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Immunocytochemical localization of phenolic compounds in pollen walls using antibodies againstp-coumaric acid coupled to bovine serum albumin

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Summary

Two different antibodies against bovine serum albumin (BSA)-p-coumaric acid-conjugates were produced and used to localize phenolic compounds in exines of pollen from different species,p-Coumaric acid (pC) was coupled to BSA either via the carboxy group (BSA-pC) or directly to the aromatic ring system (BSA-azopC). The polyclonal antibodies raised in rabbits were characterized by ELISA with homologous and heterologous antigens using turkey ovalbumin as carrier protein. The results showed that the two immune sera directed against BSA-pC and BSA-azo-pC, respectively, were specific forp-coumaric acid and structurally similar compounds. Only a very poor binding by acetic acid-ovalbumin-conjugates and no binding by turkey ovalbumin was detectable. The antibodies reacted with partially purified pollen walls and with highly purified exines. The intensity of the immune reaction was proved to be dependent upon the pollen source and the preparation of the pollen walls. Using light and electron microscopy, it was shown for the first time that, in the exines ofCucurbita maxima, antibody binding was predominantly observed in the region of the germ pore apertures, the outer foot layers, and in the micro- and macrospines. We conclude from this and other earlier published data that phenols are important structural compounds of sporopollenin.

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Abbreviations

AA:

acetic acid

BA:

benzoic acid

BSA:

bovine serum albumin

BSA-azo-pC:

p-coumaric acid coupled in meta position to BSA by a diazo reaction

BSA-azo-pC I:

immune serum against BSA-azo-pC

BSA-pC:

p-coumaric acid coupled to BSA via the COOH-group

BSA-pC I:

immune serum against BSA-pC

FA:

ferulic acid

OVA:

ovalbunin from turkey

pC:

p-coumaric acid

pHY:

p-hydroxybenzoic acid

SA:

sinapic acid

SYA:

syringic acid

TAT:

TBS-azide-Tween-buffer

TFA:

trifluoroacetic acid

VA:

vanillic acid

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Authors and Affiliations

  1. Institut für Botanik, Westfälische Wilhelms-Universität Münster, Schloßgarten 3, D-48149, Münster, Federal Republic of Germany

    C. Niester-Nyveld, A. Haubrich, H. Kampendonk, S. Gubatz, K. B. Tenberge, M. Rittscher, S. Wilmesmeier & R. Wiermann

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  1. C. Niester-Nyveld
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  2. A. Haubrich
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  3. H. Kampendonk
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  4. S. Gubatz
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  5. K. B. Tenberge
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  6. M. Rittscher
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  7. S. Wilmesmeier
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  8. R. Wiermann
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Correspondence to R. Wiermann.

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Niester-Nyveld, C., Haubrich, A., Kampendonk, H. et al. Immunocytochemical localization of phenolic compounds in pollen walls using antibodies againstp-coumaric acid coupled to bovine serum albumin. Protoplasma 197, 148–159 (1997). https://doi.org/10.1007/BF01288024

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  • DOI: https://doi.org/10.1007/BF01288024

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