Biology subjects

Scandola, S.

Publications and source records attributed to Scandola, S..

8 recordsLinked to original sources

bioRxiv2026

Quantitative proteomics uncovers a role for REVEILLE8- like clock genes in osmotic and salt stress response in Arabidopsis

The plant circadian clock governs the precise regulation of plant developmental and environmental responses within a 24 h photoperiod. Consisting of a series of interconnected transcription factor moderated feedback loops, each operating at specific times of day within the photoperiod, more than 30% of Arabidopsis genes show circadian regulation. REVEILLE (RVE) genes exhibit highest expression in the afternoon, and act as activators of afternoon-expressed clock genes, including TOC1 and PRR5. Specifically, RVE4, 6 and 8 have been identified as regulators of both plant growth and development, along with temperature responses. Previous work using Arabidopsis has implicated soybean RVE8-like proteins in drought responses, however, there remains a lack of understanding of how these RVEs control abiotic stress responses, particularly relating to proteome-level regulation. Here, using quantitative proteomics, we resolve how the rve4 6 8 proteome responds to osmotic and salt stress at end-of-day and end-of-night timepoints. Our results indicate that rve4 6 8 plants have substantially altered regulation of proteins related to osmotic stress, photosynthesis and phenylpropanoid metabolism, particularly at end-of-day. Further, we resolve new drought-responsive targets impacted by the loss of RVE4, 6, and 8 that are involved in vesicle transport, fatty acid metabolism and abscission. Overall, our data provide a new resource for understanding how RVE8-like proteins impact plant drought-like responses, offering opportunities for future chronoculture-informed breeding of climate-resilient crops.

Plant Biology↗

bioRxiv2024

Shaping Kale Morphology and Physiology Using Different LED Light Recipes

Light serves as a fundamental factor in plant development, both as an energy source and as an environmental cue. With the advent of light-emitting diode (LED) technology, light can be precisely manipulated to influence key plant traits. Here, we assess effects of light intensity and spectral composition on the growth and physiology of Kale (Brassica oleracea). Kale is known for its phenotypic plasticity and nutritional composition, making it a crop well-suited for indoor cultivation either as microgreens or as large leafy plants. Here, we employ a combination of advanced phenotyping, computer vision, gas chromatography-mass spectrometry (GC-MS) metabolomics, and liquid chromatography-mass spectrometry (LC-MS)-based quantitative proteomics to characterize the molecular changes that underpin light-dictated differences in the growth and metabolism of two different commercially grown kale cultivars under different light intensities and spectral compositions. We identify time-of-day and cultivar-specific light intensity and spectral composition-induced changes related to growth, shade avoidance, photosynthesis and several aspects of nutritional composition, including amino acids, glucosinolates and carotenoids. Our results offer a key resource to the plant community and demonstrate the translational potential of light manipulation in tailoring kale growth and nutritional content for enhanced crop productivity and/or health benefits, while simultaneously offering a more cost-effective solution for contemporary agricultural challenges. Significance StatementThe effects of light intensity and spectral composition differentially affect the diel molecular responses of Kale (Brassica oleracea). Our results demonstrate the translational potential of light manipulation in tailoring plant growth and nutritional content for enhanced crop productivity.

plant biology↗

bioRxiv2024

Quantitative proteomic analysis of soil-grown Brassica napus responses to nutrient deficiency

ABSTRACTMacronutrients such as nitrogen (N), phosphorus (P), potassium (K), and sulphur (S) are critical for plant growth and development. Field-grown canola (Brassica napus L.) is supplemented with fertilizers to maximize plant productivity, while deficiency in these nutrients can cause significant yield loss. A holistic understanding of the interplay between these nutrient deficiency responses in a single study and canola cultivar is thus far lacking, hindering efforts to increase the nutrient use efficiency of this important oil seed crop. To address this, we performed a comparative quantitative proteomic analysis of both shoot and root tissue harvested from soil-grown canola plants experiencing either nitrogen, phosphorus, potassium, or sulphur deficiency. Our data provide critically needed insights into the shared and distinct molecular responses to macronutrient deficiencies in canola. Importantly, we find more conserved responses to the four different nutrient deficiencies in canola roots, with more distinct proteome changes in aboveground tissue. Our results establish a foundation for a more comprehensive understanding of the shared and distinct nutrient deficiency response mechanisms of canola plants and pave the way for future breeding efforts.

plant biology↗

bioRxiv2024

B4 Raf-like MAPKKK RAF24 regulates Arabidopsis thaliana flowering time through HISTONE MONO-UBIQUITINATION 2

The timing of flowering is a critical agronomic trait governed by a number of external cues. Despite our genetic understanding of flowering time being well established, we have a limited understanding of how these signals are transmitted to different flowering genes through protein phosphorylation. Here, we characterize a novel B4 Raf-like MAPKKK protein kinase called RAF24, whose mutation results in an early flowering phenotype. Comparative analysis to related B4 Raf-like MAPKKKs indicates that RAF24 uniquely affects flowering time, while phosphoproteome analyses found RAF24 impacts the phosphorylation status of proteins involved in distinct flowering pathways. In particular, we found the RING-type ubiquitin ligase HISTONE MONO-UBIQUITINATION 2 (HUB2) to possess the largest phosphorylation change in raf24 deficient plants relative to wild-type Arabidopsis and that RAF24 suppresses ligase activity of HUB2 in order to maintain appropriate levels of H2Bub1. Furthermore, we found that RAF24 regulates HUB2 phosphorylation through subclass I and III SUCROSE NON-FERMENTING KINASE 2 (SnRK2) protein kinases; known substrates of B4 RAF-like MAPKKKs. Lastly, using a combination of phospho-mimetic and -ablative plant lines, we validate the importance of HUB2 phosphorylation at S314 in regulating flowering time. Collectively, our findings implicate RAF24 as a higher-order flowering regulator, while further implicating HUB2 as a centerpiece of flowering regulation.

plant biology↗

bioRxiv2023

Twilight length alters growth, flowering time, and clock function in Arabidopsis via LHY/CCA1

The plant circadian clock governs growth and development by syncing biological processes to periodic environmental changes. Decades of research has shown how the clock enables plants to respond to two environmental variables that change at different latitudes and over different seasons: photoperiod and temperature. However, a third variable, twilight length, has so far gone unstudied. The duration of twilight changes across the planet, lengthening with latitude and changing across seasons, and yet most circadian experiments are performed in lab environments with no twilight. Here, using controlled growth setups, we show that twilight length impacts plant growth and changes flowering time via the LHY/CCA1 morning module of the Arabidopsis circadian clock. Using a series of progressively truncated no-twilight photoperiods, we also found that plants are more sensitive to twilight length compared to equivalent changes in solely photoperiod. Further, transcriptome and proteome analyses indicated that twilight length alters the regulation of proteins involved in reactive oxygen species metabolism, photosynthesis, and carbon metabolism. Genetic analyses also implicated a twilight sensing pathway from phytochromes D and E, the morning elements of the circadian clock and modulating flowering time through the GI-FT pathway. Overall, our findings call for more nuanced models of daylength perception in plants and posit that twilight length is an important determinant of plant growth, development, and circadian function.

plant biology↗

bioRxiv2022

Integrated Systems-Level Proteomics and Metabolomics Reveals the Diel Molecular Landscape of Diverse Kale Cultivars

Kale are a group of diverse Brassicaceae species that are nutritious leafy greens consumed for their abundance of vitamins and micronutrients. Typified by their curly, serrated and/or wavy leaves, kale varieties have been primarily defined based on their leaf morphology and geographic origin, despite having complex genetic backgrounds. Kale is a very promising crop for vertical farming due to its high nutritional content; however, being a non-model organism, foundational, systems-level analyses of kale are lacking. Previous studies in kale have shown that time-of-day harvesting can affect its nutritional composition. Therefore, to gain a systems-level diel understanding of kale across its wide-ranging and diverse genetic landscape, we selected nine publicly available and commercially grown kale cultivars for growth under near-sunlight LED light conditions ideal for vertical farming. We then analyzed changes in morphology, growth and nutrition using a combination of phenomics, proteomics and metabolomics. As the diel molecular activities of plants drive their daily growth and development, ultimately determining their productivity as a crop, we harvested kale leaf tissue at both end-of-day (ED) and end-of-night (EN) time-points for all molecular analyses. Our results reveal that diel proteome and metabolome signatures divide the selected kale cultivars into two distinct groups, defined by their amino acid and sugar content, along with significant proteome differences involving carbon and nitrogen metabolism, mRNA splicing, protein translation and light harvesting. Together, our multi-cultivar, multi-omic analysis provides robust quantitative insights into the molecular underpinnings of the diel growth and development landscape of kale, significantly advancing our fundamental understanding of this nutritious leafy green super-food for next-generation horticulture / vertical farming applications.

plant biology↗

bioRxiv2021

Multi-omic analysis reveals the biochemical changes underpinning the varied phenotypes of the Arabidopsis long-period mutant rve 4 6 8

Plants are able to sense changes in their light environments, such as the onset of day and night, as well as anticipate these changes in order to adapt and survive. Central to this ability is the plant circadian clock, a molecular circuit that precisely orchestrates plant cell processes over the course of a day. REVEILLE proteins (RVEs) are recently discovered members of the plant circadian circuitry that activate the evening complex and PRR genes to maintain regular circadian oscillation. The RVE 8 protein and its two homologs, RVE 4 and 6, have been shown to limit the length of the circadian period, with rve 4 6 8 triple-knockout plants possessing an elongated period along with increased leaf surface area, biomass, cell size and delayed flowering relative to wild-type Col-0 plants. Here, using a multi-omics approach consisting of phenomics, transcriptomics, proteomics, and metabolomics we draw novel connections between RVE8-like proteins and a number of core plant cell processes. In particular, we reveal that loss of RVE8-like proteins results in altered carbohydrate, organic acid and lipid metabolism, including a starch excess phenotype at dawn. We further demonstrate that rve 4 6 8 plants have lower levels of 20S proteasome subunits and possess significantly reduced proteasome activity, potentially explaining the increase in cell-size observed in RVE8-like mutants. Overall, this robust, multi-omic dataset, provides substantial new insights into the far reaching impact RVE8-like proteins have on the diel plant cell environment.

plant biology↗

bioRxiv2020

Direct data-independent acquisition (direct DIA) enables substantially improved label-free quantitative proteomics in Arabidopsis

The last decade has seen significant advances in the application of quantitative mass spectrometry-based proteomics technologies to tackle important questions in plant biology. The current standard for quantitative proteomics in plants is the use of data-dependent acquisition (DDA) analysis with or without the use of chemical labels. However, the DDA approach preferentially measures higher abundant proteins, and often requires data imputation due to quantification inconsistency between samples. In this study we systematically benchmarked a recently developed library-free data-independent acquisition (directDIA) method against a state-of-the-art DDA label-free quantitative proteomics workflow for plants. We next developed a novel acquisition approach combining MS1-level BoxCar acquisition with MS2-level directDIA analysis that we call BoxCarDIA. DirectDIA achieves a 33% increase in protein quantification over traditional DDA, and BoxCarDIA a further 8%, without any changes in instrumentation, offline fractionation, or increases in mass-spectrometer acquisition time. BoxCarDIA, especially, offers wholly reproducible quantification of proteins between replicate injections, thereby addressing the long-standing missing-value problem in label-free quantitative proteomics. Further, we find that the gains in dynamic range sampling by directDIA and BoxCarDIA translate to deeper quantification of key, low abundant, functional protein classes (e.g., protein kinases and transcription factors) that are underrepresented in data acquired using DDA. We applied these methods to perform a quantitative proteomic comparison of dark and light grown Arabidopsis cell cultures, providing a critical resource for future plant interactome studies. Our results establish BoxCarDIA as the new method of choice in quantitative proteomics using Orbitrap-type mass-spectrometers, particularly for proteomes with large dynamic range such as that of plants.

plant biology↗