Characterization of an Acyltransferase Capable of Synthesizing Benzylbenzoate and Other Volatile Esters in Flowers and Damaged Leaves of <i>Clarkia breweri</i>

D’Auria, John C.; Chen, Feng; Pichersky, Eran

doi:10.1104/pp.006460

Cited by 192 publications

(163 citation statements)

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“…RNA gel blots were performed as described previously (D'Auria et al, 2002) using 2 and 5 mg of RNA in each lane for glands and leaf samples, respectively. An MKS1 probe was synthesized with the rediprime II kit (Amersham Pharmacia Biotech, Piscataway, NJ) using a PCR-amplified fragment of MKS1.…”

Section: Rna Extraction and Gel Blot Analysismentioning

confidence: 99%

Metabolic, Genomic, and Biochemical Analyses of Glandular Trichomes from the Wild Tomato SpeciesLycopersicon hirsutumIdentify a Key Enzyme in the Biosynthesis of Methylketones

et al. 2005

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Medium-length methylketones (C7-C15) are highly effective in protecting plants from numerous pests. We used a biochemical genomics approach to elucidate the pathway leading to synthesis of methylketones in the glandular trichomes of the wild tomato Lycopersicon hirsutum f glabratum (accession PI126449). A comparison of gland EST databases from accession PI126449 and a second L. hirsutum accession, LA1777, whose glands do not contain methylketones, showed that the expression of genes for fatty acid biosynthesis is elevated in PI126449 glands, suggesting de novo biosynthesis of methylketones. A cDNA abundant in the PI126449 gland EST database but rare in the LA1777 database was similar in sequence to plant esterases. This cDNA, designated Methylketone Synthase 1 (MKS1), was expressed in Escherichia coli and the purified protein used to catalyze in vitro reactions in which C12, C14, and C16 β-ketoacyl–acyl-carrier-proteins (intermediates in fatty acid biosynthesis) were hydrolyzed and decarboxylated to give C11, C13, and C15 methylketones, respectively. Although MKS1 does not contain a classical transit peptide, in vitro import assays showed that it was targeted to the stroma of plastids, where fatty acid biosynthesis occurs. Levels of MKS1 transcript, protein, and enzymatic activity were correlated with levels of methylketones and gland density in a variety of tomato accessions and in different plant organs.

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Section: Rna Extraction and Gel Blot Analysismentioning

confidence: 99%

Metabolic, Genomic, and Biochemical Analyses of Glandular Trichomes from the Wild Tomato SpeciesLycopersicon hirsutumIdentify a Key Enzyme in the Biosynthesis of Methylketones

et al. 2005

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“…Several genes encoding enzymes involved in benzenoid biosynthesis have been identified from C. breweri (Dudareva et al, 1998;D'Auria et al, 2002), snapdragon (Antirrhinum majus) (Murfitt et al, 2000), and rose (Rosa hybrida) (Lavid et al, 2002). Only two corresponding cDNAs have been identified from petunia petals, S-adenosyl-L-Met:benzoic acid carboxyl methyltransferase (BSMT) and benzoyl-CoA:benzyl alcohol/phenylethanol benzoyltransferase (BPBT) (Negre et al, 2003;Boatright et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

ODORANT1Regulates Fragrance Biosynthesis in Petunia Flowers

et al. 2005

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Floral scent is important to plant reproduction because it attracts pollinators to the sexual organs. Therefore, volatile emission is usually tuned to the foraging activity of the pollinators. In Petunia hybrida, volatile benzenoids determine the floral aroma. Although the pathways for benzenoid biosynthesis have been characterized, the enzymes involved are less well understood. How production and emission are regulated is unknown. By targeted transcriptome analyses, we identified ODORANT1 (ODO1), a member of the R2R3-type MYB family, as a candidate for the regulation of volatile benzenoids in Petunia hybrida cv W115 (Mitchell) flowers. These flowers are only fragrant in the evening and at night. Transcript levels of ODO1 increased before the onset of volatile emission and decreased when volatile emission declined. Downregulation of ODO1 in transgenic P. hybrida Mitchell plants strongly reduced volatile benzenoid levels through decreased synthesis of precursors from the shikimate pathway. The transcript levels of several genes in this pathway were reduced by suppression of ODO1 expression. Moreover, ODO1 could activate the promoter of the 5-enol-pyruvylshikimate-3-phosphate synthase gene. Flower pigmentation, which is furnished from the same shikimate precursors, was not influenced because color and scent biosynthesis occur at different developmental stages. Our studies identify ODO1 as a key regulator of floral scent biosynthesis.

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“…RNA gel blot analysis was performed as previously described by Chen et al (2003) and D'Auria et al (2002).…”

Section: Determination Of Gene Expression By Rt-pcrmentioning

confidence: 99%

An Arabidopsis thaliana gene for methylsalicylate biosynthesis, identified by a biochemical genomics approach, has a role in defense

et al. 2003

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SummaryEmission of methylsalicylate (MeSA), and occasionally of methylbenzoate (MeBA), from Arabidopsis thaliana leaves was detected following the application of some forms of both biotic and abiotic stresses to the plant. Maximal emission of MeSA was observed following alamethicin treatment of leaves. A gene (AtBSMT1) encoding a protein with both benzoic acid (BA) and salicylic acid (SA) carboxyl methyltransferase activities was identi®ed using a biochemical genomics approach. Its ortholog (AlBSMT1) in A. lyrata, a close relative of A. thaliana, was also isolated. The AtBSMT1 protein utilizes SA more ef®ciently than BA, whereas AlBSMT1 catalyzes the methylation of SA less effectively than that of BA. The AtBSMT1 and AlBSMT1 genes showed expression in leaves under normal growth conditions and were more highly expressed in the¯owers. In A. thaliana leaves, the expression of AtBSMT1 was induced by alamethicin, Plutella xylostella herbivory, uprooting, physical wounding, and methyl jasmonate. SA was not an effective inducer. Using a b-glucuronidase (GUS) reporter approach, the promoter activity of AtBSMT1 was localized to the sepals of¯owers, and also to leaf trichomes and hydathodes. Upon thrip damage to leaves, AtBSMT1 promoter activity was induced speci®cally around the lesions.

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Characterization of an Acyltransferase Capable of Synthesizing Benzylbenzoate and Other Volatile Esters in Flowers and Damaged Leaves of Clarkia breweri

Cited by 192 publications

References 50 publications

Metabolic, Genomic, and Biochemical Analyses of Glandular Trichomes from the Wild Tomato SpeciesLycopersicon hirsutumIdentify a Key Enzyme in the Biosynthesis of Methylketones

Metabolic, Genomic, and Biochemical Analyses of Glandular Trichomes from the Wild Tomato SpeciesLycopersicon hirsutumIdentify a Key Enzyme in the Biosynthesis of Methylketones

ODORANT1Regulates Fragrance Biosynthesis in Petunia Flowers

An Arabidopsis thaliana gene for methylsalicylate biosynthesis, identified by a biochemical genomics approach, has a role in defense

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