Brassica rapa L. subsp. sylvestris L. Janch. var. esculenta Hort., commonly known as Friariello Napoletano, is a traditional Italian landrace valued for its distinctive flavor, nutritional richness, and cultural relevance in Mediterranean cuisine. The present study investigates the biochemical changes during postharvest storage at two temperatures (4 °C and 10 °C) for 2 and 20 days in its inflorescences and leaves. The experiment aimed to evaluate the evolution of primary and secondary metabolites, with a focus on pigments, amino acids, antioxidants, and glucosinolates. Significant degradation of chlorophylls was observed, particularly in leaves, with reductions of over 90% after 20 days at both temperatures. Conversely, α-tocopherol content increased significantly, especially in inflorescences, indicating an antioxidant response to storage stress. Amino acid analysis revealed a sharp decline in glutamate (up to 79%) and glutamine (up to 83%) in leaves, while proline levels increased across both tissues, reflecting an osmoprotective response. Essential amino acids (EAAs) showed variable responses, with certain EAAs, such as histidine and phenylalanine, accumulating under specific storage conditions. Soluble sugars, starch, and glucosinolates also decreased significantly, with soluble sugars dropping by 87% in inflorescences and 90% in leaves after 20 days at 10 °C. Pathway analysis revealed distinct tissue-specific metabolic responses, with inflorescences exhibiting more stable antioxidant levels and greater resilience to oxidative stress compared to leaves. These findings provide insights into the metabolic adjustments during postharvest senescence and may support future strategies aimed at preserving shelf life and nutritional quality of this traditional Mediterranean vegetable.

The shoot apical meristem (SAM) is responsible for overall shoot growth by generating all aboveground structures. Recent research has revealed that the SAM displays an autonomous heat stress (HS) memory of a previous non -lethal HS event. Considering the importance of the SAM for plant growth, it is essential to determine how its thermomemory is mechanistically controlled. Here, we report that HEAT SHOCK TRANSCRIPTION FACTOR A7b (HSFA7b) plays a crucial role in this process in Arabidopsis, as the absence of functional HSFA7b results in the temporal suppression of SAM activity after thermopriming. We found that HSFA7b directly regulates ethylene response at the SAM by binding to the promoter of the key ethylene signaling gene ETHYLENE -INSENSITIVE 3 to establish thermotolerance. Moreover, we demonstrated that HSFA7b regulates the expression of ETHYLENE OVERPRODUCER 1 (ETO1) and ETO1-LIKE 1, both of which encode ethylene biosynthesis repressors, thereby ensuring ethylene homeostasis at the SAM. Taken together, these results reveal a crucial and tissue -specific role for HSFA7b in thermomemory at the Arabidopsis SAM.

Although several large-scale single-cell RNA sequencing (scRNAseq) studies addressing the root of Arabidopsis (Arabidopsis thaliana) have been published, there is still need for a de novo reference map for both root and especially above-ground cell types. As the plants' transcriptome substantially changes throughout the day, shaped by the circadian clock, we performed scRNAseq on both Arabidopsis root and above-ground tissues at defined times of the day. For the root scRNAseq analysis, we used tissue-specific reporter lines grown on plates and harvested at the end of the day (ED). In addition, we submitted above-ground tissues from plants grown on soil at ED and end of the night to scRNAseq, which allowed us to identify common cell types/markers between root and shoot and uncover transcriptome changes to above-ground tissues depending on the time of the day. The dataset was also exploited beyond the traditional scRNAseq analysis to investigate non-annotated and di-cistronic transcripts. We experimentally confirmed the predicted presence of some of these transcripts and also addressed the potential function of a previously unidentified marker gene for dividing cells. In summary, this work provides insights into the spatial control of gene expression from nearly 70,000 cells of Arabidopsis for below- and whole above-ground tissue at single-cell resolution at defined time points.

EDITORIAL article Front. Plant Sci., 31 August 2022Sec. Plant Metabolism and Chemodiversity https://doi.org/10.3389/fpls.2022.1011224

De novo fatty acid biosynthesis in plants relies on a prokaryotic-type acetyl-CoA carboxylase (ACCase) that resides in the plastid compartment. The enzyme is composed of four subunits, one of which is encoded in the plastid genome, whereas the other three subunits are encoded by nuclear genes. The plastid gene (accD) encodes the β-carboxyltransferase subunit of ACCase and is essential for cell viability. To facilitate the functional analysis of accD, we pursued a transplastomic knockdown strategy in tobacco (Nicotiana tabacum). By introducing point mutations into the translational start codon of accD, we obtained stable transplastomic lines with altered ACCase activity. Replacement of the standard initiator codon AUG with UUG strongly reduced AccD expression, whereas replacement with GUG had no detectable effects. AccD knockdown mutants displayed reduced ACCase activity, which resulted in changes in the levels of many but not all species of cellular lipids. Limiting fatty acid availability caused a wide range of macroscopic, microscopic, and biochemical phenotypes, including impaired chloroplast division, reduced seed set, and altered storage metabolism. Finally, while the mutants displayed reduced growth under photoautotrophic conditions, they showed exaggerated growth under heterotrophic conditions, thus uncovering an unexpected antagonistic role of AccD activity in autotrophic and heterotrophic growth.

Phytochrome photoreceptors are known to regulate plastic growth responses to vegetation shade. However, recent reports also suggest an important role for phytochromes in carbon resource management, metabolism, and growth. Here, we use 13CO2 labelling patterns in multiallele phy mutants to investigate the role of phytochrome in the control of metabolic fluxes. We also combine quantitative data of 13C incorporation into protein and cell wall polymers, gas exchange measurements, and system modelling to investigate why biomass is decreased in adult multiallele phy mutants. Phytochrome influences the synthesis of stress metabolites such as raffinose and proline, and the accumulation of sugars, possibly through regulating vacuolar sugar transport. Remarkably, despite their modified metabolism and vastly altered architecture, growth rates in adult phy mutants resemble those of wild-type plants. Our results point to delayed seedling growth and smaller cotyledon size as the cause of the adult-stage phy mutant biomass defect. Our data signify a role for phytochrome in metabolic stress physiology and carbon partitioning, and illustrate that phytochrome action at the seedling stage sets the trajectory for adult biomass production.

Trehalose 6-phosphate (Tre6P) is a sucrose signalling metabolite that has been implicated in regulation of shoot branching, but its precise role is not understood. We expressed tagged forms of TREHALOSE-6-PHOSPHATE SYNTHASE1 (TPS1) to determine where Tre6P is synthesized in arabidopsis (Arabidopsis thaliana), and investigated the impact of localized changes in Tre6P levels, in axillary buds or vascular tissues, on shoot branching in wild-type and branching mutant backgrounds. TPS1 is expressed in axillary buds and the subtending vasculature, as well as in the leaf and stem vasculature. Expression of a heterologous Tre6P phosphatase (TPP) to lower Tre6P in axillary buds strongly delayed bud outgrowth in long days and inhibited branching in short days. TPP expression in the vasculature also delayed lateral bud outgrowth and decreased branching. Increased Tre6P in the vasculature enhanced branching and was accompanied by higher expression of FLOWERING LOCUS T (FT) and upregulation of sucrose transporters. Increased vascular Tre6P levels enhanced branching in branched1 but not in ft mutant backgrounds. These results provide direct genetic evidence of a local role for Tre6P in regulation of axillary bud outgrowth within the buds themselves, and also connect Tre6P with systemic regulation of shoot branching via FT.

In plants, the shoot apical meristem (SAM) is essential for the growth of aboveground organs. However, little is known about its molecular responses to abiotic stresses. Here, we show that the SAM of Arabidopsis thaliana displays an autonomous heat-stress (HS) memory of a previous non-lethal HS, allowing the SAM to regain growth after exposure to an otherwise lethal HS several days later. Using RNA sequencing, we identified genes participating in establishing the SAM's HS transcriptional memory, including the stem cell (SC) regulators CLAVATA1 (CLV1) and CLV3, HEAT SHOCK PROTEIN 17.6A (HSP17.6A), and the primary carbohydrate metabolism gene FRUCTOSE-BISPHOSPHATE ALDOLASE 6 (FBA6). We demonstrate that sugar availability is essential for survival of plants at high temperature. HEAT SHOCK TRANSCRIPTION FACTOR A2 (HSFA2A) directly regulates the expression of HSP17.6A and FBA6 by binding to the heat-shock elements in their promoters, indicating that HSFA2 is required for transcriptional activation of SAM memory genes. Collectively, these findings indicate that plants have evolved a sophisticated protection mechanism to maintain SCs and, hence, their capacity to re-initiate shoot growth after stress release.

The even chain length and regular spacing of these clusters allows amylopectin to form a semi-crystalline structure, with regular layers within the starch granule. The even chain length and regular spacing of these clusters allows amylopectin to form a semi-crystalline structure, with regular layers within the starch granule. Net carbon fixation allows phosphorylated intermediates to be withdrawn from the Calvin-Benson cycle for synthesis of various products, of which sucrose and starch are usually the most abundant. Sucrose is synthesized in the cytosol using triose-phosphates exported from the chloroplast via the triose-phosphate translocator, while starch is made in the chloroplasts. Starch can be degraded hydrolytically by endo- and exo-amylases to produce malto-oligosaccharides of various sizes, or phosphorolytically by starch phosphorylase. Several additional enzymes are required for the complete degradation of glucan chains.

SUMMARY - Trehalose 6-phosphate (Tre6P) is a sucrose signalling metabolite that has been implicated in regulation of shoot branching, but its precise role is not understood. - We expressed tagged forms of TREHALOSE-6-PHOSPHATE SYNTHASE1 (TPS1) to determine where Tre6P is synthesized in arabidopsis ( Arabidopsis thaliana ), and investigated the impact of localized changes in Tre6P levels, in axillary buds or vascular tissues, on shoot branching in wild-type and branching mutant backgrounds. - TPS1 is expressed in axillary buds and the subtending vasculature, as well as in the leaf and stem vasculature. Expression of a heterologous trehalose-6-phosphate phosphatase (TPP) to lower Tre6P in axillary buds strongly delayed bud outgrowth in long days and inhibited branching in short days. TPP expression in the vasculature also delayed lateral bud outgrowth and decreased branching. Increased Tre6P in the vasculature enhanced branching and was accompanied by higher expression of FLOWERING LOCUS T ( FT) and up-regulation of sucrose transporters. Increased vascular Tre6P levels enhanced branching in branched1 but not in ft mutant backgrounds. - These results provide direct genetic evidence of a local role for Tre6P in regulation of axillary bud outgrowth within the buds themselves, and also connect Tre6P with systemic regulation of shoot branching via FT.

In flowering plants, sugars act as carbon sources providing energy for developing embryos and seeds. Although most studies focus on carbon metabolism in whole seeds, knowledge about how particular sugars contribute to the developmental transitions during embryogenesis is scarce. To develop a quantitative understanding of how carbon composition changes during embryo development, and to determine how sugar status contributes to final seed or embryo size, we performed metabolic profiling of hand-dissected embryos at late torpedo and mature stages, and dormant seeds, in two Arabidopsis thaliana accessions with medium [Columbia-0 (Col-0)] and large [Burren-0 (Bur-0)] seed sizes, respectively. Our results show that, in both accessions, metabolite profiles of embryos largely differ from those of dormant seeds. We found that developmental transitions from torpedo to mature embryos, and further to dormant seeds, are associated with major metabolic switches in carbon reserve accumulation. While glucose, sucrose, and starch predominantly accumulated during seed dormancy, fructose levels were strongly elevated in mature embryos. Interestingly, Bur-0 seeds contain larger mature embryos than Col-0 seeds. Fructose and starch were accumulated to significantly higher levels in mature Bur-0 than Col-0 embryos, suggesting that they contribute to the enlarged mature Bur-0 embryos. Furthermore, we found that Bur-0 embryos accumulated a higher level of sucrose compared to hexose sugars and that changes in sucrose metabolism are mediated by sucrose synthase (SUS), with SUS genes acting non-redundantly, and in a tissue-specific manner to utilize sucrose during late embryogenesis.

Several halophytes and a few crop plants, including Poaceae, synthesize and accumulate glycine betaine (GB) in response to environmental constraints. GB plays an important role in osmoregulation, in fact, it is one of the main nitrogen-containing compatible osmolytes found in Poaceae. It can interplay with molecules and structures, preserving the activity of macromolecules, maintaining the integrity of membranes against stresses and scavenging ROS. Exogenous GB applications have been proven to induce the expression of genes involved in oxidative stress responses, with a restriction of ROS accumulation and lipid peroxidation in cultured tobacco cells under drought and salinity, and even stabilizing photosynthetic structures under stress. In the plant kingdom, GB is synthesized from choline by a two-step oxidation reaction. The first oxidation is catalyzed by choline monooxygenase (CMO) and the second oxidation is catalyzed by NAD+-dependent betaine aldehyde dehydrogenase. Moreover, in plants, the cytosolic enzyme, named N-methyltransferase, catalyzes the conversion of phosphoethanolamine to phosphocholine. However, changes in CMO expression genes under abiotic stresses have been observed. GB accumulation is ontogenetically controlled since it happens in young tissues during prolonged stress, while its degradation is generally not significant in plants. This ability of plants to accumulate high levels of GB in young tissues under abiotic stress, is independent of nitrogen (N) availability and supports the view that plant N allocation is dictated primarily to supply and protect the growing tissues, even under N limitation. Indeed, the contribution of GB to osmotic adjustment and ionic and oxidative stress defense in young tissues, is much higher than that in older ones. In this review, the biosynthesis and accumulation of GB in plants, under several abiotic stresses, were analyzed focusing on all possible roles this metabolite can play, particularly in young tissues.

The phytochromes (phys) photoreceptors are known to be major regulators of plastic growth responses to vegetation shade. Recent reports have begun to uncover an important role for phys in carbon resource management. Our earlier work showed that phy mutants had a distinct metabolic profile with elevated levels of metabolites including TCA intermediates, amino acids and sugars. Here we show that in seedlings phy regulates the balance between glucose and starch. Multi-allele phy mutants have excess glucose and low starch levels, which is conducive to hypocotyl elongation. 13C-CO2 labelling demonstrates that metabolic flux balance in adult plants is markedly altered in phy mutants. Phytochrome reduces synthesis rates of stress metabolites, including raffinose and proline and several typical stress-induced biosynthetic genes related to these metabolites show higher expression in phy mutants. Since growth and metabolism are typically inter-connected, we investigated why phy mutants have severely reduced biomass. Quantification of carbon fixation, biomass accumulation, and 13C labelling of cell wall polysaccharides established that relative growth rate is impaired in multi allele phy mutants for the first 2.5 weeks after germination but equivalent to the WT thereafter. Mathematical modelling predicts that the altered growth dynamics and final biomass deficit can be explained by the smaller cotyledon size of the multiple phy mutants. This indicates that the established role of phy in promoting seedling establishment has enduring effects that govern adult plant biomass.

Several halophytes and a few crop plants, including Poaceae, synthesize and accumulate glycine betaine (GB) in response to environmental constraints. GB plays an important role in osmoregulation, in fact, it is one of the main nitrogen-containing compatible osmolytes found in Poaceae. It can interplay with molecules and structures, preserving the activity of macromolecules, maintaining the integrity of membranes against stresses and scavenging ROS. Exogenous GB applications have been proven to induce the expression of genes involved in oxidative stress responses, with a restriction of ROS accumulation and lipid peroxidation in cultured tobacco cells under drought and salinity, and even stabilizing photosynthetic structures under stress. In the plant kingdom, GB is synthesized from choline by a two-step oxidation reaction. The first oxidation is catalyzed by choline monooxygenase (CMO) and the second oxidation is catalyzed by NAD+-dependent betaine aldehyde dehydrogenase. Moreover, in plants, the cytosolic enzyme, named N-methyltransferase, catalyzes the conversion of phosphoethanolamine to phosphocholine. However, changes in CMO expression genes under abiotic stresses have been observed. GB accumulation is ontogenetically controlled since it happens in young tissues during prolonged stress, while its degradation is generally not significant in plants. This ability of plants to accumulate high levels of GB in young tissues under abiotic stress, is independent of nitrogen (N) availability and supports the view that plant N allocation is dictated primarily to supply and protect the growing tissues, even under N limitation. Indeed, the contribution of GB to osmotic adjustment and ionic and oxidative stress defense in young tissues, is much higher than that in older ones. In this review, the biosynthesis and accumulation of GB in plants, under several abiotic stresses, were analyzed focusing on all possible roles this metabolite can play, particularly in young tissues.

Rubisco is the central enzyme of photosynthetic carbon assimilation in the Calvin-Benson-Bassham (CBB) cycle. It catalyzes the carboxylation of one molecule of ribulose-1,5-bisphosphate (RuBP) producing two molecules of 3-phosphoglycerate, from which sugars, starch, amino acids, and fatty acids are