The olive tree (Olea europaea L.) is one of the main crops of the Mediterranean region which suffers from drought and soil salinization. We assessed the photosynthetic rate, leaf water content and antioxidative enzyme activity (APX, GPX, SOD and CAT) of three Greek olive cultivars (‘Amfisis’, ‘Mastoidis’ and ‘Lefkolia Serron’) subjected to drought and salinity stresses. Hyperspectral reflectance data were acquired using an analytical spectral device (ASD) FieldSpec^® 3 spectroradiometer, while principal component regression, partial least squares regression and linear discriminant analysis were used to estimate the relationship between spectral and physiological measurements. The photosynthetic rate and water content of stressed plants decreased, while enzyme activity had an increasing tendency. ‘Amfisis’ was more resistant to drought and salinity stress than ‘Mastoidis’ and ‘Lefkolia Serron’. The NDVI appeared to have the highest correlation with the photosynthetic rate, followed by the PRI. APX enzyme activity was the most highly correlated with the 1150–1370 nm range, with an additional peak at 1840 nm. CAT enzyme activity resulted in the highest correlation with the visible part of the spectrum with two peaks at 1480 nm and 1950 nm, while GPX enzyme activity appeared to have a strong correlation within all the available spectral ranges except for 670–1180 nm. Finally, SOD activity showed high correlation values within 1190–1850 nm. This is the first time the correlation of hyperspectral imagery with photosynthetic rate and antioxidant enzyme activities was determined, providing the background for high-throughput plant phenotyping through a drone with a hyperspectral camera. This progress would provide the possibility of early stress detection in large olive groves and assist farmers in decision making and optimizing crop management, health and productivity.

Olive tree growth and reproduction are severely affected by temperature extremes, compromising fruit yield. In that aspect, the olive varieties “Koroneiki” and “Mastoidis” were employed in a mild cold stress experiment, imitating night frost incidents to assess their biochemical, physiological and reproductive functions in relation to genotype. The physiological performance of the stressed plants was not significantly altered, suggesting that both cultivars were well adapted to mild cold night stress. The biochemical response of the plants, regarding antioxidant enzymes, H₂O₂ and TBARS accumulation, confirmed that both cultivars could cope with the stress applied. The mRNA levels of the PPO gene, which participates in hydroxytyrosol biosynthesis and plant defense, were elevated after 24-h stress at 0 °C, in both cultivars with “Mastoidis” plants exhibiting higher levels for a longer period. Three more genes involved in hydroxytyrosol biosynthesis upregulated their expression levels as a response to cold stress. The numerous plant phenology aspects measured reinforced the conclusion that both cultivars responded to the stress applied. The results of the present study may contribute to better understanding olive tree adaptive responses to low temperature events, an abiotic stress condition that is often present in an open plantation, thus assisting farmers on breeding and cultivar selection.