Genetic and physiological requirements for high-level sesquiterpene-production in tomato glandular trichomes
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PROVIDER: PRJEB58757 | ENA |
REPOSITORIES: ENA
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Project description:Glandular trichomes (GT) are specialized cell factories that have the capacity to produce large amounts of metabolites which can amount to over 10% of the leaf dry weight. The specific expression of secondary metabolite pathways in glandular trichomes has facilitated their elucidation. However, little is known about the connection between central carbon and specialized metabolism in these cells. To address this question, we used the type VI glandular trichomes from a cultivated (Solanum lycopersicum LA4024) and a wild tomato accession (Solanum habrochaites LA1777) as a model. Our study is based on metabolomics, transcriptomics, proteomics and 13C-labeling datasets of trichome and leaves samples. This comparative analysis allowed us to identify specific features of trichomes in comparison to leaves. Here, the transcript raw and RMA-normalized data is hosted.
2017-05-31 | E-MTAB-4482 | biostudies-arrayexpress
Project description:Type-VI glandular trichomes of wild tomato Solanum habrochaites PI127826 produce high levels of the sesquiterpene 7-epizingiberene and its derivatives, making the plant repellent and toxic to several pest insects and pathogens. How wild tomato trichomes achieve such high terpene production is still largely unknown. Here we show that a cross (F1) with a cultivated tomato produced only minute levels of 7-epizingiberene. In the F2-progeny, selected for the presence of the 7-epizingiberene biosynthesis genes, only three percent produced comparable amounts the wild parent, indicating this trait is recessive and multigenic. Moreover, trichome density alone did not explain the total levels of terpene levels found on the leaves. We selected F2 plants with the "high-production active-trichome phenotype" of PI127826, having trichomes producing about 150 times higher levels of terpenes than F2 individuals that displayed a "low-production lazy-trichome phenotype". Terpene quantities in trichomes of these F2 plants correlated with the volume of the storage cavity and shape of the gland. We found that trichome morphology is not a predetermined characteristic, but cavity volume rather depended on gland-cell metabolic activity. Inhibitor assays showed that the plastidial-precursor pathway (MEP) is fundamental for high-level production of both cytosolic as well as plastid-derived terpenes in tomato trichomes. Additionally, gene expression profiles of isolated secretory cells showed that key enzymes in the MEP pathway were higher expressed in active trichomes. We conclude that the MEP pathway is the primary precursor-supply route in wild tomato type-VI trichomes and that the high-production phenotype of the wild tomato trichome is indeed a multigenic trait.
| S-EPMC10020594 | biostudies-literature
Project description:Glandular trichomes (GT) are specialized cell factories that have the capacity to produce large amounts of metabolites which can amount to over 10% of the leaf dry weight. The specific expression of secondary metabolite pathways in glandular trichomes has facilitated their elucidation. However, little is known about the connection between central carbon and specialized metabolism in these cells. To address this question, we used the type VI glandular trichomes from a cultivated (Solanum lycopersicum LA4024) and a wild tomato accession (Solanum habrochaites LA1777) as a model. Our study is based on metabolomics, transcriptomics, proteomics and 13C-labeling datasets of trichome and leaves samples. This comparative analysis allowed us to identify specific features of trichomes in comparison to leaves. Tomato type VI trichomes are photosynthetic but acquire their carbon essentially from the leaves in the form of sucrose. Relatively high expression levels of photosystem genes and comparatively low expression of Calvin-cycle and photorespiration genes indicate that photosynthesis is used primarily to produce energy and reducing power to drive the high metabolic activity. High levels of polyunsaturated fatty acids, oxylipins, and of oxidized glutathione and high transcript and protein levels of associated enzymes further indicate a distinct capacity of GT to cope with oxidative stress. Enzymes which are involved in the supply of key precursors for the production of terpenoids both from the cytosol and in the chloroplasts are markedly increased. Finally, some enzymes that are typically associated with C4-photosynthesis are highly expressed in trichomes, suggesting that CO2 recycling processes may increase the efficiency of C usage in these cells. Based on this information we propose a model on how the glandular trichomes achieve high metabolic efficiency.
2017-07-04 | PXD003154 | Pride
Project description:Glandular trichomes are metabolic cell factories with the capacity to produce large quantities of secondary metabolites. Little is known about the connection between central carbon metabolism and metabolic productivity for secondary metabolites in glandular trichomes. To address this gap in our knowledge, we performed comparative metabolomics, transcriptomics, proteomics and 13C-labeling of type VI glandular trichomes and leaves from a cultivated (Solanum lycopersicum LA4024) and a wild (Solanum habrochaites LA1777) tomato accession. Specific features of glandular trichomes that drive the formation of secondary metabolites could be identified. Tomato type VI trichomes are photosynthetic but acquire their carbon essentially from leaf sucrose. The energy and reducing power from photosynthesis are used to support the biosynthesis of secondary metabolites, while the comparatively reduced Calvin-Benson-Bassham cycle activity may be involved in recycling metabolic CO2. Glandular trichomes cope with oxidative stress by producing high levels of polyunsaturated fatty acids, oxylipins, and glutathione. Finally, distinct mechanisms are present in glandular trichomes to increase the supply of precursors for the isoprenoid pathways. Particularly, the citrate-malate shuttle supplies cytosolic acetyl CoA and plastidic glycolysis and malic enzyme support the formation of plastidic pyruvate. A model is proposed on how glandular trichomes achieve high metabolic productivity.
2017-04-25 | MTBLS297 | MetaboLights
Project description:Genetic analysis of interspecific populations derived from crosses between the wild tomato species Solanum habrochaites f glabratum, which synthesizes and accumulates insecticidal methylketones (MKs) such as 2-undecanone and 2-tridecanone in glandular trichomes, and Solanum lycopersicum (cultivated tomato), which does not, demonstrated that MK metabolism in the wild species can be attributed to several loci. Comparative trascriptome analysis of the glandular trichomes of F2 segregants bulked into low- and high-MK plants identified several genes whose transcripts were either more or less abundant in the high-MK plants.
2009-06-05 | GSE16431 | GEO
Project description:Purpose: Plant trichomes are hair-like protuberances developing from epidermal cells of most terrestrial plants, serve as an excellent system for analyzing the molecular mechanisms underlying cell fate choices, cell cycle control, and cell morphogenesis. Regulation of trichomes in arabidopsis has been well characterized. However, the mechanisms of multicellular trichomes is unclear. To further identify the genes responsible for the SlCycB2-mediated phenotype change, we compared the gene expression profiles of SlCycB2-OE and RNAi plants by RNA-seq. Methods: The transcriptomes of between SlCycB2-OE and RNAi in tomato (Solanum lycopersicum) cultivar Ailsa Craig (AC) by RNA-seq analysis were evaluated using the Illumina HiSeq™ 2500 sequencing platform. Raw sequences were filtered and the resulting sets of clean reads were used for the following analysis by Hisat2 and DEGseq software. qRT-PCR validation was performed using SYBR Green assays. Results: In this study, we determined that SlCycB2 encodes a nuclear localized protein, which is highly expressed in trichomes, stigmas, hypocotyls and stems, and induced by the light, suppressed under the dark. Transgene analysis showed that suppression of SlCycB2 could promote type III and type V trichomes formation. On the contrary, overexpression of SlCycB2 could result in nearly disappearance of all non-glandular trichomes (including type III and V), as well as the glandular ones of type I and type VI. Detection of secondary metabolites indicated that the production of monoterpene and sesquiterpene were significantly decreased in SlCycB2-OE plants, which thus caused the reduction of the defense against Prodenia litura. Transcriptome profile demonstrated that the differential expressed genes mainly participate in the biosynthesis of terpenes, cutin, suberine and wax. Furthermore, we identified several homologs of this gene in many plants, SlCycB3 in tomato, NtCycB2 in tobacco, AtCycB2 in Arabidopsis, which have similar regulatory functions in trichome formation. SlCycB2 overexpression also led to abnormal flower with the unclosed stamen, shortened style and aberrant pollen. Conclusions: The results demonstrate that SlCycB2 play a critical role in multicellular trichome formation, secondary metabolite biosynthesis, Prodenia litura defense and reproductive organ development in tomato. The similar roles of its homologs in multicellular trichome formation suggest that Solanaceous species may share common regulatory pathway.
2016-12-02 | GSE90747 | GEO
Project description:Trichomes are found on the leaf surface of many plant species. They can be classified into either glandular or non-glandular types. Glandular trichomes are usually multicelluar structures and have long been known to participate in defense against pathogens through the production of specialized chemical products. Non-glandular trichomes, such as those in Arabidopsis thaliana, have been hypothesized to have roles in physical defense against insects, UV protection, detoxification, and heat dispersal. However, it has been difficult to separate a physical role from other possible functions for non-glandular trichomes. We have taken advantage of several mutants in the trichome developmental pathway and gene expression analyses to identify a set of genes expressed predominantly in Arabidopsis trichomes. Keywords: comparing gene expression between wild type and mutants
2008-04-30 | GSE9597 | GEO
Project description:Trichomes are found on the leaf surface of many plant species. They can be classified into either glandular or non-glandular types. Glandular trichomes are usually multicelluar structures and have long been known to participate in defense against pathogens through the production of specialized chemical products. Non-glandular trichomes, such as those in Arabidopsis thaliana, have been hypothesized to have roles in physical defense against insects, UV protection, detoxification, and heat dispersal. However, it has been difficult to separate a physical role from other possible functions for non-glandular trichomes. We have taken advantage of several mutants in the trichome developmental pathway and gene expression analyses to identify a set of genes expressed predominantly in Arabidopsis trichomes. Experiment Overall Design: We compared the gene expression between the wild type and three trichomeless mutants: gl1, ttg1 and gl3egl1 to see which genes' transcripts are enriched in trichomes.
2008-06-16 | E-GEOD-9597 | biostudies-arrayexpress
Project description:Background: Glandular trichomes found in vascular plants are called natural cell factories because they synthesize and store secondary metabolites in the glandular cell. In order to systematically understand the metabolic processes in the glandular cells, it is indispensable to analyze the cellular proteome dynamics. The conventional proteomics methods based on mass spectrometry have been enabled large-scale protein analysis, but requires a large number of trichome samples for in-depth analysis and is not suitable for rapid and sensitive quantification of targeted proteins.
Results: Here we present a high-throughput strategy for quantifying targeted proteins in a specific trichome glandular cell, using selected reaction monitoring (SRM) assay. SRM assay platform, targeting the proteins in tomato type VI trichome gland cells as a model study, demonstrated its effectiveness in multiple protein quantitation from a limited sample amount. A large-scale SRM assay developed using triple quadrupole mass spectrometer online connected to nanoflow liquid chromatography has enough capability to accurately measure the expression level of 221 targeted proteins contained in the glandular cell sample recovered from the 100 glandular trichomes within 120 minutes.
Conclusions: Targeted proteomics approach using established SRM assays makes it possible to quantify the proteins of interest with minimum effort in the sampling. The remarkable success of the SRM assay developments by a simple experimental workflow will increase the plant biologist's accessibility to proteomics research for glandular trichome.
2019-08-13 | PXD012250 | panorama