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SUMMARY: Plant hormones are chemical signals governing almost every aspect of a plant's life cycle and responses to environmental cues. They are enmeshed within complex signaling networks that can only be deciphered by using broad‐scale analytical methods to capture information about several plant hormone classes simultaneously. Methods used for this purpose are all based on reversed‐phase (RP) liquid chromatography and mass spectrometric detection. Hydrophilic interaction chromatography (HILIC) is an alternative chromatographic method that performs well in analyses of biological samples. We therefore developed and validated a HILIC method for broad‐scale plant hormone analysis including a rapid sample preparation procedure; moreover, derivatization or fractionation is not required. The method enables plant hormone screening focused on polar and moderately polar analytes including cytokinins, auxins, jasmonates, abscisic acid and its metabolites, salicylates, indoleamines (melatonin), and 1‐aminocyclopropane‐1‐carboxylic acid (ACC), for a total of 45 analytes. Importantly, the major pitfalls of ACC analysis have been addressed. Furthermore, HILIC provides orthogonal selectivity to conventional RP methods and displays greater sensitivity, resulting in lower limits of quantification. However, it is less robust, so procedures to increase its reproducibility were established. The method's potential is demonstrated in a case study by employing an approach combining hormonal analysis with phenomics to examine responses of three Arabidopsis ecotypes toward three abiotic stress treatments: salinity, low nutrient availability, and their combination. The case study showcases the value of the simultaneous determination of several plant hormone classes coupled with phenomics data when unraveling processes involving complex cross‐talk under diverse plant‐environment interactions. [ABSTRACT FROM AUTHOR]

We have developed and validated a novel LC-MS/MS method for simultaneously analyzing amino acids, biogenic amines, and their acetylated and methylated derivatives in plants. This method involves a one-step extraction of 2–5 mg of lyophilized plant material followed by fractionation of different biogenic amine forms, and exploits an efficient combination of hydrophilic interaction liquid chromatography (HILIC), reversed phase (RP) chromatography with pre-column derivatization, and tandem mass spectrometry (MS). This approach enables high-throughput processing of plant samples, significantly reducing the time needed for analysis and its cost. We also present a new synthetic route for deuterium-labeled polyamines. The LC-MS/MS method was rigorously validated by quantifying levels of nitrogen-related metabolites in seedlings of seven plant species, including Arabidopsis, maize, and barley, all of which are commonly used model organisms in plant science research. Our results revealed substantial variations in the abundance of these metabolites between species, developmental stages, and growth conditions, particularly for the acetylated and methylated derivatives and the various polyamine fractions. However, the biological relevance of these plant metabolites is currently unclear. Overall, this work contributes significantly to plant science by providing a powerful analytical tool and setting the stage for future investigations into the functions of these nitrogen-related metabolites in plants. [ABSTRACT FROM AUTHOR]

Currently, reforestation efforts focus primarily on deciduous species replacing coniferous monocultures. Related to this are efforts to identify their interactions in the soil. Root exudation has many functions, including plant communication with soil microorganisms and the solubilisation of nutrients. Root exudates reflect the situation in the soil, as well as refer to the plant species and health/fitness. The idea here was to reveal the typical content of low-molecular-weight phenolic acids in the soil solution that occur in the early-life stages of most typical temperate deciduous trees Quercus robur, Sorbus torminalis, Fagus sylvatica and Acer pseudoplatanus, including the fast-growing invasive Robinia pseudoacacia. To compare their initial strategies and ambient, seedlings were planted in the pot experiment for one season since emergence. The following phenolic acids were detected in detectable concentrations: vanillic, 4-benzoic, syringic, p-coumaric and salicylic. Each tree species tested showed a unique fingerprint in these acids, which can be considered species-specific, i.e., their presence differed among the species. Robinia pseudoacacia (unlike the other trees tested) showed the ability to maintain high levels of p-coumaric acid in the soil solution, indicating its potential to survive in nutrient-poor soil and achieve rapid growth. On the contrary, the levels of all phenolic acids detected in the soil solution of Quercus robur and Sorbus torminalis were very low. These fingerprints should be extended to other compounds and also to older trees. [ABSTRACT FROM AUTHOR]

The REQUIRED FOR ARBUSCULAR MYCORRHIZATION1 (RAM1) transcription factor from the GRAS family is well known for its role as a master regulator of the arbuscular mycorrhizal (AM) symbiosis in dicotyledonous and monocotyledonous species, being essential in transcriptional reprogramming for the development and functionality of the arbuscules. In tomato, SlGRAS27 is the putative orthologue of RAM1 (here named SlRAM1), but has not yet been characterized. A reduced colonization of the root and impaired arbuscule formation were observed in SlRAM1 -silenced plants, confirming the functional conservation of the RAM1 orthologue in tomato. However, unexpectedly, SlRAM1 -overexpressing (UBIL:SlRAM1) plants also showed decreased mycorrhizal colonization. Analysis of non-mycorrhizal UBIL:SlRAM1 roots revealed an overall regulation of AM-related genes and a reduction of strigolactone biosynthesis. Moreover, external application of the strigolactone analogue GR244DO almost completely reversed the negative effects of SlRAM1 overexpression on the frequency of mycorrhization. However, it only partially recovered the pattern of arbuscule distribution observed in control plants. Our results strongly suggest that SlRAM1 has a dual regulatory role during mycorrhization and, in addition to its recognized action as a positive regulator of arbuscule development, it is also involved in different mechanisms for the negative regulation of mycorrhization, including the repression of strigolactone biosynthesis. [ABSTRACT FROM AUTHOR]

Co-occurring heat and drought stresses challenge crop performance. Stomata open to promote evaporative cooling during heat stress, but close to retain water during drought stress, which resulted in complex stomatal regulation under combined heat and drought. We aimed to investigate stomatal regulation in leaves and flowers of perennial, indeterminate cultivars of tomatoes subjected to individual and combined heat and drought stress followed by a recovery period, measuring morphological, physiological, and biochemical factors involved in stomatal regulation. Under stress, stomata of leaves were predominantly affected by drought, with lower stomatal density and stomatal closing, resulting in significantly decreased photosynthesis and higher leaf temperature. Conversely, stomata in sepals seemed affected mainly by heat during stress. The differential patterns in stomatal regulation in leaves and flowers persisted into the recovery phase as contrasting patterns in stomatal density. We show that flower transpiration is regulated by temperature, but leaf transpiration is regulated by soil water availability during stress. Organ-specific patterns of stomatal development and abscisic acid metabolism mediated this phenomenon. Our results throw light on the dual role of stomata in heat and drought tolerance of vegetative and generative organs, and demonstrate the importance of considering flower surfaces in the phenotyping of stomatal reactions to stress. [ABSTRACT FROM AUTHOR]

The carnivorous plants in the order Caryophyllales co-opted jasmonate signalling from plant defence to botanical carnivory. However, carnivorous plants have at least 11 independent origins, and here we ask whether jasmonate signalling has been co-opted repeatedly in different evolutionary lineages. We experimentally wounded and fed the carnivorous plants Sarracenia purpurea (order Ericales), Cephalotus follicularis (order Oxalidales), Drosophyllum lusitanicum (order Caryophyllales), and measured electrical signals, phytohormone tissue level, and digestive enzymes activity. Coronatine was added exogenously to confirm the role of jasmonates in the induction of digestive process. Immunodetection of aspartic protease and proteomic analysis of digestive fluid was also performed. We found that prey capture induced accumulation of endogenous jasmonates only in D. lusitanicum , in accordance with increased enzyme activity after insect prey or coronatine application. In C. follicularis , the enzyme activity was constitutive while in S. purpurea was regulated by multiple factors. Several classes of digestive enzymes were identified in the digestive fluid of D. lusitanicum. Although carnivorous plants from different evolutionary lineages use the same digestive enzymes, the mechanism of their regulation differs. All investigated genera use jasmonates for their ancient role, defence, but jasmonate signalling has been co-opted for botanical carnivory only in some of them. [ABSTRACT FROM AUTHOR]

Eggplant (Solanum melongena L.) is one of the most popular common major vegetable crops worldwide. This study evaluated the nutritional content of seven commercial eggplant fruits in terms of fatty acid, mineral, sugar, organic acid, amino acid and polyamine contents. The most abundant fatty acid was linoleic acid (range, 39.14-53.81%, ave. 45%), and the most abundant mineral was K (range, 1556.2-3171.6 mg/kg fw, ave. 2331.9). The major organic acid was malic acid (range, 129.87-387.01 mg/g fw, ave. 157.49), and the major sugar was fructose (range, 1242.81- 1379.77 mg/100 g fw, ave. 1350.88). The major polyamine was putrescine (11.54 and 25.70 nmol/g fw, ave. 17.86), and the major amino acid was glutamine (148.4 and 298.75 mg/100 g fw, ave. 219.74). Overall, taking into account the export potential of eggplants, these results may contribute to further studies aiming to improve other nutrient-rich varieties of eggplant in breeding programs.

In his famous book Insectivorous plants, Charles Darwin observed that the bending response of tentacles in the carnivorous sundew plant Drosera rotundifolia was not triggered by a drop of water, but rather the application of many dissolved chemicals or mechanical stimulation. In this study, we tried to reveal this 150-years-old mystery using methods not available in his time. We measured electrical signals, phytohormone tissue level, enzyme activities and an abundance of digestive enzyme aspartic protease droserasin in response to different stimuli (water drop, ammonia, mechanostimulation, chitin, insect prey) in Cape sundew (Drosera capensis). Drops of water induced the lowest number of action potentials (APs) in the tentacle head, and accumulation of jasmonates in the trap was not significantly different from control plants. On the other hand, all other stimuli significantly increased jasmonate accumulation; the highest was found after the application of insect prey. Drops of water also did not induce proteolytic activity and an abundance of aspartic protease droserasin in contrast to other stimuli. We found that the tentacles of sundew plants are not responsive to water drops due to an inactive jasmonic acid signalling pathway, important for the induction of significant digestive enzyme activities. [ABSTRACT FROM AUTHOR]