High-throughput quantitative analysis of phytohormones in sorghum leaf and root tissue by ultra-performance liquid chromatography-mass spectrometry

• Published in: Analytical and bioanalytical chemistry 2019-07, Vol.411 (19), p.4839-4848
• Main Authors: Sheflin, Amy M., Kirkwood, Jay S., Wolfe, Lisa M., Jahn, Courtney E., Broeckling, Corey D., Schachtman, Daniel P., Prenni, Jessica E.
• Format: Article
• Language: English
• Subjects: Abscisic acid; Adaptation; Analysis; Analytical Chemistry; Auxins; Biochemistry; Brassinosteroids; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; Cytokinins; Food Science; Gene expression; Genetic factors; Genomes; Genomics; Genotypes; Gibberellins; High-throughput; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Instrument industry; Jasmonic acid; Laboratory Medicine; LC-MS/MS; Leaves; Liquid chromatography; Liquid-liquid extraction; Low concentrations; Mass spectrometry; Mass spectroscopy; Mathematical analysis; Microbiomes; Monitoring/Environmental Analysis; Organic chemistry; Physiological aspects; Phytohormones; Plant hormones; Plant tissues; Quantification; Quantitative analysis; Research Paper; Salicylates; Solid phases; Sorghum; Structure-function relationships; Young Investigators in (Bio-)Analytical Chemistry
• ISSN: 1618-2642; 1618-2650
• DOI: 10.1007/s00216-019-01658-9

Plant development, growth, and adaptation to stress are regulated by phytohormones, which can influence physiology even at low concentrations. Phytohormones are chemically grouped according to both structure and function as auxins, cytokinins, abscisic acid, jasmonates, salicylates, gibberellins, and brassinosteroids, among others. This chemical diversity and requirement for highly sensitive detection in complex matrices create unique challenges for comprehensive phytohormone analysis. Here, we present a robust and efficient quantitative UPLC-MS/MS assay for 17 phytohormones, including jasmonates, salicylates, abscisic acid, gibberellins, cytokinins, and auxins. Using this assay, 12 phytohormones were detected and quantified in sorghum plant tissue without the need for solid phase extraction (SPE) or liquid-liquid extraction. Variation of phytohormone profiles was explored in both root and leaf tissues between three genotypes, harvested at two different developmental time points. The results highlight the importance of tissue type, sampling time, and genetic factors when designing experiments that involve phytohormone analysis of sorghum. This research lays the groundwork for future studies, which can combine phytohormone profiling with other datasets such as transcriptome, soil microbiome, genome, and metabolome data, to provide important functional information about adaptation to stress and other environmental variables.