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Polyamines and transglutaminase activity are involved in compatible and self-incompatible pollination of Citrus grandis

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Abstract

Pollination of pummelo (Citrus grandis L. Osbeck) pistils has been studied in planta by adding compatible and self-incompatible (SI) pollen to the stigma surface. The pollen germination has been monitored inside the pistil by fluorescent microscopy showing SI altered morphologies with irregular depositions of callose in the tube walls, and heavy callose depositions in enlarged tips. The polyamine (PA) content as free, perchloric acid (PCA)-soluble and -insoluble fractions and transglutaminase (TGase) activity have been analyzed in order to deepen their possible involvement in the progamic phase of plant reproduction. The conjugated PAs in PCA-soluble fraction were definitely higher than the free and the PCA-insoluble forms, in both compatible and SI pollinated pistils. In pistils, pollination caused an early decrease of free PAs and increase of the bound forms. The SI pollination, showed highest values of PCA-soluble and -insoluble PAs with a maximum in concomitance with the pollen tube arrest. As TGase mediates some of the effects of PAs by covalently binding them to proteins, its activity, never checked before in Citrus, was examined with two different assays. In addition, the presence of glutamyl-PAs confirmed the enzyme assay data and excluded the possibility of a misinterpretation. The SI pollination caused an increase in TGase activity, whereas the compatible pollination caused its decrease. Similarly to bound PAs, the glutamyl-PAs and the enzyme activity peaked in the SI pollinated pistils in concomitance with the observed block of the pollen tube growth, suggesting an involvement of TGase in SI response.

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Abbreviations

BCA:

Bicinchoninic acid

BSA:

Bovine serum albumin

DMC:

N′,N′-dimethyl casein

DTT:

Dithiothreitol

EDTA:

Ethylene diamine tetraacetic acid

EGTA:

Ethylene glycol tetraacetic acid

GSI:

Gametophytic self-incompatible

HPLC:

High-performance liquid chromatography

PAs:

Polyamines

PBS:

Phosphate-buffered saline

PMSF:

Phenyl methyl sulfonyl fluoride

PCA:

Perchloric acid

PCD:

Programmed cell death

PU:

Putrescine

SI:

Self-incompatible

SD:

Spermidine

SM:

Spermine

SSI:

Sporophytic self-incompatible

TCA:

Trichloroacetic acid

TGase:

Transglutaminase

tTGase:

Tissue transglutaminase

UP:

Unpollinated

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Acknowledgments

Thanks are due to the Italian Ministero dell’Università that supported this work by a grant PRIN 2007 to Dipartimento Biologia Evoluzionistica Sperimentale, Università di Bologna (http://prin.miur.it/) and Dipartimento di Scienze delle Produzioni Agrarie e Alimentari, Università di Catania. Thanks to I. Menghetti for work she made during the training in the Dip. Biologia Evoluzionistica Sperimentale and to Nicodemo Mele for his help in figure preparation.

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Authors and Affiliations

  1. Dipartimento di Scienze delle Produzioni Agrarie e Alimentari, Università di Catania, via Valdisavoia 5, 95123, Catania, Italy

    Alessandra Gentile, Gaetano Distefano, Giuseppina Las Casas & Stefano La Malfa

  2. Dipartimento di Biologia e.s., Università degli Studi di Bologna, via Irnerio 42, 40126, Bologna, Italy

    Fabiana Antognoni, Rosa Anna Iorio, Donatella Serafini-Fracassini & Stefano Del Duca

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  1. Alessandra Gentile
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  2. Fabiana Antognoni
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  4. Gaetano Distefano
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  5. Giuseppina Las Casas
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  7. Donatella Serafini-Fracassini
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  8. Stefano Del Duca
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Correspondence to Stefano Del Duca.

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Gentile, A., Antognoni, F., Iorio, R.A. et al. Polyamines and transglutaminase activity are involved in compatible and self-incompatible pollination of Citrus grandis . Amino Acids 42, 1025–1035 (2012). https://doi.org/10.1007/s00726-011-1017-9

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