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Phytohormone profiles of sterile Douglas-fir mutants and the responses to stem-injected gibberellins

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Significant differences in phytohormone profiles existed among the normal tree and two sterile mutant trees in interior Douglas-fir. Stem injection of gibberellin A4 and A7 induced male cone formation in one of the two genotypes of sterile trees.

Abstract

Two sterile genotypes of interior Douglas-fir (Pseudotsuga menziesii var. glauca (Beissn.) Franco) were analyzed. The phenotype of these mutant trees is characterized by compact internodes, stunted vegetative growth and an inability to initiate male or female cone buds. Long shoots were sampled at the stage prior to cone bud initiation in both sterile and wild type trees. Multiple phytohormones and their selected metabolites were analyzed using high-performance liquid chromatography–electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS). Compared with concentrations in wild-type samples, levels of total quantifiable cytokinins were about half (51.9%) in samples of sterile trees. In particular, levels of both trans-zeatin riboside and isopentenyl adenosine were lower. Concentrations of abscisic acid (ABA)-related metabolites, especially ABA-glucose ester, were higher (1.4-fold) in sterile trees. Total quantifiable gibberellins were about one-third (28.3%) lower in the sterile trees: concentrations of GA34 were significantly lower. There were no significant differences in concentrations of indole-3-acetic acid (IAA) or its conjugated form, IAA-aspartate. After stem injection of GA4+7, male cone formation was induced in one of the sterile genotypes. Gibberellin injection also caused the vegetative growth phenotype to change. Branches expanded to approximately five times their previous size in years past and their internodes showed more normal spacing. With their unique phytohormone profiles, these sterile mutants of Douglas-fir could provide a valuable research system for further exploration of molecular and physiological regulations during conifer reproduction.

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Abbreviations

HPLC-ESI-MS/MS:

High-performance liquid chromatography–electrospray ionization tandem mass spectrometry

MRM:

Multiple-reaction monitoring

GA:

Gibberellin

ABA:

Abscisic acid

PA:

Phaseic acid

DPA:

Dihydrophaseic acid

7′-OH ABA:

7′-Hydroxy ABA

neoPA:

neoPhaseic acid

ABA-GE:

Abscisic acid glucose ester

IAA:

Indole-3-acetic acid

IAA-Asp:

Indole-3-acetic acid aspartate

IAA-Glu:

Indole-3-acetic acid glutamate

t-Z:

trans-Zeatin

t-ZR:

trans-Zeatinriboside

c-ZR:

cis-Zeatinriboside

t-ZOG:

trans-Zeatin-O-glucoside

c-ZOG:

cis -Zeatin-O-glucoside

dhZ:

Dihydrozeatin

dhZR:

Dihydrozeatin-riboside

2iP:

Isopentenyl adenine

iPA:

Isopentenyl adenosine

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Acknowledgements

We gratefully acknowledge the financial support of the British Columbia Ministry of Forests, Lands and Natural Resource Operations, as well as the Forest Genetics Council of British Columbia. This project was also supported by the Discovery Grant Program of the Natural Sciences and Engineering Research Council of Canada (PvA). The authors would like to express great appreciation to Olivia Goede, Meaghan Duke (University of Victoria), Dr. Xiumei Han, Monika Lafond and Vera Čekić (NRC) for their assistance in sample collection and hormone profiling.

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Authors and Affiliations

  1. Centre for Forest Biology, Department of Biology, University of Victoria, 3800 Finnerty Rd, Victoria, BC, V8W 3N5, Canada

    Lisheng Kong & Patrick von Aderkas

  2. Beijing Advanced Innovation Center for Tree Breeding by Molecular Design, College of Biological Science and Biotechnology, Beijing Forestry University, 100083, Beijing, People’s Republic of China

    Lisheng Kong

  3. Kalamalka Research Station, 3401 Reservoir Rd, Vernon, BC, V1B 2C7, Canada

    Barry Jaquish

  4. Aquatic and Crop Resource Development, National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada

    L. Irina Zaharia

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  1. Lisheng Kong
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  2. Barry Jaquish
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  3. L. Irina Zaharia
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  4. Patrick von Aderkas
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Contributions

Lisheng Kong: experiment design, treatment application, data analysis and MS preparation; Barry Jaquish: plant material preparation and MS revision; L. Irina Zaharia: phytohormone analysis and MS revision; Patrick von Aderkas: experiment design and MS preparation.

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Correspondence to Lisheng Kong.

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Communicated by Lee Kalcsits.

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Kong, L., Jaquish, B., Zaharia, L.I. et al. Phytohormone profiles of sterile Douglas-fir mutants and the responses to stem-injected gibberellins. Trees 35, 1961–1969 (2021). https://doi.org/10.1007/s00468-021-02163-2

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