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Proteo-metabolomic study of olive (cv. ‘Chondrolia Chalkidikis’) drupe development and maturation

ABSTRACT: Olive (Olea europaea L.) is one of the most economically relevant tree crops in the Mediterranean basin. In this study, a comparative proteomic along with metabolomic-wide investigation was carried out on drupes of Greek olive cultivar 'Chondrolia Chalkidikis', collected across six developmental stages (S), namely seed development (S1, S2), mesocarp development (S3, S4, S5) and full maturation (S6). These stages were first characterized through the dynamics of fruit weight, dimensions and color parameters such as lightness, redness and yellowness. Combined gas chromatography–mass spectrometry and reversed–phase liquid chromatography quadrupole–time–of–flight mass spectrometry (RPLC–QToF–MS) procedures quantified 47 primary (e.g. allose, galactose, quinic acid, sorbitol, stearic acid) and 21 secondary (e.g. elenolic acid, oleacin, rutin, luteolin, hydroxytyrosol) metabolites in mesocarp samples during development. Protein analysis via nano–LC coupled to HDAM Orbitrap mass spectrometer, identified 3258 proteins from which the 350 were differentially accumulated between the final maturation stages (S5 and S6). Olive genome-based functional annotation showed that the largest proportion of identified proteins were involved in primary metabolism [i.e. lipoxygenases (LOX1/5)], energy [i.e. ferredoxin NADP+ reductase (FNR)], signal transduction [i.e. serine/threonine kinases (SAPK2, SRK2A, STK), transcription [i.e. elongation factor 2 (EEF2)] and protein destination [i.e. serine carboxypeptidase (SCPL)]. This investigation provides a reference framework for further nutritional and breeding studies, also allowing cross comparison among other olive cultivars.

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Olea Europaea (common Olive)

TISSUE(S): Plant Cell, Fruit

SUBMITTER: Martina Samiotaki

LAB HEAD: Athanassios Molassiotis

PROVIDER: PXD021086 | Pride | 2021-11-25

REPOSITORIES: Pride

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Project description:Analysis of gene-expression profiles by microarrays can be very useful to characterize new potential candidate genes, key regulatory networks, and to define phenotypes or molecular signatures to improve the diagnosis or classification of the disease. We have used this approach in the study of one of the major causes of allergic diseases in Mediterranean countries, the olive pollen response, in order to find differential molecular markers among five clinical groups, Non-allergic, Asymptomatic, Allergic but not to olive pollen, Non-treated, olive pollen allergic patients and Olive pollen allergic patients (under specific-immunotherapy). The results of gene-expression by principal components analysis (PCA) clearly showed five clusters of samples that correlated with the five clinical groups. Analysis of differential gene-expression by multiple testing, and functional analysis by KEGG and Gene-Ontology revealed differential genes and pathways among the 5 clinical groups. The study population comprised 28 subjects, selected from a previous immunological study (Aguerri et al. Eur. J. Inflammation 2012, in press), from Andalusia, who were recruited in 2 olive pollen exposure situations: during (April-June) and outside the pollen season (October-December). We established 5 groups, and 6 subjects from each group were selected for gene-expression analysis: Group 1, non-allergic subjects; Group 2, asymptomatic subjects (diagnosed with olive pollen allergy by skin testing, with no seasonal respiratory symptoms [rhinitis and/or asthma], and who consulted for adverse reaction to drugs); Group 3, patients who were allergic, but not to olive pollen; Group 4, non-treated olive pollenM-bM-^@M-^Sallergic; and Group 5, olive pollenM-bM-^@M-^Sallergic patients (receiving olive pollenM-bM-^@M-^Sspecific immunotherapy).The subjects were unrelated and recruited at the Allergy Service of 4 hospitals in Andalusia (Granada, JaM-CM-)n, Sevilla, and MM-CM-!laga). Olive pollenM-bM-^@M-^Sallergic patients fulfilled the following criteria: seasonal rhinitis and/or asthma from April to June, a positive skin prick test result for O. europaea pollen extract (ALK AbellM-CM-3, Madrid, Spain), and no previous immunotherapy. Informed consent was obtained from each subject. Ethical approval for the study was obtained from the Ethical and Research Committee of the participating hospitals. PBMCs were isolated from heparin-containing peripheral blood samples taken during and outside pollen season, by gradient centrifugation on Lymphoprep (Comercial Rafer, Zaragoza, Spain) following the manufacturerM-bM-^@M-^Ys instructions.

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Proteo-metabolomic study of olive (cv. ‘Chondrolia Chalkidikis’) drupe development and maturation

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