Panax ginseng C.A. meyer (family: Araliaceae) is a perennial crop that has been widely used as a traditional medicine in Korea. Various P. ginseng cultivars exhibit a range of morphological and physiological traits as well as genetic diversity. To elucidate the differences of primary metabolism underlying such genetic diverstiy, we performed primary metabolite profiles in adventitious roots from five Panax ginseng cultivars using gas chromatography-mass spectrometry (GC-MS). The GC-MS analysis revealed eight primary metabolites as biomarkers and allowed us to classify the five cultivars into three groups. We selected three cultivars to represent each group and analyzed their transcriptomes by Illumina sequencing. We inspected 100 unigenes involved in seven primary metabolite biosynthesis pathways and found that 21 unigenes encoding 15 enzymes were differentially expressed among the three cultivars. Integrated analysis of transcriptomes and metabolomes revealed that the ginseng cultivars differ in primary metabolites as well as in the putative genes involved in the complex process of primary metabolic pathways. Our data derived from this integrated analysis provide insights into the underlying complexity of genes and metabolites that co-regulate flux through these pathways in ginseng.

Panax Ginseng is a perennial medicinal plant originated from North-east asia. Because of its well-known tonic effects mainly from ginsenosides, various types of processed ginseng products have been distributed around the world. Here, we analyzed secondary metabolite profiling of adventitious roots of 5 korean ginseng cultivars, Chunpoong (CP), Sunhyang (SH), Gopoong (GO), Sunun (SU), and Cheongsun (CS). At the same time, the profiles of relative gene expressions related to ginsenoside biosynthesis pathway were compared among ginseng cultivars. Secondary metabolite profiles were revealed by UPLC/Q-TOF-MS from extracts of bioreactor derived adventitious roots of five ginseng cultivars. Using principal component analysis, secondary metabolite profiles of ginseng cultivars were categorized into three groups. Metabolites with high VIP values were annotated using known database and standards compounds. Relative gene expression of ginsenoside related gene were analyzed using realtime PCR. The three groups had distinct metabolite contents. Furthermore, gene expression profiles related to ginsenoside were also different, which might contribute diverse secondary metabolite composition of ginseng cultivars. Further integrated analysis would provide a relationship between genetic background of ginseng cultivars and secondary metabolite profiles.

The generation and analysis of genomic resources information are essential to understand genomic features of crops. Even though medicinal component and its effect of Panax ginseng was well studied, the genomic study has been recently started. The ginseng genome has been known to undergo two rounds of whole genome duplication (WGD), therefore we investigated an evidence of WGD in ginseng draft sequence for understanding current ginseng genome structure. Four paralogous gene-rich genome blocks were found, consisted by eight scaffolds, using about 3.0 Gb whole genome draft sequence and 48,821 unigenes of P. ginseng generated by whole genome shotgun sequencing. The eight scaffold sequences were ordered and connected into four genomic blocks, using zig-zag extension within scaffold sequences recently duplicated. The paralogous scaffold pairs that were recently duplicated showed high sequence conservation in genic and non-genic regions. However, paleo duplicated paralogue scaffold sequences showed little conservation only in genic regions. Finally, a total of 110 paralogous gene pairs and its expression were identified from recently and paleo duplicated scaffold pairs, which were co-linear among four genomic blocks. This study provides the first insight into duplicated genome structure of ginseng and will be a valuable information for further ginseng genomics including improvement of draft sequence quality, chromosome anchoring of scaffolds, and genetic mapping.

Clubroot is a devastating disease caused by Plasmodiophora brassicae and results in severe losses of yield and quality in Brassica crops including Brassica oleracea. Therefore, it is important to identify resistance gene for CR disease and apply it to breeding of Brassica crops. In this study, we applied genotyping-by-sequencing (GBS) technique to construct high resolution genetic map and mapping of clubroot resistance (CR) genes. A total of 18,187 GBS markers were identified between two parent lines resistant and susceptible to the disease, of which 4,103 markers were genotyped in all 78 F2 plants generated from crossing of both parent lines. The markers were clustered into nine linkage groups spanning 879.9 cM, generating high resolution genetic map enough to refine reported reference genome of cabbage. In addition, through QTL analysis using 78 F2:3 progenies and mapping based on the genetic map, two and single major QTLs were identified for resistance of race 2 and race 9 of P. brassicae, respectively. These QTLs did not show collinearity with CR loci found in Chinese cabbage (Brassica rapa) but roughly overlapped with CR loci identified in cabbage for resistance to race 4. Taken together, genetic map and QTLs obtained in this study will provide valuable information to improve reference genome and clubroot resistance in cabbage.

The C-repeat/dehydration-responsive element binding transcription factors (CBF/DREBs) are involved in an important pathway for abiotic stress-response in plants. We have identified CBF/DREB1 gene family from Brassica rapa whole genome sequence and designated them as BrDREB1s. They contain conserved nucleus localization signal, AP2/EREBP domain, and CBF/DREB1 signature, as other known plant CBF/DREB1s. By comparative genomics, we found that nine of ten BrDREB1 genes were present in seven macro-synteny blocks co-linear to four Arabidopsis counterpart blocks and also genomic organizations of their flanking regions were very similar to those for co-linear Arabidopsis CBF/DREB1 genes. In particular, three genes, BrDREB1A, BrDREB1B1, and BrDREB1C1, were closely located within a 59 kb genomic sequence, which was similar to that of their Arabidopsis counterpart genes. However, the genomic regions of those BrDREB1 genes contained additional sequences, compared to their co-linear regions in A. thaliana. The expression of BrDREB1 genes under abiotic stresses were examined by searching microarray database and by RT-PCR analysis. All of eight genes tested were highly up-regulated during cold treatment and some of them were also responsive to salt, drought, and ABA treatment. Taken together, these results indicate that CBF/DREB1-mediated stress signaling pathway is also functioning in B. rapa. On the other hand, differences in genomic organization and gene number for CBF/DREB1 are thought to cause different response to stress between B. rapa and A. thaliana. In this presentation, we will introduce more detailed results for CBF/DREB1 gene family in B. rapa.

Miniature inverted- repeat transposable elements are expected to play vital role in evolution of genes and genome of major eukaryotic organisms. However, there have been little reports on MITEs in B. rapa, a polyploidy model genome. We identified 13 novel MITE families in B. rapa genome by computational approach. Out of 13 MITEs families three, eight and two were classified under stowaway-like, tourist-like and hAT super families based on their unique structural characteristics. We characterized the members of 13 MITE families from the available 256 Mbp from whole genome draft sequences of B. rapa. We found ech MITE has high copiy number ranges from 14 to 977 which are distributed randomly along all the chromosomes. We also found more than 40% of the MITE members were associated with genes and gene rich regions. Furthermore, the polymorphism due to insertion and non-insertion of MITEs analysis suggest that MITEs are active in the genome. As, such the newly identified MITEs will provide a foundation for the further analysis of roles of MITEs in gene and genome evolution.

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most important vegetables and widely cultivated in Asia countries including Korea and China. Recently, whole genome sequence and full-length cDNA information of this species became available, which are encouraging genetic studies of this species to characterize agricultural important traits. Orange-colored (Or) cultivar of Chinese cabbage has inner leaves in orange, whereas other cultivars generally cultivated have yellow (Ye)- or white-colored inner leaves. In this study, we investigated phenotypes and carotenoid biosynthesis genes related to color variation in the Or cultivar. Firstly we compared the carotenoid content and composition between the Or and Ye cultivars by HPLC analysis. The inner leaves of Or cultivar contained approximately 9-fold high β-carotene content, whereas content of both lutein and violaxanthin was decreased to less than 30%, compared to Ye cultivar. Or cultivar was segregated with ratio of 3:1 in F2 population derived from crossing between Or and Ye inbred lines, indicating that Or phenotype is controlled by single recessive gene. To identify this gene, we investigated the expression of several genes involved in carotenoid biosynthesis by RT-PCR analysis. Among genes tested, two encoding putative carotenoid isomerase (CRTISO) and phytoene desaturase (PDS) were identified to show different expression between Or and Ye cultivars. Through further analysis of genomic DNA regions of these two genes, we could expect that several mutations such as InDel and base-substitution occurred and then affected expression of these genes in Or cultivar. In this presentation, I will introduce more detailed results for Or cultivars.

To characterize CBF/DREB1-homologue in rice, nine OsDREB1 genes have been identified and characterized in this lab. Among these, it was shown that OsDREB1D was induced by drought and slightly by cold stress. We found that OsDREB1A, -1D, and -1E could up-regulate OsDhn1:LUC construct in transactivation assay using rice protoplasts. Transgenic rice plants overexpressing OsDREB1D under the maize ubiquitin promoter (Ubi:OsDREB1D) revealed an enhanced stress tolerance to drought. We also generated transgenic rice of OsDREB1D under OsPOX1 promoter (OsPOX1:OsDREB1D), which is cold stress inducible preferentially in the reproductive organs of rice. We are currently examining the mechanism of the enhanced tolerance of the transgenic plants to drought stress using both molecular physiological and biochemical techniques.