was known to be a different Capsicum clade showed the largest number of SNPs. Pepper core collection chosen to represent the genetic diversity of whole germplasm will be genotyped by high-density SNPs developed from GBS. We will perform genome-wide association study (GWAS) using genetic and phenotypic variation to identify the functional genetic loci controlling horticultural traits.

The high quality of gene set is necessary to study the functional research of genes. Although perilla is cultivated as an oil crop and as a vegetable crop in Asian countries such as Korea, Japan, northeast China and Nepal, the reference genome is absent. To assembly perilla gene set, we sequenced the various tissues of perilla (Perilla citriodora) RNA-seq with Illumina HiSeq platform, generating 548,549,314 short reads. When de novo transcriptome assembly was performed with five samples, 86,396 and 38,413 transcripts were assembled as total and representative transcripts, respectively. Using 1,917,424 proteins at Phytozome ver. 9.1, we annotated the perilla assembled transcripts, and 66,139(76.55%) and 24,030(62.55%) transcripts showed the similarity with known plant proteins (E-value < 1e-10) as total and representative transcripts, respectively. Among the diverse molecular functions, we were interested in the regulatory components, such as transcription factor and transcription regulator. Using this data, we identified 499 transcripts annotated the putative transcription factor differentially expressed transcripts. 165 putative transcription factors were significantly expressed in perilla flower and 121 putative transcription factors in both leaf and flower. This study provides the perilla reference gene set and the understanding of the molecular regulation of transcription factor dependent on the tissue.

The plant-specific transcription factor, LEAFY (LFY) is considered to be a master regulator of flower development in the model plant, Arabidopsis. This protein plays a dual role in plant growth, integrating signals from the floral inductive pathways and acting as a floral meristem identity gene by activating genes for floral organ development. Although LFY occupies an important position in flower development, the functional divergence of LFY homologues has been demonstrated in several plants including monocots and gymnosperms. In particular, the functional roles of LFY genes from orchid species such as Phalaenopsis that contain unique floral morphologies with distinct expression patterns of floral organ identity genes remain elusive. Here, PaLFY, a orthologue of Arabidopsis LFY from Phalaenopsis aphrodite subsp. formosana, a Taiwanese native monopodial orchid was isolated and characterized through analyses of expression and protein activity. PaLFY transcripts accumulated in the floral primordia of developing inflorescences and the PaLFY protein had transcriptional autoactivation activity forming as a homodimer. Furthermore, PaLFY rescues the aberrant floral phenotypes of Arabidopsis lfy mutants. Over-expression of PaLFY alone or together with PaFT1, a P. aphrodite subsp. formosana homologue of Arabidopsis FLOWERING LOCUS T (FT) in rice caused precocious heading. Consistently, higher chlorophyll content in the sepals and morphological changes in epidermal cells were observed in the floral organs of PaLFY knock-down orchids generated by virus-induced gene silencing. Taken together, these results suggest that PaLFY is functionally distinct from RICE FLORICAULA/LEAFY (RFL) but similar to Arabidopsis LFY based on phenotypes of our transgenic Arabidopsis and rice plants.

Carotenoid isomerase (CRTISO) catalyzes the isomerization of prolycopene to all-trans-lycopene in the carotenoid biosynthetic pathway. We isolated two full-length cDNA gene, CuCRTISO and CuCRTISO-like, from Citrus unshiu. To confirm whether these two genes have the function of the carotenoid isomerase, The full-length cDNA of CuCRTISO and CuCRTISO-like gene, respectively, were fused with 35S promoter and NOS terminal region and then transformed into tomato CRTISO mutant, Tangerine, which shows orange fruit due to lack of carotenoid isomerase activity. The mature fruit color of the transgenic line expressing CuCRTISO gene changed from orange to red, which was similar to the fruit color of the tomato “Money Maker”. We also carried out HPLC analysis to detect all-trans lycopene, which is produced from prolycopene by carotenoid isomerase. In the transgenic line expressing CuCRTISO the all trans lycopene was detected from mature fruit but in the tangerine mutant several prolycopenes were detected from it. On the other hand, the transgenic line expression of CuCRTISO gene retained the orange-color fruit at the mature stage as Tangerine mutant. These studies indicate that the CuCRTISO gene has a function of carotenoid isomerase and also plays a role of it in other plant species, and that the CuCRTISO-like gene might be not enough to produce the all trans lycopene or has a another unknown function(s).

We recently reported rice promoters that are active in late stages of pollen development. However, rice promoters that allow manipulation of gene expression at earlier stages of pollen development are still very limited to date. In this study, we have chosen 10 putative microspore promoters, OsMSP1 through OsMSP10, based on publicly available transcriptomic datasets in rice (Oryza sativa L.). Sequence analysis of these promoter regions revealed some cis regulatory elements involved in pollen-specific expression. We also examined promoter activities using the promoter-GUS reporter constructs in both transgenic rice and Arabidopsis. In rice, all of the 10 promoters directed GUS signals from the microspore stage throughout the all stages of pollen development. In addition, while GUS signals from 4 promoters, OsMSP2, OsMSP7, OsMSP9 and OsMSP10, seem to be expressed preferentially during pollen development, those from other six promoters were observed in vegetative tissues such as leaves, stems, and roots of seedlings. Similarly, in Arabidopsis, all of the 10 promoters directed GUS signals during pollen development. In detail, 8 promoters, OsMSP1 ~ OsMSP8 directed GUS signals from the microspore stage, whereas 2 promoters, OsMSP9 and OsMSP10, exhibited GUS signals from tricellular stage. Furthermore, seven promoters, except for OsMSP1, OsMSP2 and OsMSP10, showed GUS signals in shoot apical region or root tissues of seedlings. Furthermore, we verified microspore activity of four promoters, OsMSP1, OsMSP2, OsMSP3 and OsMSP6, by complementation analysis of the sidecar pollen (scp) mutant which displays microspore-specific defects. Currently, further analyses are underway for GUS expression of T2 generation in transgenic rice and scp complementation with remaining promoters.

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.

A freesia (Freesia hybrida Hort.) ‘Cutie Red’ was developed for the pot flower in the National Institute of Horticultural Herbal Science in 2014. This hybrid was crossed and selected from a seedling of three-way crossing the seedling of ‘Volcano’ and ‘Sailor’ with ‘Figaro’, and ‘Suzy’ in 2006 and 2010, respectively. Morphological characteristics of the selected freesia hybrid were investigated for 3 years from 2011 to 2013, and then it was named ‘Cutie Red’ in 2014. ‘Cutie Red’ has single and red petals (RHS, R45A) with yellow-orange color of center (RHS, YO21B). The average flower width is 5.3 cm and the average yield is 4. The growth of the plant is vigorous and the average height is 70.3 cm, and it is shorter than about 15.4 cm that of control cultivar ‘Red Race’. The average number of floret per stalk and stalk length, 10.7, and the stalk was 9.6 cm length, shorter than ‘Red Lace’, 13 and 11.7 cm length, respectively. The first flowering, in avarage, of ‘Cutie Red’ takes about 141 days, and it’s average vase life and yield is 9.7 days and 5 cormlets per plant, respectively.

Polyembryony in many citrus varieties is an impediment in breeding because it makes hard to identify hybrids after crossbreeding. So, it has become imperative for developing efficient methods to distinguish zygotic seedling generated from polyembryonic seed depending on citrus variety. Simple sequence repeat(SSR) marker is one of useful systems for such purpose. However, SSR markers to separate zygotic seedlings derived from the crossbreeding between ‘Marita unshiu’ (Citrus unshiu) ‘Seongjeon’ and ‘Shiranuhi mandarin’ [(C. unshiu x C. sinensis) x C. reticulata] ‘Hallabong’ have not been developed yet. In this study we tried to identify an effective SSR marker to screen zygotic seedling after crossbreeding between ‘Seongjeon’ and ‘Hallabong’. For this investigation, 387 seedlings were generated from 114 seeds produced from crossing those two varieties. A total of 116 SSR markers were tested to identify a special marker for distinguishing origin, zygotic or nucellar seedling. As a result, two markers, SSR012 and SSR093, were found to be more effective than other markers. These two SSR markers might be useful to select zygotic individuals in crossbreeing between ‘Seongjeon’ and ‘Hallabong’.

Tissue-specific promoters are a very useful tool for manipulating gene expression in a target tissue or organ; however, their range of applications in other plant species has not been determined, to date. In this study, we identified two late pollen-specific rice promoters (ProOsLPS10 and ProOsLPS11) via meta-anatomical expression analysis. We then investigated the expression of both promoters in transgenic rice (a homologous system) and Arabidopsis (a heterologous system) using ProOsLPS10 or ProOsLPS11::GFP-GUS constructs. As predicted by microarray data, both promoters triggered strong GUS expression during the late stages of pollen development in rice, with no GUS signals detected in the examined microspores and sporophytic tissues. Interestingly, these promoters exhibited different GUS expression patterns in Arabidopsis. While in Arabidopsis, the OsLPS10 promoter conferred GUS expression at the uni- and bi-cellular macrospore stages, as well as at the shoot apical region during the seedling stage, the OsLPS11 promoter was not active in the pollen at any stage, or in the examined sporophytic tissues. Furthermore, by performing a complementation analysis using a sidecar pollen (scp) mutant that displays developmental defects at the microspore stage, we found evidence that OsLPS10, which can be an applied promoter expressed in Arabidopsis, is useful for directing gene expression in the early stages of pollen development. Our results indicate that the OsLPS10 and OsLPS11 promoters can drive the expression of target genes during the late stages of pollen development in rice, but not in Arabidopsis. Our results also emphasize the necessity of confirming the applicability of an established promoter to heterologous systems.

Abeliophyllum distichum is a monotypic taxon of Oleaceae and endemic to Korea. A comprehensive study on embryogeny and fruit and seed coat ontogeny in Abeliophyllum was carried out via microtome and light microscopy. The fertilization occurs during mid– to late April and embryo matures by early July. The embryo development follows the general fashion from globular embryo – transition embryo – heart shaped embryo – torpedo embryo – walking-stick embryo to mature embryo. The pericarp clearly differentiates into three histological zones: exocarp, mesocarp, and endocarp. The young seed comprises 10-12 cells thick seed coat and the mature seed coat comprises an exotesta, 6-8 mesotesta and an endotesta. Any crystals, phenolic-like compounds, idioblasts, and the sclereids are not found in pericarp as well as seed coat. An overall development confirms Solanade type of embryogenesis in Abeliophyllum. The endocarp becomes more prominent in mature fruit

PurposeBenford'sLawisasimpleandeffectiveauditor–toolthatdetectsfraud.Thispaper’spurposeistoaudittheeffi-ciencyofBenford'slaw,whichusesasetofstrangeob-servations,certainnumbersrepeatedoverothernumbersinthedataset. Researchdesign,data,andmethodologyBenford'slawwas–appliedinnumericalanalysis.Wecansaythatinadditiontore-ducingthedurationoftheaudit,thecapacitiesoftheauditweremorerobust. vefraudandexpandtheuseofanalyticalproceduresinplanningtheaudit.Additionally,theuseoftheanalysesaspartofthecomputer'sinternalcontrolshelpedtofurtherimprovetheeffectivenessofinternalcontrolsandreinforcethem. ConclusionsBenfordanalysisshouldbecarriedoutasap–-propriate.Insubsequentstudies,itcanalsobeexaminedasatooltorevealdoubtfulaccounts.Numericalanalysisofthedataandacomputerarenecessary.Programsfordataanalysisinvariousapplicationssuchasauditing(SAS)and(ACL)and(CaseWare)and(IDEA)areavailable.

전지전능한 절대적 존재에 대한 믿음은 T. S. 엘리엇의 인생을 통하 여 드러나는 것이다. 악의 존재와 인간의 근본적인 불완전함에 대하여 날카롭게 인식하고 있었던 엘리엇은 자신이 유니테리언 교리가 상징하 는, 진보의 이상에 바탕을 둔 낙관주의에 동의하지 않았다. 이러한 낙관 주의에 대한 대안으로, 엘리엇은 종교와 문학을 통합시킬 수 있는 길을 찾으려 하였다. 문학에서 종교가 어떠한 역할을 수행하여야 하는지에 대한 그의 생각은 ｢종교와 문학｣이라는 에세이의 서두에서 잘 드러난 다. 엘리엇은 이 에세이에서 문학 비평은 윤리적, 종교적 관점을 통해 보완되어야 하며, 윤리적인 관점과 종교적인 관점이 조화를 이룰수록 문학이 더 풍부해질 수 있다고 주장한다. 엘리엇은 문학이 지니고 있는 가치를 인정하면서도, 그 가치를 바르게 평가하기 위해서는 외부에 있 는 절대적인 그 무엇이 있어야 한다고 생각한다. 그리고 종교가 문학에 대한 도덕적 또는 윤리적 판단에 대한 기준을 제공할 수 있다고 본다.

본 연구에서는 생물반응장치를 이용하여 조직 배양된 라울리아 신초에 대하여 화장품 성분으로써 응용가치를 평가하였다. 조직 배양한 라울리아 신초에 대한 항산화 및 항염 활성 효과를 연구하였다. 라울리아는 뉴질랜드나 호주에서 자생하는 국화과의 야생초본식물이다. 이미 몇몇 보고 된 논문에서 라울리아는 기관지염, 수막염 그리고 호흡기 질병 등을 유발하는 바이러스에 대한 증식 억제 활성이 있다고 보고되었다. 실험 결과 조직배양된 라울리아 신초 추출물은 자연 상태의 라울리아 추출물과 비교하여 항산화 활성 및 항염 활성 효과가 우수하였다. 조직 배양된 라울리아 신초 추출물은 자연에서 자란 라울리아 추출물보다 50μL/mL 농도에서 10~25% 항산화 활성을 증가시켰다. 또한 조직 배양된 라울리아 신초 추출물은 LPS로 유도된 대식세포에서 iNOS와 COX-2의 단백질 발현이 자연에서 자란 라울리아 추출물보다 억제되었다. 본 연구의 결과들로, 조직배양 한 라울리아 신초 추출물은 피부 보호를 위한 천연 화장품 성분으로써 우수한 가능성을 제공할 수 있을 것으로 사료된다.

The main purpose of the research is to suggest a new business model of direct distribution of organic agricultural products cultivated in suburban area. To secure competitiveness, logistics cost reduction is important through direct transaction of organic agricultural products cultivated on suburban area. The research addresses food safety, farmers' income and unemployment issues with Eco-M business model. The research methods include investigation on the followings: current domestic agriculture; related problems on the existing literature reviews; the current status of suburban agriculture; specialty store of eco-friendly agricultural products. Eco-M business model can solve various problems of suburban agriculture supply system, challenges of organic farming specialty stores, unemployment issues, and difficulties of processing verticalization. Future research should include followings. First, there should be comparative analysis between businesses of environmentfriendly specialty stores through suburban agriculture and producer's agriculture connected with organic farming market. Second, it is required to develop strategy of environment-friendly specialty stores. Finally, analysis of the potential of Sextic industries is necessary throughout production-process-service process.

Purpose – This study classified consumers' value inclination to find out ways to enhance consumers' eco-friendly product purchase intention. Further, it verified the differences among eco-friendly product purchase intentions depending upon value inclination. Research design, data, and methodology – The structured model and hypotheses were established, and 202 copies of effective questionnaires were used. In order to verify the hypotheses, we used single regression analysis, multiple regression, 3-step mediating regression, and path analysis. Results – Individualism had a positive influence upon materialism, need for uniqueness, and face wants, and collectivism had a positive influence upon materialism only. Factors of self-expressive consumption inclination had a positive influence upon eco-friendly product purchase intention, and factors of value inclination also had a positive influence. Finally, self-expressive consumption inclination mediated between value inclination and eco-friendly product purchase intention. Conclusion – Consumers with individualism inclination felt the need to connect the ownership of an eco-friendly product with their extended self and, further, it was clear that not only the government but also enterprises should build up their public image regarding eco-friendly products.