Biology subjects

Bellini, C.

Publications and source records attributed to Bellini, C..

17 recordsLinked to original sources

bioRxiv2026

Phenological plasticity provides limited resilience to climate warming in a sea turtle species with temperature-dependent sex determination

Anthropogenic climate change is threatening global biodiversity, with sea turtles particularly vulnerable as offspring sex and developmental success are strongly influenced by incubation temperature. Behavioral plasticity, including the seasonal distribution of reproductive output, may provide short-term mechanisms for mitigating these impacts. Here, we investigated season-wide hatchling sex ratios and emergence success in a small population of green turtles (Chelonia mydas), tracking individual females across their nesting seasons. Sex ratios varied markedly through the nesting season, with later nests producing a greater proportion of male hatchlings. Moreover, sex ratios were relatively consistent among nests laid by individual females. Overall, females producing more nests over a season also produced more male offspring, suggesting that both nesting phenology and reproductive output influence individual contributions to future population demographics. Mechanistic models indicate that hatchling sex ratios have trended towards female-biased ratios (>80% female) over the past 50 years, and are projected to approach complete feminization by 2100 under continued warming. Although emergence success currently remains high (>85%), it is predicted to decline sharply after mid-century, with viable hatchling production falling to [~]30% by the end of the century. Models further show that maintaining contemporary sex ratios and emergence success will require unrealistically large delays in nesting phenology, and that even extreme shifts in phenology become ineffective by 2100. Together, these findings demonstrate that individual females can increase male hatchling production by nesting later and producing more nests, but behavioral plasticity alone is unlikely to offset the accelerating impacts of climate change on this population.

ecology↗

bioRxiv2026

Structural Lung Remodeling Precedes Functional Decline After Chronic Smoldering Douglas Fir Smoke Exposure in Apoe-/- Mice

Wildland firefighters experience repeated exposure to wildfire smoke, yet the pathophysiological mechanisms underlying chronic inhalation injury remain poorly understood. Although prior studies report parenchymal destruction following prolonged woodsmoke exposure, the temporal relationship between molecular, structural, and functional decline following inhalation of smoke from needles/leaves remains unclear. To address this gap, we characterized coordinated changes in lung structure, function, and underlying molecular disruptions using a dosimetry-based murine model approximating 7-14 years of firefighter service. Male apolipoprotein E-deficient mice were exposed to smoldering Douglas fir needle smoke (40 mg/m3, 2 h/day, 5 days/week) for 8 or 16 weeks. Immunofluorescence analyses revealed an early elastolytic response at 8 weeks, with increased neutrophil elastases and matrix metalloproteinases-9 and -12, accompanied by elevated surfactant protein-D, compared to air controls. These changes were resolved by 16 weeks despite progressive tissue injury. Airspace enlargement was evident at 8 weeks, progressed by 16 weeks, and included increased alveolar blunting and septal wall thickening at the later time point. Cleaved caspase-3 was elevated at 16 weeks, indicative of advanced parenchymal damage and apoptosis. Epithelial tight-junction protein ZO-1 intensity was reduced at both evaluation points, whereas the epithelial-to-mesenchymal marker N-cadherin remained undetectable in the alveolar epithelium. Functional impairment as evident by increased static compliance and upward shifts in pressure-volume curves was only significant after 16 weeks of exposure. Findings indicate that molecular and structural injury of tissue destruction preceded measurable functional decline, underscoring the need for early biomarkers to identify smoke-induced lung injury in wildland firefighters before function loss occurs.

physiology↗

bioRxiv2026

Occupationally Relevant Wildfire Smoke Inhalation Impairs Nitric Oxide Signaling and Promotes Progressive Aortic Stiffening in Hypercholesterolemic Mice

BackgroundWildland firefighters experience repeated occupational exposure to wildfire smoke at high particulate matter (PM) concentrations, leading to elevated cardiovascular disease risk and hypertension prevalence. However, the pathophysiological processes linking cumulative smoke inhalation to vascular damage and blood pressure elevation remain poorly characterized. To evaluate these effects under controlled exposure conditions, we used a preclinical exposure model calibrated to match the cumulative PM burden deposited in wildland firefighter airways over 7-14 years of service. Male apolipoprotein E knockout (Apoe-/-) mice underwent whole-body inhalation of Douglas fir smoke or filtered air for 2 hours/day, 5 days/week, for 8 or 16 weeks at target PM concentrations of 40 mg/m3. ResultsProlonged smoke exposure induced sustained elevation of circulating tumor necrosis factor-alpha (TNF-), interleukin-1 beta (IL-1{beta}), and interleukin-6 (IL-6), coupled with diffused nuclear factor kappa B (NF-{kappa}B) activation throughout the aortic wall. Smoke inhalation disrupted endothelial adherens junctions, upregulated intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1), and promoted monocyte recruitment to aortic tissues, concurrent with enhanced monocyte chemoattractant protein-1 (MCP-1) expression. Oxidative stress was evidenced by increased nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit 2 (NOX2) expression, elevated superoxide levels, and endothelial nitric oxide synthase (eNOS) uncoupling in the aorta, leading to lipid peroxidation and accompanied by intimal apoptosis. These inflammatory and oxidative perturbations occurred alongside a pro-fibrotic phenotypic shift characterized by transforming growth factor beta 1 (TGF-{beta}1) upregulation, myofibroblast differentiation, and progressive collagen accumulation in medial and adventitial compartments of the aortic wall. Functionally, smoke exposure progressively impaired aortic cyclic distensibility through combined wall thickening and circumferential tissue stiffening, while severely attenuating endothelium-dependent and nitric oxide (NO)-mediated vasodilation. These functional and structural shifts culminated in elevated systolic and diastolic blood pressures. While endothelial dysfunction reached maximal impairment by 8 weeks, aortic stiffening continued to worsen through 16 weeks of exposure, demonstrating differential temporal progression of vascular damage. ConclusionsThese findings demonstrate that occupationally relevant wildfire smoke exposure produces convergent inflammatory, oxidative, and profibrotic vascular remodeling with progressive loss of arterial compliance and impaired endothelium-dependent vasodilation, underscoring potential vascular targets for cardiovascular health surveillance and risk mitigation in wildland firefighters.

pharmacology and toxicology↗

bioRxiv2026

Machine Learning Approach for Enumeration of Circulating Cells with Diffuse in vivo Flow Cytometry

SignificanceDiffuse in vivo flow cytometry (DiFC) is an emerging technique for enumerating rare, fluorescentlylabeled circulating tumor cells (CTCs) in small animals without drawing blood samples. DiFC uses detection of transient fluorescent peaks in time-series data. Previously, we used a simple amplitude threshold-based method for identifying peak candidates, but it ignores potentially useful information in peak shape that could reduce false-positive detections from instrument noise and increase detection efficiency of lower-amplitude peaks. AimTo develop a machine learning (ML)-integrated signal processing approach for improved CTC enumeration using DiFC by distinguishing CTC peaks from artifacts. ApproachWe developed an ML-integrated approach that incorporates a convolutional neural network (CNN) classifier. The CNN was trained to distinguish CTC peaks from artifacts by analyzing peak amplitude and temporal shape characteristics. Performance was validated on in-silico, control, and CTC-bearing mouse datasets. ResultsThe CNN classifier achieved accuracy, precision, sensitivity, and specificity exceeding 98% on test data. Compared with our previously published threshold-based approach, the ML-integrated method increased the number of correctly identified CTCs and their flow direction while reducing false detections across validation datasets. ConclusionsThe ML-integrated approach significantly improves DiFC CTC enumeration, enabling robustness against artifacts in noisy conditions.

bioengineering↗

bioRxiv2026

An Integrated Multiphoton Imaging Workflow for Quantitative Analysis of Aortic Tissue Microstructure

Quantitative, reproducible characterization of aortic microstructure is essential for advancing vascular biomechanics and mechanobiology. To address this need, we present a comprehensive image-analysis workflow that extracts quantitative descriptors of tissue microstructure from multiphoton microscopy stacks of the murine thoracic aorta. Channel-specific signals are acquired for fibrillar collagen (second harmonic generation), elastin (two-photon autofluorescence), and cell nuclei (two-photon excited fluorescence). Following reorientation into the XZ plane, individual elastic lamellae are traced to quantify lamellar thickness and interlamellar spacing using circle-based geometry (Taubin fitting). After correction for vessel wall curvature via a cylindrical transformation, segmented nuclei are assigned to medial or adventitial compartments based on visual estimates of adventitial volume fraction, and nuclear morphology is characterized via ellipsoidal fitting in terms of nuclear aspect ratio and major-axis orientation. Collagen organization is resolved in XY sections by extracting fiber centerlines to quantify straightness and amplitude; traces from serial sections are then combined to reconstruct the three-dimensional collagen network and estimate porosity and linear fiber density, while fiber orientation distributions are derived from principal component analysis-based angles and fit using a von Mises mixture model. Finally, collagen and elastin volume fractions are computed via a two-stage fixed-threshold approach calibrated on a balanced training subset. Overall, this modular and robust workflow provides an integrated framework for studying aortic wall remodeling across physiological and pathological processes. Non-Technical SummaryAs the main blood vessel in our body, the aorta needs to be both strong and flexible. This balance comes from three main parts: elastic layers that allow the aorta to stretch, strong fibers that prevent tearing, and cells that sense and respond to changes in blood pressure and other signals. When any of these components are altered, the aorta may stiffen or weaken, which can interfere with normal blood flow. In this study, we developed a clear and consistent way to measure the structure of the aortic wall using microscope images. The approach examines how thick the elastic layers are and how far apart they lie, the size and orientation of cell centers, and how straight or wavy structural fibers appear. It also estimates how much of each component is present in the aortic wall. Because the same steps are applied each time, results can be fairly compared across different conditions. Overall, this tool transforms detailed images into simple measurements, helping scientists understand how the aorta changes in health and disease.

bioengineering↗

bioRxiv2024

SUC2 sucrose transporter is required for leaf apoplasmic sucrose levels. Consequences for phloem loading strategies

Summary * The SUC/SUT sucrose transporters belong to a family of active H+/sucrose symporters, with a role of SUC2 in active apoplasmic phloem loading to drive long-distance phloem transport of sucrose in Arabidopsis. However, the cooperation with the symplasmic pathway for phloem loading remains unclear. * In this study, we explored the consequences of reducing either apoplasmic or symplasmic pathways of phloem loading. We compared a series of lines with modified expression of SUC2 gene, and we analyzed the effects on plant growth, sugar accumulation in source and sink organs, phloem transport, and gene expression. * Our data revealed that a modified expression of SUC2 impacted apoplasmic sucrose levels in source leaves but did not impact phloem transport, as might be expected, while increasing foliar storage of carbohydrates. This response differed from lines in which symplasmic communications between phloem cells was disrupted by the over-expression of a plasmodesmata-associated protein, NHL26. * Altogether, our studies indicate an unexpected effect of SUC2 for apoplasmic sucrose levels in source leaves, together with SUC1, and suggest a feedback regulation on foliar storage. This data sheds new light on the interplay between symplasmic and apoplasmic pathways for sugar loading and the consequences on leaf water flows. Summary statementThe mechanisms that coordinate apoplasmic and symplasmic loading pathways, and their effects on foliar carbon storage, remain largely unexplored. Surprisingly, the sucrose transporter SUC2 plays a significant role in maintaining sucrose levels in the apoplasm, shedding light on how apoplasmic sugar levels and water flows can interact for phloem loading.

plant biology↗

bioRxiv2024

Chronic Pod-Mod E-Cigarette Aerosol Exposure Induces Aortic Dysfunction in Hypercholesterolemic Mice: Role of Oxidative Stress and Inflammation

ObjectiveElectronic (e-)cigarettes are the most used tobacco product amongst youth, and adult smokers favor e-cigarettes over approved cessations aids. Despite the lower perceived harm of vaping compared to smoking, inhalation of e-cigarette aerosol elicits cardiovascular responses that may lead to permanent injury when repeated over time. We thus aimed to infer the long-term outcomes of vaping on the function and structure of the aorta and shed light on the underlying cellular and molecular mechanisms. Approach and ResultsWe exposed female hypercholesterolemic mice to either pod-mod e-cigarette aerosol or room air daily for 24 weeks. Chronic inhalation of e-cigarette aerosol triggered accumulation of inflam-matory signals systemically and within aortic tissues, as well as T lymphocyte accrual in the aortic wall. Reduced eNOS expression and enhanced ROS production following eNOS uncoupling and NADPH oxi-dase activation curbed nitric oxide availability in the aorta of mice exposed to e-cigarette aerosol, impairing the endothelium-dependent vasodilatation that regulates blood flow distribution. Inhalation of e-cigarette aerosol thickened and stiffened aortic tissues via collagen deposition and remodeling, hindering the storage of elastic energy and limiting the cyclic distensibility that enables the aorta to function as a pressure reservoir. These effects combined contributed to raising systolic and pulse pressure above control levels. ConclusionsChronic inhalation of aerosol from pod-mod e-cigarettes promotes oxidative stress, inflammation, and fibrosis within aortic tissues, significantly impairing passive and vasoactive aortic functions. This evidence provides new insights on the biological processes that increase the risk for adverse cardio-vascular events as a result of pod-mod e-cigarette vaping.

bioengineering↗

bioRxiv2022

Mapping the unicellular transcriptome of the ascending thoracic aorta to changes in mechanosensing and mechanoadaptation during aging

Aortic stiffening is an inevitable manifestation of chronological aging, yet the mechano-molecular programs that orchestrate region- and layer-specific adaptations along the length and through the wall of the aorta are incompletely defined. Here, we show that the decline in passive cyclic distensibility is more pronounced in the ascending thoracic (ATA) compared to distal segments of the aorta and that tissues in both the medial and adventitial compartments of the ATA stiffen during aging. Single-cell RNA sequencing of aged ATA tissues reveals altered cellular senescence, remodeling, and inflammatory responses accompanied by enrichment of T-lymphocytes and rarefaction of vascular smooth muscle cells, compared to young samples. T-lymphocytes accumulate in the adventitia and likely promote fibrosis, while activation of mechanosensitive piezo-1 enhances medial vasoconstriction. These results portray the immuno-mechanical aging of the ATA as a process that culminates in a stiffer conduit permissive to the accrual of multi-gerogenic signals priming to disease development.

bioengineering↗

bioRxiv2021

Molecular basis of differential adventitious rooting competence in poplar genotypes

O_LIRecalcitrant adventitious root (AR) development is a major hurdle in propagating commercially important woody plants. Although significant progress has been made to identify genes involved in subsequent steps of AR development, the molecular basis of differences in apparent recalcitrance to form AR between easy-to-root and difficult-to-root genotypes remains unknown. C_LIO_LITo address this, we generated cambium tissue-specific transcriptomic data from stem cuttings of hybrid aspen, T89 (difficult-to-root) and hybrid poplar OP42 (easy-to-root) and used transgenic approaches to verify the role of several transcription factors (TF) in the control of adventitious rooting. C_LIO_LIIncreased peroxidase activity is positively correlated with better rooting. We found differentially expressed genes encoding Reactive Oxygen Species (ROS) scavenging proteins to be enriched in OP42 compared to T89. A higher number of differentially expressed TF in OP42 compared to T89 cambium cells was revealed by a more intense transcriptional reprograming in the former. PtMYC2, a potential negative regulator, was less expressed in OP42 compared to T89. Using transgenic approaches, we have demonstrated that PttARF17.1 and PttMYC2.1 negatively regulate adventitious rooting. C_LIO_LIOur results provide insights into the molecular basis of genotypic differences in AR and implicate differential expression of the master regulator MYC2 as a critical player in this process. C_LI

plant biology↗

bioRxiv2021

Jasmonate inhibits adventitious root initiation through transcriptional repression of CKX1 and activation of RAP2.6L transcription factor in Arabidopsis

Adventitious rooting is a de novo organogenesis process that enables plants to propagate clonally and cope with environmental stresses. Adventitious root initiation (ARI) is controlled by interconnected transcriptional and hormonal networks, but there is little knowledge of the genetic and molecular programs orchestrating these networks. Thus, we have applied genome-wide transcriptome profiling to elucidate the profound transcriptional reprogramming events preceding ARI. These reprogramming events are associated with downregulation of cytokinin (CK) signaling and response genes, which could be triggers for ARI. Interestingly, we found that CK free-base content declined during ARI, due to downregulation of de novo CK biosynthesis and upregulation of CK inactivation pathways. We also found that MYC2-dependent jasmonate (JA) signaling inhibits ARI by downregulating expression of the CYTOKININ OXIDASE/DEHYDROGENASE1 gene. We also demonstrated that JA and CK synergistically activate expression of RELATED to APETALA2.6 LIKE (RAP2.6L) transcription factor, and constitutive expression of this transcription factor strongly inhibits ARI. Collectively, our findings reveal that previously unknown genetic interactions between JA and CK play key roles in ARI

plant biology↗

bioRxiv2020

Hexose fluxes, mediated by vacuolar SWEET transporters, are important for xylem development in the inflorescence stem of Arabidopsis thaliana.

In higher plants, the development of the vascular system is controlled by a complex network of transcription factors. However, how nutrient availability in the vascular cells affects their development remains to be addressed. At the cellular level, cytosolic sugar availability is regulated mainly by sugar exchanges at the tonoplast through active and/or facilitated transport. In Arabidopsis thaliana, among the tonoplastic transporters, SWEET16 and SWEET17 genes have been previously localized in the vascular system. Here, using a reverse genetic approach, we propose that sugar exchanges at the tonoplast, regulated by SWEET16, are important for xylem cell division as revealed in particular by the decreased number of xylem cells in the swt16 mutant and the expression of SWEET16 at the procambium-xylem boundary. In addition, we demonstrate that transport of hexoses mediated by SWEET16 and/or SWEET17 is required to sustain the formation of the xylem secondary cell wall. This result is in line with a defect in the xylem cell wall composition as measured by FTIR in the swt16swt17 double mutant and by upregulation of several genes involved in secondary cell wall synthesis. Our work therefore supports a model in which xylem development is partially dependent on the exchange of hexoses at the tonoplast of xylem-forming cells.

plant biology↗

bioRxiv2020

Light controls de novo adventitious root regeneration by modulating jasmonate and cytokinin homeostasis in Norway spruce hypocotyls

Vegetative propagation relies on the capacity of plants to regenerate de novo adventitious roots (ARs), a quantitative trait controlled by the interaction of endogenous factors such as hormones and environmental cues among which light plays a central role. However, the physiological and molecular components mediating light cues during AR initiation (ARI) remain largely elusive. We explored the effect of light spectral quality on ARI in de-rooted Norway spruce seedlings as well as on hormone metabolism with sensitive mass spectrometry-based methods. We coupled this to gene expression analysis to identify potential signaling pathways and to extensive anatomical characterization to investigate ARI at the cellular level. We showed that in contrast to white light and blue light, red light promoted ARI likely by reducing jasmonate (JA) and JA-isoleucine biosynthesis and repressing the accumulation of isopentyl-adenine-type cytokinins and abscisic acid. We confirmed that exogenously applied JA and/or CK inhibit ARI, and found that they possibly act in the same pathway. The negative effect of JA was confirmed at the histological level. We showed that JA represses the early events of ARI. In conclusion, RL promotes ARI by repressing the accumulation of the wound-induced phytohormones JA and CK. HighlightBlue and red light have an opposite effect on adventitious root initiation in Norway spruce hypocotyl, red light having a promoting effect by modulating hormone homeostasis.

plant biology↗

bioRxiv2019

Cytokinin induction by the Jasmonate-induced AP2/ERF115 represses adventitious rooting in Arabidopsis

Jasmonate (JA), an oxylipin-derived phytohormone, plays crucial roles not only in plant immunity and defense against herbivorous insects but also in plant growth and developmental processes, including regeneration and organogenesis. However, the mechanistic basis of its mode of action and precise role in integrating other signaling cues are poorly understood. Here we provide genetic evidences that JA signaling acts in both NINJA-dependent and -independent modulation of the transcriptional activity of MYC transcription factors involved in the inhibition of adventitious root initiation (ARI). Our data show that NINJA-dependent JA signaling in pericycle cells blocks early events of ARI. Moreover, transcriptomic comparison of ninja-1myc2-322B double mutant (which produce extremely few ARs) and wild type seedlings identified a novel molecular network governed by the APETALA2/ETHYLENE RESPONSE FACTOR 115 (ERF115) transcription factor. We demonstrate that JA-induced ERF115 activates the cytokinin signaling machinery and thereby represses ARI. Altogether, our results reveal a molecular network involving cooperative crosstalk between JA and CK machineries that inhibits ARI.

plant biology↗

bioRxiv2019

Conifers exhibit a characteristic inactivation of auxin to maintain tissue homeostasis

O_LIDynamic regulation of the levels of the natural auxin, indol-3-acetic acid (IAA), is essential to coordinate most of the physiological and developmental processes and responses to environmental changes. Oxidation of IAA is a major pathway to control auxin concentrations in Arabidopsis and, along with IAA conjugation, to respond to perturbation of IAA homeostasis. However, these regulatory mechanisms are still poorly investigated in conifers. To reduce this gap of knowledge, we investigated the different contribution of the IAA inactivation pathways in conifers.\nC_LIO_LIMass spectrometry-based quantification of IAA metabolites under steady state conditions and after perturbation was investigated to evaluate IAA homeostasis in conifers. Putative Picea abies GH3 genes (PaGH3) were identified by a comprehensive phylogenetic analysis including Arabidopsis and basal land plants. Auxin-inducible PaGH3 genes were identified by expression analysis and their IAA-conjugating activity was explored.\nC_LIO_LICompared to Arabidopsis, oxidative and conjugative pathways differentially contribute to reduce IAA levels in conifers. We demonstrated that the oxidation pathway plays a marginal role in controlling IAA homeostasis in spruce. On the other hand, an excess of IAA rapidly activates GH3-mediated irreversible conjugation pathways.\nC_LIO_LITaken together, these data indicate that a diversification of IAA inactivation mechanisms evolved specifically in conifers.\nC_LI

plant biology↗

bioRxiv2019

A bacterial assay for rapid screening of IAA catabolic enzymes

BackgroundPlants rely on concentration gradients of the native auxin, indole-3-acetic acid (IAA), to modulate plant growth and development. Both metabolic and transport processes participate in the dynamic regulation of IAA homeostasis. Free IAA levels can be reduced by inactivation mechanisms, such as conjugation and degradation. IAA can be conjugated via ester linkage to glucose, or via amide linkage to amino acids, and degraded via oxidation. Members of the UDP glucosyl transferase (UGT) family catalyze the conversion of IAA to indole-3-acetyl-1-glucosyl ester (IAGlc); by contrast, IAA is irreversibly converted to indole-3-acetyl-L-aspartic acid (IAAsp) and indole-3-acetyl glutamic acid (IAGlu) by Group II of the GRETCHEN HAGEN3 (GH3) family of acyl amido synthetases. DIOXYGENASE OF AUXIN OXIDATION (DAO) irreversibly oxidizes IAA to oxindole-3-acetic acid (oxIAA) and, in turn, oxIAA can be further glucosylated to oxindole-3-acetyl-1-glucosyl ester (oxIAGlc) by UGTs. These metabolic pathways have been identified based on mutant analyses, in vitro activity measurements, and in planta feeding assays. In vitro assays for studying protein activity are based on expressing Arabidopsis enzymes in a recombinant form in bacteria or yeast followed by recombinant protein purification. However, the need to extract and purify the recombinant proteins represents a major obstacle when performing in vitro assays.\n\nResultsIn this work we report a rapid, reproducible and cheap method to screen the enzymatic activity of recombinant proteins that are known to inactivate IAA. The enzymatic reactions are carried out directly in bacteria that express the recombinant protein. The enzymatic products can be measured by direct injection of a small supernatant fraction from the bacterial culture on ultrahigh-performance liquid chromatography coupled to electrospray ionization tandem spectrometry (UHPLC-ESI-MS/MS). Experimental procedures were optimized for testing the activity of different classes of IAA-modifying enzymes without the need to purify recombinant protein.\n\nConclusionsThis new method represents an alternative to existing in vitro assays. It can be applied to the analysis of IAA metabolites that are produced upon supplementation of substrate to engineered bacterial cultures and can be used for a rapid screening of orthologous candidate genes from non-model species.

plant biology↗

bioRxiv2019

Molecular framework for TIR1/AFB-Aux/IAA-dependent auxin sensing controlling adventitious rooting in Arabidopsis

In Arabidopsis thaliana, canonical auxin-dependent gene regulation is mediated by 23 transcription factors from the AUXIN RESPONSE FACTOR (ARF) family, most of which interact with 29 auxin/indole acetic acid (Aux/IAA) repressors, themselves forming, in the presence of auxin, coreceptor complexes with one of six TRANSPORT INHIBITOR1/AUXIN-SIGNALLING F-BOX PROTEINS (TIR1/AFB). Different combinations of co-receptors drive specific sensing outputs, allowing auxin to control a myriad of processes. Considerable efforts have been made to discern the temporal and spatial specificity of auxin action. However, owing to a lack of obvious phenotype in single loss-of-function mutants in Aux/IAA genes, most genetic studies have relied on gain-of-function mutants, which are highly pleiotropic. In this article, we describe a molecular framework for the role of several members of the auxin sensing machinery. Using loss-of-function mutants, we demonstrate that TIR1 and AFB2 are positive regulators, whereas IAA6, IAA9 and IAA17 are negative regulators of adventitious root (AR) formation. The three Aux/IAA proteins interact with ARF6 and/or ARF8, which we have previously shown to be positive regulators of AR formation upstream of jasmonate, and likely repress their activity. Our data also suggest a dual role for TIR1 in the control of JA biosynthesis and conjugation, as revealed by upregulation of several JA biosynthesis genes in the tir1-1 mutant. In conclusion, we propose that in the presence of auxin, TIR1 and AFB2 form specific sensing complexes with IAA6, IAA9 and/or IAA17 that modulate JA homeostasis to control AR initiation.\n\nSignificanceAdventitious root (AR) formation is a quantitative trait with high phenotypic plasticity and auxin plays a major role in its control. In previous studies, we have shown that, in Arabidopsis hypocotyls, AR formation is negatively regulated by jasmonate, whose homeostasis is controlled by a regulatory module composed of three AUXIN RESPONSE FACTORS (ARF), two of which are positive regulators of three auxin inducible Gretchen Hagen3 (GH3) genes. The present work demonstrates that the two F-box proteins, i.e., TIR1 and AFB2, together with at least three Aux/IAA proteins are likely to interact in a combinatorial manner to control the activity of the two activator ARFs upstream of JA biosynthesis and/or homeostasis, thereby controlling AR initiation in Arabidopsis hypocotyls.

plant biology↗

bioRxiv2018

Synchrotron FTIR and Raman spectroscopy provide unique spectral fingerprints for Arabidopsis floral stem vascular tissues

Cell walls are highly complex structures that are modified during plant growth and development. For example, the development of phloem and xylem vascular cells, which participate in the transport of sugars and water as well as support, can be influenced by cell-specific cell wall composition. Here, we used synchrotron radiation-based infrared (SR-FTIR) and Raman spectroscopy to analyze the cell wall composition of wild-type and double mutant sweet11-1sweet12-1, which impairs sugar transport, Arabidopsis floral stem vascular tissue. The SR-FTIR spectra showed that in addition to modified xylem cell wall composition, phloem cell walls in the double mutant line were characterized by modified hemicellulose composition. Moreover, combining Raman spectroscopy with a Classification and Regression Tree (CART) method identified combinations of Raman shifts that could distinguish xylem vessels and fibers. Additionally, the disruption of SWEET11 and SWEET12 genes impacts xylem cell wall composition in a cell-specific manner, with changes in hemicelluloses and cellulose observed at the xylem vessel interface. These results suggest that the facilitated transport of sugars by transporters that exist between vascular parenchyma cells and conducting cells is important to ensuring correct phloem and xylem cell wall composition.\n\nHighlightCombining vibrational spectroscopy techniques and multivariate analysis shows that the disruption of SWEET genes impacts phloem cell wall composition and that the effect on xylem cell wall composition is cell-specific.

plant biology↗