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

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Abstract

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.

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Acknowledgements

We thank Stephen Kresovich for providing the sorghum seed grown in this field experiment.

Funding

This research was supported by the Office of Science (BER), US Department of Energy (DE-SC0014395).

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Authors and Affiliations

  1. Department of Horticulture and Landscape Architecture, Colorado State University, 1173 Campus Delivery, Fort Collins, CO, 80523-1173, USA

    Amy M. Sheflin & Jessica E. Prenni

  2. Metabolomics Core Facility, University of California, Riverside, Riverside, CA, 92521, USA

    Jay S. Kirkwood

  3. Proteomics and Metabolomics Core Facility, Colorado State University, Fort Collins, CO, 80523, USA

    Lisa M. Wolfe & Corey D. Broeckling

  4. Department of Bioagricultural Sciences and Pest Management, Colorado State University, Fort Collins, CO, 80523, USA

    Courtney E. Jahn

  5. Department of Agronomy and Horticulture, University of Nebraska – Lincoln, Lincoln, NE, 68583-0915, USA

    Daniel P. Schachtman

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  1. Amy M. Sheflin
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Correspondence to Jessica E. Prenni.

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Published in the topical collection Young Investigators in (Bio-)Analytical Chemistry with guest editors Erin Baker, Kerstin Leopold, Francesco Ricci, and Wei Wang.

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Sheflin, A.M., Kirkwood, J.S., Wolfe, L.M. et al. High-throughput quantitative analysis of phytohormones in sorghum leaf and root tissue by ultra-performance liquid chromatography-mass spectrometry. Anal Bioanal Chem 411, 4839–4848 (2019). https://doi.org/10.1007/s00216-019-01658-9

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  • DOI: https://doi.org/10.1007/s00216-019-01658-9

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