Rational Conversion of Substrate and Product Specificity in a <i>Salvia</i> Monoterpene Synthase: Structural Insights into the Evolution of Terpene Synthase Function

Kampranis, Sotirios C.; Ioannidis, Daphne; Purvis, Alan; Mahrez, Walid; Ninga, Ederina; Katerelos, Nikolaos A.; Anssour, Samir; Dunwell, J. M.; Degenhardt, Jörg; Makris, Antonios M.; Goodenough, Peter W.; Johnson, Christopher

doi:10.1105/tpc.106.047779

Cited by 200 publications

(255 citation statements)

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“…2017). It has been demonstrated that even small differences in genetic diversity (such as single amino acid substitutions) can yield large differences in chemical profiles (Kampranis et al., 2007), and several studies have found that higher intraspecific genetic diversity reduces herbivory and disease in plant populations (Hughes, Inouye, Johnson, Underwood, & Vellend, 2008). Moreover, emerging evidence has demonstrated how variation in intraspecific chemodiversity influences community diversity among different trophic interactions (Glassmire et al., 2016; Richards et al., 2015).…”

Section: Discussionmentioning

confidence: 99%

Discovering variation of secondary metabolite diversity and its relationship with disease resistance in Cornus florida L.

Pais

Xiang

2018

Ecology and Evolution

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Understanding intraspecific relationships between genetic and functional diversity is a major goal in the field of evolutionary biology and is important for conserving biodiversity. Linking intraspecific molecular patterns of plants to ecological pressures and trait variation remains difficult due to environment‐driven plasticity. Next‐generation sequencing, untargeted liquid chromatography–mass spectrometry (LC‐MS) profiling, and interdisciplinary approaches integrating population genomics, metabolomics, and community ecology permit novel strategies to tackle this problem. We analyzed six natural populations of the disease‐threatened Cornus florida L. from distinct ecological regions using genotype‐by‐sequencing markers and LC‐MS‐based untargeted metabolite profiling. We tested the hypothesis that higher genetic diversity in C. florida yielded higher chemical diversity and less disease susceptibility (screening hypothesis), and we also determined whether genetically similar subpopulations were similar in chemical composition. Most importantly, we identified metabolites that were associated with candidate loci or were predictive biomarkers of healthy or diseased plants after controlling for environment. Subpopulation clustering patterns based on genetic or chemical distances were largely congruent. While differences in genetic diversity were small among subpopulations, we did observe notable similarities in patterns between subpopulation averages of rarefied‐allelic and chemical richness. More specifically, we found that the most abundant compound of a correlated group of putative terpenoid glycosides and derivatives was correlated with tree health when considering chemodiversity. Random forest biomarker and genomewide association tests suggested that this putative iridoid glucoside and other closely associated chemical features were correlated to SNPs under selection.

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Section: Discussionmentioning

confidence: 99%

Discovering variation of secondary metabolite diversity and its relationship with disease resistance in Cornus florida L.

Pais

Xiang

2018

Ecology and Evolution

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“…The cell lysate was centrifuged for 25 min at 12,000g, and the supernatant was subsequently used for purification of the recombinant proteins. Proteins encoded by C. forskohlii diTPS and the characterized monoterpene cineole synthase from Greek sage (Salvia fruticosa [SfCIN]; Kampranis et al, 2007) were purified on 1-mL His SpinTrap columns (GE Healthcare) using elution buffer (binding buffer with 325 mM Imidazole and 5 mM dithiothreitol [DTT]) and desalted on PD MiniTrap G-25 columns (GE Healthcare) with a desalting buffer (20 mM HEPES, pH 7.2, 350 mM NaCl, 5 mM DTT, 1 mM MgCl 2 , 5% [v/v] glycerol). In vitro TPS assays were performed by adding 15 mM GGPP or GPP and 100 mg purified CfTPS (or SfCIN) enzymes in 397 mL enzyme assay buffer (50 mM HEPES, pH 7.2, 7.5 mM MgCl 2 , 5% [v/v] glycerol, 5 mM DTT).…”

Section: Functional Characterization Of Cftps: In Vitro Assaysmentioning

confidence: 99%

Manoyl Oxide (13R), the Biosynthetic Precursor of Forskolin, Is Synthesized in Specialized Root Cork Cells in Coleus forskohlii

et al. 2014

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“…Our existing understanding of the mechanism of this and other terpene synthases would have predicted that mutations in the active site were very likely to change the proportion of products of multi-product enzymes (11)(12)(13)(14)(15)(16)(17)(18). However, we did notice anecdotally that there was a significant change in the ratio of these PaLAS-like products that depended upon the conditions and temperature of the GC-MS injector and column.…”

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The Primary Diterpene Synthase Products of Picea abies Levopimaradiene/Abietadiene Synthase (PaLAS) Are Epimers of a Thermally Unstable Diterpenol

Keeling

Madilao

Zerbe

et al. 2011

Journal of Biological Chemistry

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The levopimaradiene/abietadiene synthase from Norway spruce (Picea abies; PaLAS) has previously been reported to produce a mixture of four diterpene hydrocarbons when incubated with geranylgeranyl diphosphate as the substrate: levopimaradiene, abietadiene, neoabietadiene, and palustradiene. However, variability in the assay products observed by GC-MS of this and orthologous conifer diterpene synthases over the past 15 years suggested that these diterpenes may not be the initial enzyme assay products but are rather the products of dehydration of an unstable alcohol. We have identified epimers of the thermally unstable allylic tertiary alcohol 13-hydroxy-8(14)-abietene as the products of PaLAS. The identification of these compounds, not previously described in conifers, as the initial products of PaLAS has considerable implications for our understanding of the complexity of the biosynthetic pathway of the structurally diverse diterpene resin acids of conifer defense.

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Rational Conversion of Substrate and Product Specificity in a Salvia Monoterpene Synthase: Structural Insights into the Evolution of Terpene Synthase Function

Cited by 200 publications

References 37 publications

Discovering variation of secondary metabolite diversity and its relationship with disease resistance in Cornus florida L.

Discovering variation of secondary metabolite diversity and its relationship with disease resistance in Cornus florida L.

Manoyl Oxide (13R), the Biosynthetic Precursor of Forskolin, Is Synthesized in Specialized Root Cork Cells in Coleus forskohlii

The Primary Diterpene Synthase Products of Picea abies Levopimaradiene/Abietadiene Synthase (PaLAS) Are Epimers of a Thermally Unstable Diterpenol

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