As chemical messengers, phytohormones can enhance the tolerance of plants to stress caused by toxic elements (TEs) such as cadmium (Cd), lead (Pb), and zinc (Zn). This study investigated the combined toxicity of Cd, Pb, and Zn, and its impact on stress phytohormones (jasmonates, salicylic acid, and abscisic acid), in oat (Avena sativa L.) using anthropogenically contaminated soil in a 4-week pot experiment. The uptake of TEs by the roots increased in the multi-contaminated soil, while Zn was the only TE to be translocated to the leaves. The toxic effect of the TEs was assessed in terms of plant growth, revealing a decline in leaf dry biomass, whereas the impact on the roots was insignificant. These findings align with the levels of stress phytohormones. An increase in bioactive forms of stress phytohormones in leaves due to TEs indicates TE toxicity and leaf sensitivity. Conversely, low levels of these phytohormones, along with crosstalk between them, suggest reduced defense against TEs in the roots. The abundance of stress phytohormones declined in the following order: salicylic acid > jasmonates > abscisic acid. These results help to understand the mechanism by which plants respond to TEs, particularly their combined toxicity.

• Keywords
• abscisic acid, jasmonic acid, oat, potentially toxic elements, salicylic acid, stress response,
• Publication type
• Journal Article MeSH

BACKGROUND: Acidic phytohormones are small molecules controlling many physiological functions in plants. A comprehensive picture of their profiles including the active forms, precursors and metabolites provides an important insight into ongoing physiological processes and is essential for many biological studies performed on plants. RESULTS: A high-throughput sample preparation method for liquid chromatography-tandem mass spectrometry determination of 25 acidic phytohormones classed as auxins, jasmonates, abscisates and salicylic acid was optimised. The method uses a small amount of plant tissue (less than 10 mg fresh weight) and acidic extraction in 1 mol/L formic acid in 10% aqueous methanol followed by miniaturised purification on reverse phase sorbent accommodated in pipette tips organised in a 3D printed 96-place interface, capable of processing 192 samples in one run. The method was evaluated in terms of process efficiency, recovery and matrix effects as well as establishing validation parameters such as accuracy and precision. The applicability of the method in relation to the amounts of sample collected from distantly related plant species was evaluated and the results for phytohormone profiles are discussed in the context of literature reports. CONCLUSION: The method developed enables high-throughput profiling of acidic phytohormones with minute amounts of plant material, and it is suitable for large scale interspecies studies.

• Keywords
• 3D printing, Evolutionarily distant plant species, High-throughput, In-tip microSPE, Liquid chromatography, Mass spectrometry, Miniaturisation, Plant hormones,
• Publication type
• Journal Article MeSH

The establishment of an efficient protocol for in vitro growth and regeneration of kohlrabi (Brassica oleracea var. gongylodes) allowed us to closely examine the phytohormone profiles of kohlrabi seedlings at four growth stages (T1-T4), additionally including the effects of cytokinins (CKs)-trans-zeatin (transZ) and thidiazuron (TDZ)-and high sucrose concentrations (6% and 9%). Resulting phytohormone profiles showed complex time-course patterns. At the T2 stage of control kohlrabi plantlets (with two emerged true leaves), levels of endogenous CK free bases and gibberellin GA20 increased, while increases in jasmonic acid (JA), JA-isoleucine (JA-Ile), indole-3-acetic acid (IAA) and indole-3-acetamide (IAM) peaked later, at T3. At the same time, the content of most of the analyzed IAA metabolites decreased. Supplementing growth media with CK induced de novo formation of shoots, while both CK and sucrose treatments caused important changes in most of the phytohormone groups at each developmental stage, compared to control. Principal component analysis (PCA) showed that sucrose treatment, especially at 9%, had a stronger effect on the content of endogenous hormones than CK treatments. Correlation analysis showed that the dynamic balance between the levels of certain bioactive phytohormone forms and some of their metabolites could be lost or reversed at particular growth stages and under certain CK or sucrose treatments, with correlation values changing between strongly positive and strongly negative. Our results indicate that the kohlrabi phytohormonome is a highly dynamic system that changes greatly along the developmental time scale and also during de novo shoot formation, depending on exogenous factors such as the presence of growth regulators and different sucrose concentrations in the growth media, and that it interacts intensively with these factors to facilitate certain responses.

• Keywords
• cytokinin, in vitro development, kohlrabi, phytohormone profiling, shoot regeneration, sucrose,
• Publication type
• Journal Article MeSH

Soil salinity is severely affecting crop productivity in many countries, particularly in the Mediterranean area. To evaluate early plant responses to increased salinity and characterize tolerance markers, three important Brassica crops - Chinese cabbage (Brassica rapa ssp. pekinensis), white cabbage (B. oleracea var. capitata) and kale (B. oleracea var. acephala) were subjected to short-term (24 h) salt stress by exposing them to NaCl at concentrations of 50, 100, or 200 mM. Physiological (root growth, photosynthetic performance parameters, and Na+/K+ ratio) and biochemical parameters (proline content and lipid peroxidation as indicated by malondialdehyde, MDA, levels) in the plants' roots and leaves were then measured. Photosynthetic parameters such as the total performance index PItotal (describing the overall efficiency of PSI, PSII and the intersystem electron transport chain) appeared to be the most salinity-sensitive parameter and informative stress marker. This parameter was decreased more strongly in Chinese cabbage than in white cabbage and kale. It indicated that salinity reduced the capacity of the photosynthetic system for efficient energy conversion, particularly in Chinese cabbage. In parallel with the photosynthetic impairments, the Na+/K+ ratio was highest in Chinese cabbage leaves and lowest in kale leaves while kale root is able to keep high Na+/K+ ratio without a significant increase in MDA. Thus Na+/K+ ratio, high in root and low in leaves accompanying with low MDA level is an informative marker of salinity tolerance. The crops' tolerance was positively correlated with levels of the stress hormone abscisic acid (ABA) and negatively correlated with levels of jasmonic acid (JA), and jasmonoyl-L-isoleucine (JA-Ile). Furthermore, salinity induced contrasting changes in levels of the growth-promoting hormones brassinosteroids (BRs). The crop's tolerance was positively correlated with levels of BR precursor typhasterol while negatively with the active BR brassinolide. Principal Component Analysis revealed correlations in observed changes in phytohormones, biochemical, and physiological parameters. Overall, the results show that kale is the most tolerant of the three species and Chinese cabbage the most sensitive to salt stress, and provide holistic indications of the spectrum of tolerance mechanisms involved.

• Keywords
• Chinese cabbage, brassinosteroids, kale, photosynthetic performance, salinity stress, stress hormones, tolerance, white cabbage,
• Publication type
• Journal Article MeSH

: Jasmonic acid (JA) and its related derivatives are ubiquitously occurring compounds of land plants acting in numerous stress responses and development. Recent studies on evolution of JA and other oxylipins indicated conserved biosynthesis. JA formation is initiated by oxygenation of α-linolenic acid (α-LeA, 18:3) or 16:3 fatty acid of chloroplast membranes leading to 12-oxo-phytodienoic acid (OPDA) as intermediate compound, but in Marchantiapolymorpha and Physcomitrellapatens, OPDA and some of its derivatives are final products active in a conserved signaling pathway. JA formation and its metabolic conversion take place in chloroplasts, peroxisomes and cytosol, respectively. Metabolites of JA are formed in 12 different pathways leading to active, inactive and partially active compounds. The isoleucine conjugate of JA (JA-Ile) is the ligand of the receptor component COI1 in vascular plants, whereas in the bryophyte M. polymorpha COI1 perceives an OPDA derivative indicating its functionally conserved activity. JA-induced gene expressions in the numerous biotic and abiotic stress responses and development are initiated in a well-studied complex regulation by homeostasis of transcription factors functioning as repressors and activators.

• Keywords
• JA biosynthetic enzymes, JA bypass, JA signaling, Jasmonic acid (JA) metabolites, active JA compounds, occurrence, transcription factors,
• MeSH
• Chloroplasts metabolism   MeSH
• Cyclopentanes metabolism   MeSH
• Species Specificity MeSH
• alpha-Linolenic Acid metabolism   MeSH
• Marchantia metabolism   MeSH
• Fatty Acids metabolism   MeSH
• Bryopsida metabolism   MeSH
• Metabolic Networks and Pathways MeSH
• Fatty Acids, Unsaturated metabolism   MeSH
• Oxylipins metabolism   MeSH
• Peroxisomes metabolism   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• 12-oxophytodienoic acid MeSH Browser
• Cyclopentanes MeSH
• jasmonic acid MeSH Browser
• alpha-Linolenic Acid MeSH
• Fatty Acids MeSH
• Fatty Acids, Unsaturated MeSH
• Oxylipins MeSH

Expression takes place for most of the jasmonic acid (JA)-induced genes in a COI1-dependent manner via perception of its conjugate JA-Ile in the SCFCOI1-JAZ co-receptor complex. There are, however, numerous genes and processes, which are preferentially induced COI1-independently by the precursor of JA, 12-oxo-phytodienoic acid (OPDA). After recent identification of the Ile-conjugate of OPDA, OPDA-Ile, biological activity of this compound could be unequivocally proven in terms of gene expression. Any interference of OPDA, JA, or JA-Ile in OPDA-Ile-induced gene expression could be excluded by using different genetic background. The data suggest individual signaling properties of OPDA-Ile. Future studies for analysis of an SCFCOI1-JAZ co-receptor-independent route of signaling are proposed.

• Keywords
• 12-oxo-phytodienoic acid (OPDA), JA-Ile perception, OPDA-Ile-induced gene expression, SCFCOI1-JAZ coreceptor complex, jasmonic acid (JA), jasmonoyl-isoleucine (JA-Ile),
• MeSH
• Arabidopsis metabolism   MeSH
• Cyclopentanes metabolism   MeSH
• Isoleucine analogs & derivatives   metabolism   MeSH
• Fatty Acids, Unsaturated metabolism   MeSH
• Oxylipins metabolism   MeSH
• Arabidopsis Proteins metabolism   MeSH
• Plant Growth Regulators metabolism   MeSH
• Signal Transduction physiology   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• 12-oxophytodienoic acid MeSH Browser
• Cyclopentanes MeSH
• Isoleucine MeSH
• jasmonic acid MeSH Browser
• jasmonoyl-isoleucine MeSH Browser
• Fatty Acids, Unsaturated MeSH
• Oxylipins MeSH
• Arabidopsis Proteins MeSH
• Plant Growth Regulators MeSH

Oxylipins of the jasmonate family are active as signals in plant responses to biotic and abiotic stresses as well as in development. Jasmonic acid (JA), its precursor cis-12-oxo-phytodienoic acid (OPDA) and the isoleucine conjugate of JA (JA-Ile) are the most prominent members. OPDA and JA-Ile have individual signalling properties in several processes and differ in their pattern of gene expression. JA-Ile, but not OPDA, is perceived by the SCFCOI1-JAZ co-receptor complex. There are, however, numerous processes and genes specifically induced by OPDA. The recently identified OPDA-Ile suggests that OPDA specific responses might be mediated upon formation of OPDA-Ile. Here, we tested OPDA-Ile-induced gene expression in wild type and JA-deficient, JA-insensitive and JA-Ile-deficient mutant background. Tests on putative conversion of OPDA-Ile during treatments revealed only negligible conversion. Expression of two OPDA-inducible genes, GRX480 and ZAT10, by OPDA-Ile could be detected in a JA-independent manner in Arabidopsis seedlings but less in flowering plants. The data suggest a bioactivity in planta of OPDA-Ile.

• MeSH
• Arabidopsis drug effects   genetics   MeSH
• Cyclopentanes metabolism   MeSH
• Ecotype MeSH
• Isoleucine analogs & derivatives   metabolism   MeSH
• Plant Leaves drug effects   metabolism   MeSH
• RNA, Messenger genetics   metabolism   MeSH
• Fatty Acids, Unsaturated metabolism   pharmacology   MeSH
• Oxylipins metabolism   MeSH
• Gene Expression Regulation, Plant drug effects   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• 12-oxophytodienoic acid MeSH Browser
• Cyclopentanes MeSH
• Isoleucine MeSH
• jasmonic acid MeSH Browser
• jasmonoyl-isoleucine MeSH Browser
• RNA, Messenger MeSH
• Fatty Acids, Unsaturated MeSH
• Oxylipins MeSH