Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 131, Issue 3, pp 459–470 | Cite as

A simple and efficient method for analysis of plant growth regulators: a new tool in the chest to combat recalcitrance in plant tissue culture

  • Lauren A. E. Erland
  • Mukund R. Shukla
  • W. Broc Glover
  • Praveen K. Saxena
Original Article


This report presents a simple, rapid and accessible validated method for quantification of eight major plant growth regulators (PGR): cytokinins (6-(γ,γ-dimethylallylamino)purine (2-iP), benzylaminopurine (BA) and zeatin), auxin (indole-3-acetic acid; IAA), jasmonic acid (JA), salicylic acid (SA), gibberellic acid (GA3) and abscisic acid (ABA) by liquid chromatography mass spectrometry. This method was tested in eight species including agricultural, ornamental and medicinal species: St. John’s wort, African violet, banana, American elm, tobacco, potato, sweet wormwood, and fennel. The method has good reproducibility and good sensitivity with %RSD (percent relative standard deviation) between 1 and 10% for all matrices and recovery values of 89 to 118% for all analytes. Method detection limits were 50.65 ng/g, 203.4 ng/g, 50.65, ng/g, 50.65 ng/g, 203.4 ng/g, 12.7 ng/g, 193 pg/g and 3.08 ng/g, for SA, IAA, zeatin, JA, GA3, ABA, 2-iP, and BA, respectively. Our results with a range of plant species show that this method represents a simple, low-cost method for analysis of PGRs, and may also serve as an useful starting point for the analysis of other related PGRs, as demonstrated by inclusion of the SA derivative, acetylsalicylic acid, and the JA derivatives: 12-oxo-phytodienoic acid and JA-isoleucine. The efficiency of this method will enable its incorporation into the plant tissue culture work flow and through characterization of endogenous PGR levels, will allow for improved method development for recalcitrant species facilitating fundamental and applied studies in plant morphogenesis, propagation and conservation.


Auxin Cytokinin Abscisic acid Jasmonate Salicylate Recalcitrance 



Abscisic acid


Acetylsalicylic acid




2,4-Dichlorophenoxyacetic acid


Gibberellic acid


Indole-3-acetic acid




Jasmonic acid


Jasmonic acid isoleucine




Mass spectrometry


12-Oxo-phytodienoic acid


p-Chlorophenoxyisobutyric acid


Plant growth regulator


Percent relative standard deviation


Salicylic acid


Single ion recording


St. John’s wort


Single lab validation


2,3,5-Triiodobenzoic acid




Ultra-performance liquid chromatography



The authors gratefully acknowledge the financial support of this work by the National Sciences and Engineering Research Council (NSERC) of Canada [Grant Number 46741] and the Gosling Research Institute for Plant Preservation (GRIPP) [Grant Number 050294].

Author contributions

LAEE participated in conception and design, data acquisition, analysis and interpretation, MRS participated in conception and design, WBG participated in conception and design and data analysis and PKS participated in conception and design and data interpretation. All authors participated in manuscript preparation and gave final approval of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflict of interest.


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Copyright information

© Springer Science+Business Media B.V. 2017

Authors and Affiliations

  1. 1.Department of Plant Agriculture, Gosling Research Institute for Plant PreservationUniversity of GuelphGuelphCanada

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