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Phenolics during early development of Betula pubescens seedlings: inhibition of phenylalanine ammonia lyase

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Abstract

We studied phenolic metabolism and plant growth in birch seedlings at the beginning of their development by inhibiting phenylalanine ammonia lyase (PAL), which is the first committed step in phenylpropanoid metabolism. Betula pubescens (Ehrh.) seeds were germinated in inhibitor-free media and the seedlings were transferred to hydroponic culture at the cotyledon stage. They were 6 days old at the start of the experiment, which lasted for 3 weeks. PAL activity was inhibited by three different concentrations of 2-aminoindane-2-phosphonic acid monohydrate (AIP) in the growing media. At the end of 3 weeks, phenolics in all plant parts (roots, stem, cotyledons, first, second and third true leaves) were determined. AIP inhibited strongly the accumulation of phenolic acids, salidroside, rhododendrins, ellagitannins and their precursors, flavan-3-ols, and soluble condensed tannins. The accumulation of lignin and flavonol glycoside derivatives was moderately inhibited. The accumulation of flavonol glycosides, such as quercetin glycosides and kaempferol glycosides, was not generally inhibited, even in leaves that emerged during the experiment, while the accumulation of insoluble condensed tannins was inhibited only slightly and not in all plant parts. This suggests that flavonol glycosides, which may have a UV-B protective role, and insoluble condensed tannins, which may have structural functions, are prioritized in seedling development. Inhibition of PAL with AIP decreased seedling growth and possible reasons for this are discussed.

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Abbreviations

AIP:

2-Aminoindane-2-phosphonic acid monohydrate

PAL:

Phenylalanine ammonia lyase

Phe:

l-Phenylalanine

Trp:

l-Tryptophan

Tyr:

l-Tyrosine

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Acknowledgments

We thank Outi Nousiainen, Sinikka Sorsa and Ria Paunonen for the help in the laboratory and during sampling. We also thank Matti Rousi at Finnish Forest Research Institute for the seeds, Kenneth Meaney, Riitta Tegelberg and two anonymous reviewers for valuable comments on the manuscript, Line Nybakken for valuable discussions and Jyrki Pusenius for statistical discussions. The Academy of Finland (Finnish Centre of Excellence Program 2000–2005, project no. 64308), Finnish Cultural Foundation and Niemi Foundation provided financial support for this study.

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Authors and Affiliations

  1. Natural Product Research Laboratories, Faculty of Biosciences, University of Joensuu, Box 111, 80101, Joensuu, Finland

    Sarita Keski-Saari, Maarit Falck & Riitta Julkunen-Tiitto

  2. Finnish Forest Research Institute, Joensuu Research Station, Yliopistonkatu 6, Box 68, 80101, Joensuu, Finland

    Jaakko Heinonen

  3. Department of Organic Chemistry, Faculty of Chemistry, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370, Wrocław, Poland

    Jerzy Zoń

Authors
  1. Sarita Keski-Saari
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  2. Maarit Falck
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  3. Jaakko Heinonen
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  4. Jerzy Zoń
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  5. Riitta Julkunen-Tiitto
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Corresponding author

Correspondence to Sarita Keski-Saari.

Additional information

Communicated by P. von Aderkas.

Electronic supplementary material

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468_2006_117_MOESM1_ESM.tif

Electronic Supplementary Material S1. Schematic presentation of the shikimic acid route leading to aromatic amino acids and the phenylpropanoid route leading to different kinds of phenolics. PAL inhibition by AIP is shown. (TIF 457 kb)

468_2006_117_MOESM2_ESM.doc

Electronic Supplementary Material S2. Concentrations of individual phenolic compounds (mean ± SE, mg g−1 dry weight) in seedlings of Betula pubescens when 6-day-old, and grown in different concentrations of AIP for three weeks. Statistical significances of responses of individual compounds to AIP in all plant parts (6-day-old seedlings not included in the analyses). Under each compound, the plant parts that do not contain the compound are excluded. P is the significance obtained by ANOVA or Kruskal–Wallis test (marked with d). P adj is the corresponding significance adjusted for multiple tests using Hochberg's procedure (Hochberg 1988), *P < 0.05, **P < 0.01, ***P < 0.001. Df 3 and df error 16, except in 3rd true leaves df 1, df error 8, n = 5. a = In six day-old seedlings true leaves had not emerged yet. b = Compounds in 3rd true leaves of AIP15 and AIP30 seedlings were not analyzed. c = (+)-Catechin was present in the cotyledons. Cot = cotyledons, 1st = 1st true leaves, 2nd = 2nd true leaves, 3rd = 3rd true leaves. (DOC 1,176 kb)

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Keski-Saari, S., Falck, M., Heinonen, J. et al. Phenolics during early development of Betula pubescens seedlings: inhibition of phenylalanine ammonia lyase. Trees 21, 263–272 (2007). https://doi.org/10.1007/s00468-006-0117-8

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  • DOI: https://doi.org/10.1007/s00468-006-0117-8

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