ABSTRACT
Unlike terrestrial angiosperm plants, the freshwater aquatic angiosperm duckweed (Spirodela polyrhiza) grows directly in water and has distinct responses to heavy-metal stress. Plantlets accumulate metabolites, including lipids and carbohydrates, under heavy-metal stress, but how they balance metabolite levels is unclear, and the gene networks that mediate heavy-metal stress responses remain unknown. Here, we show that heavy-metal stress induced by flue gas desulfurization (FGD) wastewater reduces chlorophyll contents, inhibits growth, reduces membrane lipid biosynthesis, and stimulates membrane lipid degradation in S. polyrhiza, leading to triacylglycerol and carbohydrate accumulation. In FGD wastewater-treated plantlets, the degraded products of monogalactosyldiacylglycerol, primarily polyunsaturated fatty acids (18:3), were incorporated into triacylglycerols. Genes involved in early fatty acid biosynthesis, ß-oxidation, and lipid degradation were upregulated while genes involved in cuticular wax biosynthesis were downregulated by treatment. The transcription factor gene WRINKLED3 (SpWRI3) was upregulated in FGD wastewater-treated plantlets, and its ectopic expression increased tolerance to FGD wastewater in transgenic Arabidopsis (Arabidopsis thaliana). Transgenic Arabidopsis plants showed enhanced glutathione and lower malondialdehyde contents under stress, suggesting that SpWRI3 functions in S. polyrhiza tolerance of FGD wastewater-induced heavy-metal stress. These results provide a basis for improving heavy metal-stress tolerance in plants for industrial applications.
Subject(s)
Arabidopsis , Araceae , Metals, Heavy , Wastewater , Arabidopsis/genetics , Lipidomics , Metals, Heavy/toxicity , Metals, Heavy/metabolism , Plants, Genetically Modified , Gene Expression Profiling , Araceae/metabolism , Membrane Lipids/metabolismABSTRACT
Ethiopian mustard (Brassica carinata) is an ancient crop with remarkable stress resilience and a desirable seed fatty acid profile for biofuel uses. Brassica carinata is one of six Brassica species that share three major genomes from three diploid species (AA, BB, and CC) that spontaneously hybridized in a pairwise manner to form three allotetraploid species (AABB, AACC, and BBCC). Of the genomes of these species, that of B. carinata is the least understood. Here, we report a chromosome scale 1.31-Gbp genome assembly with 156.9-fold sequencing coverage for B. carinata, completing the reference genomes comprising the classic Triangle of U, a classical theory of the evolutionary relationships among these six species. Our assembly provides insights into the hybridization event that led to the current B. carinata genome and the genomic features that gave rise to the superior agronomic traits of B. carinata. Notably, we identified an expansion of transcription factor networks and agronomically important gene families. Completion of the Triangle of U comparative genomics platform has allowed us to examine the dynamics of polyploid evolution and the role of subgenome dominance in the domestication and continuing agronomic improvement of B. carinata and other Brassica species.
Subject(s)
Brassica , Brassica/genetics , Tetraploidy , Genome, Plant/genetics , Polyploidy , DiploidyABSTRACT
Genetic signature of climate adaptation has been widely recognized across the genome of many organisms; however, the eco-physiological basis for linking genomic polymorphisms with local adaptations remains largely unexplored. Using a panel of 218 world-wide Arabidopsis accessions, we characterized the natural variation in root suberization by quantifying 16 suberin monomers. We explored the associations between suberization traits and 126 climate variables. We conducted genome-wide association analysis and integrated previous genotype-environment association (GEA) to identify the genetic bases underlying suberization variation and their involvements in climate adaptation. Root suberin content displays extensive variation across Arabidopsis populations and significantly correlates with local moisture gradients and soil characteristics. Specifically, enhanced suberization is associated with drier environments, higher soil cation-exchange capacity, and lower soil pH; higher proportional levels of very-long-chain suberin is negatively correlated with moisture availability, lower soil gravel content, and higher soil silt fraction. We identified 94 putative causal loci and experimentally proved that GPAT6 is involved in C16 suberin biosynthesis. Highly significant associations between the putative genes and environmental variables were observed. Roots appear highly responsive to environmental heterogeneity via regulation of suberization, especially the suberin composition. The patterns of suberization-environment correlation and the suberin-related GEA fit the expectations of local adaptation for the polygenic suberization trait.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Arabidopsis/genetics , Arabidopsis Proteins/genetics , Genome-Wide Association Study , Plant Roots/genetics , SoilABSTRACT
The polyacetylenic lipids falcarinol, falcarindiol, and associated derivatives, termed falcarins, have a widespread taxonomical distribution in the plant kingdom and have received increasing interest for their demonstrated health-promoting properties as anti-cancer and anti-inflammatory agents. These fatty acid-derived compounds are also linked to plant pathogen resistance through their potent antimicrobial properties. Falcarin-type polyacetylenes, which contain two conjugated triple bonds, are derived from structural modifications of the common fatty acid oleic acid. In the past half century, much progress has been made in understanding the structural diversity of falcarins in the plant kingdom, whereas limited progress has been made on elucidating falcarin function in plant-pathogen interactions. More recently, an understanding of the biosynthetic machinery underlying falcarin biosynthesis has emerged. This review provides a concise summary of the current state of knowledge on falcarin structural diversity, biosynthesis, and plant defense properties. We also present major unanswered questions about falcarin biosynthesis and function.
Subject(s)
Fatty Acids , Plants , Polyacetylene PolymerABSTRACT
The alcohol- and alkane-forming pathways in cuticular wax biosynthesis are well characterized in Arabidopsis. However, potential interactions between the two pathways remain unclear. Here, we reveal that mutation of CER4, the key gene in the alcohol-forming pathway, also led to a deficiency in the alkane-forming pathway in distal stems. To trace the connection between the two pathways, we characterized two homologs of fatty alcohol oxidase (FAO), FAO3 and FAO4b, which were highly expressed in distal stems and localized to the endoplasmic reticulum. The amounts of waxes from the alkane-forming pathway were significantly decreased in stems of fao4b and much lower in fao3 fao4b plants, indicative of an overlapping function for the two proteins in wax synthesis. Additionally, overexpression of FAO3 and FAO4b in Arabidopsis resulted in a dramatic reduction of primary alcohols and significant increases of aldehydes and related waxes. Moreover, expressing FAO3 or FAO4b led to significantly decreased amounts of C18-C26 alcohols in yeast co-expressing CER4 and FAR1. Collectively, these findings demonstrate that FAO3 and FAO4b are functionally redundant in suppressing accumulation of primary alcohols and contributing to aldehyde production, which provides a missing and long-sought-after link between these two pathways in wax biosynthesis.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Alcohol Oxidoreductases , Alcohols/metabolism , Aldehyde Oxidoreductases/genetics , Aldehyde Oxidoreductases/metabolism , Alkanes/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant , Nuclear Proteins/metabolism , Plant Epidermis/metabolism , Waxes/metabolismABSTRACT
Salt stress impairs growth and yield in tomato, which is mostly cultivated in arid and semi-arid areas of the world. A number of wild tomato relatives (Solanum pimpinellifolium, S. pennellii, S. cheesmaniae and S. peruvianum) are endemic to arid coastal areas and able to withstand higher concentration of soil salt concentrations, making them a good genetic resource for breeding efforts aimed at improving salt tolerance and overall crop improvement. However, the complexity of salt stress response makes it difficult to introgress tolerance traits from wild relatives that could effectively increase tomato productivity under high soil salt concentrations. Under commercial production, biomass accumulation is key for high fruit yields, and salt tolerance management strategies should aim to maintain a favourable plant water and nutrient status. In this review, we first compare the effects of salt stress on the physiology of the domesticated tomato and its wild relatives. We then discuss physiological and energetic trade-offs for the different salt tolerance mechanisms found within the Lycopersicon clade, with a focus on the importance of root traits to sustain crop productivity.
ABSTRACT
3-Methylglutaconic (3MGC) aciduria occurs in numerous inborn errors associated with compromised mitochondrial energy metabolism. In these disorders, 3MGC CoA is produced de novo from acetyl CoA in three steps with the final reaction catalysed by 3MGC CoA hydratase (AUH). In in vitro assays, whereas recombinant AUH dehydrated 3-hydroxy-3-methylglutaryl (HMG) CoA to 3MGC CoA, free CoA was also produced. Although HMG CoA is known to undergo non-enzymatic intramolecular cyclisation, forming HMG anhydride and free CoA, the amount of free CoA generated increased when AUH was present. To test the hypothesis that the AUH-dependent increase in CoA production is caused by intramolecular cyclisation of 3MGC CoA, gas chromatography-mass spectrometry analysis of organic acids was performed. In the absence of AUH, HMG CoA was converted to HMG acid while, in the presence of AUH, 3MGC acid was also detected. To determine which 3MGC acid diastereomer was formed, immunoblot assays were conducted with 3MGCylated BSA. In competition experiments, when α-3MGC IgG was preincubated with trans-3MGC acid or cis-3MGC acid, the cis diastereomer inhibited antibody binding to 3MGCylated BSA. When an AUH assay product mix served as competitor, α-3MGC IgG binding to 3MGCylated BSA was also inhibited, indicating cis-3MGC acid is produced in incubations of AUH and HMG CoA. Thus, non-enzymatic isomerisation of trans-3MGC CoA drives AUH-dependent HMG CoA dehydration and explains the occurrence of cis-3MGC acid in urine of subjects with 3MGC aciduria. Furthermore, the ability of cis-3MGC anhydride to non-enzymatically acylate protein substrates may have deleterious pathophysiological consequences.
Subject(s)
Metabolism, Inborn Errors , Anhydrides , Energy Metabolism , Humans , Immunoglobulin GABSTRACT
Cuticular lipids consisting of cutin and wax coat aerial plant surfaces providing protection against biotic and abiotic stresses. Although much progress has been made on comprehending the regulation of plant cuticular lipid biosynthesis, their functional relevance in plant protection merits further investigation of potential regulators of their synthesis. HRD1 and DOA10 mediate two major Endoplasmic Reticulum-Associated Degradation (ERAD) pathways in yeast and also regulate common pathways during lipid metabolism. However, their roles in plant lipid metabolism are not well studied. CER9, an Arabidopsis homolog of DOA10, is known to play important roles in cuticular lipid biosynthesis. This prompted us to determine if HRD1 also plays a role in regulation cuticular lipid biosynthesis. Here we report that an Arabidopsis hrd1a hrd1b double mutant is impacted in the accumulation of both cutin and cuticular waxes including a large increase in total stem cutin with a concomitant decrease in stem wax content. We further investigated genetic relationship between HRD1A/1B- and CER9-mediated ERAD pathways with regard to cuticular lipid synthesis. Surprisingly, simultaneous mutation of HRD1 and CER9 revealed additive effects on stem wax synthesis, but not stem cutin synthesis. Collectively, our study advances our understanding of the ERAD regulatory roles in cuticular lipid synthesis identifying HRD1 as an important player in the regulated deposition of Arabidopsis stem cuticular lipids.
Subject(s)
Arabidopsis Proteins , Endoplasmic Reticulum-Associated Degradation , Ubiquitin-Protein Ligases , Waxes , Arabidopsis , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Gene Expression Regulation, Plant , Ubiquitin , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolismABSTRACT
Fatty acids (FAs) play vital roles in plants as components of lipid membranes that demarcate cells and organelles, as sources of stored energy in the form of neutral lipids, and as signaling molecules that elicit plant responses to adverse conditions. The activation of FAs through the formation of acyl-CoA intermediates by acyl-CoA synthetase (ACS) family enzymes is required for their synthesis and degradation. Long-chain ACSs (LACSs) represent a small subgroup of ACS enzymes that specifically convert long-chain or very-long-chain FAs into corresponding thioesters for multiple lipid-associated processes. Alteration of LACS activity often results in pleiotropic phenotypes such as male sterility, organ fusion, aberrant cuticular structure, delayed seed germination, altered seed oil content, and plant capacity to respond to various environmental stresses. This review provides a comprehensive analysis of LACS family enzymes including substrate specificity, tissue-specific expression patterns, and distinct subcellular localization highlighting their specific roles in lipid synthesis and degradation, the effects of altered LACS activity on plant development, the relationship between LACS activity and stress resistance, and the regulation of LACS activity. Finally, we pose several major questions to be addressed, which would advance our current understanding of LACS function in plants.
ABSTRACT
Wounding during mechanical harvesting and post-harvest handling results in tuber desiccation and provides an entry point for pathogens resulting in substantial postâ-harvest crop losses. Poor wound healing is a major culprit of these losses. Wound tissue in potato (Solanum tuberosum) tubers, and all higher plants, is composed of a large proportion of suberin that is deposited in a specialized tissue called the wound periderm. However, the genetic regulatory pathway controlling wound-induced suberization remains unknown. Here, we implicate two potato transcription factors, StMYB102 (PGSC0003DMG400011250) and StMYB74 (PGSC0003DMG400022399), as regulators of wound suberin biosynthesis and deposition. Using targeted metabolomics and transcript profiling from the wound healing tissues of two commercial potato cultivars, as well as heterologous expression, we provide evidence for the molecular-genetic basis of the differential wound suberization capacities of different potato cultivars. Our results suggest that (i) the export of suberin from the cytosol to the apoplast and ligno-suberin deposition may be limiting factors for wound suberization, (ii) StMYB74 and StMYB102 are important regulators of the wound suberization process in tubers, and (iii) polymorphisms in StMYB102 may influence cultivar-specific wound suberization capacity. These results represent an important step in understanding the regulated biosynthesis and deposition of wound suberin and provide a practical foundation for targeted breeding approaches aimed at improving potato tuber storage life.
Subject(s)
Lipids/biosynthesis , Plant Proteins/genetics , Plant Tubers/physiology , Solanum tuberosum/physiology , Gene Expression Regulation, Plant , Lipids/genetics , Phenols/metabolism , Plant Cells , Plant Tubers/genetics , Polymorphism, Genetic , Solanum tuberosum/cytology , Solanum tuberosum/genetics , Transcription Factors/genetics , Waxes/metabolismABSTRACT
3-Methylglutaconic (3MGC) aciduria is a common phenotypic feature of a growing number of inborn errors of metabolism. "Primary" 3MGC aciduria is caused by deficiencies in leucine pathway enzymes while "secondary" 3MGC aciduria results from inborn errors of metabolism that impact mitochondrial energy production. The metabolic precursor of 3MGC acid is trans-3MGC CoA, an intermediate in the leucine catabolism pathway. Gas chromatography-mass spectrometry (GC-MS) analysis of commercially available trans-3MGC acid yielded a mixture of cis and trans isomers while 1H-NMR spectroscopy of trans-3MGC acid at 25°C provided no evidence for the cis isomer. When trans-3MGC acid was incubated under conditions used for sample derivatization prior to GC-MS (but with no trimethylsilane added), 1H-NMR spectroscopy provided evidence of trans to cis isomerization. Incubation of trans-3MGC acid at 37°C resulted in time-dependent isomerization to cis-3MGC acid. Cis-3MGC acid behaved in a similar manner except that, under identical incubation conditions, less isomerization occurred. In agreement with these experimental results, molecular modeling studies provided evidence that the energy minimized structure of cis-3MGC acid is 4 kJ/mol more stable than that for trans-3MGC acid. Once generated in vivo, trans-3MGC acid is proposed to isomerize via a mechanism involving π electron delocalization with formation of a resonance structure that permits bond rotation. The data presented are consistent with the occurrence of both diastereomers in urine samples of subjects with 3MGC aciduria.
ABSTRACT
Virtually all land plants are coated in a cuticle, a waxy polyester that prevents nonstomatal water loss and is important for heat and drought tolerance. Here, we describe a likely genetic basis for a divergence in cuticular wax chemistry between Sorghum bicolor, a drought tolerant crop widely cultivated in hot climates, and its close relative Zea mays (maize). Combining chemical analyses, heterologous expression, and comparative genomics, we reveal that: 1) sorghum and maize leaf waxes are similar at the juvenile stage but, after the juvenile-to-adult transition, sorghum leaf waxes are rich in triterpenoids that are absent from maize; 2) biosynthesis of the majority of sorghum leaf triterpenoids is mediated by a gene that maize and sorghum both inherited from a common ancestor but that is only functionally maintained in sorghum; and 3) sorghum leaf triterpenoids accumulate in a spatial pattern that was previously shown to strengthen the cuticle and decrease water loss at high temperatures. These findings uncover the possibility for resurrection of a cuticular triterpenoid-synthesizing gene in maize that could create a more heat-tolerant water barrier on the plant's leaf surfaces. They also provide a fundamental understanding of sorghum leaf waxes that will inform efforts to divert surface carbon to intracellular storage for bioenergy and bioproduct innovations.
Subject(s)
Gene Expression Regulation, Plant , Plant Leaves/metabolism , Sorghum/genetics , Sorghum/metabolism , Steroids/biosynthesis , Waxes/metabolism , Adaptation, Biological , Computational Biology , Droughts , Gas Chromatography-Mass Spectrometry , Gene Expression Profiling , Genome, Plant , Molecular Structure , Phylogeny , Sorghum/classification , Steroids/chemistry , Triterpenes/metabolism , Waxes/chemistry , Zea mays/genetics , Zea mays/metabolismABSTRACT
Camelina sativa is relatively drought tolerant and requires less fertilizer than other oilseed crops. Various lipid- and phenolic-based extracellular barriers of plants help to protect them against biotic and abiotic stresses. These barriers, which consist of solvent-insoluble polymeric frameworks and solvent-extractable waxes, include the cuticle of aerial plant surfaces and suberized cell walls found, for example, in periderms and seed coats. Cutin, the polymeric matrix of the cuticle, and the aliphatic domain of suberin are fatty acid- and glycerol-based polyesters. These polyesters were investigated by base-catalyzed transesterification of C. sativa aerial and underground delipidated tissues followed by gas chromatographic analysis of the released monomer mixtures. Seed coat and root suberin had similar compositions, with 18-hydroxyoctadecenoic and 1,18-octadecenedioic fatty acids being the dominant species. Root suberin presented a typical lamellar ultrastructure, but seed coats showed almost imperceptible, faint dark bands. Leaf and stem lipid polyesters were composed of fatty acids (FA), 1,ω-dicarboxylic fatty acids (DCA), ω-hydroxy fatty acids (HFA) and hydroxycinnamic acids (HCA). Dihydroxypalmitic acid (DHP) and caffeic acid were the major constituents of leaf cutin, whereas stem cutin presented similar molar proportions in several monomers across the four classes. Unlike the leaf cuticle, the C. sativa stem cuticle presented lamellar structure by transmission electron microscopy. Flower cutin was dominated by DHP, did not contain aromatics, and presented substantial amounts (>30%) of hydroxylated 1,ω-dicarboxylic acids. We found striking differences between the lipid polyester monomer compositions of aerial tissues of C. sativa and that of its close relatives Arabidopsis thaliana and Brassica napus.
Subject(s)
Dicarboxylic Acids , Fatty Acids , Lipids , Membrane Lipids , PolyestersABSTRACT
Polyacetylenic lipids accumulate in various Apiaceae species after pathogen attack, suggesting that these compounds are naturally occurring pesticides and potentially valuable resources for crop improvement. These compounds also promote human health and slow tumor growth. Even though polyacetylenic lipids were discovered decades ago, the biosynthetic pathway underlying their production is largely unknown. To begin filling this gap and ultimately enable polyacetylene engineering, we studied polyacetylenes and their biosynthesis in the major Apiaceae crop carrot (Daucus carota subsp. sativus). Using gas chromatography and mass spectrometry, we identified three known polyacetylenes and assigned provisional structures to two novel polyacetylenes. We also quantified these compounds in carrot leaf, petiole, root xylem, root phloem, and root periderm extracts. Falcarindiol and falcarinol predominated and accumulated primarily in the root periderm. Since the multiple double and triple carbon-carbon bonds that distinguish polyacetylenes from ubiquitous fatty acids are often introduced by Δ12 oleic acid desaturase (FAD2)-type enzymes, we mined the carrot genome for FAD2 genes. We identified a FAD2 family with an unprecedented 24 members and analyzed public, tissue-specific carrot RNA-Seq data to identify coexpressed members with root periderm-enhanced expression. Six candidate genes were heterologously expressed individually and in combination in yeast and Arabidopsis (Arabidopsis thaliana), resulting in the identification of one canonical FAD2 that converts oleic to linoleic acid, three divergent FAD2-like acetylenases that convert linoleic into crepenynic acid, and two bifunctional FAD2s with Δ12 and Δ14 desaturase activity that convert crepenynic into the further desaturated dehydrocrepenynic acid, a polyacetylene pathway intermediate. These genes can now be used as a basis for discovering other steps of falcarin-type polyacetylene biosynthesis, to modulate polyacetylene levels in plants, and to test the in planta function of these molecules.
Subject(s)
Daucus carota/genetics , Daucus carota/metabolism , Enzymes/genetics , Plant Proteins/genetics , Polyacetylene Polymer/metabolism , Alkynes/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , Chromatography, Thin Layer , Diynes/metabolism , Enzymes/metabolism , Fatty Acid Desaturases/genetics , Fatty Acid Desaturases/metabolism , Fatty Alcohols/metabolism , Gas Chromatography-Mass Spectrometry , Linoleic Acid/metabolism , Oleic Acids/metabolism , Plant Proteins/metabolism , Plants, Genetically Modified , Polyacetylene Polymer/analysis , Saccharomyces cerevisiae/geneticsABSTRACT
The plant lipid barriers cuticle and suberin represent one of the largest biological interfaces on the planet. They are comprised of an insoluble polymeric domain with associated organic solvent-soluble waxes. Suberin-associated and plant cuticular waxes contain mixtures of aliphatic components that may include alkyl hydroxycinnamates (AHCs). The canonical alkyl hydroxycinnamates are comprised of phenylpropanoids, typically coumaric, ferulic, or caffeic acids, esterified with long chain to very long chain fatty alcohols. However, many related structures are also present in the plant kingdom. Although their functions remain elusive, much progress has been made on understanding the distribution, biosynthesis, and deposition of AHCs. Herein a summary of the current state of knowledge on plant AHCs is provided.
ABSTRACT
We report n-6 monounsaturated primary alcohols (C26:1, C28:1, and C30:1 homologs) in the cuticular waxes of Arabidopsis (Arabidopsis thaliana) inflorescence stem, a class of wax not previously reported in Arabidopsis. The Arabidopsis cer17 mutant was completely deficient in these monounsaturated alcohols, and CER17 was found to encode a predicted ACYL-COENZYME A DESATURASE LIKE4 (ADS4). Studies of the Arabidopsis cer4 mutant and yeast variously expressing CER4 (a predicted fatty acyl-CoA reductase) with CER17/ADS4, demonstrated CER4's principal role in synthesis of these monounsaturated alcohols. Besides unsaturated alcohol deficiency, cer17 mutants exhibited a thickened and irregular cuticle ultrastructure and increased amounts of cutin monomers. Although unsaturated alcohols were absent throughout the cer17 stem, the mutation's effects on cutin monomers and cuticle ultrastructure were much more severe in distal than basal stems, consistent with observations that the CER17/ADS4 transcript was much more abundant in distal than basal stems. Furthermore, distal but not basal stems of a double mutant deficient for both CER17/ADS4 and LONG-CHAIN ACYL-COA SYNTHETASE1 produced even more cutin monomers and a thicker and more disorganized cuticle ultrastructure and higher cuticle permeability than observed for wild type or either mutant parent, indicating a dramatic genetic interaction on conversion of very long chain acyl-CoA precursors. These results provide evidence that CER17/ADS4 performs n-6 desaturation of very long chain acyl-CoAs in both distal and basal stems and has a major function associated with governing cutin monomer amounts primarily in the distal segments of the inflorescence stem.
Subject(s)
Alcohols/metabolism , Arabidopsis Proteins/metabolism , Arabidopsis/enzymology , Fatty Acid Desaturases/metabolism , Membrane Lipids/metabolism , Waxes/metabolism , Amino Acid Sequence , Arabidopsis/metabolism , Arabidopsis/ultrastructure , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/genetics , Chromatography, Gas , Endoplasmic Reticulum/metabolism , Endoplasmic Reticulum/ultrastructure , Epistasis, Genetic , Fatty Acid Desaturases/chemistry , Fatty Acid Desaturases/genetics , Gene Expression Regulation, Plant , Inflorescence/metabolism , Mutation/genetics , Plant Epidermis/cytology , Plant Epidermis/metabolism , Plant Epidermis/ultrastructure , Plant Stems/metabolism , Plant Stems/ultrastructure , Protein TransportABSTRACT
Suberin is a complex hydrophobic polymer that acts as a barrier controlling water and solute fluxes and restricting pathogen infections. Suberin is deposited immediately outside of the plasmalemma in the cell wall of certain tissues such as endodermis of roots, aerial and underground periderms, and seed coats. Suberin consists of a variety of fatty acid derivatives polymerized with glycerol and phenolics. In this study, we show using liquid chromatography-tandem mass spectrometry and gas chromatography-mass spectrometry techniques that most of the fatty alcohols not covalently linked to the suberin polymer are in the form of alkyl hydroxycinnamates (AHCs), with alkyl caffeates predominating. Such compounds are not restricted to the periderm of mature roots but also are present in the endodermis of younger roots, where they are not extracted by rapid dipping in chloroform. Analysis of several mutants affected in key enzymes involved in the biosynthesis and export of suberin monomers suggests that the formation of the suberin polymer and associated waxes involves common pathways and occurs concomitantly in Arabidopsis (Arabidopsis thaliana) roots. Although fatty alcohols represent only minor components of the suberin polymer in Arabidopsis roots, this study demonstrates that they constitute the major aliphatics of suberin-associated waxes in the form of AHCs. Therefore, our results indicate that esterified fatty alcohols, both soluble and polymerized forms, represent major constituents of Arabidopsis root suberized barriers, being as abundant as α,ω-dicarboxylic and unsubstituted fatty acids. In addition, our results show that suberized layers represent a major sink for acyl-lipid metabolism in Arabidopsis roots.
Subject(s)
Arabidopsis/metabolism , Coumaric Acids/metabolism , Fatty Alcohols/metabolism , Plant Roots/metabolism , Aldehyde Oxidoreductases/genetics , Aldehyde Oxidoreductases/metabolism , Arabidopsis/genetics , Arabidopsis/growth & development , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Coumaric Acids/chemistry , Fatty Alcohols/chemistry , Gas Chromatography-Mass Spectrometry , Lipids/chemistry , Lipids/genetics , Mutation , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Plant Roots/chemistry , Plants, Genetically Modified , Waxes/metabolismABSTRACT
Aliphatic waxes can be found in association with suberized tissues, including roots. Non-polar lipids were isolated by rapid solvent extraction of mature regions of intact roots from eleven angiosperms, including both monocots and dicots. The majority of roots analyzed were taproots or tuberous taproots that had undergone secondary growth and thus were covered by a suberized periderm. The exceptions therein were maize (Zea mays L.) and rice (Oryza sativa L.), which present a suberized exodermis. The analysis herein focused on aliphatic waxes, with particular emphasis on alkyl hydroxycinnamates (AHCs). AHCs were widely distributed, absent from only one species, were found in both aerial and subterranean portions of tuberous taproots, and were associated with the fibrous roots of both maize and rice. Most species also contained monoacylglycerols, fatty alcohols and/or free fatty acids. Carrot (Daucus carrota L.) was the outlier, containing only free fatty acids, sterols, and polyacetylenes as identified components. Sterols were the only ubiquitous component across all roots analyzed. Monoacylglycerols of ω-hydroxy fatty acids were present in maize and rice root waxes. For species within the Brassiceae, wax compositions varied between subspecies or varieties and between aerial and subterranean portions of taproots. In addition, reduced forms of photo-oxidation products of ω-hydroxy oleate and its corresponding dicarboxylic acid (10,18-dihydroxy-octadec-8-enoate, 9,18-dihydroxy-octadec-10-enoate and 9-hydroxyoctadec-10-ene-1,18-dioate) were identified as naturally occurring suberin monomers in rutabaga (Brassica napus subsp. rapifera Metzg.) periderm tissues.
Subject(s)
Plant Roots/chemistry , Plants/chemistry , Waxes/analysis , Waxes/chemistry , Brassica napus/chemistry , Coumaric Acids/analysis , Coumaric Acids/chemistry , Daucus carota/chemistry , Fatty Alcohols/analysis , Fatty Alcohols/chemistry , Lipids/analysis , Lipids/chemistry , Microscopy, Electron, Transmission , Oryza/chemistry , Plant Roots/ultrastructure , Sterols/analysis , Sterols/chemistry , Nicotiana/chemistry , Zea mays/chemistryABSTRACT
Camelina sativa (L.) Crantz is an emerging low input, stress tolerant crop with seed oil composition suitable for biofuel and bioproduct production. The chemical compositions and ultrastructural features of surface waxes from C. sativa aerial cuticles, seeds, and roots were analyzed using gas chromatography and microscopy. Alkanes, primary fatty alcohols, and free fatty acids were common components of all analyzed organs. A particular feature of leaf waxes was the presence of alkyl esters of long-chain fatty acids and very long-chain fatty alcohols, ranging from C38 to C50 and dominated by C42, C44 and C46 homologues. Stem waxes were mainly composed of non-sterol pentacyclic triterpenes. Flowers accumulated significant amounts of methyl-branched iso-alkanes (C29 and C31 total carbon number) in addition to straight-chain alkanes. Seed waxes were mostly primary fatty alcohols of up to 32 carbons in length and unbranched C29 and C31 alkanes. The total amount of identified wax components extracted by rapid chloroform dipping of roots was 280µgg(-1) (fresh weight), and included alkyl hydroxycinnamates, predominantly alkyl coumarates and alkyl caffeates. This study provides qualitative and quantitative information on the waxes of C. sativa root, shoot, and seed boundary tissues, allowing the relative activities of wax biosynthetic pathways in each respective plant organ to be assessed. This detailed description of the protective surface waxes of C. sativa may provide insights into its drought-tolerant and pathogen-resistant properties, and also identifies C. sativa as a potential source of renewable high-value natural products.