Search bioRxiv⌕ Search

Biology subjects

Domergue, F.

Publications and source records attributed to Domergue, F..

10 recordsLinked to original sources

A single genetic innovation at the origin of plant terrestrialization

How the functional innovations that enabled plants to first colonize land 450 million years ago evolved remains puzzling. Here, we show that the gain of a single family of enzymes was pivotal in the evolution of two of these innovations, the cuticle that protect plants from dehydration and UV light, and the Arbuscular Mycorrhizal symbiosis promoting water and nutrient uptake. We show that functional phosphatase domain Glycerol-3-Phosphate Acyl Transferase (p-GPATs) evolved in the first land plants, concomitantly with the cuticle and symbiosis. Mutation of two p-GPATs from the liverwort Marchantia paleacea is sufficient to abolish both cuticle formation and AM symbiosis, leading to major developmental defects. We propose that the evolution of p-GPATs in land plants acted as a two birds-one stone innovation, diverting intracellular lipids to the extracellular space and providing a simple path to the evolution of two traits essential for the colonization of land.

plant biology↗

Unveiling the molecular identity of plant autophagic compartments: A proteo-lipidomic study in Arabidopsis thaliana

Autophagy is an intracellular degradation and recycling pathway essential for cell quality control and plant tolerance to stress. The formation and maturation of autophagosomes, the cargo-packing vesicles, rely on extensive membrane remodeling, yet little is known about the nature, dynamics and functions of lipids in these events. Here, we established a method combing cell fractionation and immuno-isolation in native conditions to purify autophagic membranes from Arabidopsis thaliana. By integrating proteomic and lipidomic analyses, we defined their molecular footprint which, coupled to colocalization analyses, revealed potent actors involved in lipid metabolism, membrane trafficking and membrane remodeling, supporting a close interplay between lipid homeostasis and autophagosome biology. Characterization of the phagophore lipid composition showed low sterol and sphingolipid content, a predominance of glycerophospholipids, including phosphoinositides, and a particular enrichment in phosphatidylcholine and phosphatidylglycerol. Comparisons with other plant endomembranes and autophagic compartments from other organisms revealed the singularity of this lipid signature and notably identified phosphatidylglycerol as a plant-specific component of autophagic membranes. Analyses of inducible phosphatidylglycerol-deficient plants showed defects in autophagy activity thereby supporting the functional relevance of the phagophore lipid composition, particularly that of phosphatidylglycerol homeostasis. Together, our findings place lipids as fundamental components of the autophagy molecular landscape and provide a framework to further investigate their contribution to autophagosome biology and functions in plant acclimation to environmental changes. Significance StatementAutophagy is catabolic pathway critical for eukaryotic life and essential for plant acclimation to stress. It hinges on the remarkable plasticity of a specialized membrane, the phagophore, to orchestrate and support intense membrane remodeling events towards the formation of the autophagic vesicle containing cargo. To resolve their elusive molecular bases, we need to integrate inputs from both components of biological membranes: proteins and lipids, yet information regarding lipids is still very limited. Here, we isolated plant phagophores, established their protein and lipid molecular footprint, revealed its singularity and showed its physiological and functional relevance for autophagy activity. Our work highlights lipids as key regulators of plant autophagy and opens new doors for investigating how membrane dynamics shape cellular stress responses.

plant biology↗

A dual component system instructs membrane hydrolysis during the final stages of plant autophagy

Autophagy is an intracellular catabolic process conserved across eukaryotes and critical for plant stress tolerance. Upon their delivery in the vacuole, how autophagic bodies containing cargo are hydrolyzed to warrant autophagy degradation remains poorly characterized. Here, we identify two Arabidopsis phospholipases as core components of the autophagy machinery. We find that LCAT3 and LCAT4 traffic to the vacuolar lumen and converge on autophagic bodies using differential pathways, placing them on the outer and inner side of the vesicle, respectively. Double knockouts lcat3,4 accumulate autophagic bodies and show reduced autophagy activity. In vivo reconstitution demonstrates that LCAT3 can hydrolyze the membrane of autophagic bodies, enabling the activity of LCAT4 to enhance this process. In sum, our work reveals that the concerted action of a multi-component system is required for the efficient and specific disruption of autophagic bodies as an obligatory step for the completion of the autophagy pathway.

plant biology↗

Genetic interaction between GL15 and FDL1 modulates juvenile cuticle deposition and leaf permeability in maize

The plant cuticle is a hydrophobic layer that serves as the primary barrier between plant surfaces and the external environment, limiting water loss and protecting against various stresses. This study investigates the genetic interaction between the maize ZmFDL1 and ZmGL15 genes and their role in modulating juvenile cuticle deposition and leaf permeability. The research was undertaken to understand how these genes influence cuticle composition and structure, which are crucial for plant protection against environmental stresses such as drought. We analysed the chemical composition of cutin and waxes, as well as cuticle-dependent leaf permeability. Our findings reveal that ZmFDL1 and ZmGL15 individually and synergically regulate the deposition of cuticular components, with significant impacts on leaf permeability. The gl15-S mutant exhibited an adult-like cuticle with higher cutin and lower wax contents, leading to reduced leaf permeability and improved water retention under drought conditions. These results highlight the importance of cutin in forming an effective water barrier and suggest that modulating cuticle composition with enhanced cutin content could be a strategy for improving drought tolerance in crops. This study provides new insights into the genetic regulation of cuticle biosynthesis and its relevance for plant adaptation to water scarcity. HighlightO_LIZmFDL1 and ZmGL15 interact to modulate cuticle composition and reduce leaf water loss. C_LIO_LIZmFDL1 and ZmGL15 genes regulate juvenile cuticle deposition in maize. C_LIO_LIThe gl15-S mutant shows an adult-like cuticle with higher cutin and lower wax contents. C_LIO_LIThe gl15-S mutation is epistatic to fdl1-1, partially rescuing the defective fdl1 cuticle phenotype. C_LIO_LIThe cuticle composition in gl15-S mutants leads to reduced leaf permeability and improved water retention under drought conditions. C_LIO_LICuticular layer with enhanced cutin content could improve drought tolerance in crops. C_LI

genetics↗

SPHINGOLIPIDS AND DELTA8-SPHINGOLIPID DESATURASE FROM THE PICOALGA O. TAURI AND INVOLVEMENT IN TEMPERATURE ACCLIMATION

Sphingolipids are crucial components of cell membranes. Sphingolipid {Delta}8-unsaturation is more specific to plants and is involved in the regulation of stress responses. The structure and functions of sphingolipids in microalgae are still poorly understood. Ostreococus tauri is a minimal microalga at the base of the green lineage, and is therefore a key organism for understanding lipid evolution. The present work reports the characterisation as well as the temperature regulation of sphingolipids and {Delta}8-sphingolipid desaturase from O. tauri. Complex sphingolipids are glycosylceramides with unique glycosyl moieties encompassing hexuronic acid residues, reminiscent of bacterial glucuronosylceramides, with up to three additional hexose residues. In contrast, the ceramide backbones show limited variety, with dihydroxylated C18/C18:1E{Delta}8 sphingoid bases and C16:0 fatty-acyl chain being the main compounds. The sphingolipid {Delta}8-desaturase from O. tauri, although phylogenetically related to plant homologues has a substrate preference similar to the diatom homologue. Both sphingolipid {Delta}8-desaturase transcripts and sphingolipid {Delta}8-unsaturation are regulated in a temperature- dependent manner being higher at 14{degrees}C than 24{degrees}C. Overexpressing the sphingolipid {Delta}8- desaturase in O. tauri at 24{degrees}C results in higher sphingolipid unsaturation and impairs the increase in cell size, structure and chlorophyll. In particular, the cell-size defect is not detected in cells acclimated to 14{degrees}C and is furthermore suppressed upon transfer from 24{degrees}C to 14{degrees}C. Our work provides the first functional evidence for the involvement of sphingolipid {Delta}8-unsaturation for temperature acclimation in microalgae, suggesting that this function is an ancestral feature in the green lineage.

biochemistry↗

Cytokinin response regulator ARR16 regulates seed coat permeability in Arabidopsis natural accessions

The seed coat that encases the embryo is constituted from multiple specialized-cell layers and their permeability significantly influences seed quality traits that have major agronomic impact. The regulatory mechanisms that modulate seed coat permeability are, however, not well understood. Here, we identified a novel regulatory gene for seed permeability through a genome wide association study (GWAS) in Arabidopsis based on image analysis of tetrazolium staining. Type-A ARABIDOPSIS RESPONSE REGULATOR 16 (ARR16) is a component of the signal transduction pathway for the plant hormone cytokinins (CKs), and in addition to less permeable seed coats, arr16 mutant seeds were larger, had a longer lifespan, and more suberin phenolics, the hydrophobic lipid biopolyester components of cell walls that act as a water-repellant. Moreover, double mutants in CK receptor ARABIDOPSIS HISTIDINE KINASE genes, notably ahk2 ahk4 and ahk3 ahk4, showed similar seed phenotypes to those of arr16. Based on naturally-occurring variation in the ARR16 gene, eight haplotypes were detected and associated with permeable or impermeable phenotypes. Permeable haplotypes had significantly lower suberin autofluorescence compared to impermeable haplotypes. RT-qPCR analysis demonstrated that ARR16 transcripts were highly abundant in developing seeds of representative accessions having permeable haplotypes but not in those of having impermeable haplotypes, indicating that these haplotypes were causal for ARR16 transcript abundance and thereby regulate seed coat permeability in natural accessions. Our findings demonstrate a new role for CKs signaling in seed coat differentiation and that this underlies natural variation for seed permeability through the modulation of suberin accumulation. Significance StatementSeed coat permeability affects key traits that impact seed quality, such as dormancy, longevity, and germination tolerance to abiotic stress. Little is currently known about how seed coat permeability is modulated. Here, we developed an original method for quantification of seed coat permeability through imaging and used this in a genome-wide association study with Arabidopsis accessions. The CKs response regulator ARR16 was identified as a causal gene thereby establishing a novel function for this phytohormone in seed coat differentiation. Moreover, this was linked to the modulation of suberin accumulation by ARR16 and for the first time implicates CKs signal transduction in the control of suberin deposition in seeds.

plant biology↗

VviMYB41 orthologs contribute to the water deficit induced suberization of grapevine fine roots

The permeability of roots to water and nutrients is controlled through a variety of mechanisms and one of the most conspicuous is the presence of structures such as the Casparian strips and suberin lamellae. Roots actively regulate the creation of these structures developmentally, along the length of the root, and in response to the environment, including abiotic stresses such as drought. In the current study, we characterized the suberin composition along the length of grapevine fine roots during development and in response to water deficit. In parallel samples we quantified changes in expression of suberin biosynthesis- and deposition-related gene families (via RNAseq) allowing the identification of drought-responsive suberin-related genes. Grapevine suberin composition did not differ between primary and lateral roots, and was similar to that of other species. Under water deficit there was a global upregulation of suberin biosynthesis which resulted in an increase of suberin specific monomers, but without changes in their relative abundances, and this upregulation took place across all the developmental stages of fine roots. These changes corresponded to the upregulation of numerous suberin biosynthesis- and deposition-related genes which included orthologs of the previously characterized AtMYB41 transcriptional factor. Functional validation of two grapevine MYB41 orthologs, VviMYB41 and VviMYB41-like, confirmed their ability to globally upregulate suberin biosynthesis and deposition. This study provides a detailed characterization of the developmental and water deficit induced suberization of grapevine fine roots and identifies important orthologs responsible for suberin biosynthesis, deposition, and its regulation in grape. One sentence summaryOur study details the biochemical changes and molecular regulation of how grapevines decrease their root permeability during drought.

plant biology↗

The drought-responsive ZmFDL1 gene regulates cuticle biosynthesis and cuticle-dependent leaf permeability

In higher plants, the outer surface of the aerial parts is covered by the cuticle, a complex lipid layer that constitutes a barrier against damages caused by environmental factors and provides protection against non-stomatal water loss. We show in this study that cuticle deposition, during the juvenile phase of in maize (Zea mays) plant development, and cuticle-dependent leaf permeability are controlled by the MYB transcription factor ZmMYB94/FUSED LEAVES1 (ZmFDL1). Biochemical analysis showed that in fdl1-1 mutant seedlings at the coleoptile stage both cutin and wax biosynthesis and deposition were altered. Among cutin compounds, {omega}-hydroxy fatty acids and polyhydroxy-fatty acids were specifically affected, while the reduction of epicuticular waxes, was mainly observed in primary long chain alcohols, and to a minor extent, long-chain wax esters. Transcriptome analysis allowed the identification of novel candidate genes involved in lipid metabolism and the assembly of a proposed pathway for cuticle biosynthesis in maize. Lack of ZmFDL1 affects the expression of genes located in different modules of the pathway and correspondence between gene transcriptional variations and biochemical defects have been highlighted. A decrease in cuticle-dependent leaf permeability was observed in maize seedlings exposed to drought as well as ABA treatment, which implies coordinated changes in the transcript levels of ZmFDL1 and associated genes. Overall, our results suggest that the response to water stress implies the activation of wax biosynthesis and the involvement of by both ZmFDL1 and ABA regulatory pathways. One-Sentence SummaryCuticle biosynthesis and cuticle-mediated drought-response during the juvenile phase of maize plant growth, are regulated by the MYB transcription factor fused leaves1 (ZmFDL1) and influenced by ABA.

plant biology↗

Plastidic Δ6 Fatty-Acid Desaturases With Distinctive Substrate Specificity Regulate The Pool Of C18-PUFAs In The Ancestral Picoalga Ostreococcus tauri

Eukaryotic {Delta}6-desaturases are microsomal enzymes which balance the synthesis of {omega}-3 and {omega}-6 C18-polyunsaturated-fatty-acids (PUFA) accordingly to their specificity. In several microalgae, including O. tauri, plastidic C18-PUFA are specifically regulated by environmental cues suggesting an autonomous control of {Delta}6-desaturation of plastidic PUFA. Sequence retrieval from O. tauri desaturases, highlighted two putative {Delta}6/{Delta}8-desaturases sequences clustering, with other microalgal homologs, apart from other characterized {Delta}-6 desaturases. Their overexpression in heterologous hosts, including N. benthamiana and Synechocystis, unveiled their {Delta}6-desaturation activity and plastid localization. O. tauri lines overexpressing these {Delta}6-desaturases no longer adjusted their plastidic C18-PUFA amount under phosphate starvation but didnt show any obvious physiological alterations. Detailed lipid analyses from the various overexpressing hosts, unravelled that the substrate features involved in the {Delta}6-desaturase specificity importantly involved the lipid head-group and likely the non-substrate acyl-chain, in addition to the overall preference for the {omega}-class of the substrate acyl-chain. The most active desaturase displayed a broad range substrate specificity for plastidic lipids and a preference for {omega}-3 substrates, while the other was selective for {omega}-6 substrates, phosphatidylglycerol and 16:4-galactolipid species specific to the native host. The distribution of plastidial {Delta}6-desaturase products in eukaryotic hosts suggested the occurrence of C18-PUFA export from the plastid. One sentence summaryOsteococcus tauri plastidic lipid C18-PUFA remodelling involves two plastid-located cytochrome-b5 fused {Delta}6-desaturases with distinct preferences for both head-group and acyl-chain.

biochemistry↗

A stress-response-related inter-compartmental signalling pathway regulates embryonic cuticle integrity in Arabidopsis

The embryonic cuticle is necessary for normal seed development and seedling establishment in Arabidopsis. Although mutants with defective embryonic cuticles have been identified, neither the deposition of cuticle material, nor its regulation, has been described during embryogenesis. Here we use electron microscopy, lipid staining and permeability assays to show that cuticle deposition initiates de novo in patches on globular embryos. By combining these techniques with genetics and gene expression analysis, we show that successful patch coalescence to form a continuous cuticle requires a signalling involving the endosperm-specific subtilisin protease ALE1 and the receptor kinases GSO1 and GSO2, which are expressed in the developing embryonic epidermis. Transcriptome analysis shows that this pathway regulates stress-related gene expression in seeds. Consistent with these findings we show genetically, and through activity analysis, that the stress-associated MPK6 protein acts downstream of GSO1 and GSO2 in the developing embryo. We propose that a stress-related signalling pathway has been hijacked in some angiosperm seeds through the recruitment of endosperm-specific components. Our work reveals the presence of an inter-compartmental dialogue between the endosperm and embryo that ensures the formation of an intact and functional cuticle around the developing embryo through an \"auto-immune\" type interaction.

plant biology↗