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ARS Home » Midwest Area » Columbia, Missouri » Plant Genetics Research » Research » Publications at this Location » Publication #413871
Research Project: Adaptation of Grain Crops to Varying Environments Including Climates, Stressors, and Human Uses

Location: Plant Genetics Research

Title: Analysis of polar and nonpolar small plant growth hormones and quantification by LC/MS

Author
item DILKES, BRIAN - Purdue University
item Best, Norman

Submitted to: Cold Spring Harbor Protocols
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/21/2025
Publication Date: 4/17/2025
Citation: Dilkes, B.P., Best, N.B. 2025. Analysis of polar and non-polar small plant growth hormones and quantification by LC/MS. Cold Spring Harbor Protocols. https://doi.org/10.1101/pdb.prot108647.
DOI: https://doi.org/10.1101/pdb.prot108647

Interpretive Summary: Plant hormones are small metabolites that regulate all aspects of plant growth and development. They accumulate at very low concentrations in maize tissue, therefore requiring sensitive protocols to measure their abundance. We have developed a method to measure six classes of plant hormones in a single experiment. This manuscript provides a step by step method to extract and measure plant hormones by liquid chromatography and mass spectrometry. We provide the masses and retention times for a number of metabolites in each of the six classes of plant hormones. This protocol provides researchers the means to measure plant hormones to further validate genetic and biochemical studies.

Technical Abstract: Plant hormones are small metabolites that regulate all aspects of plant growth and development, including plant defense. The detection and quantification of these hormones are critical to understanding the mechanism of growth regulation in plants. In maize, a wealth of genetic resources has enabled progress on elucidating the genetic mechanisms underlying plant growth. Biochemical studies of growth in maize can provide insight into the physiological mechanisms of growth control by measuring endogenous levels of plant hormones, and this knowledge would be enhanced by the development of a method to analyze several hormones in a single small sample of tissue. We provide here a simple protocol to extract and accurately quantify six classes of plant hormones in a single liquid chromatography/mass spectrometry injection run using maize tissues. Those hormones include abscisic acid (ABA), 1-aminocyclopropane-1-carboxylate (ACC), gibberellic acid (GA), 3-indoleacetic acid (IAA), jasmonic acid (JA), and salicylic acid (SA), as well as an accumulated phytoanticipin of maize, 24-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA), which influences the levels of IAA.