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Dormancy termination of western white pine (Pinus monticola Dougl. Ex D. Don) seeds is associated with changes in abscisic acid metabolism

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Abstract

Western white pine (Pinus monticola) seeds exhibit deep dormancy at maturity and seed populations require several months of moist chilling to reach their uppermost germination capacities. Abscisic acid (ABA) and its metabolites, phaseic acid (PA), dihydrophaseic acid (DPA), 7′-hydroxy ABA (7′OH ABA) and ABA-glucose ester (ABA-GE), were quantified in western white pine seeds during dormancy breakage (moist chilling) and germination using an HPLC–tandem mass spectrometry method with multiple reaction monitoring and internal standards incorporating deuterium-labeled analogs. In the seed coat, ABA and metabolite levels were high in dry seeds, but declined precipitously during the pre-moist-chilling water soak to relatively low levels thereafter. In the embryo and megagametophyte, ABA levels decreased significantly during moist chilling, coincident with an increase in the germination capacity of seeds. ABA catabolism occurred via several routes, depending on the stage and the seed tissue. Moist chilling of seeds led to increases in PA and DPA levels in both the embryo and megagametophyte. Within the embryo, 7′OH ABA and ABA-GE also accumulated during moist chilling; however, 7′OH ABA peaked early in germination. Changes in ABA flux, i.e. shifts in the ratio between biosynthesis and catabolism, occurred at three distinct stages during the transition from dormant seed to seedling. During moist chilling, the relative rate of ABA catabolism exceeded ABA biosynthesis. This trend became even more pronounced during germination, and germination was also accompanied by a decrease in the ABA catabolites DPA and PA, presumably as a result of their further metabolism and/or leaching/transport. The transition from germination to post-germinative growth was accompanied by a shift toward ABA biosynthesis. Dormant imbibed seeds, kept in warm moist conditions for 30 days (after an initial 13 days of soaking), maintained high ABA levels, while the amounts of PA, 7′OH ABA, and DPA decreased or remained at steady-state levels. Thus, in the absence of conditions required to break dormancy there were no net changes in ABA biosynthesis and catabolism.

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Abbreviations

ABA:

abscisic acid

ABA-GE:

abscisic acid glucose ester

DPA:

dihydrophaseic acid

7′OH ABA:

7′-hydroxy abscisic acid

8′OH ABA:

8′-hydroxy abscisic acid

MRM:

multiple reaction monitoring

PA:

phaseic acid

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Acknowledgements

We thank Dave Kolotelo and Dean Christianson of the BC Ministry of Forests, BC, Canada for helping obtain mature seed of western white pine and Mary Loewen and Tim Squires for their technical help at PBI, Saskatoon, Canada. We are also grateful to Marek Galka (PBI) for the synthesis of ABA-GE and 7′OH ABA and to Irina Zaharia (PBI) for the PA and DPA in labeled and unlabeled forms. This research was supported by Natural Sciences and Engineering Research Council of Canada (Strategic) and BC Ministry of Advanced Education, Training and Technology grants awarded to A.R.K and a Protein Engineering Network of Centres of Excellence (PENCE) grant awarded to A.R.K., S.R.A., A.R.S.R. and A.J.C., and other investigators. J.A.F. is a recipient of a Science Horizons Internship.

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Authors and Affiliations

  1. Department of Biological Sciences, Simon Fraser University, Burnaby, BC, V5A 1S6, Canada

    J. Allan Feurtado & Allison R. Kermode

  2. Plant Biotechnology Institute, National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK, S7N 0W9, Canada

    Stephen J. Ambrose, Adrian J. Cutler, Andrew R. S. Ross & Suzanne R. Abrams

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  1. J. Allan Feurtado
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  2. Stephen J. Ambrose
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  3. Adrian J. Cutler
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  4. Andrew R. S. Ross
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  5. Suzanne R. Abrams
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  6. Allison R. Kermode
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Correspondence to Allison R. Kermode.

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Feurtado, J.A., Ambrose, S.J., Cutler, A.J. et al. Dormancy termination of western white pine (Pinus monticola Dougl. Ex D. Don) seeds is associated with changes in abscisic acid metabolism. Planta 218, 630–639 (2004). https://doi.org/10.1007/s00425-003-1139-8

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