Characterization of Differentially Expressed Genes under Salt Stress in Olive

Climate change, currently taking place worldwide and also in the Mediterranean area, is leading to a reduction in water availability and to groundwater salinization. Olive represents one of the most efficient tree crops to face these scenarios, thanks to its natural ability to tolerate moderate salinity and drought. In the present work, four olive cultivars (Koroneiki, Picual, Royal de Cazorla and Fadak86) were exposed to high salt stress conditions (200 mM of NaCl) in greenhouse, in order to evaluate their tolerance level and to identify key genes involved in salt stress response. Molecular and physiological parameters, as well as plant growth and leaves’ ions Na+ and K+ content were measured. Results of the physiological measurements showed Royal de Cazorla as the most tolerant cultivar, and Fadak86 and Picual as the most susceptible ones. Ten candidate genes were analyzed and their complete genomic, CDS and protein sequences were identified. The expression analysis of their transcripts through reverse transcriptase quantitative PCR (RT-qPCR) demonstrated that only OeNHX7, OeP5CS, OeRD19A and OePetD were upregulated in tolerant cultivars, thus suggesting their key role in the activation of a salt tolerance mechanism.

Characterization of Polyphenol and Volatile Fractions of Californian-Style Black Olives and Innovative Application of E-nose for Acrylamide Determination

Californian-style black olives require a sterilization treatment that produces a carcinogenic contaminant, acrylamide. Thus, this compound was evaluated in two different olive cultivars using an electronic nose (E-nose). The sterilization intensity had a significant influence on the final phenol concentrations, acrylamide content, and volatile compounds. Increasing the sterilization intensity from 10 to 26 min (F0) reduced the phenol content, but it promoted acrylamide synthesis, leading to a wide range of this toxic substance. The Ester and phenol groups of volatile compounds decreased their content when the sterilization treatment increased; however, aldehyde and other volatile compound groups significantly increased their contents according to the thermal treatments. The compounds 4-ethenyl-pyridine, benzaldehyde, and 2,4-dimethyl-hexane are volatile compounds with unpleasant odours and demonstrated a high amount of influence on the differences found after the application of the thermal treatments. The “Manzanilla Cacereña” variety presented the highest amount of phenolic compounds and the lowest acrylamide content. Finally, it was found that acrylamide content is correlated with volatile compounds, which was determined using multiple linear regression analysis (R2 = 0.9994). Furthermore, the aroma of table olives was analysed using an E-nose, and these results combined with Partial Least Square (PLS) were shown to be an accurate method (range to error ratio (RER) >10 and ratio of performance to deviation (RPD) >2.5) for the indirect quantification of this toxic substance.

Prioritization of olive breeding objectives in Spain: Analysis of a producers and researchers survey

Aim of study: Trait prioritization of potential olive (Olea europaea L.) breeding objectives has been evaluated in this work from two surveys to researchers working on olive and olive producers / orchard managers. Area of study: Olive growing area in Andalusia (South of Spain) Material and methods: Twenty-five breeding objectives were associated to plant growth, fructification, oil content and composition, resistance to pest and disease and tolerance to soil and climatic conditions. Two assessment methods were applied (ranking and rating), showing similar results in both the researchers and producers surveys. Main results: Higher productivity was the objective with the highest score, followed by Verticillium wilt resistance and higher oil content. After them, tolerance to water stress and resistance to Xylella fastidiosa were among the preferred objectives. Conversely, the least preferred objectives were late harvesting, high fruit size and tolerance to water lodging and calcareous soils. In the producers’ survey, results have been consistent among the different types of orchards and farmers’ characteristics. It is also interesting to notice that more than 50% of the producers expressed their willingness to orchard enlargement or renewal in the three coming years and 25% of them would change the olive cultivar. They would be willing to pay an average 43% overprice for new cultivar fulfilling their requirements and 75% would support the use of genetic modified olives. Research highlights: These results should be considered to analyze the scope of current breeding programs and define the main criteria to be considered for future works aiming at developing new olive cultivars.

Prioritization of olive breeding objectives in Spain: Analysis of a producers and researchers survey

Aim of study: Trait prioritization of potential olive (Olea europaea L.) breeding objectives has been evaluated in this work from two surveys to researchers working on olive and olive producers / orchard managers. Area of study: Olive growing area in Andalusia (South of Spain) Material and methods: Twenty-five breeding objectives were associated to plant growth, fructification, oil content and composition, resistance to pest and disease and tolerance to soil and climatic conditions. Two assessment methods were applied (ranking and rating), showing similar results in both the researchers and producers surveys. Main results: Higher productivity was the objective with the highest score, followed by Verticillium wilt resistance and higher oil content. After them, tolerance to water stress and resistance to Xylella fastidiosa were among the preferred objectives. Conversely, the least preferred objectives were late harvesting, high fruit size and tolerance to water lodging and calcareous soils. In the producers’ survey, results have been consistent among the different types of orchards and farmers’ characteristics. It is also interesting to notice that more than 50% of the producers expressed their willingness to orchard enlargement or renewal in the three coming years and 25% of them would change the olive cultivar. They would be willing to pay an average 43% overprice for new cultivar fulfilling their requirements and 75% would support the use of genetic modified olives. Research highlights: These results should be considered to analyze the scope of current breeding programs and define the main criteria to be considered for future works aiming at developing new olive cultivars.

Phenolic Compounds and Triterpenes in Different Olive Tissues and Olive Oil By-Products, and Cytotoxicity on Human Colorectal Cancer Cells: The Case of Frantoio, Moraiolo and Leccino Cultivars (Olea europaea L.)

Phenolic and triterpenoid compounds of the olive tree are recognized as having a key role in health promotion, thanks to their multiple protective actions in humans. To expand the source of these bioactive compounds, the phenolic and triterpenoid profiles of leaf, branch, destoned fruit, destoned pomace, shell, seed, and extra virgin olive oil from the Frantoio, Leccino, and Moraiolo olive cultivars were simultaneously characterized by HPLC-DAD-MS. Overall, 43 molecules were quantitated and expressed on the obtained dry extracts. Oleuropein was mainly concentrated in branches (82.72 g/kg), fruits (55.79 g/kg), leaves (36.71 g/kg), and shells (1.26 g/kg), verbascoside (4.88 g/kg) in pomace, and nüzhenide 11-methyl oleoside (90.91 g/kg) in seeds. Among triterpenoids, which were absent in shells, the highest amount of oleanolic acid was found in olive leaves (11.88 g/kg). HCT-116 colorectal cells were chosen to assess the cytotoxicity of the dry extract, using the phytocomplex from Frantoio, which was the richest in phenols and triterpenoids. The IC50 was also determined for 13 pure molecules (phenols and terpenoids) detected in the extracts. The greatest inhibition on the cell’s proliferation was induced by the branch dry extract (IC50 88.25 μg/mL) and by ursolic acid (IC50 24 μM). A dose-dependent relationship was observed for the tested extracts.

Industrial Processing of Algerian Table Olive Cultivars Elaborated as Spanish Style

Olives from the Sigoise, Verdale, and Sevillana cultivars were elaborated as Spanish-style table olives by four Algerian factories, and the quality and food safety of the industrial table olives have been studied by the analysis of physicochemical and microbiological parameters. Differences were observed between the treatments carried out by the different factories throughout the manufacturing process, especially during the washing stage, but no significant differences were found between the analyzed samples for the concentration of sugars and polyphenols. The final pH values reached at the end of fermentation ranged between 5.04 and 4.27, and the titratable acidity was above 0.4% for all samples. Lactic and acetic acids were produced in mean concentrations of 0.68% and 0.21% as a result of lactic acid bacteria (LAB) and yeast metabolism, respectively. However, the presence of butyric, isobutyric, and propionic acids was also detected, and was related to the growth of undesirable spoilage microorganisms, responsible for secondary fermentations. The high-throughput sequencing of bacterial DNA suggested the dominance of LAB species belonging to genera Lactiplantibacillus, Leuconostoc, Pediococcus, Oenococcus, or Enterococcus. The Enterobacteriaceae family was detected during the first days of brining and in only one sample after 120 days of fermentation. Other spoilage microorganisms were found, such as Lentilactobacillus buchneri or the Pectinatus and Acetobacter genera, capable of consuming lactic acid and these played an essential role in the onset of spoilage. The Clostridium and Enterobacter genera, producers of butyric and propionic acids, were responsible for the malodorous fermentation present in the industrial samples that were analyzed. The study concluded that the safety of the table olives analyzed was compromised by the presence of undesirable microorganisms and microbial stability was not guaranteed. The elaboration process can be improved by reducing the washing steps and the time should be reduced to avoid the loss of fermentable matter, with the goal of reaching a pH < 4.0 after the fermentation and preventing the possibility of the growth of spoilage microorganisms and foodborne pathogens.