We developed an efficient protocol for transformation of balloon flower (Platycodon grandiflorum) root cultures by using cotyledon leaf explants that were infected by Agrobacterium rhizogenes. We found that four different strains of A. rhizogenes differed in their ability to transform P. grandiflorum hairy root cultures. We also found correct antibiotics concentration for selection after transformation by Agrobacterium. Our results demonstrate that use of suitable strain of A. rhizogenes and correct level of antibiotics for the hairy root culture, and may allow to study and apply of valuable metabolites like phenolic compounds from P. grandiflorum hairy root culture.

Buckwheat (Fagopyrum esculentum Moench.) is an alternative crop belonging to the Polygonaceae family. Buckwheat has been well known as a plant source of rutin, quercetin, kaempferol-3-rutinoside and other phenolic compounds. Buckwheat contains more rutin than most of the other plants, which exhibits antioxidative, antihemorrhagic and blood vessel protecting properties. Phenylalanin ammonia-lyase (PAL), chalcone synthase (CHS), flavone 3-hydroxylase (F3H), and anthocyanidin synthase (ANS) were important enzymes involved in the flavonoid biosynthesis in buckwheat. In this study, we have purified full-length PAL, CHS, F3H and ANS from buckwheat using RACE PCR with degenerated primers targeted to conserved regions of orthologous sequences PAL, CHS, F3H and ANS available. The open reading frame of FePAL was 2112 nucleotides long, encodes for 704 amino acids. FePAL gene shares 80-81% identity and 88-90% similarity with Arabidopsis thaliana PAL1, Capsicum annuum PAL, Lactuca sativa PAL, and Populus trichocarpa PAL2. The cDNA of FeCHS has 1179 nucleotides open reading frame, encoding for 393 amino acids. FeCHS has identities and similarities ranging 84-86% and 92-95%, respectively, with other CHSs from P. trichocarpa, Citrus sinensis, Gerbera hybrid, and Vitis rotundifolia, whereas this gene shares 93% identity and 98% similarity with Polygonum cuspidatum. The cDNA of FeF3H has 1101 nucleotides open reading frame, encoding for 367 amino acids. FeF3H has identities and similarities ranging 76-81% and 92-97%, respectively, with other F3Hs from Gossypium hirsutum, Dimocarpus longan, Garcinia mangostana, Camellia sinensis, Vitis vinifera, Rubus coreanus, Pyrus communis, Glycine max, and Actinidia chinensis. The open reading frame of FeANS was 1074 nucleotides long, encodes for 358 amino acids. FeANS gene shares 74-76% identity and 94-96% similarity with Gypsophila elegans, C. sinensis and G. hirsutum. These result provieded the theoretical basis for the metabolic engineering on buckwheat.

Triterpene saponins (ginsenosides) are main bioactive compounds in Panax ginseng. From mevalonic acid pathway, the isoprene units was synthesized that is precusor of triterpene saponin. This study provides the complete nucleotide sequence of the full-length cDNA involved in mevalonic acid biosynthesis pathway. We had a full-length cDNA sequences as followed; HMGS (1944bp, 1407bp ORF), HMGR (2120bp,1728bp ORF) and MDD (1760bp, 1263bp ORF) with partial poly (A). We were used real time PCR and High Performance Liquid Chromatography (HPLC) to analysis the expression level of the gene involved in MVA pathway and to ginsenoside contents, respectively. All cloned genes are constitutively expressed in all tested tissues but at different levels with the highest expression in fine roots. But ginsenoside contents is highest in leaf > fine root> lateral root > red berry> main root > petiol > stem. After treantment of methyl Jasmonate (MeJA), MVA pathway genes are highly expressed and ginsenoside contents was high at 72h.

Methyl jasmonate(MeJA) treatment promotes the plant secondary, especially triterpene saponin which is main compounds in Platycodon grandiflorum. We were performed 5,760 expressed sequence tags from MeJA treated hairy root. Total number of ESTs are 2,536 (811 contigs and 1,725 singletons) among them 2,299 are pass the annotation. The most abundant transcript were encoding fatty acid desaturase (FAD) and lots of secondary metabolite transcript were observed. Four MVA pathway genes (HMGS, HMGR, MK, MDD) and one MEP pathway gene (IDI) are identified. The unigene were classified into three major functional categories by standard GO (Gene Ontology) - cellular component, molecular function and biological process. In this study we were cloned four genes from ESTs : HMGS (1401bp ORF), MK (1167bp ORF), MDD (1254bp ORF), IDI (909bp ORF)

Allium sativum, belongs to a member of the onion family (Alliaceae) are economically important vegetables because of the culinary value and medicinal purpose. Using PCR strategy with degenerated primers targeted to conserved regions of orthologous phenylalanine ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) sequences available, full-length PAL and C4H from A. sativum. The amino acid sequence of these genes is highly conserved, particularly AsPAL and AsC4H has greater than 70% amino acid identity to other plants. AsPAL and AgC4H were most highly expressed in roots of A. sativum, whereas lowest level of transcript was detected in flower. Phenolic compounds most highly produced in flowers of A. sativum. The presented sequences and expression an alysis of PAL and C4H will provide possible material to enhance the understading of phenolic compounds synthesis in A. sativum.

The pollen grain is a unique tricellular structure suitable for the delivery of the sperm cells to the ovule. All nutrients required for microspore and pollen cell growth are derived by passage through the anther locule and secretion by the tapetum lining. During later stages the tapetum degenerates but contributes to produce pigments, waxes, lipids and proteins which form the pollen coat and function in signaling between male (pollen) and female (pistil) tissues. The development of both normal pollen and tapetum is necessary for the fertilization processes in rice and would be exploited for the induction of male-sterility which is very useful to improve economic value of crops. We aredeveloping new approaches using a conditional male-sterility for the F1 hybrid seed production in rice. The conventional three parental systems for F1 hybrid seed production requirethe following three lines: male-sterile line, maintainer line, and restorer line. In this system, a critical requirement is to maintain the male-sterile inbred lines. Here we suggest molecular approaches, in which the engineered male-sterile plants are generated by regulating endogenous hormonal balance through the loss-of-function of genes. We can expect the male-sterility can be restored by exogenous applications of hormones such as gibberellin or jasmonic acid. Based on two parental systems, we will address the answer onfollowing question: how can we maintain a male-sterile line producing 100% male-sterile progenies without a maintainer? This work was supported by grants from Crop Functional Genomics Center of the 21C Frontier Program (CG1517), RepublicKorea.

Bitter melon (Momordica charantia L.) is a tropical and subtropical vine of the family Cucurbitaceae, widely grown for edible fruit, which is among the most bitter of all vegetables. Bitter melon contains an array of biologically active plant chemicals including triterpenes, proteins, and steroids. The fruit is useful in gout, rheumatism and subacute cases of the spleen and liver diseases. It is supposed to purify blood and dissipate melancholia. It has also been shown to have hypoglycaemic properties (anti-diabetic) in animal as well as human studies. Raw materials of used in pharmaceuticals, health food, cosmetics, food, perfumery and other industries. However, bitter melon is not popular in Korea. For research, development and commercialization of bitter melon in Korea, we collected 13 varieties of bitter melon from domestic andabroad. We study biological and chemical characteristics of bitter melon. In general frui tsize and weight of Philippines and Japanese varieties was larger than that of Chinese and Korean. We are studying chemical analysis of useful compound (diosgenin and GABA) from bitter melon.