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Histological and molecular analysis of pollen–pistil interaction in clementine

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Abstract

In contrast to model species, the self-incompatibility reaction in citrus has been poorly studied. It is assumed to be gametophytically determined and genetically controlled by the S-locus, which in other species encodes for glycoproteins (S-RNases) showing ribonuclease activity. To investigate pollen–pistil interaction, the pollen tube growth of two clementine varieties, ‘Comune’ (self-incompatible) and ‘Monreal’ (a ‘Comune’ self-compatible mutation) was analysed by histological assays in self- and cross-pollination conditions. Cross-pollination assays demonstrated that the mutation leading to self-compatibility in ‘Monreal’ occurred in the stylar tissues. Similar rates of pollen germination were observed in both genotypes. However, ‘Comune’ pollen tubes showed altered morphology and arrested growth in the upper style while in ‘Monreal’ they grew straight toward the ovary. Moreover, to identify genes putatively involved in pollen–pistil interaction and self-incompatibility, research based on the complementary DNA-amplified fragment length polymorphism technique was carried out to compare the transcript profiles of unpollinated and self-pollinated styles and stigmas of the two cultivars. This analysis identified 96 unigenes such as receptor-like kinases, stress-induced genes, transcripts involved in the phenylpropanoid pathway, transcription factors and genes related to calcium and hormone signalling. Surprisingly, a high percentage of active long terminal repeat (LTR) and non-LTR retrotransposons were identified among the unigenes, indicating their activation in response to pollination and their possible role in the regulation of self-incompatibility genes. The quantitative reverse trascription-polymerase chain reaction analysis of selected gene tags showed transcriptional differences between the two genotypes during pollen germination and pollen tube elongation.

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Abbreviations

cDNA-AFLP:

Complementary DNA-amplified fragment length polymorphism

EST:

Expressed sequence tag

LTR:

Long terminal repeat

qRT-PCR:

Quantitative reverse trascription-polymerase chain reaction

SC:

Self-compatible

SI:

Self-incompatible

TDF:

Transcript derived fragment

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Acknowledgments

Funding was provided by the Italian Ministry of Agriculture and Forestry—Project RAVAGRU (Advanced research in citriculture), publication no. 27—and by Italian Ministry of University—Project PRIN “Il processo produttivo delle piante arboree da frutto: aspetti molecolari, fisiologici ed agronomici dell’incompatibilità fiorale e strategie di controllo.”

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  1. Dipartimento di OrtoFloroArboricoltura e Tecnologie Agroalimentari, University of Catania, Via Valdisavoia 5, Catania, 95123, Italy

    Gaetano Distefano, Marco Caruso, Stefano La Malfa, Alessandra Gentile & Eugenio Tribulato

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  1. Gaetano Distefano
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  2. Marco Caruso
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  3. Stefano La Malfa
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  4. Alessandra Gentile
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Correspondence to Alessandra Gentile.

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Communicated by D. Somers.

Gaetano Distefano and Marco Caruso have contributed equally to this work.

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Distefano, G., Caruso, M., La Malfa, S. et al. Histological and molecular analysis of pollen–pistil interaction in clementine. Plant Cell Rep 28, 1439–1451 (2009). https://doi.org/10.1007/s00299-009-0744-9

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