Physiological and metabolic changes in response to Boron levels are mediated by ethylene affecting tomato fruit yield.

Boron (B) is an essential nutrient for the plant, and its stress (both deficiency and toxicity) are major problems that affect crop production. Ethylene metabolism (both signaling and production) is important to plants' differently responding to nutrient availability. To better understand the connections between B and ethylene, here we investigate the function of ethylene in the responses of tomato (Solanum lycopersicum) plants to B stress (deficiency, 0 µM and toxicity, 640 µM), using ethylene related mutants, namely nonripening (nor), ripening-inhibitor (rin), never ripe (Nr), and epinastic (Epi). Our results show that B stress does not necessarily inhibit plant growth, but both B stress and ethylene signaling severely affected physiological parameters, such as photosynthesis, stomatal conductance, and chlorophyll a fluorescence. Under B toxicity, visible symptoms of toxicity appeared in the roots and margins of the older leaves through necrosis, caused by the accumulation of B which stimulated ethylene biosynthesis in the shoots. Both nor and rin (ethylene signaling) mutants presented similar responses, being these genotypes more sensitive and displaying several morphophysiological alterations, including fruit productivity reductions, in response to the B toxicity conditions. Therefore, our results suggest that physiological and metabolic changes in response to B fluctuations are likely mediated by ethylene signaling.

Regulatory mechanisms behind the phenotypic plasticity associated with Setaria italica water deficit tolerance.

Drought is one of the main environmental stresses that negatively impacts vegetative and reproductive yield. Water deficit responses are determined by the duration and intensity of the stress, which, together with plant genotype, will define the chances of plant survival. The metabolic adjustments in response to water deficit are complex and involve gene expression modulation regulated by DNA-binding proteins and epigenetic modifications. This last mechanism may also regulate the activity of transposable elements, which in turn impact the expression of nearby loci. Setaria italica plants submitted to five water deficit regimes were analyzed through a phenotypical approach, including growth, physiological, RNA-seq and sRNA-seq analyses. The results showed a progressive reduction in yield as a function of water deficit intensity associated with signaling pathway modulation and metabolic adjustments. We identified a group of loci that were consistently associated with drought responses, some of which were related to water deficit perception, signaling and regulation. Finally, an analysis of the transcriptome and sRNAome allowed us to identify genes putatively regulated by TE- and sRNA-related mechanisms and an intriguing positive correlation between transcript levels and sRNA accumulation in gene body regions. These findings shed light on the processes that allow S. italica to overcome drought and survive under water restrictive conditions.

Predictive metabolomics of multiple Atacama plant species unveils a core set of generic metabolites for extreme climate resilience.

Current crop yield of the best ideotypes is stagnating and threatened by climate change. In this scenario, understanding wild plant adaptations in extreme ecosystems offers an opportunity to learn about new mechanisms for resilience. Previous studies have shown species specificity for metabolites involved in plant adaptation to harsh environments. Here, we combined multispecies ecological metabolomics and machine learning-based generalized linear model predictions to link the metabolome to the plant environment in a set of 24 species belonging to 14 families growing along an altitudinal gradient in the Atacama Desert. Thirty-nine common compounds predicted the plant environment with 79% accuracy, thus establishing the plant metabolome as an excellent integrative predictor of environmental fluctuations. These metabolites were independent of the species and validated both statistically and biologically using an independent dataset from a different sampling year. Thereafter, using multiblock predictive regressions, metabolites were linked to climatic and edaphic stressors such as freezing temperature, water deficit and high solar irradiance. These findings indicate that plants from different evolutionary trajectories use a generic metabolic toolkit to face extreme environments. These core metabolites, also present in agronomic species, provide a unique metabolic goldmine for improving crop performances under abiotic pressure.

Physiological and metabolic alterations in basil (Ocimum basilicum L) varieties under distinct soil water levels / Alteraciones fisiológicas y metabólicas en variedades de albahaca (Ocimum basilicum L) bajo distintos niveles de agua del suelo

Basil (Ocimum basilicumL.) is a medicinal species used in several areas, such as food, medicines and cosmetics, and the understanding of its physiological behavior under environmental conditions is of paramount importance for the improvement of cultivation methods. The objective of this study was to evaluate the influence of different water availability under physiological, biochemical and metabolic characteristics, in three distinct genotypes: 'Alfavaca basilicão', 'Gennaro de menta' and 'Grecco à palla', during two different phenological stages (vegetative and reproductive). It was found that the water deficit promotes physiological changes to tolerate water stress, and the studied genotypes have different routes to achieve this physiological tolerance, which culminates in a distinct accumulation of metabolites in plants, and can be considered interesting if the final product is the production of essential oils.

La albahaca (Ocimum basilicum L.) es una planta medicinal utilizada en varias áreas: alimenticia, medicinal e industria cosmética; es de suma importancia el entendimiento de su comportamiento fisiológico bajo diferentes condiciones ambientales con el fin de mejorar los procesos del cultivo. El objetivo de este estudio fue evaluar la influencia de diferentes disponibilidades hídricas en las características fisiológicas, bioquímicas y metabólicas en tres genotipos de albahaca: "Alfavaca basilicão", "Gennaro de menta" y "Grecco à palla" durante dos etapas fenológicas (vegetativa y reproductiva). Fue encontrado que el déficit hídrico promueve cambios fisiológicos con el fin de tolerar el estrés hídrico. Los genotipos estudiados presentaron diferentes rutas para alcanzar esta tolerancia fisiológica, la cual culmina con distintas acumulaciones de metabolitos en las plantas, y puede ser considerado interesante si el producto final es la producción de aceites esenciales.

Chemophenetic Significance of Anomalocalyx uleanus Metabolites are Revealed by Dereplication Using Molecular Networking Tools.

Anomalocalyx uleanus (Pax & K. Hoffm.) Ducke (Euphorbiaceae) is a singular species in the genus and is restricted and exclusive to the Brazilian Amazon. A phytochemical study of A. uleanus leaves was performed, yielding the isolation of five major compounds: catechin/epicatechin, afzelin, quercetin 3-O-α-L-rhamnopyranoside, and astilbin. The phytochemical compositions of the methanolic extracts of leaves, roots, bark, and stem bark were determined using a dereplication approach. Forty-six compounds were annotated from the liquid chromatography-mass spectrometry (LC-MS/MS) data, while four lipids were identified using gas chromatography-mass spectrometry (GC-MS). In total, fifty compounds were detected, and they belonged to the primary metabolism and several classes of natural products such as flavonoids, flavonoids O-glycosides, flavonoids C-glycosides, biflavonoids, procyanidin, triterpene, triterpenes esterified with phenylpropanoids, phenylpropanoid derivatives, flavonolignans, coumarins, quinic acid derivatives, and benzoic acid derivatives. This is the first report on the phytochemical data of the genus Anomalocalyx, and the results of this study will contribute to the chemosystematic knowledge of the Euphorbiaceae family and justify the need for investigation of the pharmacological potential of the species A. uleanus.

Characterization of In Vivo Function(s) of Members of the Plant Mitochondrial Carrier Family.

Although structurally related, mitochondrial carrier family (MCF) proteins catalyze the specific transport of a range of diverse substrates including nucleotides, amino acids, dicarboxylates, tricarboxylates, cofactors, vitamins, phosphate and H+. Despite their name, they do not, however, always localize to the mitochondria, with plasma membrane, peroxisomal, chloroplast and thylakoid and endoplasmic reticulum localizations also being reported. The existence of plastid-specific MCF proteins is suggestive that the evolution of these proteins occurred after the separation of the green lineage. That said, plant-specific MCF proteins are not all plastid-localized, with members also situated at the endoplasmic reticulum and plasma membrane. While by no means yet comprehensive, the in vivo function of a wide range of these transporters is carried out here, and we discuss the employment of genetic variants of the MCF as a means to provide insight into their in vivo function complementary to that obtained from studies following their reconstitution into liposomes.

NADP-Dependent Malic Enzyme 1 Participates in the Abscisic Acid Response in Arabidopsis thaliana.

Arabidopsis thaliana possesses three cytosolic (NADP-ME1-3) and one plastidic (NADP-ME4) NADP-dependent malic enzymes. NADP-ME2 and -ME4 show constitutive expression, in contrast to NADP-ME1 and -ME3, which are restricted to particular tissues. Here, we show that NADP-ME1 transcript and protein were almost undetectable during normal vegetative growth, but gradually increased and reached levels higher than those of the other isoforms in the latest stages of seed development. Accordingly, in knockout nadp-me1 mature seeds the total NADP-ME activity was significantly lower than in wild type mature seeds. The phenotypic analysis of nadp-me1 plants indicated alterations of seed viability and germination. Besides, the treatment with abscisic acid (ABA), NaCl and mannitol specifically induced the accumulation of NADP-ME1 in seedlings. In line with this, nadp-me1 plants show a weaker response of primary and lateral root length and stomatal opening to the presence of ABA. The results suggest that NADP-ME1 plays a specialized role, linked to ABA signaling during the seed development as well as in the response to water deficit stress.

Structural disorder in plant proteins: where plasticity meets sessility.

Plants are sessile organisms. This intriguing nature provokes the question of how they survive despite the continual perturbations caused by their constantly changing environment. The large amount of knowledge accumulated to date demonstrates the fascinating dynamic and plastic mechanisms, which underpin the diverse strategies selected in plants in response to the fluctuating environment. This phenotypic plasticity requires an efficient integration of external cues to their growth and developmental programs that can only be achieved through the dynamic and interactive coordination of various signaling networks. Given the versatility of intrinsic structural disorder within proteins, this feature appears as one of the leading characters of such complex functional circuits, critical for plant adaptation and survival in their wild habitats. In this review, we present information of those intrinsically disordered proteins (IDPs) from plants for which their high level of predicted structural disorder has been correlated with a particular function, or where there is experimental evidence linking this structural feature with its protein function. Using examples of plant IDPs involved in the control of cell cycle, metabolism, hormonal signaling and regulation of gene expression, development and responses to stress, we demonstrate the critical importance of IDPs throughout the life of the plant.

Specific Arabidopsis thaliana malic enzyme isoforms can provide anaplerotic pyruvate carboxylation function in Saccharomyces cerevisiae.

NAD(P)-malic enzyme (NAD(P)-ME) catalyzes the reversible oxidative decarboxylation of malate to pyruvate, CO2 , and NAD(P)H and is present as a multigene family in Arabidopsis thaliana. The carboxylation reaction catalyzed by purified recombinant Arabidopsis NADP-ME proteins is faster than those reported for other animal or plant isoforms. In contrast, no carboxylation activity could be detected in vitro for the NAD-dependent counterparts. In order to further investigate their putative carboxylating role in vivo, Arabidopsis NAD(P)-ME isoforms, as well as the NADP-ME2del2 (with a decreased ability to carboxylate pyruvate) and NADP-ME2R115A (lacking fumarate activation) versions, were functionally expressed in the cytosol of pyruvate carboxylase-negative (Pyc- ) Saccharomyces cerevisiae strains. The heterologous expression of NADP-ME1, NADP-ME2 (and its mutant proteins), and NADP-ME3 restored the growth of Pyc- S. cerevisiae on glucose, and this capacity was dependent on the availability of CO2 . On the other hand, NADP-ME4, NAD-ME1, and NAD-ME2 could not rescue the Pyc- strains from C4 auxotrophy. NADP-ME carboxylation activity could be measured in leaf crude extracts of knockout and overexpressing Arabidopsis lines with modified levels of NADP-ME, where this activity was correlated with the amount of NADP-ME2 transcript. These results indicate that specific A. thaliana NADP-ME isoforms are able to play an anaplerotic role in vivo and provide a basis for the study on the carboxylating activity of NADP-ME, which may contribute to the synthesis of C4 compounds and redox shuttling in plant cells.

Carbon assimilation in Eucalyptus urophylla grown under high atmospheric CO2 concentrations: A proteomics perspective.

Photosynthetic organisms may be drastically affected by the future climate projections of a considerable increase in CO2 concentrations. Growth under a high concentration of CO2 could stimulate carbon assimilation-especially in C3-type plants. We used a proteomics approach to test the hypothesis of an increase in the abundance of the enzymes involved in carbon assimilation in Eucalyptus urophylla plants grown under conditions of high atmospheric CO2. Our strategy allowed the profiling of all Calvin-Benson cycle enzymes and associated protein species. Among the 816 isolated proteins, those involved in carbon fixation were found to be the most abundant ones. An increase in the abundance of six key enzymes out of the eleven core enzymes involved in carbon fixation was detected in plants grown at a high CO2 concentration. Proteome changes were corroborated by the detection of a decrease in the stomatal aperture and in the vascular bundle area in Eucalyptus urophylla plantlets grown in an environment of high atmospheric CO2. Our proteomics approach indicates a positive metabolic response regarding carbon fixation in a CO2-enriched atmosphere. The slight but significant increase in the abundance of the Calvin enzymes suggests that stomatal closure did not prevent an increase in the carbon assimilation rates. BIOLOGICAL SIGNIFICANCE: The sample enrichment strategy and data analysis used here enabled the identification of all enzymes and most protein isoforms involved in the Calvin-Benson-Bessham cycle in Eucalyptus urophylla. Upon growth in CO2-enriched chambers, Eucalyptus urophylla plantlets responded by reducing the vascular bundle area and stomatal aperture size and by increasing the abundance of six of the eleven core enzymes involved in carbon fixation. Our proteome approach provides an estimate on how a commercially important C3-type plant would respond to an increase in CO2 concentrations. Additionally, confirmation at the protein level of the predicted genes involved in carbon assimilation may be used in plant transformation strategies aiming to increase plant adaptability to climate changes or to increase plant productivity.

Combined effect of water loss and wounding stress on gene activation of metabolic pathways associated with phenolic biosynthesis in carrot.

The application of postharvest abiotic stresses is an effective strategy to activate the primary and secondary metabolism of plants inducing the accumulation of antioxidant phenolic compounds. In the present study, the effect of water stress applied alone and in combination with wounding stress on the activation of primary (shikimic acid) and secondary (phenylpropanoid) metabolic pathways related with the accumulation of phenolic compound in plants was evaluated. Carrot (Daucus carota) was used as model system for this study, and the effect of abiotic stresses was evaluated at the gene expression level and on the accumulation of metabolites. As control of the study, whole carrots were stored under the same conditions. Results demonstrated that water stress activated the primary and secondary metabolism of carrots, favoring the lignification process. Likewise, wounding stress induced higher activation of the primary and secondary metabolism of carrots as compared to water stress alone, leading to higher accumulation of shikimic acid, phenolic compounds, and lignin. Additional water stress applied on wounded carrots exerted a synergistic effect on the wound-response at the gene expression level. For instance, when wounded carrots were treated with water stress, the tissue showed 20- and 14-fold increases in the relative expression of 3-deoxy-D-arabino-heptulosanate synthase and phenylalanine ammonia-lyase genes, respectively. However, since lignification was increased, lower accumulation of phenolic compounds was detected. Indicatively, at 48 h of storage, wounded carrots treated with water stress showed ~31% lower levels of phenolic compounds and ~23% higher lignin content as compared with wounded controls. In the present study, it was demonstrated that water stress is one of the pivotal mechanism of the wound-response in carrot. Results allowed the elucidation of strategies to induce the accumulation of specific primary or secondary metabolites when plants are treated with water stress alone or when additional water stress is applied on wounded tissue. If the accumulation of a specific primary or secondary metabolite were desirable, it would be recommended to apply both stresses to accelerate their biosynthesis. However, strategies such as the use of enzymatic inhibitors to block the carbon flux and enhance the accumulation of specific compounds should be designed.

O efeito da biodiversidade nativa da Mata Atlântica sobre o metabolismo de amadurecimento da banana (Musa acuminata AAA, Cavendish, cv. Nanicão) / he effect of the Atlantic Coastal Forest´s native biodiversity on the metabolism of banana ripening (Musa acuminata AAA, Cavendish, cv. Nanicão).

A produção de banana no Vale do Ribeira, em São Paulo, está inserida numa região que abriga um fragmento remanescente da Mata Atlântica, considerada um dos principais repositórios de biodiversidade e a floresta mais devastada entre os biomas brasileiros. Medidas sustentáveis, como a adoção de práticas agrícolas alternativas, tem sido o caminho para a conservação da fauna e flora, como também para a redução do uso inapropriado de terras que consequentemente pode aumentar o índice de doenças que atacam as plantas. Apesar, do razoável conhecimento científico sobre a fisiologia pós-colheita da banana, ainda não foram elucidadas quais são as respostas fisiológicas do fruto diante das alterações do meio ambiente decorrentes das práticas agroecológicas. Principalmente quais são os mecanismos de ação de compostos específicos, relacionados com a resistência da planta contra os estresses bióticos e abióticos. Também, não há uma abordagem analítica integrada que identifique os reguladores das vias metabólicas e possibilite um estudo holístico a nível molecular. Neste trabalho, foi avaliado o efeito da proximidade da biodiversidade nativa da Mata Atlântica sobre o perfil de metabólitos da banana (Musa acuminata AAA, Cavendish, cv. Nanicão). Foram comparados os frutos da parcela Biodiversidade, o qual apresenta 60% de seu perímetro limítrofe à floresta com a parcela Controle, inserida em uma área de produção de banana convencional. Neste estudo, foram determinados os perfis de etileno, vida verde, amido, açúcares, textura, cor, metabólitos semi-voláteis e não-voláteis e poliaminas. Como resultado, ambas as parcelas avaliadas apresentaram diferenças significativas no metabolismo primário e secundário, nos frutos verdes e maduros, respectivamente. Os metabólitos com diferença significativa entre as parcelas experimentais apresentaram maior envolvimento no metabolismo primário, sobretudo na via de biossíntese de aminoácidos, compostos precursores de uma ampla faixa de metabólitos secundários. As alterações quanto à abundância destes compostos são uma referência de que as mudanças do meio modificam as respostas da planta a estresses e estímulos

Banana production in the Ribeira Valley, in São Paulo, is set in a region that embraces a remaining fragment of the Atlantic Coastal Forest (Mata Atlântica), considered one of the main biodiversity repositories and the most devastated forest among the Brazilians´ biomes. Sustainable measures such as the adoption of alternative farming practices has been the way for the conservation of fauna and flora, but also to reduce the inappropriate use of land which in turn may increase the disease index that attack plants. Although, reasonable scientific knowledge about banana postharvest physiology is available, the fruit´s physiologic response given the environment changes as a result agroecological practices has not been elucidated yet. Primarily what are the mechanisms of action of specific compounds, related to plant resistance against biotic and abiotic stresses. In addition, there is not an integrated analytical approach to identify regulators of metabolic pathways and enable a holistic study at the molecular level. In this study, we evaluated the effect of proximity to the native biodiversity of the Atlantic Forest on the banana metabolites profile (Musa acuminata AAA, Cavendish, cv. Nanicão). The fruits of Biodiversity group were compared, which has 60% of its perimeter adjacent to the forest with the Control group set in a conventional banana production area. In this study, we determined the ethylene profiles, green life, starch, sugars, texture, colour, semi- volatile and non- volatile metabolites and polyamines. As a result, both evaluated groups showed significant differences in primary and secondary metabolism, green and ripen fruits respectively. The metabolites with significant differences between treatments showed greater involvement in primary metabolism, especially in amino acid biosynthesis, precursors of secondary metabolites. Changes in the abundance of these compounds are indicators that alterations in the environment modify the plant responses to stresses and stimuli