Skip to main content
SpringerLink
Log in

Gas chromatography-mass spectrometry evidence for several endogenous auxins in pea seedling organs

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Qualitative analysis by gas chromatography-mass spectrometry (GC-MS) of the auxins present in the root, cotyledons and epicotyl of 3-dold etiolated pea (Pisum sativum L., cv. Alaska) seedlings has shown that all three organs contain phenylacetic acid (PAA), 3-indoleacetic acid (IAA) and 4-chloro-3-indoleacetic acid (4Cl-IAA). In addition, 3-indolepropionic acid (IPA) was present in the root and 3-indolebutyric acid (IBA) was detected in both root and epicotyl. Phenylacetic acid, IAA and IPA were measured quantitatively in the three organs by GC-MS-single ion monitoring, using deuterated internal standards. Levels of IAA were found to range from 13 to 115 pmol g-1 FW, while amounts of PAA were considerably higher (347–451 pmol g-1 FW) and the level of IPA was quite low (5 pmol g-1 FW). On a molar basis the PAA:IAA ratio in the whole seedling was approx. 15:1.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

IAA:

3-indoleacetic acid

4Cl-IAA:

4-chloro-3-indoleacetic acid

IBA:

3-indolebutyric acid

IPA:

3-indolepropionic acid

PAA:

phenylacetic acid

GC-MS:

gas chromatography-mass spectrometry

HPLC:

high-performance liquid chromatography

PFB:

pentafluorobenzyl ester

PFBBr:

pentafluorobenzyl bromide

SIM:

single-ion monitoring

TMSI:

trimethylsilyl ester

References

  • Akiyama, M., Sakurai, N., Kuraishi, S. (1983) A simplified method for the quantitative determination of indoleacetic acid by high performance liquid chromatography with a fluorimetric detector. Plant Cell Physiol. 24, 1431–1439

    Google Scholar 

  • Allen, J.R.F., Rivier, L., Pilet, P.E. (1982) Quantification of indol-3-yl acetic acid in pea and maize seedlings by gas chromatography-mass spectrometry. Phytochemistry 21, 525–530

    Google Scholar 

  • Badenoch-Jones, J., Summons, R.E. Rolfe, B.G., Letham, D.S. (1984) Phytohormones, Rhizobium mutants and nodulation in legumes. IV. Auxin metabolites in pea root nodules. J. Plant Growth Regul. 3, 23–29

    Google Scholar 

  • Bandurski, R.S., Schulze, A. (1977) Concentration of indole-3-acetic acid and its derivatives in plants. Plant Physiol. 60, 211–213

    Google Scholar 

  • Bayer, M.H. (1969) Gas chromatographic analysis of acidic indole auxins in Nicotiana. Plant Physiol. 44, 267–271

    Google Scholar 

  • Blau, K., King, G.S. (1978) Handbook of derivatives for chromatography. Heydon, London, UK

    Google Scholar 

  • Blommaert, K.L.J. (1954) Growth-and inhibiting-substances in relation to the rest period of the potato tuber. Nature 174, 970–972

    Google Scholar 

  • Caruso, J.L., Zeisler, C.S. (1983) Indole-3-acetic acid in Douglas fir seedlings: a reappraisal. Phytochemistry 22, 589–590

    Google Scholar 

  • Davis, L.A., Heinz, D.E., Addicott, F.T. (1968) Gas-liquid chromatography of trimethylsilyl derivatives of abscisic acid and other plant hormones. Plant Physiol. 43, 1389–1394

    Google Scholar 

  • Dreher, T.W., Poovaiah, B.W. (1982) Changes in auxin content during development in strawberry fruits. J. Plant Growth Regul. 1, 267–276

    Google Scholar 

  • Ehmann, A. (1977) The Van Urk-Salkowski reagent — a sensitive and specific chromogenic reagent for silica gel thin-layer chromatographic detection and identification of indole derivatives. J. Chromatogr. 132, 267–276

    Google Scholar 

  • Engvild, K.C., Egsgaard, H., Larsen, E. (1978) Gas chromatographic-mass spectrometric identification of 4-chloroindolyl-3-acetic acid methyl ester in immature green peas Physiol. Plant. 42, 365–368

    Google Scholar 

  • Engvild, K.C., Egsgaard, H., Larsen, E. (1980) Determination of 4-chloroindole-3-acetic acid methyl ester in Lathyrus, Vicia and Pisum by gas chromatography. Physiol. Plant. 48, 499–503

    Google Scholar 

  • Epstein, E., Cohen, J.D. (1981) Microscale preparation of pentafluorobenzyl esters. Electron-capture gas chromatographic detection of indole-3-acetic acid from plants. J. Chromatogr. 209, 413–420

    Google Scholar 

  • Feldman, L. (1980) Auxin biosynthesis and metabolism in isolated roots of Zea mays. Physiol. Plant. 49, 145–150

    Google Scholar 

  • Fregeau, J.A., Wightman, F. (1983) Natural occurrence and biosynthesis of auxins in chloroplast and mitochondrial fractions from sunflower leaves. Plant Sci. Lett. 32, 23–24

    Google Scholar 

  • Gianfagna, T.J., Davies, P.J. (1980) N-Benzoylaspartate and N-phenylacetylaspartate from pea seeds. Phytochemistry 19, 959–961

    Google Scholar 

  • Liu, S.-T., Katz, C.D., Knight, C.A. (1978) Indole-3-acetic acid synthesis in tumerous and nontumerous species of Nicotiana. Plant Physiol. 61, 743–747

    Google Scholar 

  • Magnus, V., Bandurski, R.S., Schulze, A. (1980) Synthesis of 4,5,6,7 and 2,4,5,6,7 deuterium-labelled indole-3-acetic acid for use in mass spectrometric assays. Plant Physiol. 66, 775–781

    Google Scholar 

  • Markham, G., Lichty, D.G., Wightman, F. (1980) Comparative study of derivatisation procedures for the quantitative determination of the auxin, phenylacetic acid, by gas chromatography. J. Chromatogr. 192, 429–433

    Google Scholar 

  • Martin, H.V., Elliott, M.C., Wangermann, E., Pilet, P.-E., (1978) Auxin gradient along the root of the maize seedling. Planta 141, 179–181

    Google Scholar 

  • Marumo, S., Hattori, H., Abe, H., Munakata, K. (1968) Isolation of 4-chlorindolyl-3-acetic acid from immature seeds of Pisum sativum. Nature 219, 959–960

    Google Scholar 

  • McDougall, J., Hillman, J.R. (1980) Derivatives of indole-3-acetic acid for SIM GC-MS studies. Z. Pflanzenphysiol. 98, 89–93

    Google Scholar 

  • Mulvena, D.P., Slaytor, M. (1982) Separation of tryptophan derivatives in Phalaris aquatica by thin-layer chromatography. J. Chromatogr. 245, 155–157

    Google Scholar 

  • Pless, T., Böttger, M., Hedden, P., Graebe, J. (1984) Occurrence of 4-Cl-indoleacetic acid in broad beans and correlation of its levels with seed development. Plant Physiol. 74, 320–323

    Google Scholar 

  • Powell, L.E. (1964) Preparation of indole extracts from plants for gas chromatography and spectrophotofluorometry. Plant Physiol. 39, 836–842

    Google Scholar 

  • Rivier, L., Pilet, P.-E. (1974) Indolyl-3-acetic acid in cap and apex of maize roots: identification and quantification by mass fragmentography. Planta 74, 107–112

    Google Scholar 

  • Sandberg, G., Jensen, E., Crozier, A. (1984) Analysis of 3-indolecarboxylic acid in Pinus sylvestris needles. Phytochemistry 23, 99–102

    Google Scholar 

  • Savidge, R.A., Heald, J.K., Wareing, P.F. (1982) Non-uniform distribution and seasonal variation of endogenous indol-3yl-acetic acid in the cambial region of Pinus contorta Dougl. Planta 155, 89–92

    Google Scholar 

  • Schlenk, H., Gellermann, J.L. (1960) Esterification of fatty acids with diazomethane on a small scale. Anal. Chem. 30, 1412–1414

    Google Scholar 

  • Segal, L.M., Wightman, F. (1982) Gas chromatographic and GC-MS evidence for the occurrence of 3-indolylpropionic acid and 3-indolylacetic acid in seedlings of Cucurbita pepo. Physiol. Plant. 56, 367–370

    Google Scholar 

  • Wightman, F. (1977) Gas chromatographic identification and quantitative estimation of natural auxins in developing plant organs. In: Plant growth regulation, pp. 77–90, Pilet, P.E., ed. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Wightman, F., Lichty, D.L. (1982) Identification of phenylacetic acid as a natural auxin in the shoots of higher plants. Physiol. Plant. 55, 17–24

    Google Scholar 

  • Wightman, F., Thimann, K.V. (1980) Hormonal factors controlling the initiation and development of lateral roots I. Sources of primordia-inducing substances in the primary root of pea seedlings. Physiol. Plant. 49, 13–20

    Google Scholar 

  • Wightman, F., Schneider, E.A., Thimann, K.V. (1980) Hormonal factors controlling the initiation and development of lateral roots. II. Effects of exogenous growth factors on lateral root formations in pea roots. Physiol. Plant. 49, 304–314

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biology Department, Carleton University, K1S 5B6, Ottawa, Ontario, Canada

    Elnora A. Schneider & Frank Wightman

  2. Ottawa-Carleton Institute for Research and Graduate Studies in Chemistry, University of Ottawa, K1N 6N5, Ottawa, Ontario, Canada

    Clement W. Kazakoff

Authors
  1. Elnora A. Schneider
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Clement W. Kazakoff
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Frank Wightman
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schneider, E.A., Kazakoff, C.W. & Wightman, F. Gas chromatography-mass spectrometry evidence for several endogenous auxins in pea seedling organs. Planta 165, 232–241 (1985). https://doi.org/10.1007/BF00395046

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00395046

Key words

Search

Navigation