Physiological and biochemical study of the drought tolerance of 14 main olive cultivars in the Mediterranean basin.

A complete study of 14 olive cultivars of great economic importance was carried out. These cultivars are Arbequina, Arbosana, Chemlali, Cornicabra, Cornezuelo de Jaén, Empeltre, Frantoio, Hojiblanca, Koroneiki, Manzanilla de Sevilla, Martina, Picual, Sikitita1 and Sikitita 2. All of them are certified by the World Olive Germplasm Bank of Córdoba (Spain). They are predominant cultivars in the olive groves of different locations throughout the Mediterranean basin, and they were subjected to total water deficit for a minimum of 14 days and a maximum of 42 days in the present study. Data such as chlorophyll content, soil moisture and specific leaf area were gathered. Photosynthetic parameters measured at the respective saturation irradiance of each cultivar were also analysed: assimilation rate, transpiration, stomatal conductance, photosynthetic efficiency, photochemical and non-photochemical quenching, photonic flux density, electron transference ratio, efficient use of water and amount of proline and malondialdehyde as indicators of oxidative stress. In addition to the control, two different experimental conditions were analysed: moderate drought, after 14 days of lack of irrigation, and severe drought, after 28-42 days of total absence of irrigation, depending on the tolerance of each cultivar. Based on the results, the cultivars were characterised and divided into four groups according to their drought tolerance: tolerant, moderately tolerant, moderately sensitive and sensitive to drought. This work represents the first contribution of drought tolerance of a considerable number of olive cultivars, with all of them being subjected to the same criteria and experimental conditions for their classification.

Olive reproductive biology: implications for yield, compatibility conundrum, and environmental constraints.

Olive (Olea europaea L.) is an important Mediterranean tree species with a longstanding history of cultivation, boasting a diverse array of local cultivars. While traditional olive orchards are valued for their cultural and aesthetic significance, they often face economic sustainability challenges in the modern context. The success of both traditional and newly introduced cultivars (e.g. those obtained by cross-breeding) is hindered by self-incompatibility, a prevalent issue for this species that results in low fruit set when limited genetic diversity is present. Further, biological, environmental, and agronomic factors have been shown to interlink in shaping fertilization patterns, hence impacting on the final yield. Climatic conditions during pollination, such as excessive rainfall or high temperatures, can further exacerbate the problem. In this work, we provide an overview of the various factors that trigger the phenomenon of suboptimal fruit set in olive trees. This work provides a comprehensive understanding of the interplay among these factors, shedding light on potential mechanisms and pathways that contribute to the observed outcomes in the context of self-incompatibility in olive.

Relationship between Height and Exposure in Multispectral Vegetation Index Response and Product Characteristics in a Traditional Olive Orchard.

The present research had two aims. The first was to evaluate the effect of height and exposure on the vegetative response of olive canopies' vertical axis studied through a multispectral sensor and on the qualitative and quantitative product characteristics. The second was to examine the relationship between multispectral data and productive characteristics. Six olive plants were sampled, and their canopy's vertical axis was subdivided into four sectors based on two heights (Top and Low) and two exposures (West and East). A ground-vehicle-mounted multispectral proximal sensor (OptRx from AgLeader®) was used to investigate the different behaviours of the olive canopy vegetation index (VI) responses in each sector. A selective harvest was performed, in which each plant and sector were harvested separately. Product characterisation was conducted to investigate the response of the products (both olives and oils) in each sector. The results of Tukey's test (p > 0.05) showed a significant effect of height for the VI responses, with the Low sector obtaining higher values than the Top sector. The olive product showed some height and exposure effect, particularly for the olives' dimension and resistance to detachment, which was statistically higher in the upper part of the canopies. The regression studies highlighted some relationships between the VIs and product characteristics, particularly for resistance to detachments (R2 = 0.44-0.63), which can affect harvest management. In conclusion, the results showed the complexity of the olive canopies' response to multispectral data collection, highlighting the need to study the vertical axis to assess the variability of the canopy itself. The relationship between multispectral data and product characteristics must be further investigated.

Unravelling Different Water Management Strategies in Three Olive Cultivars: The Role of Osmoprotectants, Proteins, and Wood Properties.

Understanding the responses of olive trees to drought stress is crucial for improving cultivation and developing drought-tolerant varieties. Water transport and storage within the plant is a key factor in drought-tolerance strategies. Water management can be based on a variety of factors such as stomatal control, osmoprotectant molecules, proteins and wood properties. The aim of the study was to evaluate the water management strategy under drought stress from an anatomical and biochemical point of view in three young Italian olive cultivars (Giarraffa, Leccino and Maurino) previously distinguished for their physiological and metabolomic responses. For each cultivar, 15 individuals in pots were exposed or not to 28 days of water withholding. Every 7 days, the content of sugars (including mannitol), proline, aquaporins, osmotins, and dehydrins, in leaves and stems, as well as the chemical and anatomical characteristics of the wood of the three cultivars, were analyzed. 'Giarraffa' reduced glucose levels and increased mannitol production, while 'Leccino' accumulated more proline. Both 'Leccino' and 'Maurino' increased sucrose and aquaporin levels, possibly due to their ability to remove embolisms. 'Maurino' and 'Leccino' accumulated more dehydrins and osmotins. While neither genotype nor stress affected wood chemistry, 'Maurino' had a higher vessel-to-xylem area ratio and a larger hydraulic diameter, which allows it to maintain a high transpiration rate but may make it more susceptible to cavitation. The results emphasized the need for an integrated approach, highlighting the importance of the relative timing and sequence of each parameter analyzed, allowing, overall, to define a "strategy" rather than a "response" to drought of each cultivar.

Disentangling the link between leaf photosynthesis and turgor in fruit growth.

Despite the importance of understanding plant growth, the mechanisms underlying how plant and fruit growth declines during drought remain poorly understood. Specifically, it remains unresolved whether carbon or water factors are responsible for limiting growth as drought progresses. We examine questions regarding the relative importance of water and carbon to fruit growth depending on the water deficit level and the fruit growth stage by measuring fruit diameter, leaf photosynthesis, and a proxy of cell turgor in olive (Olea europaea). Flow cytometry was also applied to determine the fruit cell division stage. We found that photosynthesis and turgor were related to fruit growth; specifically, the relative importance of photosynthesis was higher during periods of more intense cell division, while turgor had higher relative importance in periods where cell division comes close to ceasing and fruit growth is dependent mainly on cell expansion. This pattern was found regardless of the water deficit level, although turgor and growth ceased at more similar values of leaf water potential than photosynthesis. Cell division occurred even when fruit growth seemed to stop under water deficit conditions, which likely helped fruits to grow disproportionately when trees were hydrated again, compensating for periods with low turgor. As a result, the final fruit size was not severely penalized. We conclude that carbon and water processes are able to explain fruit growth, with importance placed on the combination of cell division and expansion. However, the major limitation to growth is turgor, which adds evidence to the sink limitation hypothesis.

Declining root water transport drives stomatal closure in olive under moderate water stress.

Efficient water transport from soil to leaves sustains stomatal opening and steady-state photosynthesis. The aboveground portion of this pathway is well-described, yet the roots and their connection with the soil are still poorly understood due to technical limitations. Here we used a novel rehydration technique to investigate changes in the hydraulic pathway between roots and soil and within the plant body as individual olive plants were subjected to a range of water stresses. Whole root hydraulic resistance (including the radial pathway from xylem to the soil-root interface) constituted 81% of the whole-plant resistance in unstressed plants, increasing to > 95% under a moderate level of water stress. The decline in this whole root hydraulic conductance occurred in parallel with stomatal closure and contributed significantly to the reduction in canopy conductance according to a hydraulic model. Our results demonstrate that losses in root hydraulic conductance, mainly due to a disconnection from the soil during moderate water stress in olive plants, are profound and sufficient to induce stomatal closure before cavitation occurs. Future studies will determine whether this core regulatory role of root hydraulics exists more generally among diverse plant species.

Biot-Granier Sensor: A Novel Strategy to Measuring Sap Flow in Trees.

The Biot-Granier (Gbt) is a new thermal dissipation-based sap flow measurement methodology, comprising sensors, data management and automatic data processing. It relies on the conventional Granier (Gcv) methodology upgraded with a modified Granier sensor set, as well as on an algorithm to measure the absolute temperatures in the two observation points and perform the Biot number approach. The work described herein addresses the construction details of the Gbt sensors and the characterization of the overall performance of the Gbt method after comparison with a commercial sap flow sensor and independent data (i.e., volumetric water content, vapor pressure deficit and eddy covariance technique). Its performance was evaluated in three trials: potted olive trees in a greenhouse and two vineyards. The trial with olive trees in a greenhouse showed that the transpiration measures provided by the Gbt sensors showed better agreement with the gravimetric approach, compared to those provided by the Gcv sensors. These tended to overestimate sap flow rates as much as 4 times, while Gbt sensors overestimated gravimetric values 1.5 times. The adjustments based on the Biot equations obtained with Gbt sensors contribute to reduce the overestimates yielded by the conventional approach. On the other hand, the heating capacity of the Gbt sensor provided a minimum of around 7 °C and maximum about 9 °C, contrasting with a minimum around 6 °C and a maximum of 12 °C given by the Gcv sensors. The positioning of the temperature sensor on the tip of the sap flow needle proposed in the Gbt sensors, closer to the sap measurement spot, allow to capture sap induced temperature variations more accurately. This explains the higher resolution and sensitivity of the Gbt sensor. Overall, the alternative Biot approach showed a significant improvement in sap flow estimations, contributing to adjust the Granier sap flow index, a vulnerability of that methodology.

Genome Wild Analysis and Molecular Understanding of the Aquaporin Diversity in Olive Trees (Olea Europaea L.).

Cellular aquaporin water channels (AQPs) constitute a large family of transmembrane proteins present throughout all kingdoms of life, playing important roles in the uptake of water and many solutes across the membranes. In olive trees, AQP diversity, protein features and their biological functions are still largely unknown. This study focuses on the structure and functional and evolution diversity of AQP subfamilies in two olive trees, the wild species Olea europaea var. sylvestris (OeuAQPs) and the domesticated species Olea europaea cv. Picual (OleurAQPs), and describes their involvement in different physiological processes of early plantlet development and in biotic and abiotic stress tolerance in the domesticated species. A scan of genomes from the wild and domesticated olive species revealed the presence of 52 and 79 genes encoding full-length AQP sequences, respectively. Cross-genera phylogenetic analysis with orthologous clustered OleaAQPs into five established subfamilies: PIP, TIP, NIP, SIP, and XIP. Subsequently, gene structures, protein motifs, substrate specificities and cellular localizations of the full length OleaAQPs were predicted. Functional prediction based on the NPA motif, ar/R selectivity filter, Froger's and specificity-determining positions suggested differences in substrate specificities of Olea AQPs. Expression analysis of the OleurAQP genes indicates that some genes are tissue-specific, whereas few others show differential expressions at different developmental stages and in response to various biotic and abiotic stresses. The current study presents the first detailed genome-wide analysis of the AQP gene family in olive trees and it provides valuable information for further functional analysis to infer the role of AQP in the adaptation of olive trees in diverse environmental conditions in order to help the genetic improvement of domesticated olive trees.

Responses of olive plants exposed to different irrigation treatments in combination with heat shock: physiological and molecular mechanisms during exposure and recovery.

MAIN CONCLUSION: A water-deficit period, leading to stomatal control and overexpression of protective proteins (sHSP and DHN), contributes to olive´s tolerance to later imposed stress episodes. Aquaporins modulation is important in olive recovery. Olive is traditionally cultivated in dry farming or in high water demanding irrigated orchards. The impact of climate change on these orchards remains to unveil, as heat and drought episodes are increasing in the Mediterranean region. To understand how young plants face such stress episodes, olive plants growing in pots were exposed to well-irrigated and non-irrigated treatments. Subsequently, plants from each treatment were either exposed to 40 °C for 2 h or remained under control temperature. After treatments, all plants were allowed to grow under well-irrigated conditions (recovery). Leaves were compared for photosynthesis, relative water content, mineral status, pigments, carbohydrates, cell membrane permeability, lipid peroxidation and expression of the protective proteins' dehydrin (OeDHN1), heat-shock proteins (OeHSP18.3), and aquaporins (OePIP1.1 and OePIP2.1). Non-irrigation, whilst increasing carbohydrates, reduced some photosynthetic parameters to values below the ones of the well-irrigated plants. However, when both groups of plants were exposed to heat, well-irrigated plants suffered more drastic decreases of net CO2 assimilation rate and chlorophyll b than non-irrigated plants. Overall, OeDHN1 and OeHSP18.3 expression, which was increased in non-irrigated treatment, was potentiated by heat, possibly to counteract the increase of lipid peroxidation and loss of membrane integrity. Plants recovered similarly from both irrigation and temperature treatments, and recovery was associated with increased aquaporin expression in plants exposed to one type of stress (drought or heat). These data represent an important contribution for further understanding how dry-farming olive will cope with drought and heat episodes.

Mapping xylem failure in disparate organs of whole plants reveals extreme resistance in olive roots.

The capacity of plant species to resist xylem cavitation is an important determinant of resistance to drought, mortality thresholds, geographic distribution and productivity. Unravelling the role of xylem cavitation vulnerability in plant evolution and adaptation requires a clear understanding of how this key trait varies between the tissues of individuals and between individuals of species. Here, we examine questions of variation within individuals by measuring how cavitation moves between organs of individual plants. Using multiple cameras placed simultaneously on roots, stems and leaves, we were able to record systemic xylem cavitation during drying of individual olive plants. Unlike previous studies, we found a consistent pattern of root > stem > leaf in terms of xylem resistance to cavitation. The substantial variation in vulnerability to cavitation, evident among individuals, within individuals and within tissues of olive seedlings, was coordinated such that plants with more resistant roots also had more resistant leaves. Preservation of root integrity means that roots can continue to supply water for the regeneration of drought-damaged aerial tissues after post-drought rain. Furthermore, coordinated variation in vulnerability between leaf, stem and root in olive plants suggests a strong selective pressure to maintain a fixed order of cavitation during drought.

Tolerance of olive (Olea europaea) cv Frantoio to Verticillium dahliae relies on both basal and pathogen-induced differential transcriptomic responses.

Verticillium wilt of olive (VWO) is one of the most serious biotic constraints for this tree crop. Our knowledge of the genetics of the tolerance/resistance to this disease is very limited. Here we show that tolerance of the cv Frantoio relies on both basal and early pathogen-induced differential transcriptomic responses. A comparative transcriptomic analysis (RNA-seq) was conducted in root tissues of cvs Frantoio (VWO-tolerant) and Picual (VWO-susceptible). RNA samples originated from roots of inoculated olive plants during the early infection stages by Verticillium dahliae (highly virulent, defoliating pathotype). A huge number of differentially expressed genes (DEGs) were found between 'Frantoio' and 'Picual' (27 312 unigenes) in the absence of the pathogen. Upon infection with V. dahliae, 'Picual' and 'Frantoio' plants responded differently too. In the early infection stages, four clusters of DEGs could be identified according to their time-course expression patterns. Among others, a pathogenesis-related protein of the Bet v I family and a dirigent-like protein involved in lignification, and several BAK1, NHL1, reactive oxygen species stress response and BAM unigenes showed noticeable differences between cultivars. Tolerance of 'Frantoio' plants to VWO is a consequence of a complex and multifaceted process which involves many plant traits.

Physiological and biochemical responses of some olive cultivars (Olea europaea L.) to water stress.

Water stress is one of the important abiotic environmental stresses that threaten the agricultural -products in the world. This experiment was carried out to determine the effect of water stress on physiological and biochemical characteristics of three commercial olive cultivars. A factorial pot experiment was conducted in the field conditions using completely randomized design in Gilan-Gharb city, Kermanshah province during 2015. One-year-old rooted cuttings of Zard, Amigdalolia and Konservolia olive cultivars were planted in 12-liter pots and subjected to three irrigation treatments. Irrigation treatments included control (100% of field capacity), 75% and 50% field capacity. Physiological and biochemical characteristics such as relative water content (RWC), electrolyte leakage (EL), calcium, potassium and sodium content, total phenol, malondialdehyde, peroxidases, catalase, a, b and total chlorophyll, proline and total carbohydrate were measured. Results showed that relative water content, K & Ca was reduced while sodium content increased by increasing water stress. Chlorophyll content was higher in Konservolia cultivar under water stress in comparison to the others. Water stress induced increasing in proline, total phenol and soluble carbohydrate in all cultivars. The highest total phenol and proline was recorded in Zard cultivar under water stress. Total carbohydrate increased significantly (P<0.05) in Konservolia in comparison to the others. Malondialdehyde content was increased as an index of oxidative stress by drought. The highest peroxidases and catalase activity were recorded under drought stress of 50% irrigation in Konservolia olive cultivar. Generally based on results Konservolia, Zard and Amigdalolia were more tolerant respectively.

Xylem resistance to embolism: presenting a simple diagnostic test for the open vessel artefact.

Xylem vulnerability to embolism represents an essential trait for the evaluation of the impact of hydraulics in plant function and ecology. The standard centrifuge technique is widely used for the construction of vulnerability curves, although its accuracy when applied to species with long vessels remains under debate. We developed a simple diagnostic test to determine whether the open-vessel artefact influences centrifuge estimates of embolism resistance. Xylem samples from three species with differing vessel lengths were exposed to less negative xylem pressures via centrifugation than the minimum pressure the sample had previously experienced. Additional calibration was obtained from non-invasive measurement of embolism on intact olive plants by X-ray microtomography. Results showed artefactual decreases in hydraulic conductance (k) for samples with open vessels when exposed to a less negative xylem pressure than the minimum pressure they had previously experienced. X-Ray microtomography indicated that most of the embolism formation in olive occurs at xylem pressures below -4.0 MPa, reaching 50% loss of hydraulic conductivity at -5.3 MPa. The artefactual reductions in k induced by centrifugation underestimate embolism resistance data of species with long vessels. A simple test is suggested to avoid this open vessel artefact and to ensure the reliability of this technique in future studies.

A possible role for flowering locus T-encoding genes in interpreting environmental and internal cues affecting olive (Olea europaea L.) flower induction.

Olive (Olea europaea L.) inflorescences, formed in lateral buds, flower in spring. However, there is some debate regarding time of flower induction and inflorescence initiation. Olive juvenility and seasonality of flowering were altered by overexpressing genes encoding flowering locus T (FT). OeFT1 and OeFT2 caused early flowering under short days when expressed in Arabidopsis. Expression of OeFT1/2 in olive leaves and OeFT2 in buds increased in winter, while initiation of inflorescences occurred i n late winter. Trees exposed to an artificial warm winter expressed low levels of OeFT1/2 in leaves and did not flower. Olive flower induction thus seems to be mediated by an increase in FT levels in response to cold winters. Olive flowering is dependent on additional internal factors. It was severely reduced in trees that carried a heavy fruit load the previous season (harvested in November) and in trees without fruit to which cold temperatures were artificially applied in summer. Expression analysis suggested that these internal factors work either by reducing the increase in OeFT1/2 expression or through putative flowering repressors such as TFL1. With expected warmer winters, future consumption of olive oil, as part of a healthy Mediterranean diet, should benefit from better understanding these factors.

Unusual positional effects on flower sex in an andromonoecious tree: Resource competition, architectural constraints, or inhibition by the apical flower?

PREMISE OF THE STUDY: Two, nonmutually exclusive, mechanisms-competition for resources and architectural constraints-have been proposed to explain the proximal to distal decline in flower size, mass, and/or femaleness in indeterminate, elongate inflorescences. Whether these mechanisms also explain unusual positional effects such as distal to proximal declines of floral performance in determinate inflorescences, is understudied. METHODS: We tested the relative influence of these mechanisms in the andromonoecious wild olive tree, where hermaphroditic flowers occur mainly on apical and the most proximal positions in determinate inflorescences. We experimentally increased the availability of resources for the inflorescences by removing half of the inflorescences per twig or reduced resource availability by removing leaves. We also removed the apical flower to test its inhibitory effect on subapical flowers. KEY RESULTS: The apical flower had the highest probability of being hermaphroditic. Further down, however, the probability of finding a hermaphroditic flower decreased from the base to the tip of the inflorescences. An experimental increase of resources increased the probability of finding hermaphroditic flowers at each position, and vice versa. Removal of the apical flower increased the probability of producing hermaphroditic flowers in proximal positions but not in subapical positions. CONCLUSIONS: These results indicate an interaction between resource competition and architectural constraints in influencing the arrangement of the hermaphroditic and male flowers within the inflorescences of the wild olive tree. Subapical flowers did not seem to be hormonally suppressed by apical flowers. The study of these unusual positional effects is needed for a general understanding about the functional implications of inflorescence architecture.

How anthropogenic changes may affect soil-borne parasite diversity? Plant-parasitic nematode communities associated with olive trees in Morocco as a case study.

BACKGROUND: Plant-parasitic nematodes (PPN) are major crop pests. On olive (Olea europaea), they significantly contribute to economic losses in the top-ten olive producing countries in the world especially in nurseries and under cropping intensification. The diversity and the structure of PPN communities respond to environmental and anthropogenic forces. The olive tree is a good host plant model to understand the impact of such forces on PPN diversity since it grows according to different modalities (wild, feral and cultivated olives). A wide soil survey was conducted in several olive-growing regions in Morocco. The taxonomical and the functional diversity as well as the structures of PPN communities were described and then compared between non-cultivated (wild and feral forms) and cultivated (traditional and high-density olive cultivation) olives. RESULTS: A high diversity of PPN with the detection of 117 species and 47 genera was revealed. Some taxa were recorded for the first time on olive trees worldwide and new species were also identified. Anthropogenic factors (wild vs cultivated conditions) strongly impacted the PPN diversity and the functional composition of communities because the species richness, the local diversity and the evenness of communities significantly decreased and the abundance of nematodes significantly increased in high-density conditions. Furthermore, these conditions exhibited many more obligate and colonizer PPN and less persister PPN compared to non-cultivated conditions. Taxonomical structures of communities were also impacted: genera such as Xiphinema spp. and Heterodera spp. were dominant in wild olive, whereas harmful taxa such as Meloidogyne spp. were especially enhanced in high-density orchards. CONCLUSIONS: Olive anthropogenic practices reduce the PPN diversity in communities and lead to changes of the community structures with the development of some damaging nematodes. The study underlined the PPN diversity as a relevant indicator to assess community pathogenicity. That could be taken into account in order to design control strategies based on community rearrangements and interactions between species instead of reducing the most pathogenic species.

Fine-scale ecological and economic assessment of climate change on olive in the Mediterranean Basin reveals winners and losers.

The Mediterranean Basin is a climate and biodiversity hot spot, and climate change threatens agro-ecosystems such as olive, an ancient drought-tolerant crop of considerable ecological and socioeconomic importance. Climate change will impact the interactions of olive and the obligate olive fruit fly (Bactrocera oleae), and alter the economics of olive culture across the Basin. We estimate the effects of climate change on the dynamics and interaction of olive and the fly using physiologically based demographic models in a geographic information system context as driven by daily climate change scenario weather. A regional climate model that includes fine-scale representation of the effects of topography and the influence of the Mediterranean Sea on regional climate was used to scale the global climate data. The system model for olive/olive fly was used as the production function in our economic analysis, replacing the commonly used production-damage control function. Climate warming will affect olive yield and fly infestation levels across the Basin, resulting in economic winners and losers at the local and regional scales. At the local scale, profitability of small olive farms in many marginal areas of Europe and elsewhere in the Basin will decrease, leading to increased abandonment. These marginal farms are critical to conserving soil, maintaining biodiversity, and reducing fire risk in these areas. Our fine-scale bioeconomic approach provides a realistic prototype for assessing climate change impacts in other Mediterranean agro-ecosystems facing extant and new invasive pests.

OeFAD8, OeLIP and OeOSM expression and activity in cold-acclimation of Olea europaea, a perennial dicot without winter-dormancy.

MAIN CONCLUSION: Cold-acclimation genes in woody dicots without winter-dormancy, e.g., olive-tree, need investigation. Positive relationships between OeFAD8, OeOSM , and OeLIP19 and olive-tree cold-acclimation exist, and couple with increased lipid unsaturation and cutinisation. Olive-tree is a woody species with no winter-dormancy and low frost-tolerance. However, cold-tolerant genotypes were empirically selected, highlighting that cold-acclimation might be acquired. Proteins needed for olive-tree cold-acclimation are unknown, even if roles for osmotin (OeOSM) as leaf cryoprotectant, and seed lipid-transfer protein for endosperm cutinisation under cold, were demonstrated. In other species, FAD8, coding a desaturase producing α-linolenic acid, is activated by temperature-lowering, concomitantly with bZIP-LIP19 genes. The research was focussed on finding OeLIP19 gene(s) in olive-tree genome, and analyze it/their expression, and that of OeFAD8 and OeOSM, in drupes and leaves under different cold-conditions/developmental stages/genotypes, in comparison with changes in unsaturated lipids and cell wall cutinisation. Cold-induced cytosolic calcium transients always occurred in leaves/drupes of some genotypes, e.g., Moraiolo, but ceased in others, e.g., Canino, at specific drupe stages/cold-treatments, suggesting cold-acclimation acquisition only in the latter genotypes. Canino and Moraiolo were selected for further analyses. Cold-acclimation in Canino was confirmed by an electrolyte leakage from leaf/drupe membranes highly reduced in comparison with Moraiolo. Strong increases in fruit-epicarp/leaf-epidermis cutinisation characterized cold-acclimated Canino, and positively coupled with OeOSM expression, and immunolocalization of the coded protein. OeFAD8 expression increased with cold-acclimation, as the production of α-linolenic acid, and related compounds. An OeLIP19 gene was isolated. Its levels changed with a trend similar to OeFAD8. All together, results sustain a positive relationship between OeFAD8, OeOSM and OeLIP19 expression in olive-tree cold-acclimation. The parallel changes in unsaturated lipids and cutinisation concur to suggest orchestrated roles of the coded proteins in the process.

Effects of different arbuscular mycorrhizal fungal backgrounds and soils on olive plants growth and water relation properties under well-watered and drought conditions.

The adaptation capacity of olive trees to different environments is well recognized. However, the presence of microorganisms in the soil is also a key factor in the response of these trees to drought. The objective of the present study was to elucidate the effects of different arbuscular mycorrhizal (AM) fungi coming from diverse soils on olive plant growth and water relations. Olive plants were inoculated with native AM fungal populations from two contrasting environments, that is, semi-arid - Freila (FL) and humid - Grazalema (GZ) regions, and subjected to drought stress. Results showed that plants grew better on GZ soil inoculated with GZ fungi, indicating a preference of AM fungi for their corresponding soil. Furthermore, under these conditions, the highest AM fungal diversity was found. However, the highest root hydraulic conductivity (Lpr ) value was achieved by plants inoculated with GZ fungi and growing in FL soil under drought conditions. So, this AM inoculum also functioned in soils from different origins. Nine novel aquaporin genes were also cloned from olive roots. Diverse correlation and association values were found among different aquaporin expressions and abundances and Lpr , indicating how the interaction of different aquaporins may render diverse Lpr values.