Dataset Information

The roles of a flavone-6-hydroxylase and 7-O-demethylation in the flavone biosynthetic network of sweet basil.

ABSTRACT: Lipophilic flavonoids found in the Lamiaceae exhibit unusual 6- and 8-hydroxylations whose enzymatic basis is unknown. We show that crude protein extracts from peltate trichomes of sweet basil (Ocimum basilicum L.) cultivars readily hydroxylate position 6 of 7-O-methylated apigenin but not apigenin itself. The responsible protein was identified as a P450 monooxygenase from the CYP82 family, a family not previously reported to be involved in flavonoid metabolism. This enzyme prefers flavones but also accepts flavanones in vitro and requires a 5-hydroxyl in addition to a 7-methoxyl residue on the substrate. A peppermint (Mentha × piperita L.) homolog displayed identical substrate requirements, suggesting that early 7-O-methylation of flavones might be common in the Lamiaceae. This hypothesis is further substantiated by the pioneering discovery of 2-oxoglutarate-dependent flavone demethylase activity in basil, which explains the accumulation of 7-O-demethylated flavone nevadensin.

SUBMITTER: Berim A

PROVIDER: S-EPMC3548489 | biostudies-literature | 2013 Jan

REPOSITORIES: biostudies-literature

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The roles of a flavone-6-hydroxylase and 7-O-demethylation in the flavone biosynthetic network of sweet basil.

Berim Anna A Gang David R DR

The Journal of biological chemistry 20121126 3

Lipophilic flavonoids found in the Lamiaceae exhibit unusual 6- and 8-hydroxylations whose enzymatic basis is unknown. We show that crude protein extracts from peltate trichomes of sweet basil (Ocimum basilicum L.) cultivars readily hydroxylate position 6 of 7-O-methylated apigenin but not apigenin itself. The responsible protein was identified as a P450 monooxygenase from the CYP82 family, a family not previously reported to be involved in flavonoid metabolism. This enzyme prefers flavones but ...[more]

PMID: 23184958

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Project description:Sweet basil (Ocimum basilicum) is an economically important herb and its global production is threatened by basil downy mildew caused by the obligate biotrophic oomycete Peronospora belbahrii. Effective tools are required for functional understanding of its genes involved in synthesis of valuable secondary metabolites in essential oil and disease resistance, and breeding for varieties with improved traits. Clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 gene editing technology has revolutionized crop breeding and functional genomics. The applicability and efficacy of this genomic tool in the allotetraploid sweet basil were tested by editing a potential susceptibility (S) gene ObDMR1, the basil homolog of Arabidopsis DMR1 (Downy Mildew Resistant 1) whose mutations conferred nearly complete resistance against Arabidopsis downy mildew pathogen, Hyaloperonospora arabidopsidis. Two single guide RNAs targeting two different sites of the ObDMR1 coding sequence were designed. A total of 56 transgenic lines were obtained via Agrobacterium-mediated stable transformation. Mutational analysis of 54 T0 transgenic lines identified 92.6% lines carrying mutations at target 1 site, while a very low mutation frequency was detected at target 2 site. Deep sequencing of six T0 lines revealed various mutations at target 1 site, with a complete knockout of all alleles in one line. ObDMR1 homozygous mutant plants with some being transgene free were identified from T1 segregating populations. T2 homozygous mutant plants with 1-bp frameshift mutations exhibited a dwarf phenotype at young seedling stage. In summary, this study established a highly efficient CRISPR/Cas9-mediated gene editing system for targeted mutagenesis in sweet basil. This system has the capacity to generate complete knockout mutants in this allotetraploid species at the first generation of transgenic plants and transgene-free homozygous mutants in the second generation. The establishment of this system is expected to accelerate basil functional genomics and breeding.

Project description:For decisions on supplemental lighting a quantitative knowledge of the plants' responses to light under varying conditions is fundamental. In this study, we developed light dose-response curves of growth and morphological traits for Ocimum basilicum L. and examined the effects of light color (blue, red, and white plus far-red) and natural environment (season) on these curves. Four greenhouse experiments were conducted throughout the year to determine the efficiencies of the light regimes on growth and their effects on plant morphology. A special aspect was the photosynthetic efficiency of far-red light. Linear and monomolecular relationships were found for the relationships between plant traits and supplemental light dose. Traits related to biomass productivity increased linearly with light dose whereas some morphological characters showed a saturation behavior. Red light and white plus far-red light were more efficient in plant dry weight production than blue light, and the plants adapted differently to the light qualities: higher biomass under red light was related to a plant architecture more favorable for light capture, i.e., taller plants and bigger leaves. White plus far-red light, on the other hand, increased leaf mass per area (LMA) and light use efficiency (LUE). Blue light resulted in lowest plant light interception and LUE. Considering photosynthetic effects of near-infrared light (PPFD800, 400-800 nm) instead of photosynthetic photon flux density (PPFD700, 400-700 nm) led to strongly reduced efficiencies. Traits related to photosynthesis such as dry weight, LMA and LUE were particularly affected by PPFD800. There were no interactions between the efficiencies of the different light colors and the seasons. Efficiencies of all light regimes were significantly lower during summer compared to spring and winter. Higher dry weight production during summer compared to winter and spring were a consequence of increased light interception rather than changes in LUE. The observed differences in seasonal efficiencies were directly linked to the amount of natural light present as indicated by changes in the ratio of supplemental to natural light.

Dataset Information

The roles of a flavone-6-hydroxylase and 7-O-demethylation in the flavone biosynthetic network of sweet basil.

Publications

The roles of a flavone-6-hydroxylase and 7-O-demethylation in the flavone biosynthetic network of sweet basil.

Similar Datasets

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