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Comparison of endogenous cytokinins, ABA and metabolites during female cone bud differentiation in low and high cone-producing genotypes of lodgepole pine

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Abstract

In lodgepole pine (Pinus contorta Dougl. ex Loud. var. latifolia Engelm.), cone bud initiation within long-shoot buds varies according to genotype. We chose to study hormone profiles of two genotypes that differed significantly in cone yield. Phytohormone profiles were established by high performance liquid chromatography–electrospray ionization tandem mass spectrometry in multiple reaction monitoring mode with samples collected from genotypes 299 and 233, the typically high and low cone producers. Generally, concentrations of trans-zeatin-O-glucoside were higher in genotype 299, whereas dihydrozeatin concentrations were higher in genotype 233. Both isopentenyl adenine and isopentenyl adenosine were present at higher concentrations in genotype 233. The ratio of total quantifiable zeatin (Z)-type cytokinins to isopentenyl (iP)-type cytokinins was approximately threefold higher in genotype 299 during female cone bud differentiation. In genotype 299, ABA concentration was significantly lower than in genotype 233 on the first sampling date, while the phaseic acid concentration was lower consistently throughout the period investigated. Dihydrophaseic acid was present in low concentrations in most samples of genotype 233, but was not quantifiable in genotype 299. Our study reveals that long-shoot buds of the high cone-producing genotype had higher ratios of Z-type cytokinins to iP-type cytokinins than were found in the low cone-producing genotype. High cone-producing buds also contained less ABA, phaseic acid and dihydrophaseic acid during female cone bud differentiation.

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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

t-Z:

Trans-zeatin

t-ZR:

Trans-zeatin riboside

t-Z-O-Glu:

Trans-zeatin-O-glucoside

dhZ:

Dihydrozeatin

dhZR:

Dihydrozeatin riboside

iP:

Isopentenyl

2iP:

Isopentenyl adenine

iPA:

Isopentenyl adenosine

References

  • Abrams SR, Nelson K, Ambrose SJ (2003) Deuterated abscisic acid analogs for mass spectrometry and metabolism studies. J Label Comp Radiopharm 46:273–283

    Article  CAS  Google Scholar 

  • Bao R-Y, Zheng C-X (2005) Content changes of several endogenous plant hormones in female-sterile Pinus tabulaeformis Carr. For Stud China 7:16–19

    Article  Google Scholar 

  • Bollmark M, Chen HJ, Moritz T, Eliason L (1995) Relations between cytokinin level, bud development and apical control in Norway spruce, Picea abies. Physiol Plant 95:563–568

    Article  CAS  Google Scholar 

  • Bonnet-Masimbert M, Zaerr JB (1987) 2. The role of plant growth regulators in promotion of flowering. Plant Growth Regul 6:13–35

    Article  CAS  Google Scholar 

  • Bravo LA, Zúñiga GE, Alberdi M, Corcuera LJ (1998) The role of ABA in freezing tolerance and cold acclimation in barley. Physiol Plant 103:17–23

    Article  CAS  Google Scholar 

  • Chen HJ, Bollmark M, Eliason L (1996) Evidence that cytokinin controls bud size and branch form in Norway spruce. Physiol Plant 98:612–618

    Article  CAS  Google Scholar 

  • Chiwocha SDS, Abrams SR, Ambrose SJ, Cutler AJ, Loewen M, Ross ARS, Kermode AR (2003) A method for profiling classes of plant hormones and their metabolites using liquid chromatography-electrospray ionization tandem mass spectrometry: analysis of hormone regulation of thermodormancy of lettuce (Lactuca sativa L.) seeds. Plant J 35:405–417

    Article  PubMed  CAS  Google Scholar 

  • Chiwocha SDS, Cutler AJ, Abrams SR, Ambrose SJ, Yang J, Ross ARS, Kermode AR (2005) The etr1-2 mutation in Arabidopsis thaliana affects the abscisic acid, auxin, cytokinin and gibberellin metabolic pathways during maintenance of seed dormancy, moist-chilling and germination. Plant J 42:35–48

    Article  PubMed  CAS  Google Scholar 

  • Corbesier L, Prinsen E, Jacqmard A, Lejeune P, Van Onckelen H, Périlleux C, Bernier G (2003) Cytokinin levels in leaves, leaf exudate and shoot apical meristem of Arabidopsis thaliana during floral transition. J Exp Bot 54:2511–2517

    Article  PubMed  CAS  Google Scholar 

  • Cutler AJ, Krochko JE (1999) Formation and breakdown of ABA. Trends Plant Sci 4:472–478

    Article  PubMed  Google Scholar 

  • Fiehn O, Kopka J, Dörmann P, Altmann T, Trethewey RN, Willmitzer L (2000) Metabolite profiling for plant functional genomics. Nat Biotech 18:1157–1161

    Article  CAS  Google Scholar 

  • Frugis G, Giannino D, Mele G, Nicolodi C, Chiappetta A, Bitonti MB, Innocenti AM, Dewitte W, Van Onckelen H, Mariotti D (2001) Overexpression of KNAT1 in lettuce shifts leaf determinate growth to a shoot-like indeterminate growth associated with an accumulation of isopentenyl-type cytokinins. Plant Physiol 126:1370–1380

    Article  PubMed  CAS  Google Scholar 

  • Hirose N, Takei K, Kuroha T, Kamada-Nobusada T, Hayashi H, Sakakibara H (2008) Regulation of cytokinin biosynthesis, compartmentalization and translocation. J Exp Bot 59:75–83

    Article  PubMed  CAS  Google Scholar 

  • Imbault N, Tardieu I, Joseph C, Zaerr JB, Bonnet-Masimbert M (1988) Possible role of isopentenyladenine and isopentenyladenosine in flowering of Pseudotsuga menziesii: endogenous variations and exogenous applications. Plant Physiol Biochem 26:289–295

    CAS  Google Scholar 

  • Kakimoto T (2003) Perception and signal transduction of cytokinins. Ann Rev Plant Biol 54:605–627

    Article  CAS  Google Scholar 

  • Kong L, von Aderkas P (2007) Genotype effects on ABA consumption and somatic embryo maturation in interior spruce (Picea glauca x engelmanni). J Exp Bot 58:1525–1531

    Article  PubMed  CAS  Google Scholar 

  • Kong L, Abrams SR, Owen S, Graham H, von Aderkas P (2008) Phytohormones and their metabolites during long shoot development in Douglas-fir following cone induction by gibberellin injection. Tree Physiol 28:1357–1364

    PubMed  CAS  Google Scholar 

  • Kong L, Abrams SR, Owen S, Van Niejenhuis A, von Aderkas P (2009) Dynamic changes in concentrations of auxin, cytokinin, ABA and selected metabolites in multiple genotypes of Douglas-fir (Pseudotsuga menziesii) during a growing season. Tree Physiol 29:183–190

    Article  PubMed  CAS  Google Scholar 

  • Kong L, von Aderkas P, Abrams SR (2011) A metabolomic plant hormone analysis of cone induction in conifers. In: Yaish MW (ed) The flowering process and its control in plants: gene expression and hormone interaction. Research Signpost, Kerala, pp 101–122

    Google Scholar 

  • Kumar S, Kaur G, Nayyar H (2008) Exogenous application of abscisic acid improves cold tolerance in chickpea (Cicer arietinum L.). J Agr Crop Sci 194:449–456

    Article  CAS  Google Scholar 

  • Lejeune P, Bernier G, Requier M-C, Kinet J-M (1994) Cytokinins in phloem and xylem saps of Sinapis alba during floral induction. Physiol Plant 90:522–528

    Article  CAS  Google Scholar 

  • Longman KA (1983) Effects of gibberellin, genotype and environment on cone initiation, shoot growth and branching in Pinus contorta. Ann Bot 50:247–257

    Google Scholar 

  • McMullan EE (1980) Effect of applied growth regulators on cone production in Douglas-fir, and relation of endogenous growth regulators to cone production capacity. Can J For Res 10:405–422

    Article  CAS  Google Scholar 

  • Menéndez V, Revilla MA, Fal MA, Fernández H (2009) The effect of cytokinins on growth and sexual organ development in the gametophyte of Blechnum spicant L. Plant Cell Tissue Org Cult 96:245–250

    Article  Google Scholar 

  • Morris JW, Doumas P, Morris RO, Zaerr JB (1990) Cytokinins in vegetative and reproductive buds of Pseudotsuga menziesii. Plant Physiol 93:67–71

    Article  PubMed  CAS  Google Scholar 

  • O’Reilly C, Owens JN (1987) Long-shoot bud development, shoot growth, and foliage production in provenances of lodgepole pine. Can J For Res 17:1421–1433

    Article  Google Scholar 

  • O’Reilly C, Owens JN (1988) Reproductive growth and development in seven provenances of lodgepole pine. Can J For Res 18:43–53

    Article  Google Scholar 

  • Pharis RP, King RW (1985) Gibberellins and reproductive development in seed plants. Ann Rev Plant Physiol 36:517–568

    Article  CAS  Google Scholar 

  • Philippe G, Baldet P, Héois B, Ginisty C (2006) Reproduction sexuée des conifères et production de semences en vergers à graines. Editeur Cemagref, Paris

    Google Scholar 

  • Pilate G, Sotta B, Maldiney R, Bonnet-Masimbert M, Miginiac E (1990) Endogenous hormones in Douglas-fir trees induced to flower by gibberellin A4/7 treatment. Plant Physiol Biochem 28:359–366

    CAS  Google Scholar 

  • Romanov GA, Lomin SN, Schmülling T (2006) Biochemical characteristics and ligand-binding properties of Arabidopsis cytokinin receptor AHK3 compared to CRE1/AHK4 as revealed by a direct binding assay. J Exp Bot 57:4051–4058

    Article  PubMed  CAS  Google Scholar 

  • Ross SD (1983) Enhancement of shoot elongation in Douglas-fir by gibberellin A4/7 and its relation to the hormonal promotion of flowering. Can J For Res 13:986–994

    Article  CAS  Google Scholar 

  • Ross ARS, Ambrose SJ, Cutler AJ, Feurtado JA, Kermode AR, Nelson K, Zhou R, Abrams SR (2004) Determination of endogenous and supplied deuterated abscisic acid in plant tissues by high performance liquid chromatography-electrospray ionization tandem mass spectrometry with multiple reaction monitoring. Anal Biochem 329:324–333

    Article  PubMed  CAS  Google Scholar 

  • Sakakibara H (2006) Cytokinins: activity, biosynthesis, and translocation. Ann Rev Plant Biol 57:431–449

    Article  CAS  Google Scholar 

  • Smith R, Greenwood M (1995) Effects of gibberellin A4/7, root pruning and cytokinins on seed and pollen cone production in black spruce (Picea mariana). Tree Physiol 15:457–465

    PubMed  CAS  Google Scholar 

  • Spíchal L, Rakova NY, Riefler M, Mizuno T, Romanov GA, Strnad M, Schmülling T (2004) Two cytokinin receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, differ in their ligand specificity in a bacterial assay. Plant Cell Physiol 45:1299–1305

    Article  PubMed  Google Scholar 

  • Tompsett PB (1977) Studies of growth and flowering in Picea sitchensis (Bong.) Carr. I. Effect of growth regulator application to mature scions on seedling rootstocks. Ann Bot 41:1171–1178

    CAS  Google Scholar 

  • Valdés AE, Centeno ML, Espinel S, Fernández B (2002) Could plant hormones be the basis of maturation indices in Pinus radiata? Plant Physiol Biochem 40:211–216

    Article  Google Scholar 

  • Valdés AE, Fernández B, Centeno ML (2003) Alterations in endogenous levels of cytokinins following grafting of Pinus radiata support ratio of cytokinins as an index of ageing and vigour. J Plant Physiol 160:1407–1410

    Article  PubMed  Google Scholar 

  • Valdés AE, Galuszka P, Fernández B, Centeno ML, Frébort I (2007) Developmental stage as a possible factor affecting cytokinin content and cytokinin dehydrogenase activity in Pinus sylvestris. Biol Plant 51:193–197

    Article  Google Scholar 

  • von Aderkas P, Kong L, Carlson M (2007) One bud, two bud, three bud, four: making lodgepole pine buds count. TICtalk 8:4–6

    Google Scholar 

  • Wakushima S (2004) Promotion of female strobili flowering and seed production in two Japanese pine species by 6-benzylaminopurine (BAP) paste application in a field seed orchard. J Plant Growth Regul 23:135–145

    Article  CAS  Google Scholar 

  • Weiss D, Ori N (2007) Mechanisms of cross talk between gibberellin and other hormones. Plant Physiol 144:1240–1246

    Article  PubMed  CAS  Google Scholar 

  • Zaharia LI, Galka MM, Ambrose SJ, Abrams SR (2005) Preparation of deuterated abscisic acid metabolites for use in mass spectrometry and feeding studies. J Label Comp Radiopharm 48:435–445

    Article  CAS  Google Scholar 

  • Zhang H, Horgan KJ, Reynolds PHS, Norris GE, Jameson PE (2001) Novel cytokinins: the predominant forms in adult buds of Pinus radiata. Physiol Plant 112:127–134

    Article  PubMed  CAS  Google Scholar 

  • Zhang H, Horgan KJ, Reynolds PHS, Jameson PE (2003) Cytokinins and bud morphology in Pinus radiate. Physiol Plant 117:264–269

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from the British Columbia Forest Investment Account, Forest Genetics Conservation and Management Program and the Natural Sciences and Engineering Research Council of Canada Discovery Program (PvA). The authors would like to express great appreciation to the project steering committee of cone induction and the following individuals for their help in this study: Jack Woods (FGCBC and SelectSeed Co. Ltd.), Tim Lee and Dan Gaudet (Vernon Seed Orchard Company) and, Samir Demdoum, Sébastien Bonthoux and Genoa Barchet (University of Victoria). The authors also wish to thank the following members of the Hormone Profiling Lab at NRC-PBI: Monika Lafond and Vera Čekić for the preparation of standard solutions and for the extraction and work-up of all samples for analysis, and Dr. Irina Zaharia for her assistance in this work.

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Authors and Affiliations

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

    Lisheng Kong & Patrick von Aderkas

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

    Stacey J. Owen & Suzanne R. Abrams

  3. Vernon Seed Orchard Company, 6555 Bench Row Road, Vernon, BC, V1H 1G2, Canada

    Tia Wagner

  4. Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA, 30602, USA

    Lisheng Kong

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  1. Lisheng Kong
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  2. Patrick von Aderkas
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  5. Suzanne R. Abrams
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Correspondence to Lisheng Kong.

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Communicated by R. Aloni.

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Kong, L., von Aderkas, P., Owen, S.J. et al. Comparison of endogenous cytokinins, ABA and metabolites during female cone bud differentiation in low and high cone-producing genotypes of lodgepole pine. Trees 25, 1103–1110 (2011). https://doi.org/10.1007/s00468-011-0585-3

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