Rice is one of the most important major food crops which provide the major food for more than half of global population. To improve the grain quality as well as grain yield has been the essential breeding goal in rice. The composition of amylopectin is the determinant of rice eating quality under certain threshold of protein content and the ratio of amylose and amylopectin. In this study, RBE 1 driven by CaMV-35S promoter was constructed and transformed using Agrobacterium tumefaciens. We selected single copy with low amylose content among transgenic lines. The mRNA expression was investigated using RT-PCR, and enzyme activity was determined using activity staining method in mid-milky stage endosperm. Also, the overexpression vectors for RBE 1 and SSS 1 driven by seed specific globulin promoter were constructed, respectively. Moreover, the RNA interference vectors for soluble starch synthase 1 and granule bound starch synthase 1 derived by CaMV35S promoter were constructed, respectively and transformed using Agrobacterium tumefaciens. The transgene has been confirmed by amplification of HPT and target gene. The transgenic plants obtained will be used to investigate the gene function of related starch pathway in plant cells using Gopumbyeo as a wild type rice, based on the gain-of-function and the loss-of-function. The development of designed site-specific endonucleases boosted the establishment of gene targeting (GT) techniques in a row of different species. However, the methods described in plants require a highly efficient transformation and regeneration procedure and, therefore, can be applied to very few species. Here, we describe a highly efficient GT system that is suitable for all transformable plants regardless of transformation efficiency. Efficient in planta GT was achieved in rice by expression of a site specific endonuclease (SSS1::ZFN) that not only cuts within the target but also the chromosomal transgenic donor, leading to an excised targeting vector.

The purposes of this research project are to identify quantitative trait loci (QTLs) associated with yield-related traits using a recombinant inbred line (RIL) population derived from a cross between a high-yield soybean genotype SS0404-T5-76 and Daewonkong and to develop high-yield soybean and lodging-resistant sprout soybean cultivars. For development of DNA markers and identification of functional sequence variations, firstly, whole genome of five soybean genotypes, Sinpaldalkong 2, SS2-2, Pungsanamulkong, SS0404-T5-76 and Daewongkong, were sequenced using Illumina Hi-Seq technology. SS2-2 is a EMS-induced mutant of Sinpadalkong 2. SS0404-T5-76 showing high-yield is a F8 RIL derived from a cross of Pungsanamulkong x SS2-2. Daewonkong is a elite cultivar with high-protein. Furthermore, to construct a genetic linkage map, we are advancing F4 lines of SS0404-T5-76 x Daewonkong by single seed-descent. Secondly, we developed high-protein and high-yield soybean lines and lodging-resistant sprout lines. Area-adaptability tests of these promising lines are performing in three different locations including Jeju, Naju, and Suwon. Based on the results of area adaptability tests, we are planing to conduct cultivar registration of the promising soybean lines.

In pepper, investigation of important traits such as disease resistances, high yield and pungency were mostly focused on a cultivated species, Capsicum annuum. This narrow breeding pool hampered to develop improved cultivars. Exploitation of wild germplasm in Capsicum has been recognized as an important issue. The construction of core collection and analysis of genetic diversity in Capsicum is the first step to make full use of germplasm. Although there have been several attempts to construct core collections in Capsicum, most of the works were limited due to handling small number of samples, relying mainly on the characterization of morphological traits and focusing on C. annuum species. Therefore, the comprehensive studies for genetic diversity and structure of Capsicum including phenotypic data, molecular marker patterns and evaluation of useful alleles are very necessary to understand the structure and patterns of genetic diversity in Capsicum. We are developing for a core collection set in Capsicum using molecular markers and phenotypic data with over 3,000 germplasm accessions.

The latest report on draft genome of Brassica rapa sequence has been published. To elucidate the functions of a large population of these genes and to search efficiently for agriculturally useful genes, the Full-length cDNA Over-eXpressor (FOX) gene hunting system was used. The FOX library was transformed into rice by Agrobacteriummediated transformation. Approximately 1,150 FOX-rice lines were generated. Genomic PCR analysis indicated that the average length of FL-cDNAs was 900∼1,200 bp with functional annotation of many unknown function (35.5%). Most of the randomly selected transgenic rice lines showed overexpression (92%) and barely mRNA expression in the wild type Gopum. Moreover, 94% of the 850 transgenic rice lines were moderately tolerant (slightly yellow) to cold and 9 lines were tolerant (seedling light green). For the salinity evaluation, most of the transgenic lines (85%) were highly susceptible whereas seven lines were tolerant. In addition, morphological evaluation of rice lines showed minimal phenotypic alteration (12%). About 25.1 and 22% were earlier in terms of days to heading and chlorophyll contents, respectively. Further, 18% of FOX rice lines showed lower chlorophyll contents. Filled grains, number of tillers, panicle length, culm and plant height were relatively less variable among the lines. These results provided useful genes for functional analyses in the mechanisms of identified transgenic tolerant lines.

The Cmr1 gene in peppers confers resistance to Cucumber mosaic virus isolate-P0 (CMV-P0). Cmr1 restricts the systemic spread of CMV-Fny, whereas this gene cannot block the spread of CMV-P1 to the upper leaves, resulting in systemic infection. To identify the virulence determinant of CMV-P1, six reassortant viruses and six chimeric viruses derived from CMV-Fny and CMV-P1 cDNA clones were used. Our results demonstrate that the helicase domain encoded by CMV-P1 RNA1 determines susceptibility to systemic infection. To identify the key amino acids determining systemic infection with CMV-P1, we then constructed amino acid substitution mutants. Of the mutants tested, amino acid residues at positions 865, 896, 957, and 980 in the 1a protein sequence of CMV-P1 affected the systemic infection. Virus localization studies with CMV-GFP clones and in situ localization of virus RNA revealed that these four amino acid residues together form the movement determinant for CMV-P1 movement from the epidermal cell layer to mesophyll cell layers. Quantitative real-time PCR revealed that CMV-P1 and a chimeric virus with four amino acid residues of CMV-P1 accumulated more genomic RNA in inoculated leaves than did CMV-Fny, indicating that those four amino acids are also involved in virus replication. These results demonstrate that the helicase domain is responsible for systemic infection by controlling virus replication and cell-to-cell movement. Whereas four amino acids are responsible for acquiring virulence in CMV-Fny, six amino acid (positions at 865, 896, 901, 957, 980 and 993) substitutions in CMV-P1 were required for complete loss of virulence in ‘Bukang’.

eIF4E family is well known for recessive resistance gene of potyvirus in many crops. And Turnip mosaic virus (TuMV) is one of the major viruses in Brassicaceae crops which belong to the genus Potyvirus. To elucidate the key amino acids in the interaction between TuMV VPg and Brassica eIF(iso)4E, amino acids of eIF(iso)4E were mutated. Seven amino acids in cap binding pocket were chose for the candidate amino acid that may play a role in the interaction of TuMV VPg. We demonstrated that a single amino acid mutation in cap binding pocket of Brassica eIF(iso)4E can abolish the interaction with TuMV VPg. eIF(iso)4E which has a mutation at each W49, W95 and K150 positions impaired in its interaction with VPg prominently according to the yeast two hybrid analysis. Complementation of an eIF4E knockout yeast strain by mutated eIF(iso)4E proteins showed that all eIF(iso)4E mutants were able to complement eIF4E of yeast. To find out if these mutations affect the susceptibility of Chinese cabbage, transformant analysis was performed. eIF(iso)4E W95L, W95L/K150E and susceptible wild type were over-expressed in susceptible Chinese cabbage. According to the TuMV screening result of T1 and T2 transformants, over-expression of the eIF(iso)4E mutants showed resistance to four TuMV strains (CHN2, 3, 4 and 5). Our results support that the mutations in eIF(iso)4E may control the broad spectrum TuMV resistance.

The antimicrobial peptide possesses defence system to virus, fungi and bacteria. To study antibiotic in plant, antimicrobial peptides were obtained by PCR analysis by primers designed from antimicrobial peptides (Gene bank accession no. NM-004345), cloned in pET28 expression vector and the vector transformed into E. coli. And this gene was inserted into Ti-plasmid VB2 vector, which contained the pGD1 promoter. The expression construction was transformed into Agrobacterium EHA105 and then plant tissues of rice (Oryza sativa). Seeds from transgenic plants (T0) were germinated on selective media containing spectinomycin 50 mg/L. Selected plants and wild type were analyzed by PCR and RT-PCR with pGD1 promoter region and transgene specific primer set. All transgenic plants showed expression pattern of similar levels. We showed that the chromobody is effective in binding GFPand antimicrobial peptide gene in tobacco leaf. Most interestingly, this can be applied to interfere with the function of GFP fusion protein and to mislocalize (trap) GFP fusions to the plant cytoplasm in order to alter the phenotype mediated by the targeted proteins. Bacterial blight disease was enhanced resistance in transgenic lines. These results showed that antibiotic peptides might show a broad-spectrum antimicrobial activity.

Kenaf (Hibiscus cannabinus) is an annual herbaceous plant of the family Malvaceae that has been planted in Africa for more than 4000 years and used as source of fiber, energy and feed stock. Also, kenaf seeds are good source for edible oil used for first class cooking oil and margarine production. The seeds can be used for lubrication, soap, paint and varnishes. This study was carried out to evaluate fatty acids variation among sixteen kenaf germplasm and gamma-ray induced mutants derived from Jinju and Auxu. Linoleic, oleic, and palmitic acid were the predominant fatty acids in all kenaf seed oils. The sixteen accessions showed a wide range of fatty acid compositions, spanning from 28.94 to 43.36% saturated, 56.64 to 71.05% total unsaturated, 15.52 to 46.85% monounsaturated, and 13.56 to 48.97% polyunsaturated fatty acids. The mutant lines derived from Jinju, significantly surpassed parental mean for all the palmitic and oleic acid. Also, the mutant lines derived from Auxu showed broad ranges of variation in oleic and linoleic acid and narrow ranges of variation in stearic and palmitic acid. The relative amount of monounsaturated fatty acids (MUFA) were increased at all the gamma-ray induced mutants. These results will provide a valuable information to assist parental selection of kenaf breeding.

Mutant populations generated by ethyl methanesulfonate (EMS) have been used for crop improvement and functional genomics. Since pepper is very recalcitrant to be transformed, EMS mutagenesis can be a very useful to generate useful alleles and to characterize the function of genes. We have been developing a mutant population aiming at 5,000 mutants by treating EMS on seeds of C. annuum ‘Yuwolcho’. A total of 4,300 M1 mutants have been developed until now. Among the 4,300 M1 population, almost 800 M2 mutant lines have been phenotyped and evaluated to confirm the effect of EMS. We categorized seven key organ development and subdivided them into twenty secondary categories. Among them, 50 and 72 families have been shown variations in plant growth and leaf development, respectively. In addition, we detected nucleotide variations using HRM analysis in eIF4E and putative aminotransferase genes. These results demonstrated that our mutant population can be very useful for study function of genes in near future.

For QTL analysis of agronomic traits based on cultivation of low and high altitude locations, BC1 F5 181 lines were developed from a cross of Tongil type Gayabyeo and japonica Chhomrong originated from Nepal. Plant materials were grown in both of low altitude area of Milyang, Korea and high altitude area of Khumaltar, Nepal. In QTLs analysis, a total of 42 QTLs were detected in days to flowering, culm length, panicle length, number of panicles/hill, panicle exertion, and spikelet ripening ratio. Although many of the QTLs were coincided between the two locations of Korea and Nepal, several QTLs were revealed as location specific in high altitude area of Khumaltar. Especially, the regions harboring marker RM489-RM14281 on chromo- some 3, RM5642-RM19049 on chromosome 5, and RM2854-RM6696 on chromosome 12 where QTLs were clustered and coincided between the two locations are considered as positive target regions for environmental independent traits. Furthermore, high rate of location specific QTLs such as panicle exertion and spikelet ripening ratio could be considered carefully for understanding the mechanism of cold tolerance based on cultivation of different altitude location.

A new species of Chrysosplenium (Saxifragaceae), C. epigealum J.W.Han & S.H.Kang is described from Mt. Seoraksan, Inje-gun, Gangwon-do, Korea. This new species is distinct from C. flaviflorum Ohwi, its closely relative species, in having calyx 2-2.5 mm long, pistils slightly shorter than calyx, filaments 2-3 times longer than anthers and stolons epigeal.

We presented fundamental stellar parameters and evolutionary statuses of six solar type detached eclipsing binaries whose masses are in the range of 0.97-1.43 M⊙. EK Cep and FL Lyr belong to the zero age main sequence. HS Hya, IT Cas and CD Tau are on the main sequence. Their ages are 1.3, 1.9 and 2.2 Gyr, respectively. Both component stars of AI Phe evolved to sub giants and its age is 4.0 Gyr. Those ages of the detached binary systems show good agreement with the time scale for synchronization and circularization of the binary systems.

The ultrastructural characteristics of germ cell differentiations during spermatogenesis and mature sperm morphology in male () were evaluated via transmission electron microscopic observation. The accessory cells, which contained a large quantity of glycogen particles and lipid droplets in the cytoplasm, are assumed to be involved in nutrient supply for germ cell development. Morphologically, the sperm nucleus and acrosome of this species are ovoid and conical in shape, respectively. The acrosomal vesicle, which is formed by two kinds of electron-dense or lucent materials, appears from the base to the tip: a thick and slender elliptical line, which is composed of electron-dense opaque material, appears along the outer part (region) of the acrosomal vesicle from the base to the tip, whereas the inner part (region) of the acrosomal vesicle is composed of electron-lucent material in the acrosomal vesicle. Two special characteristics, which are found in the acrosomal vesicle of A. () in Pinnidae (subclass Pteriomorphia), can be employed for phylogenetic and taxonomic analyses as a taxonomic key or a significant tool. The spermatozoa were approximately in length, including a sperm nucleus (about in length), an acrosome (about in length), and a tail flagellum (about ). The axoneme of the sperm tail evidences a 9+2 structure.