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Robert C Schuurink - One of the best experts on this subject based on the ideXlab platform.
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SlMYC1 Regulates Type VI Glandular Trichome Formation and Terpene Biosynthesis in Tomato Glandular Cells
The Plant Cell, 2018Co-Authors: Jiesen Xu, Zeger Otto Van Herwijnen, Dorthe Bettina Drager, Michel A. Haring, Robert C SchuurinkAbstract:Tomato (Solanum lycopersicum) glandular Trichomes function as biochemical factories that synthesize a diverse array of specialized metabolites. Terpenoids are the most diverse class of plant specialized metabolites, with volatile mono- and sesquiterpenes playing important roles in plant defense. Although the biosynthetic pathways of volatile terpenes in tomato glandular Trichomes have been well described, little is known about their regulation. Here, we demonstrate that SlMYC1, a basic helix-loop-helix transcription factor, differentially regulates mono- and sesquiterpene biosynthesis in the type VI glandular Trichomes of tomato leaves and stems. SlMYC1 functions as a positive regulator of monoterpene biosynthesis in both leaf and stem Trichomes but as a negative regulator of sesquiterpene biosynthesis in stem Trichomes. SlMYC1 is also essential for type VI glandular Trichome development, as knocking down SlMYC1 led to the production of smaller type VI glandular Trichomes at lower densities, and knocking out this gene led to their absence. Our findings reveal a role for SlMYC1 not only in type VI glandular Trichome development but also in the regulation of terpene biosynthesis in tomato.
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engineering of tomato glandular Trichomes for the production of specialized metabolites
Methods in Enzymology, 2016Co-Authors: Ruy W J Kortbeek, Aldana M Ramirez, Eleni A Spyropoulou, Paul Johan Diergaarde, I Ottenbruggeman, M T J De Both, Axel Schmidt, Raimund Nagel, J. Xu, Robert C SchuurinkAbstract:Glandular Trichomes are specialized tissues on the epidermis of many plant species. On tomato they synthesize, store, and emit a variety of metabolites such as terpenoids, which play a role in the interaction with insects. Glandular Trichomes are excellent tissues for studying the biosynthesis of specialized plant metabolites and are especially suitable targets for metabolic engineering. Here we describe the strategy for engineering tomato glandular Trichomes, first with a transient expression system to provide proof of Trichome specificity of selected promoters. Using microparticle bombardment, the Trichome specificity of a terpene-synthase promoter could be validated in a relatively fast way. Second, we describe a method for stable expression of genes of interest in Trichomes. Trichome-specific expression of another terpene-synthase promoter driving the yellow-fluorescence protein-gene is presented. Finally, we describe a case of the overexpression of farnesyl diphosphate synthase (FPS), specifically in tomato glandular Trichomes, providing an important precursor in the biosynthetic pathway of sesquiterpenoids. FPS was targeted to the plastid aiming to engineer sesquiterpenoid production, but interestingly leading to a loss of monoterpenoid production in the transgenic tomato Trichomes. With this example we show that Trichomes are amenable to engineering though, even with knowledge of a biochemical pathway, the result of such engineering can be unexpected.
Alain Tissier - One of the best experts on this subject based on the ideXlab platform.
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qtl mapping of the shape of type vi glandular Trichomes in tomato
Frontiers in Plant Science, 2018Co-Authors: Stefan Bennewitz, Nick Bergau, Alain TissierAbstract:Glandular Trichomes contribute to the high resistance of wild tomato species against insect pests not only thanks to the metabolites they produce but also because of morphological and developmental features which support the high production of these defense compounds. In Solanum habrochaites, type VI Trichomes have a distinct spherical shape and a large intercellular storage cavity where metabolites can accumulate and are released upon breaking off of the glandular cells. In contrast, the type VI Trichomes of S. lycopersicum have a four-leaf clover shape corresponding to the four glandular cells and a small internal cavity with limited capacity for storage of compounds. To better characterize the genetic factors underlying these Trichome morphological differences we created a back-cross population of 116 individuals between S. habrochaites LA1777 and S. lycopersicum var. cerasiformae WVa106. A Trichome score that reflects the shape of the type VI Trichomes allowing the quantification of this trait was designed. The scores were distributed normally across the population, which was mapped with a total of 192 markers. This resulted in the identification of 6 QTLs on chromosomes I, VII, VII and XI. The QTL on chromosome I with the highest LOD score was confirmed and narrowed down to a 500 gene interval in an advanced population derived from one of the back-cross lines. Our results provide the foundation for the genetic dissection of type VI Trichome morphology and the introgression of these Trichome traits into cultivated tomato lines for increased insect resistance.
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Isoprenoid and Metabolite Profiling of Plant Trichomes
Methods of Molecular Biology, 2014Co-Authors: Gerd Ulrich Balcke, Sebastian Zabel, Stefan Bennewitz, Alain TissierAbstract:Plant glandular Trichomes are specialized secretory structures located on the surface of the aerial parts of plants with large biosynthetic capacity, often with terpenoids as output molecules. The collection of plant Trichomes requires a method to separate Trichomes from leaf epidermal tissues. For metabolite profiling, Trichome tissue needs to be rapidly quenched in order to maintain the indigenous state of intracellular intermediates. Appropriate extraction and chromatographic separation methods must be available, which address the wide-ranging polarity of metabolites. In this chapter, a protocol for Trichome harvest using a frozen paint brush is presented. A work flow for broad-range metabolite profiling using LC-MS(2) analysis is described, which is applicable to assess very hydrophilic isoprenoid precursors as well as more hydrophobic metabolites from Trichomes and other plant tissues.
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Trichome Specific Expression: Promoters and Their Applications
Transgenic Plants - Advances and Limitations, 2012Co-Authors: Alain TissierAbstract:As often reminded to the readers in articles or reviews which deal with plant adaptation to their environment, higher plants are sessile organisms, a life habit which does not allow them to escape danger or to move to avoid adverse conditions. This environmental pressure has led to a myriad of adaptations, which are reflected in the vast diversity of plant habitats, morphologies, life cycles and physiological adaptations among others. The surface of the aerial parts of plants is a major interaction domain between the plant and its environment and as such is the site of many adaptations, be they chemical or anatomical. Among those adaptations, the leaf hairs or Trichomes, which cover the surface of a large number of plant species, play a prominent role. Plant Trichomes constitute a world of their own, so great is their diversity. In a review published in 1978 and entitled “A glossary of plant hair terminology”, Payne compiles a comprehensive list of more than 490 terms used to describe Trichome morphology (Payne, 1978). Despite this extensive diversity, two major classes of Trichome may be distinguished on the basis of their capacity to produce and secrete or store significant quantities of secondary metabolites, namely glandular or non-glandular. Nonglandular Trichomes, or leaf hairs, are poorly metabolically active and provide protection mainly through physical means, for example by restricting access to insects, but also by preventing water losses, or protecting against UV radiation. Arabidopsis thaliana has been a model for the study of non-glandular Trichome development and many genes involved in non-glandular Trichome initiation and development could be identified and characterized (Uhrig and Hulskamp, 2010). The metabolic activity of these non-glandular Trichomes is however fairly limited and offers little potential for metabolic engineering. A particular class of hairs is the fibers which are present in various species. Cotton seed Trichomes are the most economically important since they are the basis of the cotton fiber, but other species such as cottonwood also have fiber hairs. Glandular Trichomes are present in many different plant families and can also be divided in two main classes. The capitate Trichomes typically have 1 to 10 glandular cells located at the tip of the Trichome stalk, and the secretion is directly exuded from the top cells. The secreted material is in general fairly viscous, and in many cases it makes the leaves sticky. Those Trichomes are encountered for example in the Solanaceae (tobacco, tomato, potato, etc.) and in some Lamiaceae species (e.g. Salvia). Peltate Trichomes have the capacity to synthesize and store volatile compounds (monoand sesquiterpenes, phenylpropenes) in a subcuticular cavity. Typical representative examples
Jiesen Xu - One of the best experts on this subject based on the ideXlab platform.
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SlMYC1 Regulates Type VI Glandular Trichome Formation and Terpene Biosynthesis in Tomato Glandular Cells
The Plant Cell, 2018Co-Authors: Jiesen Xu, Zeger Otto Van Herwijnen, Dorthe Bettina Drager, Michel A. Haring, Robert C SchuurinkAbstract:Tomato (Solanum lycopersicum) glandular Trichomes function as biochemical factories that synthesize a diverse array of specialized metabolites. Terpenoids are the most diverse class of plant specialized metabolites, with volatile mono- and sesquiterpenes playing important roles in plant defense. Although the biosynthetic pathways of volatile terpenes in tomato glandular Trichomes have been well described, little is known about their regulation. Here, we demonstrate that SlMYC1, a basic helix-loop-helix transcription factor, differentially regulates mono- and sesquiterpene biosynthesis in the type VI glandular Trichomes of tomato leaves and stems. SlMYC1 functions as a positive regulator of monoterpene biosynthesis in both leaf and stem Trichomes but as a negative regulator of sesquiterpene biosynthesis in stem Trichomes. SlMYC1 is also essential for type VI glandular Trichome development, as knocking down SlMYC1 led to the production of smaller type VI glandular Trichomes at lower densities, and knocking out this gene led to their absence. Our findings reveal a role for SlMYC1 not only in type VI glandular Trichome development but also in the regulation of terpene biosynthesis in tomato.
Richard A. Dixon - One of the best experts on this subject based on the ideXlab platform.
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Trichome a comparative omics database for plant Trichomes
Plant Physiology, 2010Co-Authors: Guodong Wang, Dong Sik Yang, Pierre Broun, Yuhong Tang, Lloyd W Sumner, David M Marks, Richard A. Dixon, Patrick X ZhaoAbstract:Plant secretory Trichomes have a unique capacity for chemical synthesis and secretion and have been described as biofactories for the production of natural products. However, until recently, most Trichome-specific metabolic pathways and genes involved in various Trichome developmental stages have remained unknown. Furthermore, only a very limited amount of plant Trichome genomics information is available in scattered databases. We present an integrated “omics” database, Trichome, to facilitate the study of plant Trichomes. The database hosts a large volume of functional omics data, including expressed sequence tag/unigene sequences, microarray hybridizations from both Trichome and control tissues, mass spectrometry-based Trichome metabolite profiles, and Trichome-related genes curated from published literature. The expressed sequence tag/unigene sequences have been annotated based upon sequence similarity with popular databases (e.g. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, and Transporter Classification Database). The unigenes, metabolites, curated genes, and probe sets have been mapped against each other to enable comparative analysis. The database also integrates bioinformatics tools with a focus on the mining of Trichome-specific genes in unigenes and microarray-based gene expression profiles. Trichome is a valuable and unique resource for plant Trichome research, since the genes and metabolites expressed in Trichomes are often underrepresented in regular non-tissue-targeted cDNA libraries. Trichome is freely available at http://www.plantTrichome.org/.
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transcriptome analysis of arabidopsis wild type and gl3 sst sim Trichomes identifies four additional genes required for Trichome development
Molecular Plant, 2009Co-Authors: David M Marks, Jonathan P Wenger, Ross Jilk, Edward K. Gilding, Richard A. DixonAbstract:Transcriptome analyses have been performed on mature Trichomes isolated from wild-type Arabidopsis leaves and on leaf Trichomes isolated from the gl3-sst sim double mutant, which exhibit many attributes of immature Trichomes. The mature Trichome profile contained many highly expressed genes involved in cell wall synthesis, protein turnover, and abiotic stress response. The most highly expressed genes in the gl3-sst sim profile encoded ribosomal proteins and other proteins involved in translation. Comparative analyses showed that all but one of the genes encoding transcription factors previously found to be important for Trichome formation, and many other Trichome-important genes, were preferentially expressed in gl3-sst sim Trichomes. The analysis of genes preferentially expressed in gl3-sst sim led to the identification of four additional genes required for normal Trichome development. One of these was the HDG2 gene, which is a member of the HD-ZIP IV transcription factor gene family. Mutations in this gene did not alter Trichome expansion, but did alter mature Trichome cell walls. Mutations in BLT resulted in a loss of Trichome branch formation. The relationship between blt and the phenotypically identical mutant, sti, was explored. Mutations in PEL3, which was previously shown to be required for development of the leaf cuticle, resulted in the occasional tangling of expanding Trichomes. Mutations in another gene encoding a protein with an unknown function altered Trichome branch formation.
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a new method for isolating large quantities of arabidopsis Trichomes for transcriptome cell wall and other types of analyses
Plant Journal, 2008Co-Authors: David M Marks, Jonathan P Wenger, Edward K. Gilding, Lissete Betancur, Richard A. Dixon, Stefan Bauer, Fang Chen, Candace H HaiglerAbstract:A new procedure has been developed for the isolation of wild-type and mutant Arabidopsis Trichomes. The isolated Trichomes maintained enzymatic activity and were used for DNA, protein, and RNA isolation. The RNA was used to generate probes suitable for Affymetrix analysis. The validity of the Affymetrix results was confirmed by quantitative PCR analysis on a subset of genes that are preferentially expressed in Trichomes or leaves. Sufficient quantities of Trichomes were isolated to probe the biochemical nature of Trichome cell walls. These analyses provide evidence for the presence of lignin in Arabidopsis Trichome cell walls. The monosaccharide analysis and positive staining with ruthenium red indicates that the walls also contain a large portion of pectin. The 2.23-fold ratio of pectin-related sugars compared with potential cellulosic glucose suggests that the polysaccharides of the Trichome cell walls are more like those of typical primary walls even though the wall becomes quite thick. Overall, these analyses open the door to using the Arabidopsis Trichome cell wall as an excellent model to probe various questions concerning plant cell wall biosynthesis.
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Transcriptome analysis of alfalfa glandular Trichomes
Planta, 2005Co-Authors: Naveed Aziz, Nancy L. Paiva, Richard A. DixonAbstract:Glandular Trichomes are a major site of plant natural product synthesis and accumulation for protection against insect predation. However, to date few studies have attempted to obtain a global view of Trichome gene expression. Two contrasting approaches have been adopted to investigate genes expressed in glandular Trichomes from alfalfa ( Medicago sativa L.). In the first approach, 5,674 clones from an alfalfa glandular Trichome cDNA library were sequenced. The most highly abundant expressed sequence tag (EST) corresponded to a lipid transfer protein. The presence of ESTs corresponding to enzymes for all steps in the biosynthesis of flavonoids suggests that these are important metabolites in alfalfa Trichome biology, as confirmed by histochemistry and metabolite profiling. No ESTs corresponded to enzymes of cyclized terpenoid biosynthesis. In a second approach, microarray analysis was used to compare levels of alfalfa transcripts corresponding to 16,086 Medicago truncatula A17 genes in stems with and without Trichomes. This revealed over 1,000 genes with strong preferential expression in the Trichome fraction of the stem, 70% of which are of unknown function. These define a class of genes that are not Trichome-specific, since M. truncatula A17 does not itself have glandular Trichomes, but has potential importance for Trichome function within the stem.
Xiaoya Chen - One of the best experts on this subject based on the ideXlab platform.
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temporal control of Trichome distribution by microrna156 targeted spl genes in arabidopsis thaliana
The Plant Cell, 2010Co-Authors: Wenjuan Cai, Lingjian Wang, Shucai Wang, Chunmin Shan, Xiaoya ChenAbstract:The production and distribution of plant Trichomes is temporally and spatially regulated. After entering into the flowering stage, Arabidopsis thaliana plants have progressively reduced numbers of Trichomes on the inflorescence stem, and the floral organs are nearly glabrous. We show here that SQUAMOSA PROMOTER BINDING PROTEIN LIKE (SPL) genes, which define an endogenous flowering pathway and are targeted by microRNA 156 (miR156), temporally control the Trichome distribution during flowering. Plants overexpressing miR156 developed ectopic Trichomes on the stem and floral organs. By contrast, plants with elevated levels of SPLs produced fewer Trichomes. During plant development, the increase in SPL transcript levels is coordinated with the gradual loss of Trichome cells on the stem. The MYB transcription factor genes TrichomeLESS1 (TCL1) and TRIPTYCHON (TRY) are negative regulators of Trichome development. We show that SPL9 directly activates TCL1 and TRY expression through binding to their promoters and that this activation is independent of GLABROUS1 (GL1). The phytohormones cytokinin and gibberellin were reported to induce Trichome formation on the stem and inflorescence via the C2H2 transcription factors GIS, GIS2, and ZFP8, which promote GL1 expression. We show that the GIS-dependent pathway does not affect the regulation of TCL1 and TRY by miR156-targeted SPLs, represented by SPL9. These results demonstrate that the miR156-regulated SPLs establish a direct link between developmental programming and Trichome distribution.
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promoter of a cotton fibre myb gene functional in Trichomes of arabidopsis and glandular Trichomes of tobacco
Journal of Experimental Botany, 2008Co-Authors: Xiaoxia Shangguan, Zongxia Yu, Lingjian Wang, Bing Xu, Xiaoya ChenAbstract:Cotton fibres are unicellular seed Trichomes. Our previous study suggested that the cotton R2R3 MYB transcript factor GaMYB2 is a functional homologue of the Arabidopsis Trichome regulator GLABRA1 (GL1). Here, the GaMYB2 promoter activity is reported in cotton (Gossypium hirsutum), tobacco (Nicotiana tabacum), and Arabidopsis plants. A 2062 bp promoter of GaMYB2 was isolated from G. arboreum, and fused to a b-glucuronidase (GUS) reporter gene. In cotton, the GaMYB2 promoter exhibited activities in developing fibre cells and Trichomes of other aerial organs, including leaves, stems and bracts. In Arabidopsis the promoter was specific to Trichomes. Different from Arabidopsis and cotton that have unicellular nonglandular simple Trichomes, tobacco plants contain more than one type of Trichome, including multicellular simple and glandular secreting Trichomes (GSTs). Interestingly, in tobacco plants the GaMYB2 promoter directed GUS expression exclusively in glandular cells of GSTs. A series of 5#-deletions revealed that a 360 bp fragment upstream to the translation initiation codon was sufficient to drive gene expression. A putative ciselement of the T/G-box was located at -233 to -214; a yeast one-hybrid assay showed that Arabidopsis bHLH protein GLABRA3 (GL3), also a Trichome regulator, and GhDEL65, a GL3-like cotton protein, had high binding activities to the T/G-box motif. Overexpression of GL3 or GhDEL65 enhanced the GaMYB2 promoter activity in transgenic Arabidopsis plants. A comparison of GaMYB2 promoter specificities in Trichomes of different plant species with different types of Trichomes provides a tool for further dissection of plant Trichome structure and development.
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Promoter of a cotton fibre MYB gene functional in Trichomes of Arabidopsis and glandular Trichomes of tobacco
Journal of experimental botany, 2008Co-Authors: Xiaoxia Shangguan, Lingjian Wang, Xiaoya ChenAbstract:Cotton fibres are unicellular seed Trichomes. Our previous study suggested that the cotton R2R3 MYB transcript factor GaMYB2 is a functional homologue of the Arabidopsis Trichome regulator GLABRA1 (GL1). Here, the GaMYB2 promoter activity is reported in cotton (Gossypium hirsutum), tobacco (Nicotiana tabacum), and Arabidopsis plants. A 2062 bp promoter of GaMYB2 was isolated from G. arboreum, and fused to a beta-glucuronidase (GUS) reporter gene. In cotton, the GaMYB2 promoter exhibited activities in developing fibre cells and Trichomes of other aerial organs, including leaves, stems and bracts. In Arabidopsis the promoter was specific to Trichomes. Different from Arabidopsis and cotton that have unicellular non-glandular simple Trichomes, tobacco plants contain more than one type of Trichome, including multicellular simple and glandular secreting Trichomes (GSTs). Interestingly, in tobacco plants the GaMYB2 promoter directed GUS expression exclusively in glandular cells of GSTs. A series of 5'-deletions revealed that a 360 bp fragment upstream to the translation initiation codon was sufficient to drive gene expression. A putative cis-element of the T/G-box was located at -233 to -214; a yeast one-hybrid assay showed that Arabidopsis bHLH protein GLABRA3 (GL3), also a Trichome regulator, and GhDEL65, a GL3-like cotton protein, had high binding activities to the T/G-box motif. Overexpression of GL3 or GhDEL65 enhanced the GaMYB2 promoter activity in transgenic Arabidopsis plants. A comparison of GaMYB2 promoter specificities in Trichomes of different plant species with different types of Trichomes provides a tool for further dissection of plant Trichome structure and development.
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control of plant Trichome development by a cotton fiber myb gene
The Plant Cell, 2004Co-Authors: Shui Wang, Lingjian Wang, Jiawei Wang, Bin Luo, Jinying Gou, Xiaoya ChenAbstract:Cotton (Gossypium spp) plants produce seed Trichomes (cotton fibers) that are an important commodity worldwide; however, genes controlling cotton fiber development have not been characterized. In Arabidopsis thaliana the MYB gene GLABRA1 (GL1) is a central regulator of Trichome development. Here, we show that promoter of a cotton fiber gene, RD22-like1 (RDL1), contains a homeodomain binding L1 box and a MYB binding motif that confer Trichome-specific expression in Arabidopsis. A cotton MYB protein GaMYB2/Fiber Factor 1 transactivated the RDL1 promoter both in yeast and in planta. Real-time PCR and in situ analysis showed that GaMYB2 is predominantly expressed early in developing cotton fibers. After transferring into Arabidopsis, GL1::GaMYB2 rescued Trichome formation of a gl1 mutant, and interestingly, 35S::GaMYB2 induced seed-Trichome production. We further demonstrate that the first intron of both GL1 and GaMYB2 plays a role in patterning Trichomes: it acts as an enhancer in Trichome and a repressor in nonTrichome cells, generating a Trichome-specific pattern of MYB gene expression. Disruption of a MYB motif conserved in intron 1 of GL1, WEREWOLF, and GaMYB2 genes affected Trichome production. These results suggest that cotton and Arabidopsis use similar transcription factors for regulating Trichomes and that GaMYB2 may be a key regulator of cotton fiber development.
