Synthetic and analytical routes to the L-amino acid conjugates of cis-OPDA and their identification and quantification in plants.

Cis-(+)-12-oxophytodienoic acid (cis-(+)-OPDA) is a bioactive jasmonate, a precursor of jasmonic acid, which also displays signaling activity on its own. Modulation of cis-(+)-OPDA actions may be carried out via biotransformation leading to metabolites of various functions. This work introduces a methodology for the synthesis of racemic cis-OPDA conjugates with amino acids (OPDA-aa) and their deuterium-labeled analogs, which enables the unambiguous identification and accurate quantification of these compounds in plants. We have developed a highly sensitive liquid chromatography-tandem mass spectrometry-based method for the reliable determination of seven OPDA-aa (OPDA-Alanine, OPDA-Aspartate, OPDA-Glutamate, OPDA-Glycine, OPDA-Isoleucine, OPDA-Phenylalanine, and OPDA-Valine) from minute amount of plant material. The extraction from 10 mg of fresh plant tissue by 10% aqueous methanol followed by single-step sample clean-up on hydrophilic-lipophilic balanced columns prior to final analysis was optimized. The method was validated in terms of accuracy and precision, and the method parameters such as process efficiency, recovery and matrix effects were evaluated. In mechanically wounded 30-day-old Arabidopsis thaliana leaves, five endogenous (+)-OPDA-aa were identified and their endogenous levels were estimated. The time-course accumulation revealed a peak 60 min after the wounding, roughly corresponding to the accumulation of cis-(+)-OPDA. Our synthetic and analytical methodologies will support studies on cis-(+)-OPDA conjugation with amino acids and research into the biological significance of these metabolites in plants.

New Water-Soluble Cytokinin Derivatives and Their Beneficial Impact on Barley Yield and Photosynthesis.

Solubility of growth regulators is essential for their use in agriculture. Four new cytokinin salts─6-benzylaminopurine mesylate (1), 6-(2-hydroxybenzylamino)purine mesylate (2), 6-(3-hydroxybenzylamino)purine mesylate (3), and 6-(3-methoxybenzylamino)purine mesylate (4)─were synthesized, and their crystal structures were determined to clarify structural influence on water solubility. The mesylates were several orders of magnitude more water-soluble than the parent CKs. The new salts significantly reduced chlorophyll degradation and impairment of photosystem II functionality in barley leaf segments undergoing artificial senescence and had pronounced effects on the leaves' endogenous CK pools, maintaining high concentrations of functional metabolites for several days, unlike canonical CKs. A foliar treatment with 1 and 3 increased the harvest yield of spring barley by up to 8% when compared to treatment with the parent CKs while also increasing the number of productive tillers. This effect was attributed to the higher bioavailability of the mesylate salts and the avoidance of dimethyl sulfoxide exposure.

Plant hormone cytokinin at the crossroads of stress priming and control of photosynthesis.

To cope with biotic and abiotic stress conditions, land plants have evolved several levels of protection, including delicate defense mechanisms to respond to changes in the environment. The benefits of inducible defense responses can be further augmented by defense priming, which allows plants to respond to a mild stimulus faster and more robustly than plants in the naïve (non-primed) state. Priming provides a low-cost protection of agriculturally important plants in a relatively safe and effective manner. Many different organic and inorganic compounds have been successfully tested to induce resistance in plants. Among the plethora of commonly used physicochemical techniques, priming by plant growth regulators (phytohormones and their derivatives) appears to be a viable approach with a wide range of applications. While several classes of plant hormones have been exploited in agriculture with promising results, much less attention has been paid to cytokinin, a major plant hormone involved in many biological processes including the regulation of photosynthesis. Cytokinins have been long known to be involved in the regulation of chlorophyll metabolism, among other functions, and are responsible for delaying the onset of senescence. A comprehensive overview of the possible mechanisms of the cytokinin-primed defense or stress-related responses, especially those related to photosynthesis, should provide better insight into some of the less understood aspects of this important group of plant growth regulators.

Non-invasive monitoring of hydraulic surge propagation in a wounded tobacco plant.

BACKGROUND: When a plant is wounded, a rapid hydraulic surge, acting probably as a systemic signal, spreads from the site of injury throughout the plant and leads to small transient deformation of tissues. So far, the propagation of hydraulic surge has been monitored by contact and thus potentially invasive methods. RESULTS: Here we present a non-invasive optical method, which allows simultaneous monitoring of micrometric shift of two opposite stem margins. The usefulness of this method was demonstrated by the measurement of the hydraulic surge propagation in a tobacco (Nicotiana tabacum (L.) cv. Samsun) after burning of its upper leaf. We have observed transient narrowing the stem below the burned leaf, which started within a few minutes after local burning. The comparison of the shift of the stem margin following vascular trace of the burned leaf and the margin on the opposite side of the stem has revealed that the stem deformation is highly asymmetric. CONCLUSIONS: This optical method represents a novel tool to investigate the mechanism of systemic response of plants to local damage. Our results points out the complexity of the relationship between hydraulic surge propagation and stem deformation.