The field of plant hormonomics focuses on the qualitative and quantitative analysis of the hormone complement in plant samples, akin to other omics sciences. Plant hormones, alongside primary and secondary metabolites, govern vital processes throughout a plant's lifecycle. While active hormones have received significant attention, studying all related compounds provides valuable insights into internal processes. Conventional single-class plant hormone analysis employs thorough sample purification, short analysis and triple quadrupole tandem mass spectrometry. Conversely, comprehensive hormonomics analysis necessitates minimal purification, robust and efficient separation and better-performing mass spectrometry instruments. This review summarizes the current status of plant hormone analysis methods, focusing on sample preparation, advances in chromatographic separation and mass spectrometric detection, including a discussion on internal standard selection and the potential of derivatization. Moreover, current approaches for assessing the spatiotemporal distribution are evaluated. The review touches on the legitimacy of the term plant hormonomics by exploring the current status of methods and outlining possible future trends.

• Klíčová slova
• Hormonomics, Internal standard, Liquid chromatography, Mass spectrometry, Matrix effect, Metabolomics, Omics, Plant hormone, Solid phase extraction,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

BACKGROUND: Strigolactones represent the most recently described group of plant hormones involved in many aspects of plant growth regulation. Simultaneously, root exuded strigolactones mediate rhizosphere signaling towards beneficial arbuscular mycorrhizal fungi, but also attract parasitic plants. The seed germination of parasitic plants induced by host strigolactones leads to serious agricultural problems worldwide. More insight in these signaling molecules is hampered by their extremely low concentrations in complex soil and plant tissue matrices, as well as their instability. So far, the combination of tailored isolation-that would replace current unspecific, time-consuming and labour-intensive processing of large samples-and a highly sensitive method for the simultaneous profiling of a broad spectrum of strigolactones has not been reported. RESULTS: Depending on the sample matrix, two different strategies for the rapid extraction of the seven structurally similar strigolactones and highly efficient single-step pre-concentration on polymeric RP SPE sorbent were developed and validated. Compared to conventional methods, controlled temperature during the extraction and the addition of an organic modifier (acetonitrile, acetone) to the extraction solvent helped to tailor strigolactone isolation from low initial amounts of root tissue (150 mg fresh weight, FW) and root exudate (20 ml), which improved both strigolactone stability and sample purity. We have designed an efficient UHPLC separation with sensitive MS/MS detection for simultaneous analysis of seven natural strigolactones including their biosynthetic precursors-carlactone and carlactonoic acid. In combination with the optimized UHPLC-MS/MS method, attomolar detection limits were achieved. The new method allowed successful profiling of seven strigolactones in small exudate and root tissue samples of four different agriculturally important plant species-sorghum, rice, pea and tomato. CONCLUSION: The established method provides efficient strigolactone extraction with aqueous mixtures of less nucleophilic organic solvents from small root tissue and root exudate samples, in combination with rapid single-step pre-concentration. This method improves strigolactone stability and eliminates the co-extraction and signal of matrix-associated contaminants during the final UHPLC-MS/MS analysis with an electrospray interface, which dramatically increases the overall sensitivity of the analysis. We show that the method can be applied to a variety of plant species.

• Klíčová slova
• Phosphate starvation, Phytohormones, Quantitative analysis, Solid phase extraction (SPE), Strigolactones, UHPLC–MS/MS,
• Publikační typ
• časopisecké články MeSH

Strigolactones (SLs) are important plant hormones that are produced via the carotenoid biosynthetic pathway and occur at extremely low concentrations in various plant species. They regulate root development, play important roles in symbioses between higher plants and mycorrhizal fungi, and stimulate germination of plant-parasitic Orobanche and Striga species. Chemical analysis is central to research on the biochemistry of SLs and their roles in developmental biology and plant physiology. Here we summarize key issues relating to the identification and quantification of SLs isolated from plant tissues and exudates. The advantages and drawbacks of different protocols used for strigolactone analysis are discussed, and guidelines for selecting a procedure that will minimize losses during isolation and purification prior to final analysis are proposed. Hyphenated techniques suitable for SL analysis such as GC-MS and LC-MS/MS are also discussed, and newer ambient techniques such as HR-DART-MS and DESI-MS are highlighted as tools with considerable potential in SL research. A key advantage of these methods is that they require only simply sample preparation.

• Klíčová slova
• DESI-MS, Determination, GC–MS, HR-DART-MS, Isolation, LC–MS/MS, Strigolactones,
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH