A methodology based on Fourier transform infrared (FTIR) spectroscopy, combined with multivariate analysis methods, was applied in order to monitor extra virgin olive oils produced from three distinct cultivars on different maturation stages. For the first time, this kind of methodology is used for the simultaneous discrimination of the maturation stage, and different cultivars. Principal component analysis and discriminant analysis were utilised to create a model for the discrimination of olive oil samples. Partial least squares regression was employed to design calibration models for the determination of chemical parameters. The performance of these models was based on the multiple coefficient of determination (R(2)), the root mean square error of calibration (RMSEC) and root mean square error of cross validation (RMSECV). The prediction models for the chemical parameters resulted in a R(2) ranged from 0.93 to 0.99, a RMSEC ranged from 1% to 4% and a RMSECV from 2% to 5%. It has been shown that this kind of approach allows to distinguish the different cultivars, and to clearly discern the different maturation stages, in each one of these distinct cultivars. Furthermore, the results demonstrated that FTIR spectroscopy in tandem with chemometric techniques allows the creation of viable and accurate models, suitable for correlating the data collected by FTIR spectroscopy, with the chemical composition of the EVOOs, obtained by standard methods.
Colletotrichum acutatum is a cosmopolitan and damaging plant pathogen of temperate, subtropical, and tropical fruits and causes anthracnose on olive (Olea europaea L.). Three olive cultivars showing a variable response to infection by C. acutatum were selected to a preliminary study of pathogen development. Fruit samples, from susceptible and tolerant cultivars, were taken at 0, 24, 48, 72, and 192 h after inoculation for a microscopic and histological study of the infection and colonization process. The aim of this study was to compare the infection process: conidial germination, germ tube and appressorium formation, hyphal growth, and mesocarp colonization in susceptible and tolerant olive cultivars as a condition for further exploration of disease development, which is required to develop cultivars with improved resistance to anthracnose. The rate of mesocarp colonization differed between the susceptible and tolerant cultivars, and both intracellular hemibiotrophy and subcuticular intramural necrotrophy were observed. Hemibiotrophic infection predominated in the moderately tolerant cultivar.
The chemical composition and organoleptic characteristics of olive oils may be influenced by genotype and some agronomic factors like olive drupe harvesting date. The aim of this work was to study the influence of olive ripening stage on chemical parameters and the organoleptic profile of fruits and oils of one of the main olive Portuguese cultivars (Cobrançosa). Six different Cobrançosa clones, which were harvested at two different ripening stages in the same olive growing area, were analyzed to evaluate phenolic compounds, ortho-diphenols, flavonoids and antioxidant activity and were characterized in terms of sensorial parameters. These clones have also been classified according to the yield of production. The lowest values in phenolic content, ortho-diphenols, flavonoids and antioxidant activity occurred always in the same clone (110) in both ripening stages in olive fruits and olive oil. This clone has been previously identified as low yield. Also, the results revealed significant correlations between total phenolics and antioxidant activity (R 2 = 0.932, P \ 0.0001), between ortho-diphenols and antioxidant activity (R 2 = 0.9445, P \ 0.0001) and between flavonoids and antioxidant activity (R 2 = 0.9263, P \ 0.0001) on olive oil samples. The sensorial parameters that have been affected in olive oils were herbaceous aroma and flavor, and bitter taste that increased with the ripening process.
Olive anthracnose is a very common and severe disease caused by diverse species of fungi belonging to Colletotrichum acutatum and Colletotrichum gloeosporioides complexes. To understand aspects of the Colletotrichum colonization and primary infection in olives, Colletotrichum spp. were isolated from the interior of 2-year stems, flower buds, and immature fruits of three important olive cultivars, Galega vulgar, Cobrançosa, and Azeiteira, from different sites within Alentejo, a major olive-producing region in Portugal. A total of 270 trees was sampled, and 68 Colletotrichum spp. isolates were obtained from 46 olive trees. DNA extraction and amplification of β-tubulin and GADPH genes through PCR revealed that the vast majority of the isolates showed high similarity to Colletotrichum nymphaeae, and only three isolates showed high similarity to Colletotrichum godetiae. The highest number of Colletotrichum spp. isolates was detected in olive trees from Galega vulgar and in immature fruits. No significant differences in the number of Colletotrichum spp. isolates were found in trees from different sites. The highest percentages of infected immature fruits were obtained in trees that also presented a high percentage of 2-year stem infections, which may indicate that 2-year stems serve as important sources of inoculum, and the fungus may travel from the stems to other parts of the plant. Another indication of such possibility is that one isolate of C. nymphaeae (C. nymphaeae 2), characterized by a unique nucleotide mutation within the beta tubulin gene, was present in different organs of the same tree, both in 2-year stems and in recently formed vegetative organs as flower buds and immature fruits, which seem to suggest that it may be the same isolate, which has moved systemically inside the plant. The results presented here can play an important role in working out strategies for the effective and timely management of the disease and in reducing the number of unnecessary fungicide applications.
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