Biology subjects

Wightman, R.

Publications and source records attributed to Wightman, R..

9 recordsLinked to original sources

bioRxiv2025

An ancient plant symbiotic fungus with distinct features identified through advanced fluorescence and Raman imaging

Mycorrhizal associations between fungi and plants are a fundamental aspect of terrestrial ecosystems. The occurrence of mycorrhizae is now well established in c. 85% of extant plants, yet the geological record of these associations is sparse. Fossil evidence from early Paleozoic terrestrial environments provides rare insights into these ancient symbioses, but imaging micro-scale fossil fungi in surrounding plant tissues is challenging. We employed brightfield microscopy, confocal laser scanning microscopy (CLSM), fluorescence lifetime imaging microscopy (FLIM), and Raman spectroscopy to investigate a newly identified endomycorrhizal fungus within a 407-million-year-old plant from the Windyfield Chert. An exquisite preservation and our advanced imaging approach enabled the description of the fungal structures to unprecedented detail. The fungus, Rugosomyces lavoisieriae gen. nov., & sp. nov., exhibits structural features resembling modern Glomeromycotina arbuscular mycorrhizal fungi. Combining CLSM with FLIM provides extra information by separating features based upon fluorescence lifetime. Raman spectroscopy indicates that fungal arbuscules and plant water-conducting cells have undergone geological alterations, resulting in a similar chemical composition. These findings enhance our understanding of ancient plant-fungal symbioses and demonstrate the potential of advanced imaging techniques in paleobotanical research. Significance statementMost living plants form mycorrhizal associations with fungi, but how this ancient evolutionary relationship developed is poorly understood. Exceptional fossil materials preserve direct evidence of endomycorrhizae, but imaging micro-scale fossil fungi inside plant cells is challenging. Here we use confocal microscopy combined with fluorescence lifetime imaging and Raman spectroscopy to study a new endomycorrhizal fungus inside a 400-million-year-old plant. We document structures resembling modern Glomeromycotina. We show that the residual organic matter of the fungus and the plant cell walls has a very short fluorescence lifetime, which provides a new means of imaging these fossils. Raman spectroscopy indicates that both arbuscules and water-conducting cells were geologically altered by their diagenetic history and that they now have a similar chemical composition.

plant biology↗

bioRxiv2024

A receptor kinase complex refines cambium activity in Arabidopsis

In plant development, key receptor kinases are often active in disparate cell types, with each requiring vastly different signalling outputs. The ERECTA (ER) receptor kinase and its homologs ERL1 and ERL2 exemplify this pleiotropy. In Arabidopsis, they influence stomatal patterning, shoot meristem function, ovule morphogenesis, xylem fiber differentiation, and cell division in the vascular cambium1-6. Such diverse expression and functionality raises the question of how ER signalling can specify such distinct cell behaviours. One mechanism is via cell-type specific interactions with co-receptors, ligands, or other proteins that modulate signalling. However, little is known about ER interactors in the vascular cambium, a bifacial stem cell niche that generates phloem and xylem (Figure 1A). Combinatorial mutations between ER, ERL1 and ERL2 and receptor kinases of a second family, PXY, PXL1, and PXL2, show severe cambial defects5,7, but the mechanism underpinning these phenotypes is not known. Here we discovered that PXY proteins form protein complexes with ER and ERL2. PXY signalling can be manipulated by altering levels of its cognate ligand, TDIF. In genetic analysis, plant lines in which TDIF levels were altered had dramatic phenotypic changes that required the presence of ER or ERL2. Our results demonstrate that PXY signalling mediated cambium regulation depends on ER signalling and explains ER function in the cambium. Because the cambium produces xylem, which constitutes the wood in vascular plants, our findings position PXY-ER complexes at the centre of the accumulation of this versatile biomaterial and essential carbon sink. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=168 SRC="FIGDIR/small/594489v1_fig1.gif" ALT="Figure 1"> View larger version (79K): org.highwire.dtl.DTLVardef@289bb6org.highwire.dtl.DTLVardef@6d0fdorg.highwire.dtl.DTLVardef@1829836org.highwire.dtl.DTLVardef@d70b3b_HPS_FORMAT_FIGEXP M_FIG O_FLOATNOFigure 1.C_FLOATNO PXY forms heterodimers with ER and ERL2 (A) Transverse sections of a wild type hypocotyl showing the region of secondary growth. Black rectangle marks the area of cambium shown in the diagrammatic representation on the lower left. (B-D) Localisation of PXY-CFP (B), ER-YFP (C) and overlap (D) in Nicotiana epidermal cells. (E) FRET signal. (F) Graph showing FRET efficiencies of PXY-CFP and ER/ERL1/ERL2-YFP pairs. (G-H) Coimmunoprecipitation of PXY-FLAG with ER-HA or ERL2-HA and vice versa in Arabidopsis cells. Scales (A) are 50 M. C_FIG

plant biology↗

bioRxiv2024

HtDCR-like1 regulates the development of structurally coloured cuticle by modulating cuticle chemistry and mechanical properties in Hibiscus trionum

O_LIStructural colours are a unique trait present in some animals, plants and even bacteria. In Hibiscus trionum flowers, they arise from nano-scaled cuticular ridges and may improve pollinator foraging efficiency. These ridges result from the buckling of the cuticle which is thought to be controlled by multiple parameters including anisotropic cell growth, chemical differentiation of the cuticle and formation of two layers of different mechanical properties. C_LIO_LIHere we investigate further the molecular and physical mechanisms by which structural colours are achieved in Hibiscus trionum. We produced two transgenic lines overexpressing HtDCR-like1, encoding a BAHD acyltransferase involved in cuticle synthesis. Transgenic lines showed impairment of the production of cuticle striations. Cuticle thickness, cuticle chemistry, cell elongation and cuticle stiffness were then investigated to identify the cause of failed buckling. C_LIO_LIWe found that HtDCR-like1 overexpression leads to modification of cuticle chemistry, including a decrease in detectable C16H32O4 (10,16-DHP) and a change to the measured Youngs moduli of the cuticle layers without alteration of the cell growth pattern. C_LIO_LIThis work demonstrates the mechanisms by which gene regulation may control complex physical phenomena such as cuticle buckling via alteration of material properties of living tissues. C_LI

plant biology↗

bioRxiv2023

Altering the substitution and crosslinking of glucuronoarabinoxylans affects cell wall porosity and assembly in Brachypodium distachyon.

O_LIThe Poaceae family of plants provides cereal crops that are critical for human and animal nutrition and also they are an important source of biomass. Interacting plant cell wall components give rise to recalcitrance to digestion, thus understanding the wall molecular architecture is important to improve biomass properties. Xylan is the main hemicellulose in grass cell walls. Recently, we reported structural variation in grass xylans, suggesting functional specialisation and distinct interactions with cellulose and lignin. Here, we investigated the functions of these xylans by perturbing the biosynthesis of specific xylan types. C_LIO_LIWe generated CRISPR/Cas9 knockout mutants in Brachypodium distachyon XAX1 and GUX2 genes involved in xylan biosynthesis. Using carbohydrate gel electrophoresis we identified biochemical changes in different xylan types. Saccharification, cryo-SEM, subcritical water extraction and ssNMR were used to study wall architecture. C_LIO_LIBdXAX1A and BdGUX2 enzymes modify different types of grass xylan. Brachypodium mutant walls are more porous, suggesting the xylan substitutions directed by both BdXAX1A andGUX2 enzymes influence xylan-xylan and/or xylan-lignin interactions. C_LIO_LISince xylan substitutions influence wall architecture and digestibility, our findings open new avenues to improve cereals for food and to use grass biomass for feed and the production of bioenergy and biomaterials. C_LI

biochemistry↗

bioRxiv2022

Eudicot primary cell wall glucomannan is related in synthesis, structure and function to xyloglucan

The functional differences between plant cell wall hemicelluloses such as glucomannan, xylan and xyloglucan (XyG) remain unclear. These polysaccharides influence assembly and properties of the wall, perhaps by interacting with cellulose to affect the deposition and bundling of the fibrils. As the most abundant hemicellulose, XyG is considered important in eudicot primary cell walls (PCWs), but plants devoid of XyG show relatively mild phenotypes. We report here that a patterned {beta}-galactoglucomannan ({beta}-GGM) is widespread in PCW of eudicots and shows remarkable similarities to XyG. The sugar linkages forming the backbone and side chains of {beta}-GGM are analogous to those that make up XyG, and moreover, these linkages are formed by glycosyltransferases from the same CAZy families. Solid-state NMR indicated that {beta}-GGM shows low mobility in the cell wall, consistent with interaction with cellulose. Although Arabidopsis {beta}-GGM synthesis mutants show no obvious growth defects, genetic crosses between {beta}-GGM and XyG mutants produce exacerbated phenotypes compared to XyG mutants. These findings demonstrate a related role of these two similar but distinct classes of hemicelluloses in PCWs. This work will provide new avenues to study the roles of both {beta}-GGM and XyG in PCWs. One sentence summaryPatterned {beta}-GGM resembles xyloglucan in structure, biosynthesis and function.

plant biology↗

bioRxiv2020

Formation of flavone-based wooly fibres by glandular trichomes of Dionysia tapetodes.

Dionysia tapetodes, a small cushion-forming mountainous evergreen in the Primulaceae, possesses a vast surface-covering of long silky fibres forming the characteristic "wooly" farina. This contrasts with some related Primula which instead possess a powdery farina. Using a combination of cell biology and analytical chemical techniques, we provide a detailed insight of wooly farina formation by glandular trichomes that produce a mixture of flavone and substituted flavone derivatives, including hydroxyflavones. Conversely, our analysis show that the powdery form consist almost entirely of flavone. The wooly farina in D. tapetodes is extruded through specific sites at the surface of the glandular head cell, characterised by a small complete gap in the plasma membrane, cell wall and cuticle. The data is consistent with formation and thread elongation occurring from within the cell. The putative mechanism of wool thread formation and its stability is discussed.

plant biology↗

bioRxiv2020

A multiscale analysis of early flower development in Arabidopsis provides an integrated view of molecular regulation and growth control

The link between gene regulation and morphogenesis of multicellular organisms is a fundamental problem in biology. We address this question in the floral meristem of Arabidopsis, which generates new tissues and organs through complex changes in growth patterns. Starting from high-resolution time-lapse images, we generated a comprehensive 4-D atlas of early flower development including cell lineage, cellular growth rates and the expression patterns of 28 regulatory genes. This information was introduced in MorphoNet, a web-based open-access platform. The application of mechanistic computational models indicated that the molecular network based on the literature only explained a minority of the expression patterns. This was substantially improved by adding single regulatory hypotheses for individual genes. We next used the integrated information to correlate growth with the combinatorial expression of multiple genes. This led us to propose a set of hypotheses for the action of individual genes in morphogenesis, not visible by simply correlating gene expression and growth. This identified the central transcription factor LEAFY as a potential regulator of heterogeneous growth, which was supported by quantifying growth patterns in a leafy mutant. By providing an integrated, multiscale view of flower development, this atlas should represent a fundamental step towards mechanistic multiscale-scale models of flower development.

developmental biology↗

bioRxiv2019

Structural imaging of native cryo-preserved secondary cell walls reveals presence of macrofibrils composed of cellulose, lignin and xylan.

The woody secondary cell walls of plants are the largest repository of renewable carbon biopolymers on the planet. These walls are made principally from cellulose and hemicelluloses and are impregnated with lignin. Despite their importance as the main load bearing structure for plant growth, as well as their industrial importance as both a material and energy source, the precise arrangement of these constituents within the cell wall is not yet fully understood. We have adapted low temperature scanning electron microscopy (cryo-SEM) for imaging the nanoscale architecture of angiosperm and gymnosperm cell walls in their native hydrated state. Our work confirms that cell wall macrofibrils, cylindrical structures with a diameter exceeding 10 nm, are a common feature of the native hardwood and softwood samples. We have observed these same structures in Arabidopsis thaliana secondary cell walls, enabling macrofibrils to be compared between mutant lines that are perturbed in cellulose, hemicellulose and lignin formation. Our analysis indicates that the macrofibrils in Arabidopsis cell walls are composed, at least partially, of cellulose, xylan and lignin. This study is a useful additional approach for investigating the native nanoscale architecture and composition of hardwood and softwood secondary cell walls and demonstrates the applicability of Arabidopsis genetic resources to relate fibril structure with wall composition and biosynthesis.

plant biology↗

bioRxiv2017

Cell Cycle Control by Nuclear Sequestration of CDC20 and CDH1 mRNA in Plant Stem Cells

In eukaryotic cells, most RNA molecules are exported into the cytoplasm after being transcribed in the nucleus. Long noncoding RNAs (lncRNAs) have been found to reside and function primarily inside the nucleus, but nuclear localization of protein-coding messenger RNAs (mRNAs) has been considered rare in both animals and plants. Here we show that two mRNAs, transcribed from the CDC20 and CCS52B (plant orthologue of CDH1) genes, are specifically sequestered inside the nucleus during the cell cycle. CDC20 and CDH1 both function as coactivators of the anaphase-promoting complex or cyclosome (APC/C) E3 ligase to trigger cyclin B (C YCB) destruction. In the Arabidopsis thaliana shoot apical meristem (SAM), we find CDC20 and CCS52B are co-expressed with CYCBs in mitotic cells. CYCB transcripts can be exported and translated, whereas CDC20 and CCS52B mRNAs are strictly confined to the nucleus at prophase and the cognate proteins are not translated until the redistribution of the mRNAs to the cytoplasm after nuclear envelope breakdown (NEBD) at prometaphase. The 5 untranslated region (UTR) is necessary and sufficient for CDC20 mRNA nuclear localization as well as protein translation. Mitotic enrichment of CDC20 and CCS52B transcripts enables the timely and rapid activation of APC/C, while their nuclear sequestration at prophase appears to protect cyclins from precocious degradation.

cell biology↗