To identify novel genes engaged in plant epidermal development, we characterized the phenotypic variability of rosette leaf epidermis of 310 sequenced Arabidopsis thaliana accessions, focusing on trichome shape and distribution, compositional characteristics of the trichome cell wall, and histologically detectable metal ion distribution. Some of these traits correlated with cLimate parameters of our accession's locations of origin, suggesting environmental selection. A novel metal deposition pattern in stomatal guard cells was observed in some accessions. Subsequent GWAS analysis identified 1546 loci with protein sequence-altering SNPs associated with one or more traits, including 5 genes with previously reported relevant mutant phenotypes and 80 additional genes with known or predicted roles in relevant developmental and cellular processes. Some candidates, including GFS9/TT9, exhibited environmentally correlated allele distribution. Several large gene famiLies, namely DUF674, DUF784, DUF1262, DUF1985, DUF3741, cytochrome P450, receptor-Like kinases, Cys/His-rich C1 domain proteins and formins were overrepresented among the candidates for various traits, suggesting epidermal development-related functions. A possible participation of formins in guard cell metal deposition was supported by observations in available loss of function mutants. Screening of candidate gene lists against the STRING interactome database uncovered several predominantly nuclear protein interaction networks with possible novel roles in epidermal development.
- Keywords
- Arabidopsis thaliana, BioClim, GWAS, guard cell, metal accumulation, phenotypic variability, trichome,
- Publication type
- Journal Article MeSH
Plasma membrane (PM) lipid composition and domain organization are modulated by polarized exocytosis. Conversely, targeting of secretory vesicles at specific domains in the PM is carried out by exocyst complexes, which contain EXO70 subunits that play a significant role in the final recognition of the target membrane. As we have shown previously, a mature Arabidopsis trichome contains a basal domain with a thin cell wall and an apical domain with a thick secondary cell wall, which is developed in an EXO70H4-dependent manner. These domains are separated by a cell wall structure named the Ortmannian ring. Using phospholipid markers, we demonstrate that there are two distinct PM domains corresponding to these cell wall domains. The apical domain is enriched in phosphatidic acid (PA) and phosphatidylserine, with an undetectable amount of phosphatidylinositol 4,5-bisphosphate (PIP2), whereas the basal domain is PIP2-rich. While the apical domain recruits EXO70H4, the basal domain recruits EXO70A1, which corresponds to the lipid-binding capacities of these two paralogs. Loss of EXO70H4 results in a loss of the Ortmannian ring border and decreased apical PA accumulation, which causes the PA and PIP2 domains to merge together. Using transmission electron microscopy, we describe these accumulations as a unique anatomical feature of the apical cell wall-radially distributed rod-shaped membranous pockets, where both EXO70H4 and lipid markers are immobilized.
- Keywords
- EXO70, cell wall, exocyst complex, phosphatidic acid, phosphatidylinositol 4,5-bisphosphate, phospholipids, plasma membrane domains, polar exocytosis, trichome,
- MeSH
- Arabidopsis chemistry genetics MeSH
- Cell Membrane chemistry genetics MeSH
- Exocytosis genetics MeSH
- Phosphatidylinositol 4,5-Diphosphate chemistry metabolism MeSH
- Phosphatidylserines chemistry genetics MeSH
- Membrane Lipids genetics metabolism MeSH
- Arabidopsis Proteins chemistry genetics MeSH
- Trichomes chemistry genetics MeSH
- Vesicular Transport Proteins chemistry genetics MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- EXO70A1 protein, Arabidopsis MeSH Browser
- EXO70H4 protein, Arabidopsis MeSH Browser
- Phosphatidylinositol 4,5-Diphosphate MeSH
- Phosphatidylserines MeSH
- Membrane Lipids MeSH
- Arabidopsis Proteins MeSH
- Vesicular Transport Proteins MeSH
Exocyst component of 70-kDa (EXO70) proteins are constituents of the exocyst complex implicated in vesicle tethering during exocytosis. MILDEW RESISTANCE LOCUS O (MLO) proteins are plant-specific calcium channels and some MLO isoforms enable fungal powdery mildew pathogenesis. We here detected an unexpected phenotypic overlap of Arabidopsis thaliana exo70H4 and mlo2 mlo6 mlo12 triple mutant plants regarding the biogenesis of leaf trichome secondary cell walls. Biochemical and Fourier transform infrared spectroscopic analyses corroborated deficiencies in the composition of trichome cell walls in these mutants. Transgenic lines expressing fluorophore-tagged EXO70H4 and MLO exhibited extensive colocalization of these proteins. Furthermore, mCherry-EXO70H4 mislocalized in trichomes of the mlo triple mutant and, vice versa, MLO6-GFP mislocalized in trichomes of the exo70H4 mutant. Expression of GFP-marked PMR4 callose synthase, a known cargo of EXO70H4-dependent exocytosis, revealed reduced cell wall delivery of GFP-PMR4 in trichomes of mlo triple mutant plants. In vivo protein-protein interaction assays in plant and yeast cells uncovered isoform-preferential interactions between EXO70.2 subfamily members and MLO proteins. Finally, exo70H4 and mlo6 mutants, when combined, showed synergistically enhanced resistance to powdery mildew attack. Taken together, our data point to an isoform-specific interplay of EXO70 and MLO proteins in the modulation of trichome cell wall biogenesis and powdery mildew susceptibility.
- MeSH
- Arabidopsis * metabolism MeSH
- Cell Wall metabolism MeSH
- Plant Diseases microbiology MeSH
- Disease Resistance genetics MeSH
- Protein Isoforms genetics metabolism MeSH
- Arabidopsis Proteins * genetics metabolism MeSH
- Plant Proteins metabolism MeSH
- Trichomes genetics metabolism MeSH
- Vesicular Transport Proteins metabolism MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- EXO70H4 protein, Arabidopsis MeSH Browser
- Protein Isoforms MeSH
- Arabidopsis Proteins * MeSH
- Plant Proteins MeSH
- Vesicular Transport Proteins MeSH
In Utricularia, the flower spur is a nectary and in this organ, nectar is produced and stored. This study aimed to examine the structure of the nectary trichomes in four Utricularia species (Utricularia vulgaris L., U. australis R.Br., U. bremii Heer and U. foliosa L.) from the generic section Utricularia. We have investigated whether species with different spur morphology had similar spur anatomy and nectary trichome structure. In Utricularia flowers, nectar is produced by spur capitate trichomes (sessile or stalked). Our results showed that regardless of the various spur morphology, trichomes have similar architecture and ultrastructure. Head cells of these trichomes are transfer cells with an eccrine nectar secretion. Examined species differed in the micromorphology of papillae in spurs. The fly Eristalis tenax was found to be a pollinator of U. vulgaris. Small Halictidae bees seem to be pollinators of U. foliosa.
- Keywords
- Bladderwort, Carnivorous plant, Entomophily, Floral micromorphology, Lentibulariaceae, Nectar, Nectary ultrastructure, Pollination, Sect. Utricularia, Spur, Trichomes,
- MeSH
- Flowers chemistry MeSH
- Pollination immunology MeSH
- Plant Nectar chemistry MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Plant Nectar MeSH
BACKGROUND AND AIMS: Root hemiparasites from the rhinanthoid clade of Orobanchaceae possess metabolically active glandular trichomes that have been suggested to function as hydathode trichomes actively secreting water, a process that may facilitate resource acquisition from the host plant's root xylem. However, no direct evidence relating the trichomes to water secretion exists, and carbon budgets associated with this energy-demanding process have not been determined. METHODS: Macro- and microscopic observations of the leaves of hemiparasitic Rhinanthus alectorolophus were conducted and night-time gas exchange was measured. Correlations were examined among the intensity of guttation, respiration and transpiration, and analysis of these correlations allowed the carbon budget of the trichome activity to be quantified. We examined the intensity of guttation, respiration and transpiration, correlations among which indicate active water secretion. KEY RESULTS: Guttation was observed on the leaves of 50 % of the young, non-flowering plants that were examined, and microscopic observations revealed water secretion from the glandular trichomes present on the abaxial leaf side. Night-time rates of respiration and transpiration and the presence of guttation drops were positively correlated, which is a clear indicator of hydathode trichome activity. Subsequent physiological measurements on older, flowering plants indicated neither intense guttation nor the presence of correlations, which suggests that the peak activity of hydathodes is in the juvenile stage. CONCLUSIONS: This study provides the first unequivocal evidence for the physiological role of the hydathode trichomes in active water secretion in the rhinanthoid Orobanchaceae. Depending on the concentration of organic elements calculated to be in the host xylem sap, the direct effect of water secretion on carbon balance ranges from close to neutral to positive. However, it is likely to be positive in the xylem-only feeding holoparasites of the genus Lathraea, which is closely related to Rhinanthus. Thus, water secretion by the hydathodes might be viewed as a physiological pre-adaptation in the evolution of holoparasitism in the rhinanthoid lineage of Orobanchaceae.
- Keywords
- Ecophysiology, Lathraea, Rhinanthus alectorolophus, Triticum aestivum, holoparasite, hydathode trichome, orobanche, parasitic plant, respiration, rhinanthoid Orobanchaceae, root hemiparasite, transpiration, water regime, water secretion, xylem,
- MeSH
- Biological Evolution * MeSH
- Cell Respiration MeSH
- Host-Parasite Interactions physiology MeSH
- Plant Roots parasitology MeSH
- Linear Models MeSH
- Plant Leaves metabolism MeSH
- Orobanchaceae physiology MeSH
- Gases metabolism MeSH
- Plant Transpiration MeSH
- Trichomes physiology MeSH
- Water metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Gases MeSH
- Water MeSH
The hop plant (Humulus lupulus L.) produces several valuable secondary metabolites, such as prenylflavonoid, bitter acids, and essential oils. These compounds are biosynthesized in glandular trichomes (lupulin glands) endowed with pharmacological properties and widely implicated in the beer brewing industry. The present study is an attempt to generate exhaustive information of transcriptome dynamics and gene regulatory mechanisms involved in biosynthesis and regulation of these compounds, developmental changes including trichome development at three development stages, namely leaf, bract, and mature lupulin glands. Using high-throughput RNA-Seq technology, a total of 61.13, 50.01, and 20.18 Mb clean reads in the leaf, bract, and lupulin gland libraries, respectively, were obtained and assembled into 43,550 unigenes. The putative functions were assigned to 30,996 transcripts (71.17%) based on basic local alignment search tool similarity searches against public sequence databases, including GO, KEGG, NR, and COG families, which indicated that genes are principally involved in fundamental cellular and molecular functions, and biosynthesis of secondary metabolites. The expression levels of all unigenes were analyzed in leaf, bract, and lupulin glands tissues of hop. The expression profile of transcript encoding enzymes of BCAA metabolism, MEP, and shikimate pathway was most up-regulated in lupulin glands compared with leaves and bracts. Similarly, the expression levels of the transcription factors and structural genes that directly encode enzymes involved in xanthohumol, bitter acids, and terpenoids biosynthesis pathway were found to be significantly enhanced in lupulin glands, suggesting that production of these metabolites increases after the leaf development. In addition, numerous genes involved in primary metabolism, lipid metabolism, photosynthesis, generation of precursor metabolites/energy, protein modification, transporter activity, and cell wall component biogenesis were differentially regulated in three developmental stages, suggesting their involvement in the dynamics of the lupulin gland development. The identification of differentially regulated trichome-related genes provided a new foundation for molecular research on trichome development and differentiation in hop. In conclusion, the reported results provide directions for future functional genomics studies for genetic engineering or molecular breeding for augmentation of secondary metabolite content in hop.
- Keywords
- Humulus lupulus, RNA sequencing, bitter acids, lupulin glands, prenylflavonoids, terpenoids, trichome,
- MeSH
- Flavonoids biosynthesis chemistry metabolism MeSH
- Gene Ontology MeSH
- Humulus chemistry metabolism MeSH
- Plant Leaves genetics metabolism MeSH
- Propiophenones chemistry metabolism MeSH
- Gene Expression Regulation, Plant MeSH
- Plant Proteins genetics metabolism MeSH
- RNA-Seq MeSH
- Terpenes chemistry metabolism MeSH
- Transcription Factors metabolism MeSH
- Transcriptome genetics MeSH
- Trichomes genetics metabolism ultrastructure MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Flavonoids MeSH
- Propiophenones MeSH
- Plant Proteins MeSH
- Terpenes MeSH
- Transcription Factors MeSH
- xanthohumol MeSH Browser
Arabidopsis (Arabidopsis thaliana) leaf trichomes are single-cell structures with a well-studied development, but little is understood about their function. Developmental studies focused mainly on the early shaping stages, and little attention has been paid to the maturation stage. We focused on the EXO70H4 exocyst subunit, one of the most up-regulated genes in the mature trichome. We uncovered EXO70H4-dependent development of the secondary cell wall layer, highly autofluorescent and callose rich, deposited only in the upper part of the trichome. The boundary is formed between the apical and the basal parts of mature trichome by a callose ring that is also deposited in an EXO70H4-dependent manner. We call this structure the Ortmannian ring (OR). Both the secondary cell wall layer and the OR are absent in the exo70H4 mutants. Ecophysiological aspects of the trichome cell wall thickening include interference with antiherbivore defense and heavy metal accumulation. Ultraviolet B light induces EXO70H4 transcription in a CONSTITUTIVE PHOTOMORPHOGENIC1-dependent way, resulting in stimulation of trichome cell wall thickening and the OR biogenesis. EXO70H4-dependent trichome cell wall hardening is a unique phenomenon, which may be conserved among a variety of the land plants. Our analyses support a concept that Arabidopsis trichome is an excellent model to study molecular mechanisms of secondary cell wall deposition.
- MeSH
- Arabidopsis metabolism radiation effects ultrastructure MeSH
- Cell Wall metabolism ultrastructure MeSH
- Cytokinesis radiation effects MeSH
- Fluorescence MeSH
- Glucans metabolism MeSH
- Copper metabolism MeSH
- Mutation genetics MeSH
- Protein Subunits metabolism MeSH
- Arabidopsis Proteins metabolism MeSH
- Trichomes metabolism radiation effects ultrastructure MeSH
- Ultraviolet Rays MeSH
- Vesicular Transport Proteins metabolism MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- callose MeSH Browser
- EXO70H4 protein, Arabidopsis MeSH Browser
- Glucans MeSH
- Copper MeSH
- Protein Subunits MeSH
- Arabidopsis Proteins MeSH
- Vesicular Transport Proteins MeSH
BACKGROUND: Hop (Humulus lupulus L.) bitter acids are valuable metabolites for the brewing industry. They are biosynthesized and accumulate in glandular trichomes of the female inflorescence (hop cone). The content of alpha bitter acids, such as humulones, in hop cones can differentiate aromatic from bitter hop cultivars. These contents are subject to genetic and environmental control but significantly correlate with the number and size of glandular trichomes (lupulin glands). RESULTS: We evaluated the expression levels of 37 genes involved in bitter acid biosynthesis and morphological and developmental differentiation of glandular trichomes to identify key regulatory factors involved in bitter acid content differences. For bitter acid biosynthesis genes, upregulation of humulone synthase genes, which are important for the biosynthesis of alpha bitter acids in lupulin glands, could explain the higher accumulation of alpha bitter acids in bitter hops. Several transcription factors, including HlETC1, HlMYB61 and HlMYB5 from the MYB family, as well as HlGLABRA2, HlCYCB2-4, HlZFP8 and HlYABBY1, were also more highly expressed in the bitter hop cultivars; therefore, these factors may be important for the higher density of lupulin glands also seen in the bitter hop cultivars. CONCLUSIONS: Gene expression analyses enabled us to investigate the differences between aromatic and bitter hops. This study confirmed that the bitter acid content in glandular trichomes (lupulin glands) is dependent on the last step of alpha bitter acid biosynthesis and glandular trichome density.
- Keywords
- Bitter acids, Differential gene expression, Glandular trichome development, Hop, Humulus lupulus, Lupulin gland,
- MeSH
- Humulus metabolism MeSH
- Transcription Factors metabolism MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Transcription Factors MeSH
Data on plant herbivore damage as well as on herbivore performance have been previously used to identify key plant traits driving plant-herbivore interactions. The extent to which the two approaches lead to similar conclusions remains to be explored. We determined the effect of a free-living leaf-chewing generalist caterpillar, Spodoptera littoralis (Lepidoptera: Noctuidae), on leaf damage of 24 closely related plant species from the Carduoideae subfamily and the effect of these plant species on caterpillar growth. We used a wide range of physical defense leaf traits and leaf nutrient contents as the plant traits. Herbivore performance and leaf damage were affected by similar plant traits. Traits related to higher caterpillar mortality (higher leaf dissection, number, length and toughness of spines and lower trichome density) also led to higher leaf damage. This fits with the fact that each caterpillar was feeding on a single plant and, thus, had to consume more biomass of the less suitable plants to obtain the same amount of nutrients. The key plant traits driving plant-herbivore interactions identified based on data on herbivore performance largely corresponded to the traits identified as important based on data on leaf damage. This suggests that both types of data may be used to identify the key plant traits determining plant-herbivore interactions. It is, however, important to carefully distinguish whether the data on leaf damage were obtained in the field or in a controlled feeding experiment, as the patterns expected in the two environments may go in opposite directions.
- Keywords
- Lepidoptera, cost of defense, folivory, knapweed, selection pressure, thistle, trait evolution, trichome,
- Publication type
- Journal Article MeSH
The ARP2/3 complex and formins are the only known plant actin nucleators. Besides their actin-related functions, both systems also modulate microtubule organization and dynamics. Loss of the main housekeeping Arabidopsis thaliana Class I membrane-targeted formin FH1 (At3g25500) is known to increase cotyledon pavement cell lobing, while mutations affecting ARP2/3 subunits exhibit an opposite effect. Here we examine the role of FH1 and the ARP2/3 complex subunit ARPC5 (At4g01710) in epidermal cell morphogenesis with focus on pavement cells and trichomes using a model system of single fh1 and arpc5, as well as double fh1 arpc5 mutants. While cotyledon pavement cell shape in double mutants mostly resembled single arpc5 mutants, analysis of true leaf epidermal morphology, as well as actin and microtubule organization and dynamics, revealed a more complex relationship between the two systems and similar, rather than antagonistic, effects on some parameters. Both fh1 and arpc5 mutations increased actin network density and increased cell shape complexity in pavement cells and trichomes of first true leaves, in contrast to cotyledons. Thus, while the two actin nucleation systems have complementary roles in some aspects of cell morphogenesis in cotyledon pavement cells, they may act in parallel in other cell types and developmental stages.
- Keywords
- ARP2/3, At3g25500, At4g01710, actin nucleation, cytoskeleton, formin, pavement cell, trichome,
- Publication type
- Journal Article MeSH
