@article {Willits:2004p8030,
	title = {Bio-fermentation of modified flavonoids: an example of in vivo diversification of secondary metabolites},
	journal = {Phytochemistry},
	volume = {65},
	number = {1},
	year = {2004},
	month = {Jan},
	pages = {31{\textendash}41},
	abstract = {A bio-fermentation technique was used for the in vivo diversification of flavonoid structures based on expression in Escherichia coli of six O-methyltransferases (OMTs) from Mentha x piperita and one O-glucosyltransferase (GT) each from Arabidopsis thaliana and Allium cepa. Enzymes were shown to be regio-specific in in vitro experiments and modified a broad range of flavonoid substrates at various positions. Using the flavonol quercetin as a model substrate, we show that the product spectrum produced with the in vivo approach is identical to that found in vitro. Additionally, using mixed cultures of E. coli expressing different classes of modifying genes (OMTs and GTs), the production of polymethylated flavonoid glucosides was observed. This report demonstrates the potential to increase the structural diversity of plant secondary metabolites using a multi-enzyme, bio-fermentation approach.},
	keywords = {Amino Acid Sequence, Arabidopsis, Cloning: Molecular, Consensus Sequence, Escherichia coli, Fermentation, Flavonoids, Glucosyltransferases, Mentha piperita, Methyltransferases, Molecular Sequence Data, Onions, Phylogeny, Recombinant Proteins, Sequence Alignment, Sequence Homology: Amino Acid, Stereoisomerism, Substrate Specificity},
	author = {Michael G Willits and Ma{\"{\i}t{\'e} Giovanni and Rogerio T N Prata and Catherine M Kramer and Vincenzo De Luca and John C Steffens and Gerson Graser}
}
@article {Ringer:2005p8013,
	title = {Monoterpene metabolism. Cloning, expression, and characterization of (-)-isopiperitenol/(-)-carveol dehydrogenase of peppermint and spearmint},
	journal = {Plant Physiol},
	volume = {137},
	number = {3},
	year = {2005},
	month = {Mar},
	pages = {863{\textendash}72},
	abstract = {The essential oils of peppermint (Mentha x piperita) and spearmint (Mentha spicata) are distinguished by the oxygenation position on the p-menthane ring of the constitutive monoterpenes that is conferred by two regiospecific cytochrome P450 limonene-3- and limonene-6-hydroxylases. Following hydroxylation of limonene, an apparently similar dehydrogenase oxidizes (-)-trans-isopiperitenol to (-)-isopiperitenone in peppermint and (-)-trans-carveol to (-)-carvone in spearmint. Random sequencing of a peppermint oil gland secretory cell cDNA library revealed a large number of clones that specified redox-type enzymes, including dehydrogenases. Full-length dehydrogenase clones were screened by functional expression in Escherichia coli using a recently developed in situ assay. A single full-length acquisition encoding (-)-trans-isopiperitenol dehydrogenase (ISPD) was isolated. The (-)-ISPD cDNA has an open reading frame of 795 bp that encodes a 265-residue enzyme with a calculated molecular mass of 27,191. Nondegenerate primers were designed based on the (-)-trans-ISPD cDNA sequence and employed to screen a spearmint oil gland secretory cell cDNA library from which a 5{\textquoteright}-truncated cDNA encoding the spearmint homolog, (-)-trans-carveol-dehydrogenase, was isolated. Reverse transcription-PCR amplification and RACE were used to acquire the remaining 5{\textquoteright}-sequence from RNA isolated from oil gland secretory cells of spearmint leaf. The full-length spearmint dehydrogenase shares >99\% amino acid identity with its peppermint homolog and both dehydrogenases are capable of utilizing (-)-trans-isopiperitenol and (-)-trans-carveol. These isopiperitenol/carveol dehydrogenases are members of the short-chain dehydrogenase/reductase superfamily and are related to other plant short-chain dehydrogenases/reductases involved in secondary metabolism (lignan biosynthesis), stress responses, and phytosteroid biosynthesis, but they are quite dissimilar (approximately 13\% identity) to the monoterpene reductases of mint involved in (-)-menthol biosynthesis. The isolation of the genes specifying redox enzymes of monoterpene biosynthesis in mint indicates that these genes arose from different ancestors and not by simple duplication and differentiation of a common progenitor, as might have been anticipated based on the common reaction chemistry and structural similarity of the substrate monoterpenes.},
	keywords = {Alcohol Oxidoreductases, Amino Acid Sequence, Cloning: Molecular, Mentha piperita, Mentha spicata, Molecular Sequence Data, Molecular Structure, Monoterpenes, Multigene Family, Oils: Volatile, Phylogeny},
	doi = {10.1104/pp.104.053298},
	author = {Kerry L Ringer and Edward M Davis and Rodney Croteau}
}
@article {Yamazaki:1999p8037,
	title = {Molecular cloning and biochemical characterization of a novel anthocyanin 5-O-glucosyltransferase by mRNA differential display for plant forms regarding anthocyanin},
	journal = {J Biol Chem},
	volume = {274},
	number = {11},
	year = {1999},
	month = {Mar},
	pages = {7405{\textendash}11},
	abstract = {UDP-glucose: anthocyanin 5-O-glucosyltransferase (5-GT) is responsible for the modification of anthocyanins to more stable molecules in complexes for co-pigmentation, supposedly resulting in a purple hue. The cDNA encoding 5-GT was isolated by a differential display applied to two different forms of anthocyanin production in Perilla frutescens var. crispa. Differential display was carried out for mRNA from the leaves of reddish-purple and green forms of P. frutescens, resulting in the isolation of five cDNA clones predominantly expressed in the red form. The cDNA encoded a polypeptide of 460 amino acids, exhibiting a low homology with the sequences of several glucosyltransferases including UDP-glucose: anthocyanidin 3-O-glucosyltransferase. By using this cDNA as the probe, we also isolated a homologous cDNA clone from a petal cDNA library of Verbena hybrida. To identify the biochemical function of the encoded proteins, these cDNAs were expressed in Saccharomyces cerevisiae cells. The recombinant proteins in the yeast extracts catalyzed the conversion of anthocyanidin 3-O-glucosides into the corresponding anthocyanidin 3,5-di-O-glucosides using UDP-glucose as a cofactor, indicating the identity of the cDNAs encoding 5-GT. Several biochemical properties (optimum pH, Km values, and sensitivity to inhibitors) were similar to those reported previously for 5-GTs. Southern blot analysis indicated the presence of two copies of 5-GT genes in the genome of both red and green forms of P. frutescens. The mRNA accumulation of the 5-GT gene was detected in the leaves of the red form but not in those of the green form and was induced by illumination of light, as observed for other structural genes for anthocyanin biosynthesis in P. frutescens.},
	keywords = {Amino Acid Sequence, Anthocyanins, Cloning: Molecular, DNA: Complementary, Glucosyltransferases, Lamiaceae, Molecular Sequence Data, Phylogeny, Recombinant Proteins, RNA: Messenger, Saccharomyces cerevisiae, Sequence Homology: Amino Acid},
	author = {M Yamazaki and Z Gong and M Fukuchi-Mizutani and Y Fukui and Y Tanaka and T Kusumi and K Saito}
}
@article {Kikuchi:2005p8019,
	title = {Centromere-specific repetitive sequences from Torenia, a model plant for interspecific fertilization, and whole-mount FISH of its interspecific hybrid embryos},
	journal = {Cytogenet Genome Res},
	volume = {109},
	number = {1-3},
	year = {2005},
	month = {Jan},
	pages = {228{\textendash}35},
	abstract = {Torenia fournieri is a good model plant to study fertilization in plants because it is easy to observe the fertilization process due to the protruding nature of the embryo sac. To study the location and movement of chromosomes and their centromeres in early embryogenesis of interspecific hybrid plants, we isolated two families of centromere-specific tandem repetitive sequences from T. fournieri and T. bailonii, and named them the "TCEN-family" and "BCEN-family", respectively. Both sequences consisted of a repeat unit of 52 bp located in the pericentric and centric heterochromatins. All signals of both sequences were prominent, but their intensity varied among the chromosomes. DNA-blot hybridization indicated the presence of similar sequences of TCEN-family in T. concolor, N. caerulea, and "Summer Wave", whereas the BCEN-family was found only in T. bailonii, thus indicating the wide or specific distribution of their repetitive families observed. We also applied whole-mount FISH to the interspecific hybrid embryos by using TCEN- and BCEN-family sequences as probes. Our results suggest that whole-mount FISH with the species-specific centromere sequences as probes is an ideal method to analyze the dynamics of chromosomes and centromeres in interspecific fertilization and early embryogenesis.},
	keywords = {Centromere, Chromosomes: Plant, Cloning: Molecular, DNA: Plant, In Situ Hybridization: Fluorescence, Lamiaceae, Phylogeny, Repetitive Sequences: Nucleic Acid, Reproduction},
	doi = {10.1159/000082405},
	author = {S Kikuchi and M Kishii and M Shimizu and H Tsujimoto}
}
@article {TapernouxLuthi:2004p8016,
	title = {Cloning, functional expression, and characterization of the raffinose oligosaccharide chain elongation enzyme, galactan:galactan galactosyltransferase, from common bugle leaves},
	journal = {Plant Physiol},
	volume = {134},
	number = {4},
	year = {2004},
	month = {Apr},
	pages = {1377{\textendash}87},
	abstract = {Galactan:galactan galactosyltransferase (GGT) is a unique enzyme of the raffinose family oligosaccharide (RFO) biosynthetic pathway. It catalyzes the chain elongation of RFOs without using galactinol (alpha-galactosyl-myoinositol) by simply transferring a terminal alpha-galactosyl residue from one RFO molecule to another one. Here, we report the cloning and functional expression of a cDNA encoding GGT from leaves of the common bugle (Ajuga reptans), a winter-hardy long-chain RFO-storing Lamiaceae. The cDNA comprises an open reading frame of 1215 bp. Expression in tobacco (Nicotiana plumbaginifolia) protoplasts resulted in a functional recombinant protein, which showed GGT activity like the previously described purified, native GGT enzyme. At the amino acid level, GGT shows high homologies (>60\%) to acid plant alpha-galactosidases of the family 27 of glycosylhydrolases. It is clearly distinct from the family 36 of glycosylhydrolases, which harbor galactinol-dependent raffinose and stachyose synthases as well as alkaline alpha-galactosidases. Physiological studies on the role of GGT confirmed that GGT plays a key role in RFO chain elongation and carbon storage. When excised leaves were exposed to chilling temperatures, levels of GGT transcripts, enzyme activities, and long-chain RFO concentrations increased concomitantly. On a whole-plant level, chilling temperatures induced GGT expression mainly in the roots and fully developed leaves, both known RFO storage organs of the common bugle, indicating an adaptation of the metabolism from active growth to transient storage in the cold.},
	keywords = {Ajuga, Amino Acid Sequence, Cloning: Molecular, Cold Temperature, DNA: Complementary, Galactosyltransferases, Gene Expression Regulation: Enzymologic, Gene Expression Regulation: Plant, Molecular Sequence Data, Phylogeny, Plant Leaves, Plant Proteins, Raffinose, Sequence Analysis: DNA, Sequence Homology: Amino Acid, Tobacco},
	doi = {10.1104/pp.103.036210},
	author = {Esther M Tapernoux-L{\"u}thi and Andreas B{\"o}hm and Felix Keller}
}
@article {Matsuno:2002p8029,
	title = {CYP98A6 from Lithospermum erythrorhizon encodes 4-coumaroyl-4{\textquoteright}-hydroxyphenyllactic acid 3-hydroxylase involved in rosmarinic acid biosynthesis},
	journal = {FEBS Lett},
	volume = {514},
	number = {2-3},
	year = {2002},
	month = {Mar},
	pages = {219{\textendash}24},
	abstract = {Rosmarinic acid is the dominant hydroxycinnamic acid ester accumulated in Boraginaceae and Lamiaceae plants. A cytochrome P450 cDNA was isolated by differential display from cultured cells of Lithospermum erythrorhizon, and the gene product was designated CYP98A6 based on the deduced amino acid sequence. After expression in yeast, the P450 was shown to catalyze the 3-hydroxylation of 4-coumaroyl-4{\textquoteright}-hydroxyphenyllactic acid, one of the final two steps leading to rosmarinic acid. The expression level of CYP98A6 is dramatically increased by addition of yeast extract or methyl jasmonate to L. erythrorhizon cells, and its expression pattern reflected the elicitor-induced change in rosmarinic acid production, indicating that CYP98A6 plays an important role in regulation of rosmarinic acid biosynthesis.},
	keywords = {Acetates, Cell Extracts, Cells: Cultured, Cinnamates, Cloning: Molecular, Cyclopentanes, Cytochrome P-450 Enzyme System, Depsides, DNA: Complementary, Gene Expression, Gene Expression Profiling, Lithospermum, Mixed Function Oxygenases, Molecular Sequence Data, Oxylipins, Phenylalanine Ammonia-Lyase, Phenylpropionates, Phylogeny, Plant Proteins, RNA: Messenger},
	author = {Michiyo Matsuno and Akito Nagatsu and Yukio Ogihara and Brian E Ellis and Hajime Mizukami}
}
@article {Hirotani:2000p8040,
	title = {Cloning and expression of UDP-glucose: flavonoid 7-O-glucosyltransferase from hairy root cultures of Scutellaria baicalensis},
	journal = {Planta},
	volume = {210},
	number = {6},
	year = {2000},
	month = {May},
	pages = {1006{\textendash}13},
	abstract = {A cDNA encoding UDP-glucose: baicalein 7-O-glucosyltransferase (UBGT) was isolated from a cDNA library from hairy root cultures of Scutellaria baicalensis Georgi probed with a partial-length cDNA clone of a UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT) from grape (Vitis vinifera L.). The heterologous probe contained a glucosyltransferase consensus amino acid sequence which was also present in the Scutellaria cDNA clones. The complete nucleotide sequence of the 1688-bp cDNA insert was determined and the deduced amino acid sequences are presented. The nucleotide sequence analysis of UBGT revealed an open reading frame encoding a polypeptide of 476 amino acids with a calculated molecular mass of 53,094 Da. The reaction product for baicalein and UDP-glucose catalyzed by recombinant UBGT in Escherichia coli was identified as authentic baicalein 7-O-glucoside using high-performance liquid chromatography and proton nuclear magnetic resonance spectroscopy. The enzyme activities of recombinant UBGT expressed in E. coli were also detected towards flavonoids such as baicalein, wogonin, apigenin, scutellarein, 7,4{\textquoteright}-dihydroxyflavone and kaempferol, and phenolic compounds. The accumulation of UBGT mRNA in hairy roots was in response to wounding or salicylic acid treatments.},
	keywords = {Amino Acid Sequence, Blotting: Northern, Chromatography: High Pressure Liquid, Cloning: Molecular, Escherichia coli, Flavonoids, Glucosyltransferases, Lamiaceae, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Phylogeny, Plant Roots, Recombinant Proteins},
	author = {M Hirotani and R Kuroda and H Suzuki and T Yoshikawa}
}
