Microsatellite is one of the most suitable markers for variety identification as it has great discrimination power for varieties with narrow genetic variation. The relationship between marker genotypes and 58 melon varieties was analyzed. Of the 400 pairs of simple sequence repeat (SSR) primers screened against 11 melon varieties. 28 pairs showed highly polymorphism on the basis of PIC (Polymorphism Information Contents) value. These markers were applied to constructing DNA profile data base of 58 major melon varieties through multiplex PCR and fluorescence based automatic detection system. A total of 111 polymorphic amplified fragments were obtained by using 28 SSR markers. The average of PIC value was 0.643 ranging from 0.401 to 0.846. One hundred eleven SSR loci were used to calculate Jaccard’s distance coefficients for UPGMA cluster analysis. A clustering group of varieties, based on the results of SSR analysis, were categorized into plant shape and fruit type. Almost all of the varieties were discriminated by marker genotypes. This information may be used to select comparative variety through comparison of genetic relationship between existing variety and candidate varieties in distinctive tests and protection of plant breeders’ intellectual property rights through variety identification.

For purposes of studying intron structures and predicting consensus splice motifs, a total of 102 legume species were used to isolate introns across the family. Of 196 gene-targeted PCR primer pairs, we successfully amplified 118 intron-containing genes (60.2%) and obtained a total of 1,870 introns with an average size of 143 nucleotides. Species-based compilation of 5’- and 3’-splicing motifs showed lineage-specific conservation in each splicing motif. Compilation of the entire intron set permitted prediction of the consensus sequences of splicing signal motifs in legumes, AYGWGTABABGH and TVNC/TAGGHTV for the 5’- and 3’-splicing motifs, respectively. Interestingly, these consensus motifs are very similar to the corresponding splicing signals of two model systems, Arabidopsis and rice. This result is suggestive of conservation of pre-mRNA splicing mechanisms in higher plants. Multiple alignments of CALTL introns demonstrated that the region from the branch point to 3’ splice site was relatively more conserved than the region from5’ splice site to the branch point. Phylogenetic analysis demonstrated that each of three splicing motifs, 5’-splice sites, 3’-splice sits, and branch site, was relevant to evolutionary divergence of species and phylogenetically informative, suggesting that splice signal sequences would be useful as a potential tool for the molecular phylogenetic analysis.

A core genetic map of the legume Medicago truncatula has been established by analyzing the segregation of 288 sequence-characterized genetic markers in an F2 population composed of 93 individuals. These molecular markers correspond to 141 ESTs, 80 BAC end sequence-tags, and 67 resistance gene analogs, covering 513 cM. In the case of EST-based markers we used an intron-targeted marker strategy, with primers designed to anneal in conserved exon regions and amplify across intron regions. Polymorphisms were significantly more frequent in intron vs exon regions, thus providing an efficient mechanism to map transcribed genes. Genetic and cytogenetic analysis produced eight well-resolved linkage groups, which have been previously correlated with eight chromosomes by means of FISH with mapped BAC clones. We anticipated that mapping of conserved coding regions would have utility for comparative mapping among legumes; thus 60 of the EST-based primer pairs were designed to amplify orthologous sequences across a range of legume species. As an initial test of this strategy, we used primers designed against M. truncatula exon sequences to rapidly map genes in Medicago sativa. The resulting comparative map, which includes 68 bridging markers, indicates that the two Medicago genomes are highly similar, and establishes the basis for a “Medicago” composite map.

Microsatellite marker is one of the most suitable markers for variety identification as it has great discrimination power for varieties with limited genetic variation. The suitability of simple sequence repeat (SSR) markers for varietal identification and genetic diversity were evaluated for 325 rice varieties. Four hundred thirty six pairs of SSR primers were screened against 11 rice varieties. Twenty six primer pairs were highly polymorphic and reproducible. These primer sets were used to construct a DNA profile database containing 325 rice varieties grown in Korea through automatic detection system (ABI 3130XL). Microsatellite polymorphism was showed to be high. A total of 331 polymorphic amplified fragments were obtained by using 26 microsatellite markers. The number of alleles per locus ranged from 9 to 18 with an average of 12.73 alleles per locus. The average polymorphism information content (PIC) was 0.734 ranging from 0.555 to 0.880. Three hundred thirty one SSR loci were used to calculate Jaccard’s distance coefficients for UPGMA cluster analysis. These varieties were separated into several distinctive groups corresponding to varietal types. Almost all of the varieties were discriminated by SSR marker genotypes. This information may be useful to select comparative variety through comparison of genetic relationship analysis between existing variety and candidate varieties in distinctive tests.

Cross-species translation of genomic information may play a pivotal role in applying biological knowledge gained from one species to other genomes. Abiotic stress-responsive genes in Arabidopsis have been translated to a legume model system, Medicago truncatula. A total of 1,370 Arabidopsis genes were identified by searching TAIR database, expression profiling data and literatures. For purposes of cross-genome identification of orthologous genes, tBlastX or BlastP were employed between these two model systems. Candidate genes potentially associated with abiotic stress responses were classified into 18 functional criteria and corresponding genomic locations were analyzed by Circos program. To do this, user-friendly bioinformatic analysis platform was established. In order to discover abiotic stress-associated genes, gene network and/or interactome analyses were conducted using a combination of AraNet web-based platform and CytoScape program. As a result, we could identify 240 key genes that appeared to play an important role within central gene networks. We anticipate that these genes may impact molecular breeding programs by developing them into genetic markers and discovering trait-associated nucleotide variations.

The need to develop work on conservation of crop genetic diversity is emphasized in change of agricultural environment, such as climate change, turn of dietary habits, use of cereal to bioenergy production. To secure provide the germplasm and to derive improve quality, customers demands was surveyed in 2011 and the results are as follows. In the questionnaire on genetic resources necessary in the future, Korean landrace was the highest as 23.8%, followed by genetic resources of international agricultural research institutions as 21.4% and genetic resources of foreign countries of 21.4%. Followed by 42.8% for genetic resources from foreign countries, response ratio were 10.7%, 9.5% and 9.5% for wild or wild-relative species, advanced cultivar or breeders’ line from foreign countries, and advanced cultivar or breeders’ line from Korea, respectively. In addition, consumers desired different genetic resources according to their institutes. Satisfaction on seed purity, germination rate and expected characteristic expression with the genetic resources was higher than normal by 87.5%, 88.9%, 84.2% of respondents, respectively. In order to improve the quality of conserved resources, it is necessary to select the resources having useful characteristics from the time of introduction, and to facilitate the evaluation of the preserved accessions in the near future. Efforts for expansion and opening of information on genetic resources are required.

Most forage crops growing under field conditions are often being exposed to various environmental stresses such as drought, freezing, high temperature, waterlogging and climate change. A combination of grass breeding approaches will likely be needed to improve significantly the environmental stresses tolerance of forage crops in the field. Attempts have been taken by grass breeders to develop tolerant varieties of different crops for environmental stress. A new tall fescue variety (Festuca arundinacea Schreb.) named ‘Purumi’ was developed by the National Institute of Animal Science, Rural Development Administration from 1999 to 2007. For synthetic seed production of this new variety, 5 superior clones, EFa9108, EFa0010, EFa0020, EFa0108, and EFa0202 were selected and polycrossed. The agronomic growth characteristics and forage production capability of the seeds were studied at Cheonan from 2004 to 2005, and regional trials were conducted in Cheonan, Pyungchang, Jeju, and Jinju from 2008 to 2010. ‘Purumi’ showed enhanced winter hardiness, disease resistance, and regrowth ability as compared to ‘Fawn’. The dry matter yield of ‘Purumi’ was about 5.6% higher as 16,821kg/ha than that of ‘Fawn’. However, the nutritive value of both varieties was similar. When this new variety of tall fescue, Purumi, has been developed and distributed with its most remarkable adaptability for Korean climates and superior value as a livestock feed, it is expected to play an important role for a new restoration of the pasture industry in Korea.

Effects of nitrogen (N), phosphorous (P) and potassium (K) on the shoot and bulb growth of wild garlic (Allium victorialis var. platyphyllum) were studied by adopting in vitro culture. These macronutrients influenced the growth of both the shoot and bulb of garlic depending upon their application doses. A minimum of 3% potassium nitrate (KNO3) as a source of nitrogen was found to be critical for shoot elongation while higher concentrations were inhibitory. Garlic bulb growth was profuse on the usual KNO3 strength and sucrose (7%), followed by KNO3 (9.4 mM) supplement. On providing 41.22 mM ammonium nitrate (NH4NO3) as nitrogen source highest shoot growth was observed while 82.45 mM NH4NO3 as a source of nitrogen supported high bulb growth. With regard to potassium a good shoot growth was observed in medium that contained 0.31 mM KH2PO4 and 3% sucrose, while bulb growth was high on 2.5 mM KH2PO4 and 7% sucrose. These experiments may thus direct the development of excellent growth conditions for the commercial production of edible wild garlic.

The aim of this field experiment was to investigate the possible effects of mepiquat chloride (TE) and trinexapac-ethyl (MC) on oil composition, seed yield and endogenous gibberellins content of flax cultivar. Foliar application of plant growth retardants mepiquat chloride (0.897, 1.794 and 2.691 kg a.i. ha-1) and trinexapac-ethyl (0.756, 1.512 and 2.668 kg a.i. ha-1) had significantly increased seeds ripening rate and seed yield. In contrast, plant height was decreased by foliar application of MC and TE. The application of MC significantly increased seed oil yield (730 kg ha-1 by 27.0%) compared to the control. Seed and oil yield, and unsaturated fatty acids (oleic acid, linoleic acid and linolenic acid) were increased by foliar application of MC.

Lhx8 is a member of the LIM-homeobox transcription factor family expressed in the mouse ovary. We discovered that Lhx8 knockout females lose oocytes within 7 days after birth. Lhx8–/–ovaries fail to maintain the primordial follicles and growing follicles. Lhx8–/–ovaries misexpress numerous oocyte-specific genes such as H1foo and Nlrp14. The molecular mechanism of there gulation of Lhx8 in the oocyte has not been described. We examined to characterize Lhx8 DNA binding elements and to identify its direct target genes in the oocyte. CAST was performed using glutathione transferase Lhx8 homeodomain fusion protein (GST-LHX8HD). A 15-bp random sequence flanked by 20-bp of fixed sequences were incubated with purified GST-LHX8HD protein. Unbound DNA was washed with binding reaction buffer. Bound DNA was eluted and re-amplified by PCR for the next round of CAST. Final PCR products were cloned and sequenced to derive consensus binding sequence. EMSA was performed using 32P-labeled oligomers. Binding reactions were conducted by incubating 32P-labeled probes with purified protein. Dual luciferase assays were carried out with extracts of total HEK293 cell which was transfected by the pGL4-promoter vector containing three artificial repeats of LBE(3xLBE-Luc) and overexpression vector carrying the Lhx8 homeodomain as recommended by Promega. We identified several cis-acting sites, TGATTG as Lhx8 DNA binding elements (LBE) using a library of randomly generated oligonucleotides by CAST. EMSA reslut shows that Lhx8 preferentially binds to the oligomer including Lhx8 binding element (TGATTG) with high affinity. In addition, we found that the relative luciferase activity of reporter construct containing three copies of TGATTG was increased by 2.3-fold with Lhx8 overexpression. These results suggest that Lhx8 preferentially binds Lhx8 DNA binding element, TGATTG, and can transactivate reporter genes through the LBE. The transcription of Lhx8 target gene in oocytes directly might be regulated by its during early folliculogenesis.

Human embryonic stem cells (hESCs) have the potential for use in regenerative medicine and in the field of basic research. Therefore, effective cryopreservation and storage of hESCs are important for preservation of newly established cell line for various purposes. Despite poor survival and slow recovery after thawing, the conventional slow freezing method is most commonly used for cryopreservation of hESCs due to its simplicity and ease of use for freezing a large number of hESCs appropriate to clinical applications. Here we controlled the clump size (Group Ⅰ; 400～450 ㎛, Group Ⅱ; 800～900 ㎛, and Group Ⅲ; 1500～1700 ㎛) of hESCs at 5 days after plating using a glass pipette during cryopreservation in order to obtain a larger amount of hESCs after thawing. Attachment rates differed significantly (P<0.05) in each of the three groups and the average of attachment rate of GroupⅡ was highest in SNUhES4 and H1. In particular, the attachment rate of Group Ⅱ in SNUhES3 showed a significant improvement with ROCK inhibitor Y-27632. These results indicate that clump size and cell-cell adhesions of GroupⅡ are appropriate for cryopreservation compared to the Group Ⅰ and Group Ⅲ. This method increased cell viability and reduced the recovery time leading to various experiments, and therefore has an advantage for use with hESCs like newly established in particular. We demonstrated that use of this effective cryopreservation method with control of the clump size of hESCs can effectively improve the attachment rate and survival of post-thaw hESCs with and without Y-27632.

본 연구는 꽃장식에 있어서 침봉이나 플로랄 폼만큼 고정능력이 뛰어남과 동시에 환경에 무해하면서 미적 가치가 뛰어난 꽃 고정재료를 제시함으로써 친환경적인 꽃꽂이 방법을 제시하고자 하였다. 연구방법은 2008년 8월 독일 GBF(Grunbergr Bildungszentrum Floristik)의 플로리스트 연수과정에서 제시된 작품을 분석하였다. 고정재료는 자연에서 얻을 수 있는 재료와 재활용으로 사용되어진 재료로 나누어 분석하였다. 식물의 기관, 즉 잎, 줄기, 뿌리, 열매를 꽃 고정재료로 이용하여 장식해 본 결과 플로랄 폼 이상으로 고정 능력이 뛰어났으며 세우는 기능까지도 탁월하다는 것을 알 수 있었다. 그리고 재활용으로사용되어지는 재료들도 자연물만큼 고정능력이 뛰어나지는 않지만 꽃 고정재료가 가지고 있는 빈공간과 지탱 할 수 있는 힘을 잘 파악한 뒤에 적합한 꽃을 선택한다면 고정기능과 세우는 기능이 가능한 것을 볼 수 있었다. 이와 같이 친환경적 꽃 고정재료로써 사용되어진 식물을 포함한 자연물과 생활 속 재활용품은 침봉이나 플로랄 폼과 비슷한 고정능력과 세우는 능력을 가지고 있었다. 따라서 가급적 플로랄 폼의 사용을 자제하고 환경에 피해를 주지 않는 친환경적 꽃장식을 하는 것이 바람직할 것이다.

This study carried out development of a natural antimicrobial agent from Schima wallichii ssp. liukiuensis. Compound I exhibiting potent antimicrobial activity against Candida spp. was isolated from the methanol extracts of Schima wallichii ssp. liukiuensis. The structure of I identified as a sterol glycoside consisted of a trisaccharide and α1-sitosterol. Trisaccharide composed of L-rhamnose, D-galactose and D-glucose residues. The antimicrobial activity of I was selective on yeast rather than bacteria or other fungi. Compound I was demonstrated to be ineffective against toxicity to mouse liver cells where as protective to human dermal fibroblast cells at low concentrations. Thus, it is reasonable to expect a sterol glycoside (I) as a valuable alternative for synthetic antifungal.

최근 건강에 대한 관심이 높아지면서 과일 가공제품의 소비가 급증하고 있다. 포도는 페놀화합물의 주요 급원의 하나로 포도주스는 항산화, 항염증, 항혈소판 작용 등의 기능을 가지는 다수의 플라보노이드를 포함하고 있다. 일반적으로 캠벨 포도는 농가에서 주스나 와인으로 가장 널리 가공되고 있으나 다양한 포도 품종을 이용하여 브렌딩한 포도주스에 대한 연구는 거의 전무한 실정이다. 본 연구에서는 캠벨과 타품종 포도주스를 종류 및 비율을 달리하여 브렌딩 한

벼의 질소시비량 및 수확시기에 변화에 따른 도정특성 및 쌀 품질변화를 구명하기 위하여 일품벼와 추청벼를 시험품종으로 질소시비량과 수확시기를 다르게 하여서 시험하였다. 질소시비량은 무처리를 대조구로 하여 5 kg, 9 kg, 13 kg, 17 kg, 21 kg/10a의 5수준으로 하였고 질소시비방법은 기비-분얼비-수비를 5:2:3으로 분시하였으며, 인산과 칼리의 시비량 및 시비방법은 표준재배법에 준하여 실시하였다.수확시기는 출수 후40일, 50일, 60일, 70일, 80일로 하였다. 두 품종 모두 질소시비량과 수확시기가 달라짐에 따라 도정률에 큰 변화가 없었다. 그렇지만 질소시비량은 정현비율과 정의 상관관계를 나타내었지만 현백비율과는 부의 상관관계를 나타내었다. 품종간에는 추청벼가 일품벼보다 평균적으로 모든 시비 수준에서 높은 도정률을 나타내었다. 질소시비량과 완전미율은 부의 상관관계를 나타내었다. 품종간에는 13 kg/10a 이하의 시비수준에서는 추청벼가 일품벼보다 높은 완전미율을 나타내었지만 그보다 높은 수준에서는 일품벼가 추청벼보다 약간 높았다. 두 품종 모두 질소시비량이 증가할수록 분상질미와 싸라기 발생은 높아지는 경향이었다. 질소시비량과 수확시기에 따른 쌀의 단백질함량 변화는 질소시비량이 증가할수록 단백질함량 또한 증가하였으나 수확시기에 따라서는 차이가 없었다.