UHPLC–MS/MS based target profiling of stress-induced phytohormones

Floková, Kristýna; Tarkowská, Danuše; Miersch, Otto; Strnad, Miroslav; Wasternack, Claus; Novák, Ondřej

doi:10.1016/j.phytochem.2014.05.015

Cited by 209 publications

(167 citation statements)

References 39 publications

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“…Levels of the CKs, auxins, and JAs together with SA and ABA were determined by the isotope dilution method using ultra-HPLC tandem electrospray mass spectrometry with stable isotope-labeled internal standards used as a reference (Novák et al, 2012;Svačinová et al, 2012;Floková et al, 2014).…”

Section: Plant Hormone Analysis By Ultra-hplc Tandem Electrospray Masmentioning

confidence: 99%

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

et al. 2017

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Arbuscular mycorrhizas (AM) are the most common symbiotic associations between a plant's root compartment and fungi. They provide nutritional benefit (mostly inorganic phosphate [P i ]), leading to improved growth, and nonnutritional benefits, including defense responses to environmental cues throughout the host plant, which, in return, delivers carbohydrates to the symbiont. However, how transcriptional and metabolic changes occurring in leaves of AM plants differ from those induced by P i fertilization is poorly understood. We investigated systemic changes in the leaves of mycorrhized Medicago truncatula in conditions with no improved P i status and compared them with those induced by high-P i treatment in nonmycorrhized plants. Microarray-based genome-wide profiling indicated up-regulation by mycorrhization of genes involved in flavonoid, terpenoid, jasmonic acid (JA), and abscisic acid (ABA) biosynthesis as well as enhanced expression of MYC2, the master regulator of JA-dependent responses. Accordingly, total anthocyanins and flavonoids increased, and most flavonoid species were enriched in AM leaves. Both the AM and P i treatments corepressed iron homeostasis genes, resulting in lower levels of available iron in leaves. In addition, higher levels of cytokinins were found in leaves of AM-and P i -treated plants, whereas the level of ABA was increased specifically in AM leaves. Foliar treatment of nonmycorrhized plants with either ABA or JA induced the up-regulation of MYC2, but only JA also induced the up-regulation of flavonoid and terpenoid biosynthetic genes. Based on these results, we propose that mycorrhization and P i fertilization share cytokinin-mediated improved shoot growth, whereas enhanced ABA biosynthesis and JA-regulated flavonoid and terpenoid biosynthesis in leaves are specific to mycorrhization.

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Section: Plant Hormone Analysis By Ultra-hplc Tandem Electrospray Masmentioning

confidence: 99%

Enhanced Secondary- and Hormone Metabolism in Leaves of Arbuscular Mycorrhizal Medicago truncatula

et al. 2017

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“…Endogenous levels of CK metabolites and free IAA were detected by liquid chromatography-tandem mass spectrometry methods (Floková et al, 2014;Antoniadi et al, 2015). Samples (20 mg fresh weight) were homogenized and extracted in 1 mL of modified Bieleski buffer (60% methanol, 10% HCOOH and 30% water) together with a cocktail of stable isotope-labeled internal standards (0.25 pmol of CK bases, ribosides, and N-glucosides, 0.5 pmol of CK O-glucosides and nucleotides, and 5 pmol of [ 2 H 5 ]IAA per sample added).…”

Section: Measurements Of Endogenous Phytohormonesmentioning

confidence: 99%

Cytokinins Are Initial Targets of Light in the Control of Bud Outgrowth

et al. 2016

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Bud outgrowth is controlled by environmental and endogenous factors. Through the use of the photosynthesis inhibitor norflurazon and of masking experiments, evidence is given here that light acts mainly as a morphogenic signal in the triggering of bud outgrowth and that initial steps in the light signaling pathway involve cytokinins (CKs). Indeed, in rose (Rosa hybrida), inhibition of bud outgrowth by darkness is suppressed solely by the application of CKs. In contrast, application of sugars has a limited effect. Exposure of plants to white light (WL) induces a rapid (after 3-6 h of WL exposure) up-regulation of CK synthesis (RhIPT3 and RhIPT5), of CK activation (RhLOG8), and of CK putative transporter RhPUP5 genes and to the repression of the CK degradation RhCKX1 gene in the node. This leads to the accumulation of CKs in the node within 6 h and in the bud at 24 h and to the triggering of bud outgrowth. Molecular analysis of genes involved in major mechanisms of bud outgrowth (strigolactone signaling [RwMAX2], metabolism and transport of auxin [RhPIN1, RhYUC1, and RhTAR1], regulation of sugar sink strength [RhVI, RhSUSY, RhSUC2, and RhSWEET10], and cell division and expansion [RhEXP and RhPCNA]) reveal that, when supplied in darkness, CKs up-regulate their expression as rapidly and as intensely as WL. Additionally, up-regulation of CKs by WL promotes xylem flux toward the bud, as evidenced by Methylene Blue accumulation in the bud after CK treatment in the dark. Altogether, these results suggest that CKs are initial components of the light signaling pathway that controls the initiation of bud outgrowth.

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“…Both eluates were combined and dried under vacuum. Quantification of ABA metabolites was achieved by liquid chromatography coupled to tandem mass spectometry using an Acquity UPLC system (Waters) coupled with a Xevo TQ-S triple quadrupole mass spectrometer (Waters) as described by Floková et al (2014), with modifications. Purified samples were reconstructed in 200 mL of mobile phase, filtered with 0.45-mm PTFE membrane filter (Phenomenex), and injected onto Acquity UPLC CSH C18 column (100 3 2.1 mm, 1.7 mm; Waters).…”

Section: Aba Analysismentioning

confidence: 99%