In this study, we investigated the effect of heat exposure during olive inflorescence on reproductive development. We hypothesized that male development processes during this period are particularly sensitive to high temperatures. We characterized the development of flowers and inflorescences under natural conditions in the cultivars 'Arbequina' and 'Koroneiki', and defined eight phenological stages, beginning with the emergence of reproductive buds until anthesis (international standard BBCH scale of the phenological inflorescence development phases 53-60). In addition, we examined the response of olive inflorescence and flowering, to two heat scenarios: prolonged exposure to moderately high temperatures and a two-hour concentrated heat shock treatment. Analysis of our data revealed that both treatments had a detrimental effect on the development and functioning of the tapetum, the innermost layer of the anther, which is essential for the development and functioning of pollen. Additionally, we found that the 'Arbequina' cultivar displayed relative tolerance to heat compared to 'Koroneiki' in terms of pollen germinability after heat stress. In contrast to the response of the above cultivars, the 'Souri' exhibited an exceptionally high sensitivity to heat. An assessment of pollen viability after in vitro heat shock treatment of 54 cultivars revealed that pollen from seven varieties, 'Niedda de Oliana', 'Lech & iacute;n de Sevilla', 'Tonda Oliana', 'Tamir', 'Sigoise', 'Taggiasca', and 'Carrasquenha', showed exceptional tolerance to heat. These cultivars show a greater degree of suitability for cultivation in warm climates, (or in areas susceptible to extreme hot-spells). This study demonstrated that heat has a negative impact on the male reproductive system in olives, during early flower development and in mature pollen grains, which are relatively resistant to heat. Heat stress impaired pollen tube elongation capacity, and germination rate. Additionally, heat stress affected the ultrastructure of the external wall of the pollen grain, the exine, potentially impairing its ability to adhere to the stigma and fertilize the ovule. The sensitivity of olive reproductive capacity to heat is a genotype-dependent trait, allowing for the identification of heat-resistant cultivars for use as pollen donors and for development of new heat-resistant cultivars.

The major reasons for developing mechanical technologies for olive harvesting are the chronic shortage of workers for manual harvesting and increasing labor costs. To enable these technologies to operate, new table olive cultivars suitable for mechanical harvesting are necessary. The two major factors required for the shift from manual to mechanical harvesting of table olives are improved harvesting efficiency and prevention of fruit injury. Improved harvesting efficiency requires suitable pretreatment to enable fruit abscission with minimal defoliation, even when the harvesting is performed by a trunk shaker. The second requirement is prevention of external fruit color change or browning as a result of fruit injury, by development of olive cultivars with firm skin and higher resistance to the bruising caused by mechanical harvesting. This genetic adaptation to mechanical harvesting must be accompanied by efficient post-harvest processing of the olives. In this chapter, we will review the published studies regarding mechanical harvest of table olives, and attempt to identify the main issues, which still need to be studied in order to facilitate the transition from hand to mechanical harvest of table olives.

Climate change, with elevated temperatures throughout the year, affects many stages of the reproductive growth and development of olives as well as oil quality at harvest. Although olive (Olea europaea L.) is well adapted to the environmental conditions of the Mediterranean Basin, agricultural techniques and breeding through selection programs will have to adapt to these climate change, threatening to worsen in the near future. Defining the pathways controlling high fruit productivity and oil quantity and quality, despite elevated temperatures and sub-optimal growing conditions, is important for coping with current and predicted climate changes. As breeding programs aiming to address these crucial changes may take several decades, an urgent need to designate specific olive cultivars that are more resistant to high temperatures emerges.

Global warming is predicted to have a negative effect on plant growth due to the damaging effect of high temperatures. In order to address the effect of high temperature environments on olive oil yield and quality, we compared its effect on the fruit development of five olive cultivars placed in a region noted for its high summer temperatures, with trees of the same cultivars placed in a region of relatively mild summers. We found that the effects of a high temperature environment are genotype dependent and in general, high temperatures during fruit development affected three important traits: fruit weight, oil concentration and oil quality. None of the tested cultivars exhibited complete heat stress tolerance. Final dry fruit weight at harvest of the 'Barnea' cultivar was not affected by the high temperature environment, whereas the 'Koroneiki', 'Coratina', 'Souri' and 'Picholine' cultivars exhibited decreased dry fruit weight at harvest in response to higher temperatures by 0.2, 1, 0.4 and 0.2 g respectively. The pattern of final oil concentration was also cultivar dependent, 'Barnea', 'Coratina' and 'Picholine' not being affected by the high temperature environment, whereas the 'Koroneiki' and 'Souri' cultivars showed a decreased dry fruit oil concentration at harvest under the same conditions by 15 and 8% respectively. Regarding the quality of oil produced, the 'Souri' cultivar proved more tolerant to a high temperature environment than any other of the cultivars analyzed in this study. These results suggest that different olive cultivars have developed a variety of mechanisms in dealing with high temperatures. Elucidation of the mechanism of each of these responses may open the way to development of a variety of olives broadly adapted to conditions of high temperatures.

The "Souri" olive cultivar, which is autochthonous to the eastern Mediterranean region, has been the major olive variety cultivated traditionally under rain-fed conditions in northern Israel. The aim of this study was to determine the optimal pollen donor for the olive cultivar Souri in order to maximize Souri fruit set. Artificial cross pollination of Souri flowers with several local varieties has identified the "Nabali" as the most efficient pollinizer of the Souri. However, further experiments using artificial cross pollination conducted with cultivars not common to this region have revealed the "Arbequina" as a more efficient pollinizer of the Souri cultivar than the Nabali. Based on a preliminary paternity analysis, the Nabali was identified as the dominant pollinizer of Souri trees in traditional olive orchards in the north of Israel. However, in a multi-variety orchard, molecular paternity analysis has shown Arbequina to be the most frequent pollinizer. We then tested, during two consecutive years, whether the presence of a pollen-producing Arbequina tree adjoining Souri trees in the field will increase their fruit set. We found that Souri fruit set was 8.36% when pollinized by an Arbequina tree in close proximity to them, significantly higher than the fruit set of 5.6% for Souri trees without the nearby Arbequina cultivar. On the basis of these trials, we expect that the yield of Souri orchards will improve if Arbequina trees are planted.

Climate change has been shown to have a substantial impact on agriculture and high temperatures and heat stress are known to have many negative effects on the vegetative and reproductive phases of plants. In a previous study, we addressed the effects of high temperature environments on olive oil yield and quality, by comparing the fruit development and oil accumulation and quality of five olive cultivars placed in high temperature and moderate temperature environments. The aim of the current study was to explore the molecular mechanism resulting in the negative effect of a high temperature environment on oil quantity and quality. We analyzed the transcriptome of two extreme cultivars, 'Barnea', which is tolerant to high temperatures in regard to quantity of oil production, but sensitive regarding its quality, and 'Souri', which is heat sensitive regarding quantity of oil produced, but relatively tolerant regarding its quality. Transcriptome analyses have been carried out at three different time points during fruit development, focusing on the genes involved in the oil biosynthesis pathway. We found that heat-shock protein expression was induced by the high temperature environment, but the degree of induction was cultivar dependent. The 'Barnea' cultivar, whose oil production showed greater tolerance to high temperatures, exhibited a larger degree of induction than the heat sensitive 'Souri'. On the other hand, many genes involved in olive oil biosynthesis were found to be repressed as a response to high temperatures. OePDCT as well as OeFAD2 genes showed cultivar dependent expression patterns according to their heat tolerance characteristics. The transcription factors OeDof4.3, OeWRI1.1, OeDof4.4 and OeWRI1.2 were identified as key factors in regulating the oil biosynthesis pathway in response to heat stress, based on their co-expression characteristics with other genes involved in this pathway. Our results may contribute to identifying or developing a more heat tolerant cultivar, which will be able to produce high yield and quality oil in a future characterized by global warming.

Currently, table olives, unlike oil olives, are harvested manually. Shortage of manpower and increasing labor costs are the main incentives to mechanizing the harvesting of table olives. One of the major limiting factors in adopting mechanical harvest of table olives is the injury to fruit during mechanical harvest, which lowers the quality of the final product. In this study, we used the Israeli germplasm collection of olive cultivars at the Volcani Institute to screen the sensitivity of many olive cultivars to browning in response to injury. The browning process after induced mechanical injury was characterized in 106 olive cultivars. The proportional area of brown coloring after injury, compared to the total fruit surface area, ranged from 0 to 83.61%. Fourteen cultivars were found to be resistant to browning and did not show any brown spot 3 h after application of pressure. Among them, there are some cultivars that can serve as table olives. The different response to mechanical damage shown by the cultivars could be mainly due to genetic differences. Mesocarp cells in the fruits of the sensitive cultivars were damaged and missing the cell wall as a result of the applied pressure. The cuticles of resistant cultivars were thicker compared to those of susceptible cultivars. Finally, we showed that the browning process is enzymatic. We suggest cuticle thickness as an indicator of table olive cultivars suitable for mechanical harvest. A shift to browning-resistant cultivars in place of the popular cultivars currently in use will enable the mechanical harvest of table olive without affecting fruit quality.

Isohydric plants maintain constant water potential through rapid stomatal closure, whereas anisohydric plants only close their stomata at very low water potentials. However, distinctions between isohydric and anisohydric behaviors among different cultivars of the same species are unclear. This study compared the physiological response to prolonged drought stress in the isohydric Grenache and the anisohydric Shiraz cultivars of the Vitis vinifera species. Plants were exposed to 60-day periods of deficit irrigation (25% of plant water consumption under well-watered conditions) during the summers of 2011 and 2012. Physiological measurements, water potential, leaf gas exchange, canopy area, leaf senescence, stem characteristics, and morphological characteristics were analyzed. Stomatal conductance was consistently lower in Grenache than in Shiraz at all values of midday stem and predawn leaf water potentials, respectively. The Shiraz plants exhibited greater vegetative growth and less defoliation than the Grenache plants in response to water deficit. Anatomical architecture analyses revealed that Grenache plants had greater xylem vessel diameter, hydraulic conductivity, and stomatal density than the Shiraz plants. These results suggest isohydric and anisohydric behaviors may be well-defined, time-regulated responses rather than distinct mechanisms that plants use to cope with drought stress. The rapid response to water deficit exhibited by isohydric plants may be because they are more vulnerable to fatal xylem embolisms than anisohydric plants. Thus, the accelerated response allows isohydric plants to avoid drought stress and minimize risk of xylem cavitation, but may lower the plant's ability to survive moderate stress of prolonged drought.

The olive (Olea europaea L.) was domesticated in the Mediterranean area over 6000 years ago and is currently one of the area's most important oleaginous crops. Due to its economic, cultural and ecological importance, breeding programs aimed at obtaining new olive cultivars have been developed in most olive-producing countries. An efficient breeding program requires a large and genetically variable germplasm collection. In this study, we used next-generation sequencing technology for the identification of 145,974 single nucleotide polymorphism (SNPs) loci. A subset of 138 SNPs was then used to analyze the genetic relationships between 119 cultivars making up most of the Israeli germplasm collection. The various cultivars did not cluster according to their geographic origin but rather showed a high correlation with their function (oil, table or dual purpose). Comparison of genetic diversity between 15 cultivars using SSRs and SNPs revealed that for the purposes of analyzing genetic variation between olive cultivars, the SSR marker seems more suitable. However, based on the analysis of several trees of the same cultivar sampled from different nurseries, the SNP marker proved to be a more reliable criterion for cultivar identification. This study presents the most comprehensive SNP analysis of olive phylogeny to date. Based on the rapid development of SNP genotyping methods over the last few years, we believe that in the near future, we will be able to genotype many sample genomes using the appropriate SNPs at a reasonably low cost. Therefore, we can expect that in the future, SNPs will definitely be the marker of choice for biodiversity analysis as well as for gene cloning and QTL identification in olives.

The performance of five cv. Souri selections designated as “clones” were compared in two consecutive plots for ≈20 years each and three additional ones for 18 years. Fruit morphology, yield, oil production, and tree growth were monitored. The major results are presented as mean values of five sequential “on” years as well as the characteristic production of young and mature trees. The DNA identity was determined using both simple sequence repeats (SSRs) and single nucleotide polymorphisms (SNPs). The performance of two “clones,” SLO and SBU, showed stable diverse production characteristics and could be designated as defined clones, although no clear differences of the DNA between them or the other “clones” were apparent. Another “clone,” SGS, was also designated as a true clone as a result of both unique fruit morphology and DNA markers. The differences between the other “clones” were minute in fruit production morphology and oil production as well as on a DNA basis and their designation as being true clones is doubtful.

Previous studies using ‘Hass’ avocado cultivar showed that its small-fruit (SF) phenotype is limited by cell number. To explore the molecular components affecting avocado cell production, we isolated four cDNAs encoding: an ICK/KRP protein, known to play cell cycle-regulating roles through modulation of CDK function; two CDK proteins and a D-type cyclin, and monitored their expression patterns, comparing NF (normal fruit) versus SF profiles. The accumulation of PaKRP gradually deceased during growth in both fruit populations. Despite these similarities, SF exhibited higher levels of PaKRP accumulation at early stages of growth. Moreover, in NF, augmented PaKRP expression coincided with a decrease in CDK and PaCYCD1 levels, whereas in SF, enhanced PaKPR expression was coupled with an earlier decline of CDK and PaCYCD1 levels. For both NF and SF, enhanced mesocarp PaKRP transcript accumulation, was associated with elevated abscisic acid (ABA) and ABA catabolites content. Nevertheless, the collective ABA levels, including catabolites, were substantially higher in SF tissues, as compared with NF tissues. Finally, additional expression analysis revealed that in cultured cells, PaKRP could be induced by ABA. Together, our data links PaKRP with exit from the fruit cell cycle and suggest a role for ABA in controlling its expression.

Molecular control mechanisms for abiotic stress tolerance are based on the activation and regulation of specific stress-related genes. The phytohormone abscisic acid (ABA) is a key endogenous messenger in a plant's response to such stresses. A novel ABA binding mechanism which plays a key role in plant cell signaling cascades has recently been uncovered. In the absence of ABA, a type 2C protein phosphatase (PP2C) interacts and inhibits the kinase SnRK2. Binding of ABA to the PYR/PYLs receptors enables interaction between the ABA receptor and the PP2C protein, and abrogates the SnRK2 inactivation. The active SnRK2 is then free to activate the ABA-responsive element Binding Factors which target ABA-dependent gene expression. We used the grape as a model to study the ABA perception mechanism in fruit trees. The grape ABA signaling cascade consists of at least seven ABA receptors and six PP2Cs. We used a yeast two-hybrid system to examine physical interaction in vitro between the grape ABA receptors and their interacting partners, and found that twenty-two receptor-PP2C interactions can occur. Moreover, quantifying these affinities by the use of the LacZ reporter enables us to show that VvPP2C4 and VvPP2C9 are the major binding partners of the ABA receptor. We also tested in vivo the root and leaf gene expression of the various ABA receptors and PP2Cs in the presence of exogenic ABA and under different abiotic stresses such as high salt concentration, cold and drought, and found that many of these genes are regulated by such abiotic environmental factors. Our results indicate organ specificity in the ABA receptor genes and stress specificity in the VvPP2Cs. We suggest that VvPP2C4 is the major PP2C involved in ABA perception in leaves and roots, and VvRCAR6 and VvRCAR5 respectively, are the major receptors involved in ABA perception in these organs. Identification, characterization and manipulation of the central players in the ABA signaling cascades in fruit trees is likely to prove essential for improving their performance in the future.

Mango malformation disease (MMD) caused by various Fusaria, including the fungus Fusarium mangiferae , is difficult to diagnose and cannot be controlled effectively. In this study a PCR diagnostic tool was developed for detection of F. mangiferae by generating primers which flank fungus-specific sequences. The assay is suitable for both in-vitro and in-vivo tests, and is capable of detecting 10 pg of the fungal DNA. Relatively high genetic variation was discovered between various isolates of the pathogen based on amplified fragment length polymorphism (AFLP) analyses. In addition, some isolates of F. mangiferae (including the accession numbers FMS-123 and NRRL 25226), were found to be quite distinct from most F. mangiferae accessions. Likewise, representative MMD-associated Fusaria from the Asian and American clades of the Gibberella fujikuroi species complex were further differentiated by AFLP analyses.

The olive (Olea europaea) is one of the most important oleaginous crops of the Mediterranean basin. Increased demand for olive oil creates a need for new olive varieties to help meet the requirements of the global market. However, olive breeding has been handicapped by such varied challenges as a prolonged juvenile period, agrotechnical problems and insufficient genetic knowledge. The use of DNA markers has the potential to overcome these problems and increase the effectiveness of classical breeding programmes. In this study, co-dominant polymorphic simple sequence repeats (SSRs) were used as markers to analyse the genetic relationships between several local and other non-native olive cultivars. Cluster analysis revealed four major groups among the 15 cultivars examined in this study. Table and oil cultivars were clustered in different groups. However, the clusters did not differentiate between cultivars of different geographical origins. In addition, we used the data gathered to analyse genetic relationships to evaluate the effects of heterosis in agricultural traits. Genetic distances between cultivars were determined based on the SSR genotype data and were used for evaluating the possible effects of heterosis in various F1 populations. Interestingly, phenotypic data of F1 progenies from crosses between different cultivars indicated the potential effects of heterosis as expressed in several traits. Genetic distance between parents was significantly correlated to F1 performance for three traits: percentage of dry fruit weight, oil content and commercial oil production. Thus, crosses between olive cultivars exhibiting relatively extensive genetic distances one from the other are expected to result in better progeny performance in future Olea breeding programmes. Our study linked assessment of biodiversity of commercial olive cultivars with the application of this information in olive breeding programmes for selection of specific parents to generate superior new cultivars.

The plant hormone abscisic acid (ABA) regulates many key processes in plants including the response to abiotic stress. ABA signal transduction consists of a double-negative regulatory mechanism, whereby ABA-bound PYR/RCARs inhibit PP2C activity, and PP2Cs inactivate SnRK2s. We studied and analyzed the various genes participating in the ABA signaling cascade of the grape (Vitis vinifera). The grape ABA signal transduction consists of at least six SnRK2s. Yeast two-hybrid system was used to test direct interactions between core components of grape ABA signal transduction. We found that a total of forty eight interactions can occur between the various components. Exogenous abscisic acid (ABA) and abiotic stresses such as drought, high salt concentration and cold, were applied to vines growing in a hydroponic system. These stresses regulated the expression of various grape SnRK2s as well as ABFs in leaves and roots. Based on the interactions between SnRK2s and its targets and the expression pattern, we suggest that VvSnRK2.1 and VvSnRK2.6, can be considered the major VvSnRK2 candidates involved in the stomata response to abiotic stress. Furthermore, we found that the expression pattern of the two grape ABF genes indicates organ specificity of these genes. The key role of ABA signaling in response to abiotic stresses makes the genes involve in this signaling potential candidates for manipulation in programs designed to improve fruit tree performance in extreme environments.