Plants produce a vast array of specialized metabolites, many of which are used as pharmaceuticals, flavors, fragrances, and other high-value fine chemicals. However, most of these compounds occur in non-model plants for which genomic sequence information is not yet available. The production of a large amount of nucleotide sequence data using next-generation technologies is now relatively fast and cost-effective, especially when using the latest Roche-454 and Illumina sequencers with enhanced base-calling accuracy. To investigate specialized metabolite biosynthesis in non-model plants we have established a data-mining framework, employing next-generation sequencing and computational algorithms, to construct and analyze the transcriptomes of 75 non-model plants that produce compounds of interest for biotechnological applications. After sequence assembly an extensive annotation approach was applied to assign functional information to over 800,000 putative transcripts. The annotation is based on direct searches against public databases, including RefSeq and InterPro. Gene Ontology (GO), Enzyme Commission (EC) annotations and associated Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway maps are also collected. As a proof-of-concept, the selection of biosynthetic gene candidates associated with six specialized metabolic pathways is described. A web-based BLAST server has been established to allow public access to assembled transcriptome databases for all 75 plant species of the PhytoMetaSyn Project (www.phytometasyn.ca).

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Pinus densiflora Siebold et Zucc. is the major green canopy species in the mountainous area of Korea. To assess the response of resin acid biosynthetic genes to mechanical and chemical stimuli, we cloned cDNAs of genes encoding enzymes involved in the 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway (1-deoxy-d-xylulose 5-phosphate synthase (PdDXS), 1-deoxy-d-xylulose 5-phosphate reductoisomerase (PdDXR) and 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase (PdHDR)) by the rapid amplification of cDNA ends (RACE) technique. In addition, we cloned the gene encoding abietadiene synthase (PdABS) as a marker for the site of pine resin biosynthesis. PdHDR and PdDXS occurred as two gene families. In the phylogenetic trees, PdDXSs, PdDXR and PdHDRs each formed a separate clade from their respective angiosperm homologs. PdDXS2, PdHDR2 and PdDXR were most actively transcribed in stem wood, whereas PdABS was specifically transcribed. The abundance of PdDXS2 transcripts in wood in the resting state was generally 50-fold higher than the abundance of PdDXS1 transcripts, and PdHDR2 transcripts were more abundant by an order of magnitude in wood than in other tissues, with the ratio of PdHDR2 to PdHDR1 transcripts in wood being about 1. Application of 1 mM methyl jasmonate (MeJA) selectively enhanced the transcript levels of PdDXS2 and PdHDR2 in wood. The ratios of PdDXS2 to PdDXS1 and PdHDR2 to PdHDR1 reached 900 and 20, respectively, on the second day after MeJA treatment, whereas the transcript level of PdABS increased twofold by 3 days after MeJA treatment. Wounding of the stem differentially enhanced the transcript ratios of PdDXS2 to PdDXS1 and PdHDR2 to PdHDR1 to 300 and 70, respectively. The increase in the transcript levels of the MEP pathway genes in response to wounding was accompanied by two orders of magnitude increase in PdABS transcripts. These observations indicated that resin acid biosynthesis activity, represented by PdABS transcription, was correlated with the selective transcriptions of PdDXS2 and PdHDR2. Introduction of PdDXS2, PdHDR1 and PdHDR2 rescued their respective knockout Escherichia coli mutants, confirming that at least these three genes were functionally active. Intracellular targeting of the green fluorescent protein fused to the N-terminal 100 amino acid residues of these genes in the Arabidopsis transient expression system showed that the proteins were all targeted to the chloroplasts. Our results suggest that the MEP pathway regulates resin biosynthesis in the wood of P. densiflora by differential transcription of the multiple PdDXS and PdHDR genes.

4-(Cytidine 5'-diphospho)-2-C-methyl-D-erythritol kinase (CMK or YchB), the fourth enzyme of the 2-C-methyl-D-erythritol 4-phosphate pathway, phosphorylates the 2-hydroxyl group of 4-(cytidine 5'-diphospho)-2-C-methyl-D-erythritol in the presence of ATP. Two isogenes encoding CMK (GbCMK1 and GbCMK2) were cloned and characterized from Ginkgo biloba. The activities of both isozymes were confirmed by complementation assay using Escherichia coli NMW29, a ychB knock-out mutant. The transcript profiles of GbCMKs in the radicles and the cotyledons of the cultured Ginkgo biloba embryos demonstrated that the transcript levels of GbCMK1 were similar in both organs, whereas that of GbCMK2 was predominantly high in the ginkgolide-synthesizing radicles. Selective increases in the transcript abundance of GbCMK2 in the radicles, induced by light and methyl jasmonate treatments, were observed. These differential induction patterns of the transcripts imply GbCMK1 and GbCMK2 respectively have high correlations with the primary and the secondary metabolisms. The transit peptides of both isozymes delivered the fused green fluorescent protein (GFP) into the chloroplast in the Arabidopsis and the Nicotiana transient expression systems; interestingly, the transit peptide of GbCMK1 delivered the GFP protein into the cytosol and the nucleus in addition to the chloroplasts.

A new Rehmannia glutinosa cultivar 'Tokang' was derived from 'Jihwang 1' seedlings by medicinal crop breeding team of National Institute of Horticulture and Herbal Science, RDA in 2009. It has pink flower, dark brown seed coat, light yellow root cortex. The plant type was some rising from the ground. The content of catalpol and extract were higher than the 'Jihwang 1'(check variety). The content of catalpol was 4.55% and the extract was 71.2%. It showed stronger resistance to root rot compare with the 'Jihwang 1'. The regional yield trials were conducted at three different locations from 2007 to 2009. The average yield of 'Tokang' was 21.1ton/ha, which was 12% more than the 'Jihwang 1'. This cultivar is adaptable to the whole of Korea except for mountain areas. (Registration No. 4725