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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

Abstract

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.

Keywords

Auxin Cytokinin Abscisic acid Jasmonate Salicylate Recalcitrance 

Abbreviations

ABA

Abscisic acid

ASA

Acetylsalicylic acid

BA

6-Benzylaminopurine

2,4-D

2,4-Dichlorophenoxyacetic acid

GA3

Gibberellic acid

IAA

Indole-3-acetic acid

2-iP

6-(γ,γ-dimethylallylamino)purine

JA

Jasmonic acid

JA-Ile

Jasmonic acid isoleucine

MeOH

Methanol

MS

Mass spectrometry

OPDA

12-Oxo-phytodienoic acid

PCIB

p-Chlorophenoxyisobutyric acid

PGR

Plant growth regulator

%RSD

Percent relative standard deviation

SA

Salicylic acid

SIR

Single ion recording

SJW

St. John’s wort

SLV

Single lab validation

TIBA

2,3,5-Triiodobenzoic acid

TDZ

Thidiazuron

UPLC

Ultra-performance liquid chromatography

Notes

Acknowledgements

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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