Dynamic shifts in primary metabolism across fruit development stages in Capsicum chinense (cv. Habanero).

The development of fleshy fruits involves changes in size and mass, followed by cell differentiation, which is associated with anatomical and histological changes. Parallel to these changes, metabolic alterations lead to the production of osmolytes and energy that modify cell turgor pressure, thereby promoting cell expansion and fruit growth. Detailed information is known about these processes in climacteric fruits (e.g. tomato); however, the regulation of metabolism and its association with anatomical changes in non-climacteric fruit development are poorly understood. In this study, we used detailed anatomical and histological analyses to define three developmental phases of chili pepper (Capsicum chinense cv. Habanero): cell division, cell expansion, and ripening. We showed that each was marked by distinct metabolic profiles, underpinning the switches in energy metabolism to support cellular processes. Interestingly, mitochondrial activity was high in the early stages of development and declined over time, with a modest increase in O2 consumption by pericarp tissues at the beginning of the ripening stage. This respiratory-like burst was associated with the degradation of starch and malate, which are the sources of energy and carbon required for other processes associated with fruit maturation.

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.

Spatio-temporal characterization of the fruit metabolism in contrasting accessions of Macauba (Acrocomia aculeata).

Although Macauba (Acrocomia aculeata) has been highlighted by its high-quality oil to fit edible and nonedible purposes, data addressing carbon and nitrogen metabolism underlying development and ripening of fruits remain scarce. In addition, accessions of Macauba exibit varied oil yield in fruits, including during the fruit development stages. Here, we monitored contents of carbohydrates, proteins, amino acids and lipids in the mesocarp and endosperm of Macauba fruits until ripening. We selected three accessions from different Brazilian regions (southeast, MG; northeast, PE; and central-west, MS) that differ in the mesocarp lipid content of ripe fruits. Despite the anatomical differences, mesocarp and endosperm exhibited similar trends of metabolite accumulation for most of the analyzed compounds. In the mesocarp, total soluble protein, free amino acids, sucrose, starch and total lipids accumulate towards ripening, while glucose and fructose declined in all accessions. Endosperm differed from mesocarp solely in the amino acid content, which decreased in ripe fruits. In the endosperm, accessions accumulated carbohydrates differently. Accession PE showed comparable fructose and starch contents in the endosperm between the beginning of fruit development and ripening, while in accessions MG and MS, both compounds decreased and increased, respectively, towards ripening. Accession MG was highlighted by its highest lipid content in the two tissues indicating its potential for energy and cosmetic industries. Our results provide novel insights into metabolic changes underlying development and ripening of Macauba fruits and variability in oil content among accessions, indicating new targets for breeding programs.

Metabolic shifts during fruit development in pungent and non-pungent peppers.

Fruit pungency is caused by the accumulation of capsaicinoids, secondary metabolites whose relation to primary metabolism remains unclear. We have selected ten geographically diverse accessions of Capsicum chinense Jacq with different pungency levels. A detailed metabolic profile was conducted in the fruit placenta and pericarp at 20, 45, and 60 days after anthesis aiming at increasing our understanding of the metabolic changes in these tissues across fruit development and their potential connection to capsaicin metabolism. Overall, despite the variation in fruit pungency among the ten accessions, the composition and metabolite levels in both placenta and pericarp were uniformly stable across accessions. Most of the metabolite variability occurred between the fruit developmental stages rather than among the accessions. Interestingly, different metabolite adjustments in the placenta were observed among pungent and non-pungent accessions, which seem to be related to differences in the genetic background. Furthermore, we observed high coordination between metabolites and capsaicin production in C. chinense fruits, suggesting that pungency in placenta is adjusted with primary metabolism.

Differential mechanisms between traditionally established and new highbush blueberry (Vaccinium corymbosum L.) cultivars reveal new insights into manganese toxicity resistance.

In acid soils, manganese (Mn) concentration increases, becoming toxic to plants. Mn toxicity differentially affects physiological processes in highbush blueberry (Vaccinium corymbosum L.) cultivars. However, the mechanisms involved in Mn toxicity of the new and traditionally established cultivars are unknown. To understand Mn toxicity mechanisms, four traditionally established (Legacy, Brigitta, Duke, and Star) cultivars and two recently introduced to Chile (Camellia and Cargo) were grown under hydroponic conditions subjected to control Mn (2 µM) and Mn toxicity (1000 µM). Physiological, biochemical, and molecular parameters were evaluated at 0, 7, 14, and 21 days. We found that the relative growth rate was reduced in almost all blueberry cultivars under Mn toxicity, except Camellia, with Star being the most affected. The photosynthetic parameters were reduced only in Star by Mn treatment. Leaf Mn concentrations increased in all cultivars, exhibiting the lowest levels in Camellia and Cargo. Brigitta and Duke exhibited higher ß-carotene levels, while Cargo exhibited a reduction under toxic Mn. In Legacy, lutein levels increased under Mn toxicity. Traditionally established cultivars exhibited higher antioxidant activity than the new cultivars under Mn toxicity. The Legacy and Duke cultivars increased VcMTP4 expression with Mn exposure time. A multivariate analysis separated Legacy and Duke from Camellia; Star and Cargo; and Brigitta. Our study demonstrated that Mn toxicity differentially affects physiological, biochemical, and molecular features in the new and traditionally established cultivars, with Legacy, Duke, Camellia, and Cargo as the Mn-resistant cultivars differing in their Mn-resistance mechanisms and Star as the Mn-sensitive cultivar.

Aluminum stress differentially affects physiological performance and metabolic compounds in cultivars of highbush blueberry.

Aluminum (Al) toxicity is one of the major factors that limit the growth and production of crops in acid soils. Highbush blueberry (Vaccinium corymbosum L.) cultivars differing in resistance to Al toxicity regarding root growth and photosynthetic performance were used. In this study, we compared the physiological and metabolic strategies to cope with Al toxicity among the highbush blueberry cultivars [two new ones (Camellia and Cargo) and three established ones (Brigitta (Al-resistant), Star and Duke)]. Aluminum concentration in roots and leaves increased in all cultivars after 24 and 48 h of exposure to Al, but less so in roots of cultivar Camellia and leaves of cultivar Cargo. These two cultivars displayed minor effects of Al exposure in terms of photosynthetic activity in comparison with the established cultivars. Furthermore, Cargo did not vary fluorescence parameters, whereas Camellia exhibited a decrease in effective quantum yield (ΦPSII) and electron transport rate (ETR) and a change in non-photochemical quenching (NPQ) and maximum quantum yield (Fv/Fm) under Al after 48 h. The Al treatment increased total phenols in leaves of Brigitta, Cargo, and Camellia, whereas antioxidant activity increased in Star and Cargo after 48 h. Aluminum exposure decreased malate concentration in roots of all cultivars, but no change was noted in fumarate concentration. The antioxidant activity correlated with photosynthetic performance and the total phenol concentration in the leaves of new cultivars exposed to Al, suggesting enhanced resistance in the short-term experiment. The principal component analysis separated the new from the established cultivars. In conclusion, the new cultivars appear to be more Al-resistant than the established ones, with Star being most Al-sensitive. Regarding the Al-resistance mechanisms of the new cultivars, it is suggested that Camellia could have a root Al-exclusion mechanism under Al toxicity. This mechanism could be explained by low Al concentration in roots, suggesting that this cultivar could exude organic acid, allowing to chelate Al in the rhizosphere. Nonetheless, further researches are needed to confirm this assumption.

Eucalypt plants are physiologically and metabolically affected by infection with Ceratocystis fimbriata.

Ceratocystis wilt, caused by Ceratocystis fimbriata, is currently one of the most important disease in eucalypt plantations. Plants infected by C. fimbriata have lower volumetric growth, lower pulp yields and reduced timber values. The physiological bases of infection induced by this pathogen in eucalypt plant are not known. Therefore, this study aims to assess the physiological and metabolic changes in eucalypt clones that are resistant and susceptible to C. fimbriata. Once, we evaluated in detail their leaf gas exchange, chlorophyll a fluorescence, water potential, metabolite profiling and growth-related parameters. When inoculated, the susceptible clone displayed reduced water potential, CO2 assimilation rate, stomatal conductance, transpiration rate, photochemical quenching coefficient, electron transport rate, and root biomass. Inoculated resistant and susceptible clones both presented higher respiration rates than healthy plants. Many compounds of primary and secondary metabolism were significantly altered after fungal infection in both clones. These results suggest that, C. fimbriata interferes in the primary and secondary metabolism of plants that may be linked to the induction of defense mechanisms and that, due to water restrictions caused by the fungus in susceptible plants, there is a partial closure of the stomata to prevent water loss and a consequent reduction in photosynthesis and the transpiration rate, which in turn, leads to a decrease in the plant's growth-related. These results combined, allowed for a better understanding of the physiological and metabolic changes following the infectious process of C. fimbriata, which limit eucalypt plant growth.

Measurement of Tricarboxylic Acid Cycle Enzyme Activities in Plants.

Mitochondria are vital cytoplasmic organelle of eukaryotic cells responsible for oxidative energy metabolism and the synthesis of intermediates utilized in various other metabolic pathways. The functions of mitochondrion are the oxidation of organic acids by the tricarboxylic acid (TCA) cycle and the synthesis of ATP by the oxidative phosphorylation in the mitochondrial electron transport chain. The TCA cycle is composed by a set of enzymes that are essential for optimal functioning of the primary carbon metabolism in plants. The activity of each TCA cycle enzyme in plants may vary according to cell type, plant tissue, stage of plant development, and the environment. Here, we describe current methods used for the determination of the TCA cycle enzyme activities in different plant tissues.