Biology subjects

Fernie, A. R.

Publications and source records attributed to Fernie, A. R..

At least 37 records · Page 2Linked to original sources

bioRxiv2024

Sub-functionalization and epigenetic regulation of a biosynthetic gene cluster in Solanaceae

Biosynthetic gene clusters (BGCs) are sets of often heterologous genes that are genetically and functionally linked. Among eukaryotes, BGCs are most common in plants and fungi, where they ensure the co-expression of the different enzymes coordinating the biosynthesis of specialised metabolites. Here, we report on a newly identified withanolide BGC in Physalis grisea (ground-cherry), a member of the nightshade family (Solanaceae). A combination of transcriptomic, epigenomic and metabolic analyses, revealed that, following a duplication event, this BGC evolved two tissue-specifically expressed sub-clusters, containing eight pairs of paralogs that contribute to related but distinct biochemical processes; this sub- functionalisation is tightly associated with epigenetic features and the local chromatin environment. The two sub-clusters appear strictly isolated from each other at the chromatin structural level, each forming a highly self-interacting chromatin domain with tissue-dependent levels of condensation. This correlates with gene expression exclusively in either above- or below-ground tissue, thus spatially separating the production of different withanolide compounds. By comparative phylogenomic analysis, we show that the withanolide BGC most likely evolved before the diversification of the Solanaceae family and underwent lineage- specific diversifications and losses. The tissue-specific sub-functionalisation is common to species of the Physalideae tribe but distinct from other, independent duplication events outside of this clade. In sum, our study reports on the first instance of an epigenetically modulated sub-functionalisation within a BGC and sheds light on the biosynthesis of withanolides, a highly diverse group of steroidal triterpenoids important in plant defence and amenable to pharmaceutical applications due to their anti-inflammatory, antibiotic and anti- cancer properties.

plant biology↗

bioRxiv2024

Joint operation of the CO2 concentrating mechanism and photorespiration in green algae during acclimation to limiting CO2

Due to low availability of CO2 in aquatic environment, microalgae have evolved a CO2 concentrating mechanism (CCM). It has long been thought that operation of CCM would suppress photorespiration by increasing the CO2 concentration at the Rubisco active site, but experimental evidence is scarce. To better explore the function of photorespiration in algae, we first characterized a Chlamydomonas reinhardtii mutant defected in low-CO2 inducible 20 (LCI20) and show that LCI20 is a chloroplast-envelope glutamate/malate transporter playing a role in photorespiration. By monitoring growth and glycolate excretion in mutants deficient in either CCM or photorespiration, we conclude that: i. CCM induction does not depend on photorespiration, ii. glycolate excretion protects algal cells from the toxicity of unmetabolized photorespiratory intermediates, iii. photorespiration is active at low CO2 when the CCM is operational. This work provides a foundation for a better understanding of the carbon cycle in the ocean where significant glycolate concentrations have been found.

plant biology↗

bioRxiv2024

Genomic imprinting in an early-diverging angiosperm reveals ancient mechanisms for seed initiation in flowering plants

The evolution of the seed habit marks a pivotal innovation of the spermatophytes. Angiosperms further refined this trait by coupling the development of seed accessory structures to fertilization, optimizing resource allocation. Here, we demonstrate that post-fertilization auxin production is an evolutionarily conserved mechanism for seed initiation in angiosperms. We also provide evidence that this pathway likely emerged from a switch from maternal to paternal control after the divergence of angiosperms from their gymnosperm ancestors. Our study thus brings new insights into the evolutionary origins of the endosperm, which was a determining feature for the rapid rise to dominance of flowering plants.

plant biology↗

bioRxiv2024

Multi-omics of a rice population identifies genes and genomic regions in rice that bestow low glycemic index and high protein content

To address the growing incidences of increased diabetes and to meet the daily protein requirements, we developed low glycemic index (GI) rice varieties with protein yield exceeding 14%. In the development of recombinant inbred lines using Samba Mahsuri and IR36 amylose extender as parental lines, we identified quantitative trait loci (QTLs) and genes associated with low GI, high amylose content (AC), and high protein content (PC). By integrating genetic techniques with classification models, this comprehensive approach identified candidate genes on chromosome 2 (qGI2.1/qAC2.1 spanning the region from 18.62Mb to 19.95Mb), exerting influence on low GI and high amylose. Notably, the phenotypic variant with high value was associated with the recessive allele of the starch branching enzyme 2b (sbeIIb). The genome-edited sbeIIb line confirmed low GI phenotype in milled rice grains. Further, combinations of alleles from the highly significant SNPs from the targeted associations and epistatically interacting genes showed ultra-low GI phenotypes with high amylose and high protein. Metabolomics analysis of rice with varying AC, PC, and GI revealed that the superior lines of high AC and PC, and low GI were preferentially enriched in glycolytic and amino acid metabolism, whereas the inferior lines of low AC and PC and high GI were enriched with fatty acid metabolism. The high amylose high protein RIL (HAHP_101) was enriched in essential amino acids like lysine. Such lines may be highly relevant for food product development to address diabetes and malnutrition. Significance StatementThe increasing global incidence of diabetes calls for the development of diabetic friendly healthier rice. In this study, we developed recombinant inbred rice lines with milled rice exhibiting ultra-low to low glycemic index and high protein content from the cross between Samba Mahsuri and IR36 amylose extender. We performed comprehensive genomics and metabolomics complemented with modeling analyses emphasizing the importance of OsSbeIIb along with additional candidate genes whose variations allowed us to produce target rice lines with lower glycemic index and high protein content in a high-yielding background. These lines represent an important breeding resource to address food and nutritional security.

systems biology↗

bioRxiv2024

Chloroplastic ascorbate acts as a regulatory hub in plant metabolism regardless of oxidative stress

Ascorbate is a major plant metabolite that plays crucial roles in various processes, from reactive oxygen scavenging to epigenetic regulation. However, to what extent and how ascorbate modulates metabolism is largely unknown. To address this, we investigated the consequences of chloroplastic and total cellular ascorbate-deficiencies by studying chloroplastic ascorbate-transporter pht4;4 mutant lines, and the ascorbate-deficient vtc2-4 mutant of Arabidopsis thaliana. Under regular growth conditions, both ascorbate-deficiencies caused minor alterations in photosynthesis, with no apparent signs of oxidative damage. In contrast, metabolomics analysis revealed a global and largely overlapping metabolome rewiring in both ascorbate-deficiencies, suggesting that chloroplastic ascorbate modulates plant metabolism. We observed significant alterations in amino acid metabolism, particularly in arginine metabolism, activation of nucleotide salvage pathways, and changes in secondary metabolism. In addition, proteome-wide analysis of thermostability revealed that ascorbate may interact with enzymes involved in arginine metabolism, the Calvin-Benson cycle, and several photosynthetic electron transport components. Overall, our results suggest that, independently of oxidative stress, chloroplastic ascorbate interconnects and coordinates diverse metabolic pathways in vascular plants and thus acts as a regulatory hub.

plant biology↗

bioRxiv2024

The transcription factor ATML1 maintains giant cell identity by inducing synthesis of its own (very) long-chain fatty acid-containing ligands

During development, cells not only adopt specialized identities but also maintain those identities. Endoreduplication is thought to maintain cell identity. High concentrations of ARABIDOPSIS THALIANA MERISTEM LAYER1 (ATML1) specify giant cell identity and induce endoreduplication in sepals. How different concentrations of ATML1 can specify different identities remains unclear. Here, we show that high concentrations of ATML1 induce the biosynthesis of both long-chain and very long-chain fatty acids (LCFAs/VLCFAs), and these fatty acids are required for the maintenance of giant cell identity. Inhibition of VLCFA biosynthesis causes endoreduplicated giant cells to resume division and lose their identity, indicating that endoreduplication is not sufficient to maintain cell identity. Structural predictions suggest that LCFA-containing lipids bind to the START domain 2 of ATML1, causing ATML1 dimerization and its auto-activation. Our data and modeling imply that ATML1 induces biosynthesis of its own lipid ligands in a positive feedback loop, shedding light on the intricate network dynamics that specify and maintain giant cell identity. Teaser: Endoreduplicated cells in Arabidopsis thaliana sepals divide and de-differentiate in the absence of VLCFA biosynthesis.

plant biology↗

bioRxiv2024

Symplasmic phloem loading and sub-cellular transport in storage roots are key factors for carbon allocation in cassava

Cassava is a deciduous woody perennial shrub that stores large amounts of carbon and water in its storage roots. Previous studies have shown that assimilate unloading into storage roots happens symplasmically once secondary anatomy is established. However, mechanisms controlling phloem loading and overall carbon partitioning to different cassava tissues remain unclear. Here we used a combination of histological, transcriptional, and biochemical analyses on different cassava tissues and timepoints to better understand source-sink carbon allocation. We find that cassava likely utilizes a predominantly passive symplasmic phloem loading strategy, indicated by the lack of expression of genes coding for key players of sucrose transport, the existence of branched plasmodesmata in the companion cell/bundle sheath interface of minor leaf veins, and very high leaf sucrose concentrations. Furthermore, we show that tissue-specific changes in anatomy and NSC contents are associated with tissue-specific modification in gene expression for sucrose cleavage/synthesis, as well as subcellular compartmentalization of sugars. Overall, our data suggest that carbon allocation during storage root filling is mostly facilitated symplasmically, and is likely mostly regulated by local tissue demand and subcellular compartmentalization.

plant biology↗

bioRxiv2024

Environmentally-informed functional characterization of a plastid diatom metabolic bridge of mitochondrial origin

Withdrawal statementThe authors have withdrawn this manuscript due to a duplicate posting of manuscript number BIORXIV/2022/507166. Therefore, the authors do not wish this work to be cited as reference for the project. If you have any questions, please contact the corresponding author. The correct preprint can be found at doi: 10.1101/2022.09.08.507166

evolutionary biology↗

bioRxiv2024

Comparative analysis of the plasma metabolome of migrating passerines during stopover: Novel insights into flight metabolism

During long distance migration, many birds may experience periods of either prolonged fasting, during endurance flights, or extensive feeding during stopovers. It was previously shown that habitat selection during stopover can largely affect the migration outcome of an individual. Despite decades of research of the avian metabolism during stopover and migration, many questions have remained unanswered, as such research mainly focused on targeted metabolites and fat metabolism. Here, we examined the plasma-metabolome of migrating passerines prior to their crossing the Sahara Desert. Birds were sampled at two sites populated by Pistacia trees, bearing fat-rich fruits, and at an additional site dominated by blooming Eucalyptus trees. The blood samples were analyzed using both GC-MS and LC-MS, using an untargeted approach. We found that birds from one of the sites had a distinguish metabolic profile, suggesting recent landing. Examination of metabolic pathways activated during stopovers indicated a crucial role for cycling glucose through the Cori and Cahill cycles in resting and recovery processes. This novel perspective, conducted on free-ranging birds, suggests the evolution of avian insulin resistance due to factors such as endurance exercise, fasting, and a preference for fatty acid oxidation during migration, akin to cell trauma recovery. Additionally, we investigated inter-site variations in birds metabolic profiles. Significant variations were observed in both polar and lipophilic metabolites among the sites. Differences in polar metabolites were primarily attributed to variations in the physiological state of the birds between sites, while distinctions in the lipophilic profiles of rested birds were linked to variations in their primary food sources. This study underscores the challenge of interpreting commonly used indicators for assessing migrating birds physiological states and site quality, which are predominantly derived from lipid metabolism, in complex ecological systems.

ecology↗

bioRxiv2023

Natural variation in Arabidopsis thaliana identifies GlyoxalaseI;2 as key for the detoxification of glucose-derived reactive carbonyl species

Reactive carbonyl species (RCS) are toxic byproducts of normal metabolism that become more prevalent under oxidative stress. Here, we show that Arabidopsis thaliana ecotypes exhibit natural variation in their ability to detoxify glucose-derived RCS. We identified the IP-Pal-0 ecotype showing enhanced tolerance to glucose-derived RCS via upregulation of the glyoxalase system. In particular, the Viridiplantae-specific isoforms GLXI;2, GLXII;4, and GLXII;5 are highly expressed in IP-Pal-0 when plants are grown in the presence of 2-keto-D-glucose (KDG; glucosone) or methylglyoxal, and protein extracts from these plants display enhanced GLXI activity on KDG than other ecotypes. We identified specific motif/cis-regulatory elements in the GLXI;2 promoter regions of Col-0 and IP-Pal-0 that may underlie the differential expression of GLXI;2 associated with KDG detoxification. IP-Pal-0 GLXI;2 contains two different amino acids compared to Col-0, but these do not affect the basic kinetics of the protein. Interestingly, we found that the simultaneous change of these amino acids also occurs together in the GLXI proteins of some other organisms, suggesting a convergence in the simultaneous change of both amino acid residues. Our findings underscore the importance of Viridiplantae-specific glyoxalase isoforms in detoxifying glucose-derived RCS, particularly KDG, and highlight the promise of harnessing natural genetic diversity in the glyoxalase pathway to enhance plant stress tolerance.

plant biology↗

bioRxiv2023

Carbon usage in yellow-fleshed Manihot esculenta storage roots shifts from starch biosynthesis to cell wall and raffinose biosynthesis via the myo-inositol pathway

Cassava is a crucial staple crop for smallholder farmers in tropical Asia and Sub-Saharan Africa. Although high yield remains the top priority for farmers, the significance of nutritional values has increased in cassava breeding programs. A notable negative correlation between provitamin A and starch accumulation poses a significant challenge for breeding efforts. The negative correlation between starch and carotenoid levels in conventional and genetically modified cassava plants implies the absence of a direct genomic connection between the two traits. The competition among various carbon pathways seems to account for this relationship. In this study, we conducted a thorough analysis of 49 African cassava genotypes with varying levels of starch and provitamin A. Our goal was to identify factors contributing to differential starch accumulation. With the carotenoid levels of the varieties considered as a confounding effect on starch production, we found that yellow and white-fleshed storage roots did not differ significantly in most measured components of starch or de novo fatty acid biosynthesis. However, genes and metabolites associated with myo-inositol synthesis and cell wall component production were substantially enriched in high provitamin A genotypes. These results indicate that yellow-fleshed cultivars, in comparison to their white-fleshed counterparts, direct more carbon towards the synthesis of raffinose and cell wall components, a finding that is supported by a significant rise in the starch-free residue to total dry yield ratio in yellow storage roots versus white storage roots. Our findings enhance comprehension of the biosynthesis of starch and carotenoids in the storage roots of cassava.

plant biology↗

bioRxiv2023

Characterization of tomato canal-1 mutant using a multi-omics approach

The recently described canal-1 tomato mutant, which has a variegated leaf phenotype, has been shown to affect canalization of yield. The corresponding protein is orthologous to AtSCO2 - SNOWY COTYLEDON2, which has suggested roles in thylakoid biogenesis. Here we characterize the canal-1 mutant through a multi-omics approach, by comparing mutant to wild-type tissues. While white canal-1 leaves are devoid of chlorophyll, green leaves of the mutant appear wild-type-like, despite an impaired protein function. Transcriptomic data suggest that green mutant leaves compensate for this impaired protein function by upregulation of transcription of photosystem assembly and photosystem component genes, thereby allowing adequate photosystem establishment, which is reflected in their wild-type-like proteome. White canal-1 leaves, however, likely fail to reach a certain threshold enabling this overcompensation, and plastids get trapped in an undeveloped state, while additionally suffering from high light stress, indicated by the overexpression of ELIP homolog genes. The metabolic profile of white and to a lesser degree also green tissues revealed upregulation of amino acid levels, that was at least partially mediated by transcriptional and proteomic upregulation. These combined changes are indicative of a stress response and suggest that white tissues behave as carbon sinks. In summary, our work demonstrates the relevance of the SCO2 protein in both photosystem assembly and as a consequence in the canalization of yield. Significance statementThe variegated canalized-1 tomato mutant was recently described and the underlying gene SCO2 suggested to be a yield canalization gene. Through a multi-omics approach we show that mutants require a transcriptional upregulation of photosystem components and assembly components, likely as overcompensation for partially impaired SCO2 function, to produce a wild type-like proteome and functional photosynthetic tissue Our data, furthermore, suggest that variation of green to white leaf area from plant to plant leads to the yield variation.

plant biology↗

bioRxiv2023

Degradation of Fatty Acid Export Protein1 by Rhomboid-Like Protease11 Contributes to Cold Tolerance in Arabidopsis

Plants need to adapt to different stresses to optimize growth under unfavorable conditions. The abundance of the chloroplast envelope located Fatty Acid Export Protein1 (FAX1) decreases after the onset of low temperatures. However, it was unclear how FAX1 degradation occurs and whether altered FAX1 abundance contributes to cold tolerance in plants. The rapid cold-induced increase in rhomboid-like protease11 (RBL11) transcript, the physical interaction of RBL11 with FAX1, the specific FAX1 degradation after RBL11 expression, and the absence of cold-induced FAX1 degradation in rbl11 loss-of-function mutants suggest that this enzyme is responsible for FAX1 degradation. Proteomic analyses showed that rbl11 mutants have higher levels of FAX1 and other proteins involved in membrane lipid homeostasis, suggesting that RBL11 is a key element in the remodeling of membrane properties during cold. Consequently, in the cold, rbl11 mutants show a shift in lipid biosynthesis towards the eukaryotic pathway, which coincides with impaired cold tolerance. To demonstrate that cold sensitivity is due to increased FAX1 levels, FAX1 overexpressors were analyzed. rbl11 and FAX1 overexpressor mutants show superimposable phenotypic defects upon exposure to cold temperatures. Our results show that the cold-induced degradation of FAX1 by RBL11 is critical for Arabidopsis to survive cold and freezing periods. One sentence summaryDegradation of the inner envelope protein Fatty Acid Export1 via Rhomboid Like Protease11 represents a critical process to achieve cold and frost tolerance in Arabidopsis

plant biology↗

bioRxiv2023

The genetic architecture of the pepper metabolome provides insights into the regulation of capsianoside biosynthesis

Capsicum (pepper) is among the most economically important species worldwide, the fruit accumulates specialized metabolites with essential roles in plant environmental interaction and potential health benefits. However, the underlying genetic basis of their biosynthesis remains largely unknown. In this study, we developed and assessed both wild genetic variance and a bespoke mapping population to determine the genetic architecture of the pepper metabolome. The genetic analysis provided over 30 metabolic quantitative trait loci (mQTL) for over 1100 metabolites. We identified 92 candidate genes involved in various mQTL. Among the identified loci, we described and validated by transient overexpression a domestication gene cluster of eleven UDP-glycosyltransferases involved in monomeric capsianoside biosynthesis. We additionally constructed the biosynthetic reactions and annotated the genes involved in capsianoside biosynthesis in pepper. Given that differential glycosylation of acyclic diterpenoid glycosides contributes to plant resistance and acts as anticancer agents in humans, our data provide new insight, and resources for better understanding the biosynthesis of beneficial natural compounds to improve human health.

genetics↗

bioRxiv2023

FCS-Like Zinc finger 14 (FLZ14) mediates the crosstalk between TORC and SnRK1 in response to sugar availability.

Matching resource availability to growth is crucial for plant fitness. We identified that FCS- Like Zinc finger 14 (FLZ14) gauges carbon (C) status to adjust growth by inhibiting the TARGET OF RAPAMYCIN (TOR) activity in short days via REGULATORY- ASSOCIATED PROTEIN OF TARGET OF RAPAMYCIN 1B (RAPTOR1B). Besides being transcriptionally induced by sugars, the amplitude of FLZ14 induction correlates to the diel sugar status, indicating that FLZ14 responses to sugar oscillations in planta. Genetic evidence elucidated that FLZ14 is involved in the crosstalk between TORC and SUCROSE NON-FERMENTING RELATED KINASE 1 (SnRK1). In short photoperiods, FLZ14 and KIN10 genetically interact to set the pace of TORC activity. By contrast, under long day photoperiods, FLZ14 and RAPTOR1B genetically interact, most likely to downregulate SnRK1 signalling. This report highlights the function of a sugar-responsive gene in mediating the crosstalk between TORC and SnRK1 to fine-tune growth.

plant biology↗

bioRxiv2023

PLANT UNCOUPLING MITOCHONDRIAL PROTEIN 2 localizes to the Golgi

Mitochondria act as cellular hubs of energy transformation and metabolite conversion in most eukaryotes. Plant mitochondrial electron transport chains are particularly flexible, featuring alternative components, such as ALTERNATIVE NAD(P)H DEHYDROGENASES and ALTERNATIVE OXIDASES (AOXs), that can bypass proton translocation steps. PLANT UNCOUPLING MITOCHONDRIAL PROTEINS (named PUMPs or plant UCPs) have been identified in plants as homologues of mammalian Uncoupling Proteins (UCPs), and their biochemical and physiological roles have been investigated in the context of mitochondrial energy metabolism. To dissect UCP function in Arabidopsis, the two most conserved (UCP1 and UCP2) have been targeted in recent work by combining mutant lines to circumvent potential functional redundancy in vivo. Such approaches rely on the assumption that both proteins reside in the inner mitochondrial membrane as a prerequisite for functional redundancy. Yet, contradicting results have been reported on UCP2 localization in plants. Here we provide evidence that, conversely to UCP1, which is an abundant inner mitochondrial membrane protein, UCP2 localizes to the Golgi rather than to mitochondria. Based on multiple lines of new and prior evidence, we summarize the consensus view that we have reached and provide an example of how open, critical exchange within the research community is able to constructively address ambiguities. Our observations and considerations provide direction to the ongoing discussion about the functions of UCP proteins. They further offer new perspectives for the study of Golgi membrane transport and subcellular targeting principles of membrane proteins. Since 20 to 30 % of genes in plant genomes are predicted to encode transmembrane proteins and the function of most of those proteins has not been experimentally investigated, we highlight the importance of using independent evidence for localization as a prerequisite for understanding physiological function of membrane proteins.

plant biology↗

bioRxiv2023

MetPlast: an R package to evaluate Metabolic Plasticity using Information Theory statistical framework

MetPlast, is an R package that provides the functionalities necessary to analyze biological samples metabolic plasticity. This package takes advantage of the statistical framework provided by the Information Theory to quantify and defined metabolic plasticity parameters. Using previous implemented formula based on Shannon entropy we automatize the calculation and visualization of a set of metabolic plasticity indexes including metabolic diversity, metabolome specialization, and metabolite specialization. We use a publicly available metabolic data set on tomato domestication to demonstrate the power of the present tool to evaluate changes in metabolic plasticity parameters. Thus, MetPlast represents a new and invaluable member of the computational metabolomics toolbox, that will certainly help scientists to unveil hidden information in cell metabolomic landscape. Availability and implementationFreely available at https://github.com/danlucio86/MetPlast

bioinformatics↗

bioRxiv2023

The genetic and physiological basis of Arabidopsis thaliana tolerance to Pseudomonas viridiflava

O_LIThe opportunistic pathogen Pseudomonas viridiflava colonizes more than fifty agricultural crop species and is the most common Pseudomonas in the phyllosphere of European Arabidopsis thaliana populations. Belonging to the P. syringae complex, it is genetically and phenotypically distinct from well-characterized P. syringae sensu stricto. Despite its prevalence, we lack knowledge of how A. thaliana responds to its native isolates at the molecular level. Here, we characterize the host response in an A. thaliana - P. viridiflava pathosystem. C_LIO_LIWe measured host and pathogen growth in axenic infections, and used immune mutants, transcriptomics, and metabolomics to determine defense pathways influencing susceptibility to P. viridiflava infection. C_LIO_LIInfection with P. viridiflava increased jasmonic acid (JA) levels and the expression of ethylene defense pathway marker genes. The immune response in a susceptible host accession was delayed compared to a tolerant one. Mechanical injury rescued susceptibility, consistent with an involvement of JA. C_LIO_LIThe JA/ethylene pathway is important for suppression of P. viridiflava, yet suppression capacity varies between accessions. Our results shed light on how A. thaliana can suppress the ever-present P. viridiflava, but further studies are needed to understand how P. viridiflava evades this suppression to spread broadly across A. thaliana populations. C_LI

plant biology↗