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Controlling for genetic identity of varieties, pollen contamination and stigma receptivity is essential to characterize the self-incompatibility system of Olea europaea L.

ABSTRACT: Bervillé et al. express concern about the existence of the diallelic self-incompatibility (DSI) system in Olea europaea, mainly because our model does not account for results from previous studies from their group that claimed to have documented asymmetry of the incompatibility response in reciprocal crosses. In this answer to their comment, we present original results based on reciprocal stigma tests that contradict conclusions from these studies. We show that, in our hands, not a single case of asymmetry was confirmed, endorsing that symmetry of incompatibility reactions seems to be the rule in Olive. We discuss three important aspects that were not taken into account in the studies cited in their comments and that can explain the discrepancy: (i) the vast uncertainty around the actual genetic identity of vernacular varieties, (ii) the risk of massive contamination associated with the pollination protocols that they used and (iii) the importance of checking for stigma receptivity in controlled crosses. These studies were thus poorly genetically controlled, and we stand by our original conclusion that Olive tree exhibits DSI.

SUBMITTER: Saumitou-Laprade P

PROVIDER: S-EPMC5680419 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Controlling for genetic identity of varieties, pollen contamination and stigma receptivity is essential to characterize the self-incompatibility system of <i>Olea europaea</i> L.

Saumitou-Laprade Pierre P Vernet Philippe P Vekemans Xavier X Castric Vincent V Barcaccia Gianni G Khadari Bouchaïb B Baldoni Luciana L

Evolutionary applications 20170629 9

Bervillé et al. express concern about the existence of the diallelic self-incompatibility (DSI) system in <i>Olea europaea</i>, mainly because our model does not account for results from previous studies from their group that claimed to have documented asymmetry of the incompatibility response in reciprocal crosses. In this answer to their comment, we present original results based on reciprocal stigma tests that contradict conclusions from these studies. We show that, in our hands, not a single ...[more]

PMID: 29151877

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Project description:Background and aimsA pollen grain contains a number of esterases, many of which are released upon contact with the stigma surface. However, the identity and function of most of these esterases remain unknown. In this work, esterases from olive pollen during its germination were identifided and functionally characterized.MethodsThe esterolytic capacity of olive (Olea europaea) pollen was examined using in vitro and in-gel enzymatic assays with different enzyme substrates. The functional analysis of pollen esterases was achieved by inhibition assays by using specific inhibitors. The cellular localization of esterase activities was performed using histochemical methods.Key resultsOlive pollen showed high levels of non-specific esterase activity, which remained steady after hydration and germination. Up to 20 esterolytic bands were identified on polyacrylamide gels. All the inhibitors decreased pollen germinability, but only diisopropyl fluorophosphate (DIFP) hampered pollen tube growth. Non-specific esterase activity is localized on the surface of oil bodies (OBs) and small vesicles, in the pollen intine and in the callose layer of the pollen tube wall. Acetylcholinesterase (AChE) activity was mostly observed in the apertures, exine and pollen coat, and attached to the pollen tube wall surface and to small cytoplasmic vesicles.ConclusionsIn this work, for the first time a systematic functional characterization of esterase enzymes in pollen from a plant species with wet stigma has been carried out. Olive pollen esterases belong to four different functional groups: carboxylesterases, acetylesterases, AChEs and lipases. The cellular localization of esterase activity indicates that the intine is a putative storage site for esterolytic enzymes in olive pollen. Based on inhibition assays and cellular localization of enzymatic activities, it can be concluded that these enzymes are likely to be involved in pollen germination, and pollen tube growth and penetration of the stigma.

Project description:The main objective of this research was to understand the effect of different solvents (ethanol, ethyl acetate, and hexane) on the extraction of bioactive compounds in addition to the antioxidant activity of eight Tunisian and Algerian olive leaf (Olea europaea L) cultivars. The different extracts were evaluated for their content of total phenols, flavonoids, and condensed tannins. The antioxidant capacity was determined by ABTS (2,2-azinobis (3 ethylbenzothiazoline-6-sulfonic acid) and DPPH (2,2-diphenyl-1 picrylhydrazyl) methods. In addition, qualitative and quantitative analyses of phenolic compounds by HPLC were performed. In terms of phenolic content, ethanol was found to be the best solvent to extract phenolic compounds with antioxidant capacity. The ethanolic extract of Sigoise and Rougette varieties were the richest in total phenols 161.54 ± 0.99, and 160.53 ± 1.17 mg GAE/g DE respectively. The highest levels of flavonoids were detected in ethyl acetate extracts of Rougette (87.58 ± 6.99 mg QE/g DE) and Verdal varieties (86.77 ± 1.75 mg QE/g DE), whereas the hexanolic extracts of Gerboui, Sofiana, and Chemlali exhibited the higher amount of tannins (67.99 ± 5.41, 65.52 ± 1.94, 64.64 ± 1.51 mg CE/g DE, respectively). Furthermore, in the DPPH radical scavenging activity assay, the Sofiana extract exhibited the highest antioxidant activity (23.85 ± 0.58 mg/L). In another case, for the ABTS radical scavenging assay, the Verdal extract showed high antioxidant potential (30.22 ± 1.35 mg/L). HPLC analysis of ethanolic extracts of the different varieties allowed the identification of 19 organic compounds, with oleuropein (2604.01-7991.14 mg/100g DM) being the most abundant, followed by hydroxytyrosol (250.74-687.54 mg/100g DM) and verbascoside (216.12-672.64 mg/100g DM) being the major compounds in the olive leaves in all studied ethanolic extracts, with concentrations varying significantly according to the cultivars and geographical origin. Principal component analysis (PCA) performed on phenolic profiles and antioxidant activities revealed the influence of bioactive compounds on antioxidant activity, with high concentrations of phenols and flavonoids in olive leaf extracts having a strong antioxidant activity.

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Controlling for genetic identity of varieties, pollen contamination and stigma receptivity is essential to characterize the self-incompatibility system of Olea europaea L.

Publications

Controlling for genetic identity of varieties, pollen contamination and stigma receptivity is essential to characterize the self-incompatibility system of <i>Olea europaea</i> L.

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