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Background and aim In vitro activity evaluation of Egyptian Olea europaea leaves extracts, and in vivo healing activity assessment of the newly prepared ointment of Olea europaea leaves extracts mingled with Shea butter. Experimental procedure Different extraction methods and solvents were used to extract Egyptian Olea europaea bioactive agent(s). Antibacterial, scavenging activity and in-vivo evaluation of wound repair potentiality of Olea europaea extract were examined in normal and diabetic experimental rat models with induced circular excisions. Results and conclusion Olive leaves extract of Tanta was selected as the most active agent against Methicillin-resistant S. aureus (MRSA), with MIC value 15.6 μg/ml. Moreover, checkerboard dilution technique approved that the interaction between Tanta LEM crude extract and Ciprofloxacin was synergistic. Scavenging activity of the extract against DPPH free radicals was 87.55% at concentration of 50 μg/ml. In vivo normal and diabetic experimental rats treated with Shea butter: Tanta LEM extract (1:3 w/v) showed the maximum wound contraction and healing activity., Graphical abstract Image 1, Highlights • This is the first work to assess Egyptian olive leaves extract as an effective treatment against MRSA. • The novel combination between shea butter and Tanta LEM showed promising wound healing activity in normal and diabetic rats. • The combination between Tanta LEM extract and Ciprofloxacin was found to be synergistic.

Survival, germination, olive colonization, and water-use efficiency (WUE) impairments by Verticillium dahliae could be influenced by cultivar susceptibility or irrigation, and this could modify the irrigation–pathogen–disease relationship. In this study, the combined effects of irrigation and cultivar susceptibility on Verticillium wilt (VW) development were modelled by the temporary assessment of V. dahliae propagules (total inoculum density, density of micropropagules, and sclerotia in wet and air-dried soil; ID, MpD, SwD, and SdD, respectively), root (RCI) and shoot (SCI) colonization indexes, and WUE. The relationship of disease severity to the measured parameters was then explored. Under controlled conditions, plants of cultivars ‘Picual’ and ‘Frantoio’ were irrigated to a high and low rate by varying drip-irrigation frequencies: daily, twice weekly, and a combination of daily for 11 days and then twice weekly. Disease severity and colonization parameters were higher in ‘Picual’, while WUE was higher in ‘Frantoio’. However, high rate and twice weekly and combination treatments significantly increased disease incidence and reduced time-to-symptoms-onset only in ‘Picual’, while high rate reduced WUE and increased relative ID, MpD, and SwD in both cultivars. Irrigation did not affect SCI, but a higher RCI was found at high rate during the development of symptoms in ‘Picual’. By using classification trees to examine parameters—disease severity relationships, it was possible to determine the degree to which VW was affected by irrigation and/or cultivar susceptibility. MpD was the best indicator for VW detection at any time, WUE was best before symptoms developed, and RCI, total ID, and SdD after symptoms developed.

Flavonoids are a class of natural chemicals with variable phenolic structures that have long been recognized for their health advantages, they have recently attracted researchers’ attention for treating diabetes and hyperglycemia. The goal of this research is to develop a novel antihyperglycemic formulation using a combination of three plant flavonoids: Linum usitatissimum L. seeds (FLU), Coriandrum sativum L. seeds (FCS), and Olea europaea var. sylvestris leaves (FOE) based on a mixture design experiment approach which generates the most effective ratio of each component in a mixture instead of the trial-and-error method. Prior to the test, sub-acute toxicity research was conducted to establish a safe and effective dosage. The Oral Glucose Tolerance Test (OGTT) was used to assess the antihyperglycemic impact of these extracts and their combinations in Swiss albino mice. The dose that showed efficacy and safety was 25 mg/kg, which was utilized in all formulations. According to the results, the binary and ternary combinations showed the most significant synergetic effects. The optimum combination with the most potent effect was 37% FLU, 20% FCS, and 43% FOE. This study’s mixture design and prediction model for glycemic variation (GV) may be utilized at an industrial level to develop a novel antidiabetic and antihyperglycemic formulation that is safe and effective.

La olivicultura está afectada por un síndrome que genera marchitez en la copa del árbol. Uno de los microorganismos responsables es el hongo Verticillium dahliae, que provoca la enfermedad denominada verticilosis del olivo (VO). Según la severidad de los síntomas, se distinguen dos patotipos, defoliante (D) y no defoliante (ND). Su diagnóstico emplea la reacción en cadena de la polimerasa (PCR). Nuestro objetivo fue comparar la eficiencia de dos juegos de cebadores para detectar en planta el patotipo ND, el único presente en Argentina, mediante PCR anidada. Para ello se utilizaron los juegos de cebadores NDf/NDr - INTND2f/INTND2r (PCR-I), empleados comúnmente, y una nueva combinación, INTNDf/INTNDr - INTND2f/INTND2r (PCR-II). Se analizaron muestras de olivos sintomáticos de 39 fincas de la provincia de La Rioja. PCR-II tuvo mayor eficiencia en la detección, 57,8 % frente a 16,7 % de PCR-I. PCR-II amplificó en casi todas las variedades, edades de plantas y niveles de severidad de la enfermedad, mientras que PCR-I detectó el hongo sólo en variedades muy susceptibles y con niveles de severidad leve. El uso de PCR-II para detectar a V. dahliae en olivo permitirá estudios más precisos sobre su incidencia que contribuirán al manejo adecuado de la verticilosis. Olive growing is affected by a syndrome that causes wilt in the tree top. One of the main microorganisms accountable for this disease is the fungus Verticillium dahliae, which causes verticillium wilt (VW). Two pathotypes are distinguished within V. dahliae, defoliating (D) and non-defoliating (ND), depending on symptom severity. The pathogen is generally diagnosed by polymerase chain reaction (PCR). Our objective was to compare the efficiency of two sets of primers in detecting the ND pathotype in olive plants, the only pathotype found in Argentina, by nested PCR. For this purpose, the commonly used primer combination NDf/NDr - INTND2f/INTND2r (PCR-I) and a new combination, INTNDf/INTNDr - INTND2f/INTND2r (PCR-II), were evaluated. Samples of symptomatic olive trees from 39 commercial fields from La Rioja province were analyzed. PCR-II had higher detection efficiency than PCR-I (57.8 % and 16.7 %, respectively). PCR-II amplified in almost all the olive cultivars, plant ages and VW disease severity levels, whereas PCR-I detected the fungus only in very susceptible cultivars and in trees with low VW severity levels. The use of PCR-II for the detection of V. dahliae in olive plants will provide more accurate incidence results and will contribute to a proper management of VW. Instituto de Patología Vegetal Fil: Rattalino, Donna. Universidad Nacional de Chilecito. Instituto de Ambientes de Montaña y Regiones Áridas. Departamento de Ciencias Básicas y Tecnológicas; Argentina Fil: Otero, Maria Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Otero, Maria Laura. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Fitopatología y Modelización Agrícola (UFyMA); Argentina Fil: Moriconi, D. N. Universidad Nacional de Chilecito. Instituto de Ambientes de Montaña y Regiones Áridas. Departamento de Ciencias Básicas y Tecnológicas; Argentina Fil: Rivera, P. C. Universidad Nacional de Chilecito. Instituto de Ambientes de Montaña y Regiones Áridas. Departamento de Ciencias Básicas y Tecnológicas; Argentina Fil: Rivera, P. C. Consejo Nacional de Investigaciones Científicas y Técnicas. Universidad Nacional de Córdoba. Instituto de Diversidad y Ecología Animal; Argentina

Verticillium wilt (caused by Verticillium dahliae Kleb.) is an important disease affecting olive (Olea europaea L.) production. Effective control of this disease relies on integrated management strategies. In vitro antifungal activity of two hydroxytyrosol (HTyr) enriched extracts (HTE1 and HTE2) obtained from olive mill waste products (OMWP) was assessed against V. dahliae. Inhibitory effects of pure HTyr as a standard, and HTE1 and HTE2 at different concentrations, were evaluated on mycelium growth and conidium germination of V. dahliae. Chemical characterization of HTE1 and HTE2 allowed identification and quantification of HTyr as the main constituent in both extracts along with other low molecular weight phenols. HTE1 showed a higher inhibitory activity in both growth and conidium germination of V. dahliae. At the tested concentrations, low antifungal effects of HTyr were observed. After 3 d, 1 mg mL-1 of HTE1 gave greater inhibition of mycelium growth than HTE2 or HTyr. After 24 h, HTE1 gave 55% inhibition of conidium germination, and HTyr and HTE2 both gave 37% inhibition. This study has demonstrated that phenolic compounds derived from OMWP have antifungal activity against V. dahliae, indicating the potential of these compounds for eco-friendly control of Verticillium wilt.

Key message: All olive cultivars survived high salinity, yet, the question was how they orchestrated antioxidative defense, mannitol accumulation and ion sequestration with different efficiencies due to their genetic makeup. Abstract: A Mediterranean plant, Olea europaea (olive) is well adapted to high temperature, high light intensity and dry soil conditions hence accepted to be moderately salt tolerant. Control of Na+ and Cl? uptake, accumulation of osmoprotectants and high antioxidative capacity were all attributed to salt stress tolerance of olive, however, there is a great discrepancy between the efficiency and onset of these processes among different cultivars. In present study, three olive cultivars namely Ayvalık, Gemlik and Kilis which are well adapted to different climatic regions extending from cool and high precipitation receiving regions to long hot and dry periods with high light intensity through the growth season were tested for their relative salt-tolerance mechanisms. For this aim, 1-year-old cuttings of these cultivars were treated with 0, 100, 200 and 300 mM NaCl for 30 days. Several physiological and biochemical processes along with their growth performance were analyzed during the experimental period. In general, we found discrepancies in salt stress-coping strategies of these cultivars. Efficient osmoprotection in Gemlik cv. lead to a better leaf water status and photosynthetic performance while, active ion sequestration and high levels of SOD and POX enabled Kilis cv. to be protected from the negative impact of salinity. Moreover, Ayvalık cv. accumulated high levels of mannitol that probably served as an antioxidative molecule to cope with salt stress. © 2021, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature., TAGEM/TSK AD/14/A13/P.02/09, The research was supported by a grant from Agricultural Research and Policies Council of the Turkish Ministry of Agriculture and Forestry (TAGEM/TSK AD/14/A13/P.02/09).

6 Figuras, BACKGROUND Hydrolysis of the fruit phenolic glucosides occurring during the oil extraction process is the main biochemical reaction affecting the biosynthesis and accumulation of secoiridoid compounds in virgin olive oil. An integrated approach at the molecular, biochemical, and metabolic level was used to study the olive β‐glucosidase gene family in seven olive cultivars selected by their different phenolic profiles. RESULTS Eight β‐glucosidase genes have been identified by in silico analysis of an olive transcriptome. Their expression levels were analyzed by reverse transcription quantitative polymerase chain reaction in olive fruits at different ripening stages: I, green fruits, 16–19 weeks after flowering (WAF); II, yellow–green fruits, 22–25 WAF; III, turning fruits, 28–31 WAF; and IV, fully ripe fruits, 35–40 WAF. Gene expression was compared with the level of β‐glucosidase activity in the fruit and with the phenolic composition of fruits and oils from different olive cultivars. Phylogenetic analysis of the encoded proteins and differences found among the β‐glucosidase genes based on Gene Ontology enrichment analysis data suggests maximum involvement of two genes, OeBGLU1A and OeBGLU1B, in the phenolic composition of virgin olive oil. Positive correlation coefficients were found within each olive cultivar between OeBGLU1A and OeBGLU1B gene expression data and the phenolic content of the oil. CONCLUSION The results obtained suggest that the expression pattern of specific β‐glucosidase genes may be an accurate predictor for the phenolic content of virgin olive oil that could be used in olive breeding programs. © 2021 Society of Chemical Industry., This research was funded by the research project AGL2015‐67652‐R from the ‘Programa Estatal de I+D+i Orientada a los Retos de la Sociedad’ funded by the Government of Spain. We are grateful to Mar Pascual for her technical assistance.

Purpose of Review The olive tree (Olea europaea L.) has featured as a significant part of medicinal history, used to treat a variety of ailments within folk medicine. The Mediterranean diet, which is rich in olive products, is testament to Olea europaeas positive effects on health, associated with reduced incidences of cancer and cardiovascular disease. This review aims to summarise the current literature regarding the therapeutic potential of Olea europaea products in cancer, detailing the possible compounds responsible for its chemotherapeutic effects. Recent Findings Much of the existing research has focused on the use of cell culture models of disease, demonstrating Olea europaea extracts, and specific compounds within these extracts, have efficacy in a range of in vitro and in vivo cancer models. The source of Olea europaeas cytotoxicity is yet to be fully defined; however, compounds such as oleuropein and verbascoside have independent cytotoxic effects on animal models of cancer. Summary Initial results from animal models are promising but need to be translated to a clinical setting. Treatments utilising these compounds are likely to be well tolerated and represent a promising direction for future research.

The cellular response to stress also includes the accumulation of reactive oxygen species (ROS). When in excess, these second messengers can cause significant detrimental biochemical changes, such as stress-induced lipid peroxidation. In plants, a powerful endogenous antioxidant system is based on the enzymatic modification of redox-sensitive cysteines, carried out for example, by thioredoxin proteins. In this work, we report the cloning and characterization of a thioredoxin gene from olive (Olea europaea L.), which was named Oe-Trx h I. We also performed an expression analysis in drupes following biotic stress, as a first step towards the definition of its possible defensive role against the olive fruit fly [Bactrocera oleae (Rossi)]. The data indicate that the cloned thioredoxin h is a potential member of the biotic stress response in drupes and suggest that Oe-Trx h I may be a component active towards an increased resistance against oxidative stress. This work opens the way to further studies to assess the protective role of this gene against ROS accumulation and lipid peroxidation in drupes following larval feeding.

Rhizospheric soils from cultivated olive (Olea europaea) trees of Chemlali, Chetoui, Quaissi, and Djalat cultivars were assessed for their bacterial abundance and diversity and were further screened for production of exopolysaccharides and exoenzymes (cellulase, chitinase, amylase, protease, lipase, and peroxidase). The results of the present study indicate that Chetoui cultivar revealed higher diversity, followed by Chemlali > Quaissi > Djalat, wherein, bacilli, enteric bacteria, and pseudomonads were abundantly present as specific bacterial groups associated with the Chetoui rhizosphere. Moreover, the exopolysaccharide (EPS)-producing bacteria of Chetoui cultivar (68.4%) presented the highest efficiency, followed by Djalat (23.5%) > Chemlali (7 %) > Quaissi (1%). These results revealed that the Chetoui cultivar presented highest enzyme activities, followed by Chemlali > Djalat > Quaissi, with a distinct abundance of peroxidase- and chitinase-producing bacteria, which may play a pivotal role in adapting olives to the environmental stresses. From this preliminary study, we confirmed that olive rhizosphere microbial diversity is essentially driven by the geographical origin and genotype of olive cultivars. Furthermore, we recommended the Chetoui olive cultivar rhizosphere as a potential reservoir for exoenzyme- and EPS-producing bacteria useful for future biotechnological applications.

Although bioactivities of Olea europaea (OE) have been widely described, most of them were related to its methanolic extracts or its essential oils, While data related to aqueous extracts still very scarce. Thus, in this study, the phytochemical composition, the antioxidant activity, the cytotoxic potential, and the allelopathic potential of aqueous leaf extracts from two varieties of Olea europaea were investigated and compared. High‐performance liquid chromatography (HPLC) was used to identify and quantify the constituents of the tested plants, and spectrophotometric methods to evaluate antioxidant activities. The cytotoxic potential was investigated using murine oligodendrocytes (158N) while germination seeds’ test was used for allelopathic activity. HPLC analysis showed the presence of 10 phenolic compounds in both extracts. Chemlali variety showed the highest antioxidant and allelopathic activities. Regarding the cytotoxicity effect, a significant increase in cell viability was observed with both of our extracts compared to untreated cells. These results confirm that aqueous extracts from OE produce a range of substances with potential antioxidant, antifungal, and allelopathic effects without toxic effects. Thus, they could be used as an alternative of chemical compounds.

Маслина европейская (Olea europaea L.) – одно из древнейших культурных растений на Земле. Помимо пищевой ценности плодов большое значение имеют препараты из различных частей маслины, проявляющие широкий спектр фармакологического действия. Цель работы: сравнительный анализ содержания фенольных соединений в листьях и плодах сортов и подвида маслины европейской, произрастающих в климатических условиях Южного берега Крыма. Суммарное содержание фенольных веществ и флавоноидов определяли спектрофотометрически. Компонентный состав – методом ВЭЖХ с диодно-матричным детектированием. В результате сравнительного исследования фенольных соединений листьев и плодов четырех сортов и одного подвида маслины установлено, что в листьях маслины содержится 740–980 мг/100 г фенольных веществ (в пересчете на сырой вес), в плодах – 840–1260 мг/100 г. В листьях содержится 190–312 мг/100 г флавоноидов, в плодах – 59–990 мг/100 г. Среди фенольных соединений листьев и плодов идентифицированы лютеолин-7-О-глюкозид, апигенин-7-О-глюкозид, рутин и олеуропеин. Выявлено, что доминирующим компонентом как листьев, так и плодов четырех сортов маслины является олеуропеин, а плодов подвида O. europaea subsp. cuspidata – рутин. Содержание флавоноидов и олеуропеина в листьях и плодах наиболее морозостойкого сорта Никитская оказалось самым низким из всех исследованных образцов, что может быть связано с участием данных веществ в подготовке этого генотипа к воздействию отрицательных температур. Показано, что содержание фенольных соединений листьев и плодов O. europaea зависит от сортовой принадлежности и климатических условий выращивания., European olive (Olea europaea L.) is one of the oldest cultivated plants on Earth. In addition to the nutritional value of the fruit, medications made from different parts of the olive are of great importance cause they exhibit a wide range of pharmacological action. The main objective of the present study was a comparative analysis of the content of phenolic compounds in the leaves and fruits of varieties and subspecies of European olives growing in the climatic conditions of the Southern Coast of Crimea. Total phenolics and flavonoids were determined spectrophotometrically. HPLC with diode-array detection was applied to determine individual components. As a result of the comparative study of the phenolic compounds of leaves and fruits in four varieties and one subspecies of olives it was found that total phenolics in the leaves of olives are in the range of 740–980mg/100 g (on a wet weight basis) and total phenolics in fruits are in the range of 840–1260 mg/100 g (on a wet weight basis). Flavonoids content in the leaves is in the range of 190–312 mg/100 g and in the range of 59–990 mg/100 g in the fruits. Luteolin-7-O-glucoside, apigenin-7-O-glucoside, rutin and oleuropein were identified among the phenolics of leaves and fruits. It was found that the oleuropein was a majour component either in leaves or in fruits of four varieties of olives but rutin was a majour component in the fruits of the O. europaea subsp. cuspidata subspecies. The content of flavonoids and oleuropein in the leaves and fruits of the most frost-resistant variety Nikitskaya turned out to be the lowest among all the studied samples. It may be related to the participation of these substances during preparation of the variety to the effects of below-freezing temperatures. It has been shown that the content of phenolic compounds in leaves and fruits of O. europaea depends on the variety and agroclimatic growing conditions.

Olive is considered one of the oldest and the most important cultivated fruit trees in Albania. In the present study, the genetic diversity and structure of Albanian olive germplasm is represented by a set of 194 olive genotypes collected in-situ in their natural ecosystems and in the ex-situ collection. The study was conducted using 26 microsatellite markers (14 genomic SSR and 12 Expressed Sequence Tag microsatellites). The identity analysis revealed 183 unique genotypes. Genetic distance-based and model-based Bayesian analyses were used to investigate the genetic diversity, relatedness, and the partitioning of the genetic variability among the Albanian olive germplasm. The genetic distance-based analysis grouped olives into 12 clusters, with an average similarity of 50.9%. Albanian native olives clustered in one main group separated from introduced foreign cultivars, which was also supported by Principal Coordinate Analysis (PCoA) and model-based methods. A core collection of 57 genotypes representing all allelic richness found in Albanian germplasm was developed for the first time. Herein, we report the first extended genetic characterization and structure of olive germplasm in Albania. The findings suggest that Albanian olive germplasm is a unique gene pool and provides an interesting genetic basis for breeding programs.

The use of high throughput sequencing (HTS) for the analysis of Spanish olive trees showing leaf yellowing discoloration, defoliation, and/or decline has provided new insights into the olive viruses present in Spain and has opened discussions about the pros and cons of these technologies for diagnostic purposes. In this study, we report for the first time in Spanish orchards the presence of olive leaf yellowing-associated virus (OLYaV), for which the second full coding sequence has been determined. This virus has also been detected in a putative vector, the psyllid Euphyllura olivina. In addition, the presence in Spain of Olea europaea geminivirus (OEGV), recently reported in Italy, has been confirmed, and the full-length sequence of two isolates was obtained by HTS and Sanger sequencing. These results, as well as the detection of other viral sequences related to olive latent virus 3 (OLV-3) and olive viral satellite RNA, raises questions on the biological significance of the findings, about the requirement of standardization on the interpretation of HTS results, and the necessity of additional tests to confirm the relevance of the HTS detection of viral sequences.

10 Páginas.-- 5 Figuras.-- 2 Tablas, The C6 aldehydes, alcohols, and the corresponding esters are the most important compounds of virgin olive oil aroma. These C6 volatile compounds are synthesized via the 13-hydroperoxide lyase (13-HPL) branch of the lipoxygenase pathway. In this investigation, a functional analysis of the olive (Olea europaea L.) 13-HPL gene by its overexpression and silencing in olive transgenic lines was carried out. With this aim, sense and RNAi constructs of the olive 13-HPL gene were generated and used for the transformation of embryogenic olive cultures. Leaves from overexpressing lines showed a slight increase in 13-HPL gene expression, whereas RNAi lines exhibited a strong decrease in their transcript levels. Quantification of 13-HPL activity in two overexpressing and two RNAi lines showed a positive correlation with levels of transcripts. Interestingly, RNAi lines showed a high decrease in the content of C6 volatiles linked to a strong increase of C5 volatile compounds, altering the volatile profile in the leaves. In addition, the silencing of the 13-HPL gene severely affected plant growth and development. This investigation demonstrates the role of the 13-HPL gene in the biogenesis of olive volatile compounds and constitutes a functional genomics study in olive related to virgin olive oil quality., We acknowledge Prof. Manuel Mancha for his strong support at the beginning of the olive transformation studies, and for his continuous encouragement, which made it possible to undertake this collaborative research. This work was supported by the OLEAGEN Project funded by the Fundación Genoma España, Junta de Andalucía through Instituto de Investigación y Formación Agraria y Pesquera (IFAPA) and Corporación Tecnológica de Andalucía (CTA).

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.

Simple Summary Olive lace bugs feed on wild and cultivated Olea europaea, causing a negative impact on plant vitality and development. These insects are known to affect olive orchards in South Africa, the country where most of the olive and olive products on the continent are produced. However, the diversity of species of these pests is not clear. Morphological analysis and DNA barcoding showed the presence of Cysteochila lineata, Plerochila australis, Neoplerochila paliatseasi and Neoplerochila sp. Further analyses of genetic divergence and phylogenetic clustering in 30 species in 18 genera of Tingidae using new and publicly available DNA barcodes showed that the majority of sequences deposited on BOLD Systems were correctly assigned to species. The complete mitochondrial genomes of the four species found in South Africa were sequenced to assess their phylogenetic position within Tingidae. The four olive lace bugs formed one cluster of species, and the genus Cysteochila was not monophyletic as C. lineata grouped with the other three olive lace bugs but C. chiniana was placed in a different cluster. This result suggests that lace bug species that feed on olive trees may have a common ancestor and calls for further research on potential adaptations to O. europaea. Abstract Olive lace bugs (Hemiptera: Tingidae) are small sap-sucking insects that feed on wild and cultivated Olea europaea. The diversity of olive lace bug species in South Africa, the most important olive producer on the continent, has been incompletely surveyed. Adult specimens were collected in the Western Cape province for morphological and DNA-based species identification, and sequencing of complete mitogenomes. Cysteochila lineata, Plerochila australis, Neoplerochila paliatseasi and Neoplerochila sp. were found at 12 sites. Intra- and interspecific genetic divergences and phylogenetic clustering in 30 species in 18 genera of Tingidae using new and publicly available DNA barcodes showed high levels of congruity between taxonomic and genetic data. The phylogenetic position of the four species found in South Africa was inferred using new and available mitogenomes of Tingidae. Notably, olive lace bugs formed a cluster of closely related species. However, Cysteochila was non-monophyletic as C. lineata was recovered as a sister species to P. australis whereas Cysteochila chiniana, the other representative of the genus, was grouped with Trachypeplus jacobsoni and Tingis cardui in a different cluster. This result suggests that feeding on O. europaea may have a common origin in Tingidae and warrants future research on potential evolutionary adaptations of olive lace bugs to this plant host.

Specialized metabolism is an evolutionary answer that fortifies plants against a wide spectrum of (a) biotic challenges. A plethora of diversified compounds can be found in the plant kingdom and often constitute the basis of human pharmacopeia. Olive trees (Olea europaea) produce an unusual type of secoiridoids known as oleosides with promising pharmaceutical activities. Here, we transiently silenced oleuropein β-glucosidase (OeGLU), an enzyme engaged in the biosynthetic pathway of secoiridoids in the olive trees. Reduction of OeGLU transcripts resulted in the absence of both upstream and downstream secoiridoids in planta, revealing a regulatory loop mechanism that bypasses the flux of precursor compounds toward the branch of secoiridoid biosynthesis. Our findings highlight that OeGLU could serve as a molecular target to regulate the bioactive secoiridoids in olive oils.

Simple Summary The phenomenon of climate change affects the entire world, especially the most vulnerable areas such as the Mediterranean. Since the olive growing is one of the main economic sources for the Mediterranean countries, investigations on olive pests under global heating are necessary. Nowadays, knowledge on the topic is scarce, and nothing is known about the effects of climate change on olive pest parasitoids and predators. This information could be fundamental to understand the phenomena of pest outbreaks that are spreading in the Mediterranean olive orchards. The use of prevention tools (e.g., monitoring, prediction models) may help in controlling olive pests under a climate change scenario. Abstract Evidence of the impact of climate change on natural and agroecosystems is nowadays established worldwide, especially in the Mediterranean Basin, an area known to be very susceptible to heatwaves and drought. Olea europaea is one of the main income sources for the Mediterranean agroeconomy, and it is considered a sensitive indicator of the climate change degree because of the tight relationship between its biology and temperature trend. Despite the economic importance of the olive, few studies are nowadays available concerning the consequences that global heating may have on its major pests. Among the climatic parameters, temperature is the key one influencing the relation between the olive tree and its most threatening parasites, including Bactrocera oleae and Prays oleae. Therefore, several prediction models are based on this climatic parameter (e.g., cumulative degree day models). Even if the use of models could be a promising tool to improve pest control strategies and to safeguard the Mediterranean olive patrimony, they are not currently available for most O. europaea pests, and they have to be used considering their limits. This work stresses the lack of knowledge about the biology and the ethology of olive pests under a climate change scenario, inviting the scientific community to focus on the topic.

Phosphorus (P) availability significantly impacts olive tree reproductive development and consequential fruit production. However, the importance of P fertilization in olive cultivation is not clear, and P application is usually recommended only after P deficiency is identified. In order to determine the long-term impacts of continuous P fertilization in intensive irrigated olive cultivation, the growth and production of trees in an intensive orchard with or without P fertilization were evaluated over six consecutive seasons. Withholding of P resulted in significant reduction in soil P quantity and availability. Under lower P availability, long-term fruit production was significantly impaired due to reduced flowering and fruit set. In addition, trees under conditions of low P were characterized by higher alternate bearing fluctuations. Olive tree vegetative growth was hardly affected by P fertilizer level. The impairment of tree productivity was evident in spite of the fact that leaf P content in the treatment without P fertilization did not decrease below commonly reported and accepted thresholds for P deficiency. This implies that the leaf P content sufficiency threshold for intensive olive orchards should be reconsidered. The results demonstrate the negative impact of insufficient P fertilization and signify the need for routine P fertilization in intensive olive cultivation.

Selenium is an essential micronutrient that provides important benefits to plants and humans. At proper concentrations, selenium increases plant growth, pollen vitality, the shelf life of fresh products, and seems to improve stress resistance, these effects can certainly be attributed to its direct and indirect antioxidant capacity. For these reasons, in the present work, the effects of selenium at different dosages on in vitro cultivated olive explants were investigated to observe possible positive effects (in terms of growth and vigor) on the proliferation phase. The work was carried out on four different olive cultivars: “San Felice”, “Canino”, “Frantoio”, and “Moraiolo”. The explants were cultured in aseptic conditions on olive medium (OM), with the addition of 4 mg·L−1 of zeatin, 30 g·L−1 of sucrose, and 7 g·L−1 of agar. The experimental scheme included a comparison between explants grown with five different concentrations of Na2SeO4 (0, 10, 20, 40, and 80 mg L−1) added to the medium during three successive subcultures. Interesting information has emerged from the results and all varieties responded to different concentrations of Selenium. The optimal Se dosages varied for each cultivar, but in general, Se concentration between 10 and 40 mg L−1 increased fresh and dry weight of the explants and shoot lengths. Se treatment induced in all cultivars and for all dosages used an increase in total Se content in proliferated explants. Furthermore, as the subcultures proceeded, the ability of the explants to absorb Se did not diminish. The Se content ranged from 8.55 to 114.21 µg kg−1 plant DW in ‘Frantoio’, from 9.83 to 94.85 µg kg−1 plant DW in ‘Moraiolo’, from 19.84 to 114.21 µg kg−1 plant DW in ‘Canino’, and from 20.97 to 95.54 µg kg−1 plant DW in ‘San Felice’. In general, the effect of selenium tends to decrease with the progress of subcultures and this suggests a sort of “adaptation” effect of the explants to its presence. The present study highlights for the first time the possibility of using in vitro cultures as biotechnological support to study supplementation with selenium and its effects on in vitro olive plant growth.

Olive (Olea europaea L.) is an economically important crop because of its fruit and oil. Successful olive micropropagation is highly dependence on cultivar, shoot proliferation rate, which is generally low, the rooting of micropropagated olive plantlets is difficult, and the rate of post-transplanting losses is high. in addition, hyperhydricity, a common problem in vitro culture was found to be prevalent. the aim of this study was to establish a micropropagation system for the Turkish O. europaea L. cv. Gemlik. Initially, five different basal media were tested to determine appropriate medium for establishment of in vitro culture and Woody Plant Medium (WPM) was the most efficient. Nodal explants were cultured on WPM containing different plant growth regulators (PGRs) for shoot regeneration. Maximum regeneration frequency and number of shoots per explant were achieved from nodal explants cultured on WPM supplemented with 4.0 mg/L 6-benzyladenine (BA). However, all cultures showed high hyperhydricity and an experimentation was also conducted to resolve the hyperhydricity problem. Hyperhydricity was prevented by changing the gelling agent to Agar-Agar. the shoots regenerated from nodal explants and still attached to initial woody nodal explant were transferred to four different medium formulations each containing 2.0 mg/L zeatin (ZEA) for shoot elongation. Modified Olive Medium (MOM2: OM with three times the concentrations of KNO3) fortified with 2.0 mg/L ZEA was found to be the best for shoot elongation. the elongated shoots were rooted on Olive medium (OM) containing 160 mg/L Putrescine, 1.5 mg/L naphthaleneacetic acid (NAA), 30 g/L mannitol and solidified with 0.65% (w/v) Agar-Agar. Finally, all plantlets were successfully acclimatized in a climate chamber and the plants were transferred to greenhouse conditions. (C) 2019 SAAB. Published by Elsevier B.V. All rights reserved., Ege University Scientific Research Projects Coordination UnitEge University [09-MUH-010], This work was supported by Ege University Scientific Research Projects Coordination Unit [grant number: 09-MUH-010]. All of the experiments were carried out at the Bioengineering Department of Ege University. the authors are grateful to Mark Smedley for critically reviewing the manuscript.

The concomitant use of olive leaves (OL) and glyburide (GLB) is a possible therapy for diabetic patients. However, there is no report about the effect of OL on the antidiabetic effect of GLB till now. In the current study, the possible interaction of olive leaves extract (OLE) with GLB was assessed to determine if there was any pharmacological benefit over GLB alone. Seven groups of male Sprague Dawley rats were used. Normal rats of the 1st group treated with 2 mL/kg of 3% Tween 80 (vehicle). The 2nd–5th groups were diabetic rats received vehicle, GLB (5 mg/kg), OLE low dose and OLE high dose respectively, while the 6th–7th groups administered combinations of GLB plus OLE low dose and GLB plus OLE high dose, respectively. All treatments were administered orally once daily for 8 weeks.The use of GLB+OLE-500 obviously improved fasting blood glucose (FBG), insulin and glycated hemoglobin (HbA1c) in diabetic rats (95.5 ± 5.55 mg/dL, 6.8 ± 0.16 mg/dL and 6.1 ± 0.29%, respectively) compared to those treated with GLB monotherapy (140.0 ± 6.36 mg/dL, 5.4 ± 0.19 mg/dL and 7.0 ± 0.20%, respectively). The lipid profile [triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C)] was significantly improved in diabetic rats exposed to GLB+OLE-500 (35.6 ± 1.51 mg/dL, 48.5 ± 2.74 mg/dL, 25.1 ± 1.21 mg/dL and 17.0 ± 0.82 mg/dL, respectively) in comparison with diabetic group exposed to GLB alone (43.2 ± 2.15 mg/dL, 56.8 ± 2.14 mg/dL, 18.6 ± 0.96 mg/dL, 23.0 ± 1.26 mg/dL, respectively). Additionally, the benefit impacts of GLB+OLE-500GLB+OLE-500 therapy on the antioxidant and lipid peroxidation parameters in the pancreatic tissues of diabetic rats were higher than those of GLB monotherapy. Moreover, GLB plus OLE-500 combination had the greatest effect on restoration of the insulin content of Beta (β) cells and reduction of the glucagon and somatostatin of Alpha (α) and Delta (δ) endocrine cells in the pancreatic islets among the different treatment. The current study suggests that OL and GLB combination could cause herb-drug interactions through modulation of insulin receptor (INR), glucose transporter 2 (Slc2a2) and peroxisome proliferator-activated receptor α (PPAR-α) genes expression in the liver of diabetic rats. Keywords: Olea europaea, Streptozotocin, Glyburide, INR, Slc2a2, PPAR-α, Rat

Olive culture is a recent activity in Rio Grande do Sul, Brazil with little information on its scale insects. Abundance and seasonality of armored scale insects (Hemiptera, Diaspididae) and their parasitism was evaluated in olive orchard, Olea europaea L., cultivar Arbequina, in Caçapava do Sul (30°30′43″ S, 53°29′27″ W), RS, Brazil. Samples were monthly collected from April 2012 to March 2013. At each occasion 20 trees were drawn and, from each quadrant of their canopies (northern, southern, eastern and western) two branches (20 - 30 cm - length and contained at least 20 leaves) were randomly removed, respectively from the inner part and the outer part of the canopy. We found five species of Diaspididae, Aonidiella aurantii (Maskell), Hemiberlesia cyanophylli (Signoret), Acutaspis paulista (Hempel), Aspidiotus nerii Bouché and Melanaspis obscura (Comstock). It was not found an unique pattern of abundance, indicating that these species respond differently to temperature variations between seasons. The armored scale insects were evenly distributed among quadrants. Only H. cyanophylli showed differences in abundance between the inner and outer branches. We registered parasitism in all Diaspididae species; the total parasitism rate was 9.78%. A olivicultura é uma atividade recente no Rio Grande do Sul, Brasil, com pouca informação sobre as cochonilhas associadas a esta cultura. Abundância e sazonalidade das cochonilhas-com-escudo (Hemiptera, Diaspididae) e seu parasitismo foram avaliados em olival, Olea europaea L., cultivar Arbequina, em Caçapava do Sul (30º30'43" S, 53°29'27" O), RS, Brasil. As amostras foram coletadas mensalmente, a partir de abril de 2012 a março de 2013. Em cada ocasião, 20 árvores foram sorteadas e, de cada quadrante de suas copas (norte, sul, leste e oeste) dois ramos (20 - 30 cm de comprimento e contendo pelo menos 20 folhas) foram retirados aleatoriamente, respectivamente, da parte interna e a parte externa da copa. Foram encontradas cinco espécies de Diaspididae, Aonidiella aurantii (Maskell), Hemiberlesia cyanophylli (Signoret), Acutaspis paulista (Hempel), Aspidiotus nerii Bouché e Melanaspis obscura (Comstock). Não foi encontrado um padrão único de abundância, o que indica que essas espécies respondem diferentemente a variações de temperatura entre as estações. Verificou-se que as cochonilhas se distribuem uniformemente entre os quadrantes. Apenas H. cyanophylli mostrou diferenças na abundância entre os ramos internos e externos. Foi registrado o parasitismo em todas as espécies de Diaspididae, a taxa de parasitismo total foi de 9,78%.

An hydroxytyrosol (HTyr) enriched extract (HTE), obtained from olive mill wastewaters (OMWW) through an eco-friendly, patented process was tested on two olive tree pathogens, Pseudomonas savastanoi pv. savastanoi (Pss) and Agrobacterium tumefaciens (At). Pss, in particular, is a feared treat for olive tree cultivation being the etiological agent of the olive knot disease responsible of severe production losses. Chemical characterization allowed to identify and quantify HTyr as the main constituent along with other low molecular weight phenols. HTE has proven potent antioxidant activity and significant antimicrobial activity against Pss and At in vitro, in both cases higher than hydroxytyrosol alone, suggesting an important role also of the minor phenolic components, which act synergistically with HTyr. Alternatives to the molecules currently present on the market are needed, but only the use of standardized extraction technologies would allow to conduct solid field studies and enable this circular approach to find a real application in the olive groves.

application/pdf, Objective: To evaluate the antipiroplasmic activities of methanolic extract of Olea europaea (MOE) and acetonic extract of Acacia laeta (AAL) against Babesia and Theileria parasites in vitro and evaluate the chemotherapeutic effects of these extracts against Babesia (B.) microti in vivo. Methods: Fluorescence assay using SYBR Green 1 nucleic acid stain was used to detect inhibitory effects of the two extracts as well as the combination effects of the two extracts with diminazene aceturate and atovaquone on four Babesia species and Theileria equi in vitro while for in vivo experiments, 8-weekold female BALB/c mice were injected intraperitoneally with 1 × 107B. microti-iRBCs and treated orally at a dose of 150 mg/kg of both extracts. Results: The half maximal inhibitory concentration (IC50) values of AAL against B. bovis, B. bigemina, B. divergens, B. caballi, and Theileria equi were lower than those of MOE extracts. Toxicity assay on Madin-Darby bovine kidney, mouse embryonic fibroblast (MH/3T3), and human foreskin fibroblast cell lines showed that MOE and AAL affected only the viability of Madin-Darby bovine kidney cell line with half maximal effective concentrations (EC50) of (794.7±41.9) and (873.9±17.5) μg/mL, respectively. The oral treatments of MOE and AAL at 150 mg/kg inhibited the growth of B. microti in mice by 80.4% and 64.4%, respectively. The MOE and diminazene aceturate combination showed a higher chemotherapeutic effect than that of monotherapy. Conclusions: MOE and AAL have the potential to be an alternative remedy for treating piroplasmosis. Furthermore, the combination therapy of MOE + DA was more potent against B. microti infection in mice than their monotherapies.

In countries new to producing ‘Manzanillo’ olive trees (Olea europaea), free cross-pollination is often insufficient to obtain high levels of fruit set. An appropriate pollination design is therefore essential to ensure a timely, abundant, and compatible pollen supply. With a view to determining whether a pollination deficit exists in a nontraditional olive area such as the northern Mexico, pollination experiments were carried out in two consecutive seasons in both a monovarietal and a multivarietal Manzanillo orchard, where Sevillano, Barouni, Picual, Pendolino, Mission, Nevadillo, and Frantoio trees were growing nearby. The pollination treatments were self-, open, and cross-pollination with ‘Barouni’ and ‘Sevillano’ pollen, the latter only in the multivarietal orchard. The results confirmed the full self-incompatible condition of ‘Manzanillo’. Open-pollination did not improve fruit set in the monovarietal orchard, but it did so significantly in the multivarietal plot, where fruit set levels under open-pollination matched those of cross-pollination. Lower pollen adhesion, as well as occasional decreased germination, and reduced and delayed pollen tube growth were observed under self-pollination, highlighting self-incompatibility reactions. The reduction in fertilization rates led to low fruit set under self-pollination. Positive effects of open- and cross-pollination treatments were also noted on fruit weight (despite higher crop loads) and pulp-to-pit ratios. A strategic plantation design, including appropriate pollinizers in the right number and position, is therefore suggested for increasing ‘Manzanillo’ fruit quality and yield in Mexico. Both ‘Barouni’ and ‘Sevillano’ served as efficient pollinizers for ‘Manzanillo’, although we recommend ‘Barouni’ as a more efficient because the bloom periods of them matched that of ‘Manzanillo’

Abscisic acid (ABA) is the key phytohormone modeling the stomata behavior under drought conditions. However, stomata closure is not correlated with leaf ABA content in mycorrhizal olive plant. The aim of the present study is to evaluate the impact of arbuscular mycorrhizal (AM) symbiosis on the control of stomata functioning by ABA in two olive tree (Olea europaea L.) cultivars ‘Zarrazi’ and ‘Meski’ subjected to dehydration–rehydration treatment. AM-inoculated (Myc +) and non-inoculated (Myc-) olive plants were subjected to water stress and then rewatered (recovery). Leaf ABA content, stomatal conductance and transpiration rate were measured in: (1) irrigated control, (2) moderately and severely stressed and (3) recovered plants. In both Zarrazi and Meski Myc– plants, ABA content increased in parallel with drought intensity and stomatal closure. However, an intra-specific variability appeared in inoculated plants; in Meski Myc + plants ABA content was not influenced by drought and recovery treatments, whereas in Zarrazi Myc + plants the ABA amount increased under moderate water stress and even further after water relief, but decreased under severe water stress. However, in Myc + plants of both cultivars, stomatal closure was not correlated with leaf ABA content. The results reveal that the ABA is not the key factor controlling the stomatal closure in AM-inoculated olive plants under drought conditions. In fact, other AM-related factors are involved in the control of stomata regulation in mycorrhizal olive plants exposed to severe drought. These factors act specifically in the drought-resistant cultivar ‘Zarrazi’ permitting a suitable stomata behavior.

The fruits of Olea europaea L. is widely consumed as food, and olive pits are utilized in folk medicine to relieve gastric disturbances. In the present study, the possible anti-inflammatory, analgesic and antioxidant activities of aqueous extracts of black (BP) and green olive (GP) pit prepared at gastric fed state pH were evaluated in vitro. Moreover, the bioactive compound, hydroxytyrosol (HT), was isolated from the extracts for the first time. According to results, GP extract (62.5 to 1000 μg/mL) showed significant anti-inflammatory activity in a dose-dependent manner and HT displayed significant nitrite inhibition at 100 μM with slight analgesic activity. Extracts and HT showed a significant antioxidant activity according to Total Antioxidant Capacity (TOAC), cupric ion reducing antioxidant capacity (CUPRAC), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assays. As a conclusion, a proper formulation containing HT might be a potential remedy to relieve gastric disturbances and olive pits, can be utilized as a valuable industrial tool for the low-cost production of HT. How to cite this article: Reis R, Sipahi H, Zeybekoglu G, Celik N, Kirmizibekmez H, Kaklikkaya N, Aydin A. Hydroxytyrosol: The Factor Responsible for Bioactivity of Traditionally used Olive Pits. Euroasian J Hepatogastroenterol, 2018;8(2):126-132.

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.

La verticilosis causada por el hongo habitante de suelo Verticillium dahliae Kleb. es una enfermedad que afecta el cultivo del olivo (Olea europaea L.) y algodón (Gossypium hirsutum L.), entre otros. De acuerdo a su patogenicidad, los aislados del patógeno pueden clasificarse como patotipos defoliantes (D) y no defoliantes (ND). Los objetivos de este estudio fueron: i) determinar los patotipos de una colección de 42 aislados de V. dahliae provenientes de plantas de olivo de las provincias de Catamarca, Córdoba, La Rioja y Mendoza existente en el IPAVE-INTA Córdoba, y ii) evaluar la virulencia de ocho cepas recientemente aisladas pertenecientes a dicha colección. Se utilizó la técnica de PCR anidado para el análisis molecular de los aislados del hongo y la evaluación de la virulencia se realizó mediante inoculaciones artificiales en plantines de algodón con un diseño completamente aleatorizado. El 100 % de los aislados fueron caracterizados como ND. Dos cepas provenientes de La Rioja, dos de Catamarca y una de Mendoza, fueron significativamente más virulentas (p< 0,05) para el área bajo la curva de progreso de la enfermedad relativa, severidad media final, porcentaje de severidad media e índice de severidad. Verticillium wilt caused by the soil-borne fungus Verticillium dahliae Kleb. is a disease that affects olive (Olea europaea L.) and cotton crops, among others. According to their pathogenicity, V. dahliae isolates can be classified as defoliating (D) and non-defoliating (ND). The objectives of this study were: i) to determine the pathotypes of a collection of 42 V. dahliae isolates from olive plants in the provinces of Catamarca, Córdoba, La Rioja and Mendoza kept at IPAVE-INTA Córdoba, and (ii) to evaluate the virulence of eight recently isolated isolates belonging to this collection. Nested PCR was used for molecular analysis of the 42 fungal isolates and virulence evaluation was performed by artificial inoculations on cotton seedlings with a completely randomized design. The total of the isolates was characterized as ND. Two strains from La Rioja, two from Catamarca and one from Mendoza, were significantly more virulent (p

Olea europaea germplasm is constituted by a huge number of cultivars, each one characterized by specific features. In this context, endemic cultivars evolved for a very long period in a precise local area, developing very specific traits. These characteristics include the production and accumulation of phytochemicals, many of which are also responsible for the nutraceutical value of the drupes and of the oils therefrom. With the aim of obtaining information on the phytochemical profile of drupes of autochthonous cultivars of Cilento, Vallo di Diano and Alburni National Park, a metabolomics-based study was carried out on 19 selected cultivars. Multivariate data analysis of 1H-NMR data and 2D NMR analyses allowed the rapid identification of metabolites that were qualitatively and/or quantitatively varying among the cultivars. This study allowed to identify the cultivars Racioppella, Guglia, Pizzulella, Oliva amara, and Racioppa as the richest in health-promoting phenolic compounds. Furthermore, it showed a significant variability among the different cultivars, suggesting the possibility of using metabolic fingerprinting approaches for cultivar differentiation, once that further studies aimed at assessing the influence of growing conditions and environmental factors on the chemical profiles of single cultivars are carried out.

Maximizing nitrogen uptake efficiency is a sustainable objective to reduce the amount of nitrogen fertilizers applied in olive orchards. Many factors affect this parameter, but nothing is known about whether there are differences among cultivars related to nitrogen uptake efficiency in the olive, and if there are interactions with soil type. To clarify these questions, two groups of experiments were conducted. In the first group, four experiments were carried out in which 20 olive cultivars were compared. In the second group, three experiments were carried out to study the interaction with the type of soil. In all experiments, mist-rooted olive cuttings were growing in 1.1 L pots containing a mixture of sand and peat (2:1 by volume). Depending on the experiment, plants were placed in a shadehouse or in growth chambers at 25/15 °C (day/night) with a 14 h photoperiod and 70% humidity. Plants received weekly applications of 100 ppm N throughout the irrigation water, alternating with one or two applications of 150 mL of tap water. Some plants of each cultivar did not receive nitrogen and served as a control for NUE calculation. A nutrient solution without nitrogen was applied every four weeks to prevent nutritional deficiencies. Results indicated that there are differences among cultivars in the efficiency of uptake nitrogen. The Spanish cultivars ‘Picual’, ‘Arbequina’ and ‘Manzanilla de Sevilla’, together with the Greek cultivar ‘Koroneiki’ and the Italian ‘Frantoio’, were the cultivars with high NUE in this work. Soil type did not affect NUE, and no interaction between soil type and cultivar was observed.

This research focuses on the exploration, recovery and valorization of some minor Italian olive cultivars, about which little information is currently available. Autochthonous and unexplored germplasm has the potential to face unforeseen changes and thus to improve the sustainability of the whole olive system. A pattern of nine minor genotypes cultivated in three Italian regions has been molecularly fingerprinted with 12 nuclear microsatellites (SSRs), that were able to unequivocally identify all genotypes. Moreover, some of the principal phenolic compounds were determined and quantified in monovarietal oils and the expression levels of related genes were also investigated at different fruit developmental stages. Genotypes differed to the greatest extent in the content of oleacein (3,4-DHPEA-EDA) and total phenols. Thereby, minor local genotypes, characterized by stable production and resilience in a low-input agro-system, can provide a remarkable contribution to the improvement of the Italian olive production chain and can become very profitable from a socio-economic point of view.

Olea europaea ‘Galega vulgar’ variety is a blend of West and Central Mediterranean germplasm with cultivated-wild admixture characteristics. ‘Galega vulgar’ is known for its high rusticity and superior-quality olive oil, being the main Portuguese variety with high impact for bioeconomy. Nevertheless, it has been replaced by higher-yielding and more adapted to intensive production foreign varieties. To clarify the potential ancestral origin, genetic diversity evolution, and existing genetic relationships within the national heritage of ‘Galega vulgar’, 595 trees, belonging to ancient and centenary age groups and prospected among ten traditional production regions, were characterized using 14 SSR markers after variety validation by endocarp measurements. Ninety-five distinguishable genets were identified, revealing the presence of a reasonable amount of intra-genetic and morphological variability. A minimum spanning tree, depicting the complete genealogy of all identified genets, represented the ‘Galega vulgar’ intra-varietal diversity, with 94% of the trees showing only a two-allele difference from the most frequent genet (C001). Strong correlations between the number of differentiating alleles from C001, the clonal size, and their net divergence suggested an ancestral monoclonal origin of the ‘Galega vulgar’, with the most frequent genet identified as the most likely origin of all the genets and phenotypic diversification occurring through somatic mutations. Genetic erosion was detected through the loss of some allele combinations across time. This work highlights the need to recover the lost diversity in this traditional olive variety by including ancient private genets (associated with potential adaptation traits) in future breeding programs and investing in the protection of these valuable resources in situ by safeguarding the defined region of origin and dispersion of ‘Galega vulgar’. Furthermore, this approach proved useful on a highly diverse olive variety and thus applicable to other diverse varieties due either to their intermediate nature between different gene pools or to the presence of a mixture of cultivated and wild traits (as is the case of ‘Galega vulgar’).

The olive fruit fly is worldwide considered a major harmful pest of the olive agroecosystem. In Italy, the fruit fly infestation is traditionally countered by spraying chemical insecticides (e.g., dimethoate), but due to the recent ban of dimethoate by the Reg EU2019/1090 and the increasing awareness of consumers of food sustainability, the interest in developing chemical-free alternatives to pesticides, such as the use of particle-films, is rising. A field experiment was conducted to assess the effect of different particle films (kaolin-base and zeolitite-base) on leaf gas exchanges and leaf optical properties. Results showed that with the dust accumulation on the leaves’ surface, photosynthesis, stomatal conductance, transpiration and water use efficiency were significantly lower in kaolin-treated olive trees compared to those treated with zeolitite and to the control, while olive trees treated with zeolitite showed physiological parameters similar to the untreated plants. Microstructural differences of different particle film on the leaf and olive surfaces emerged by ESEM observations also influenced leaf optical properties. Oils produced by zeolitite-treated plants show higher intensities of gustatory and olfactory secondary flavors compared to kaolin and test oils.

The effect of commercial mycorrhizal fungi on pre-rooted olive cuttings was assessed. The study consisted of two experiments, the first arranged as a factorial design with three cultivars (Cobrançosa, Madural, and Verdeal Transmontana) and three soil treatments (commercial mycorrhizal fungi, zeolites, and control) and the second as a completely randomized design with three treatments (commercial mycorrhizal fungi, sterilized soil, and control). Cobrançosa grew better than the other cultivars, showing good rooting and initial growth features. Mycorrhizal plants showed reduced growth in comparison to those of the untreated control. This result was explained by competition for photosynthates between plant growth and the expansion of fungi hyphae. Cuttings of reduced leaf area and a twin-wall polycarbonate cover of the greenhouse, somewhat opaque to photosynthetic active radiation, may also have contributed to limit the maximum photosynthetic rate and delay the growth of the inoculated plants. Accordingly, in the mycorrhizal pots, the soil organic carbon (C) increased, probably due to the presence of fungi hyphae in soil samples. Zeolites reduced plant dry matter (DM) yield and tissue phosphorus (P) concentration compared to the control, while increasing the availability in the soil of the cations present in their initial composition. Soil sterilization seems to have reduced soil P bioavailability by inactivating soil enzymes. This study showed that the inoculation with mycorrhizal fungi of pre-rooted cuttings can delay their initial growth. Although these plants may be better prepared to grow in the field, in the short term, their lower development can be a problem for the nurseryman. This research was funded by the Foundation for Science and Technology (FCT, Portugal) and FEDER under Programme PT2020 for financial support to CIMO (UIDB/00690/2020) and CITAB (UIDB/04033/2020). The research was integrated in the activities of the operational group “Novas práticas em olivais de sequeiro: estratégias de mitigação e adaptação às alterações climáticas”, funded by PT2020 and EAFRD (European Agricultural Fund for Rural Development). info:eu-repo/semantics/publishedVersion

The olive tree (Olea europaea L. subsp. europaea) is the most important perennial crop in the Mediterranean region, producing table olives and oil, both appreciated for their nutraceutical value. Although olive oil quality traits have been extensively studied, much less attention has been paid to olive drupe. Olive drupe ripening is an extremely complex process involving numerous physiological and molecular changes that are unique in this fruit crop species. This review underlines the contribution of “-omics” techniques and of the recent advances in bioinformatics and analytical tools, notably next-generation sequencing and mass spectrometry, for the characterization of the olive ripening syndrome. The usage of high-dimensional datasets, such as transcriptomics, proteomics, and metabolomics, will provide a systematical description of the molecular-specific processes regulating olive fruit development and ripening. However, the incomplete sequence of the O. europaea L. reference genome has largely hampered the utilization of omics tools towards olive drupe research. Due to this disadvantage, the most reported -omics studies on fruit trees concern metabolomics and only a few transcriptomics and proteomics. In this review, up-to-date applications of -omics technologies towards olive drupe biology are addressed, and future perspectives in olive fruit research are highlighted.

The purpose of this study was to evaluate the relationship between two types of vigor and architectural parameters (quantitative and qualitative) and the duration of juvenile period in olive and to test the effectiveness of three different strategies for an early elimination of seedlings with long juvenile period. Architectural parameters were evaluated in 794 one-year-old olive seedlings, and first flowering time for each seedling was then recorded the following three years. Among the quantitative parameters, ‘Plant height’ and ‘Secondary shoots insertion angle’ were those most correlated with the juvenile period duration. Among the qualitative parameters, ‘Main vertical axis’, ‘Branching system’, ‘Preferential distribution of long shoots’, ‘Extreme leaf size’, ‘Branch orientation’ and ‘Branch bending’ were also able to distinguish seedlings with different juvenile period duration. Four architectural types (defined on the basis of the qualitative parameters evaluated) were found in higher percentages in seedlings with short juvenile period. Finally, three new selection strategies based on the ‘Branch orientation’ (strategy I), the established architectural types (strategy II), and the association of ‘Plant height’ with architectural type (strategy III) all provided greater capacity for discarding seedlings with long juvenile period than did using only ‘Plant height’ method, the early selection criterion currently used for that purpose in breeding programs. The strategy based on combining a quantitative criterion (i.e. ‘Plant height’) and the qualitative architectural types, provided the most promising results from a practical point of view considering both ease of measurement and selection effectiveness.

The olive tree (Olea europaea L.) is a typical Mediterranean crop, important for olive and oil production. The high tendency to bear fruits in an uneven manner, defined as irregular or alternate bearing, results in a significant economic impact for the high losses in olives and oil production. Buds from heavy loaded (‘ON’) and unloaded (‘OFF’) branches of a unique olive tree were collected in July and the next March to compare the transcriptomic profiles and get deep insight into the molecular mechanisms regulating floral induction and differentiation. A wide set of DEGs related to ethylene TFs and to hormonal, sugar, and phenylpropanoid pathways was identified in buds collected from ‘OFF’ branches. These genes could directly and indirectly modulate different pathways, suggesting their key role during the lateral bud transition to flowering stage. Interestingly, several genes related to the flowering process appeared as over-expressed in buds from March ‘OFF’ branches and they could address the buds towards flower differentiation. By this approach, interesting candidate genes related to the switch from vegetative to reproductive stages were detected and analyzed. The functional analysis of these genes will provide tools for developing breeding programs to obtain olive trees characterized by more constant productivity over the years.

Extreme climate events are increasingly frequent, and the 2017 summer was particularly critical in the Mediterranean region. Olive is one of the most important species of this region, and these climatic events represent a threat to this culture. However, it remains unclear how olive trees adjust the antioxidant enzymatic system and modulate the metabolite profile under field stress con-ditions. Leaves from two distinct adjacent areas of an olive orchard, one dry and the other hydrated, were harvested. Tree water status, oxidative stress, antioxidant enzymes, and phenolic and lipo-philic metabolite profiles were analyzed. The environmental conditions of the 2017 summer caused a water deficit in olive trees of the dry area, and this low leaf water availability was correlated with the reduction of long†chain alkanes and fatty acids. Hydrogen peroxide (H2O2) and superoxide radical (O2•–) levels increased in the trees collected from the dry area, but lipid peroxidation did not augment. The antioxidant response was predominantly marked by guaiacol peroxidase (GPOX) activity that regulates the H2O2 harmful effect and by the action of flavonoids (luteolin†7†O†glucu-ronide) that may act as reactive oxygen species scavengers. Secoiridoids adjustments may also con-tribute to stress regulation. This work highlights for the first time the protective role of some metabolite in olive trees under field drought conditions. Foundation for Science and Technology (FCT, Portugal) and Ministério da Educação e Ciência supported this work through national funds and the co-funding by the FEDER within the PT2020 Partnership Agreement and COMPETE 2010. Within the projects of the CEF research unit UI0183– UID/BIA/04004/2020, QOPNA research unit UID/QUI/00062/2019, LAQV-REQUIMTE UIDB/50006/2020, and CIMO UIDB/00690/2020. FCT supported the doctoral fellowship of MAraújo (SFRH/BD/116801/2016), N. Mariz-Ponte (SFRH/BD/138187/2018), and the research contract of MC Dias (SFRH/BPD/100865/2014) through POCH/FSE. info:eu-repo/semantics/publishedVersion

Leaves from Olea europaea represent one of the main by-products of the olive oil industry, containing a plethora of bioactive compounds with several promising activities for human health. An organic solvent-free extraction method was developed for the recovery of olive leaf phenols, which obtained an extract containing oleuropein in high amounts. A comparison of various extraction media is reported, together with the total phenolic content, DPPH (2,2-Diphenyl-1-picrylhydrazyl) content, ORAC (oxygen radical absorbance capacity), and polyphenol oxidase activity of the corresponding extracts. The polyphenol profiles and content of the most representative extracts have also been studied. Extraction solvent and temperature significantly influenced the phenolic content and antioxidant activity of the extracts, with hot water representing the solvent of election for the extraction of bioactive compounds from this matrix. All the extracts obtained showed reasonably high total phenol content (TPC) and good DPPH radical scavenging activity, among them, the water extract is characterized by desirable traits and could be used for many industrial applications and human consumption.

Possessing a variety of medicinal functions, Olea europaea L. is widely cultivated across the world. However, the anti-inflammatory mechanism of Olea europaea is not yet fully elucidated. In this study, how the methanol extract of the leaves of Olea europaea (Oe-ME) can suppress in vitro inflammatory responses was examined in terms of the identification of the target protein. RAW264.7 and HEK293T cells were used to study macrophage-mediated inflammatory responses and to validate the target protein using PCR, immunoblotting, nuclear fraction, overexpression, and cellular thermal shift assay (CETSA) under fixed conditions. Oe-ME treatment inhibited the mRNA expression levels of cyclooxygenase (COX)-2, matrix metallopeptidase (MMP)-9, and intercellular adhesion molecule-1 (ICAM-1) in activated RAW264.7 cells. Oe-ME diminished the activation of activator protein (AP)-1 and the phosphorylation of its upstream signaling cascades, including extracellular signal regulated kinase (ERK), mitogen-activated protein kinase kinase 1/2 (MEK1/2), c-Jun N-terminal kinase (JNK), mitogen-activated protein kinase kinase 3/6 (MKK3/6), p38, MKK7, and transforming growth factor-β-activated kinase 1 (TAK1), in stimulated-RAW264.7 cells. Overexpression and CETSA were carried out to verify that TAK1 is the target of Oe-ME. Our results suggest that the anti-inflammatory effect of Oe-ME could be attributed to its control of posttranslational modification and transcription of TAK1.

In this work, hybrid zinc oxide–iron oxide (ZnOFe) magnetic nanoparticles were synthesized employing Olea europaea leaf aqueous extract as a reducing/chelating and capping medium. The resulting magnetic nanoparticles were characterized by basic spectroscopic and microscopic techniques, namely, X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), fourier-transform infrared (FTIR) and atomic force microscopy (AFM), exhibiting a spherical shape, average size of 15–17 nm, and a functionalized surface. Lipase from Thermomyces lanuginosus (TLL) was efficiently immobilized on the surface of ZnOFe nanoparticles through physical absorption. The activity of immobilized lipase was found to directly depend on the enzyme to support the mass ratio, and also demonstrated improved pH and temperature activity range compared to free lipase. Furthermore, the novel magnetic nanobiocatalyst (ZnOFe-TLL) was applied to the preparation of hydroxytyrosyl fatty acid esters, including derivatives with omega-3 fatty acids, in non-aqueous media. Conversion yields up to 90% were observed in non-polar solvents, including hydrophobic ionic liquids. Different factors affecting the biocatalyst performance were studied. ZnOFe-TLL was reutilized for eight subsequent cycles, exhibiting 90% remaining esterification activity (720 h of total operation at 50 °C). The green synthesized magnetic nanoparticles, reported here for the first time, are excellent candidates as nanosupports for the immobilization of enzymes with industrial interest, giving rise to nanobiocatalysts with elevated features.

Cryopreservation is considered the best technique for the safe, long-term conservation of embryogenic cultures. However, before integrating it into a somatic embryogenesis system, the influence of cryopreservation on the final production of plants should be investigated. The objective of this investigation was to evaluate the effect of cryopreservation on the regeneration performance of olive embryogenic cultures as well as on the quality of the plants obtained and their response to ex vitro establishment. In order to analyze the influence of the genotype, all the investigations were carried out in two genetically distinct embryogenic lines. The results obtained revealed no variation in the regeneration potential or the quality of the regenerated plants due to cryopreservation. The subsequent multiplication, rooting, and acclimatization steps were not influenced by cryopreservation either, although a significant genotype × cryopreservation interaction was found for shoot length during the multiplication step. The genotype played an important role, determining the quality of the regenerated plants and some aspects of the multiplication and rooting phases. This investigation revealed that the droplet-vitrification procedure optimized for the cryopreservation of olive somatic embryos can be efficiently used for the long-term conservation of olive embryogenic lines.

The olive is a fruit tree species economically very important in countries of the Mediterranean basin. Somatic embryogenesis is a powerful in vitro technique with multiple uses in different fields, including breeding programs performed by both classical and innovative procedures. Thus, somatic embryogenesis enables the application of biotechnological methods such as genetic transformation, somaclonal variation, somatic hybridization, germplasm cryopreservation, in vitro mutagenesis or in vitro selection. This editorial paper presents a special issue focused on “Somatic embryogenesis in olive”. In this manuscript, the conceptual framework of the special issue is established and the contributions are summarized and put into context. Finally, the main bottlenecks limiting the practical applicability of somatic embryogenesis in this species are identified and the future research prospects are discussed.

Olive leaf spot (OLS) caused by Fusicladiumoleagineum is mainly controlled using copper fungicides. However, the replacement of copper-based products with eco-friendly alternatives is a priority. The use of plant resistance-inducers (PRIs) or biological control agents (BCAs) could contribute in this direction. In this study we investigated the potential use of three PRIs (laminarin, acibenzolar-S-methyl, harpin) and a BCA (Bacillus amyloliquefaciens FZB24) for the management of OLS. The tested products provided control efficacy higher than 68%. In most cases, dual applications provided higher (p &lt, 0.05) control efficacies compared to that achieved by single applications. The highest control efficacy of 100% was achieved by laminarin. Expression analysis of the selected genes by RT-qPCR revealed different kinetics of induction. In laminarin-treated plants, for most of the tested genes a higher induction rate (p &lt, 0.05) was observed at 3 days post application. Pal, Lox, Cuao and Mpol were the genes with the higher inductions in laminarin-treated and artificially inoculated plants. The results of this study are expected to contribute towards a better understanding of PRIs in olive culture and the optimization of OLS control, while they provide evidence for potential contributions in the reduction of copper accumulation in the environment.

Proteins and peptides are minor components of vegetal oils. The presence of these compounds in virgin olive oil was first reported in 2001, but the nature of the olive oil proteome is still a puzzling question for food science researchers. In this paper, we have compiled for a first time a comprehensive proteomic dataset of olive fruit and fungal proteins that are present at low but measurable concentrations in a vegetable oil from a crop of great agronomical relevance as olive (Olea europaea L.). Accurate mass nLC-MS data were collected in high definition direct data analysis (HD-DDA) mode using the ion mobility separation step. Protein identification was performed using the Mascot Server v2.2.07 software (Matrix Science) against an ad hoc database made of olive protein entries. Starting from this proteomic record, the impact of these proteins on olive oil stability and quality could be tested. Moreover, the effect of olive oil proteins on human health and their potential use as functional food components could be also evaluated. In addition, this dataset provides a resource for use in further functional comparisons across other vegetable oils, and also expands the proteomic resources to non-model species, thus also allowing further comparative inter-species studies. The data presented here are related to the research article of Castro et al. [1] .

To maximize orchard production and tree crop efficiency, optimization of both maximum orchard light interception and radiation distribution within the tree canopy are important strategies. To study the influence of planting density and fruit position within the canopy on oil quality from &lsquo, Cerasuola&rsquo, and &lsquo, Koroneiki&rsquo, olive (Olea europaea L.), fruits were harvested from the upper and lower canopy layers of trees in hedgerow planting systems at two densities: High at 1000 trees ha&minus, 1 (HD) and Medium at 500 trees ha&minus, 1 (MD). Tree crop efficiency and fruit weight, water and fat content were measured together with olive oil standard quality parameters, phenolic and volatile composition. Fruits in the upper layers of the canopy always showed a higher maturity index, 6% more fat content, and 4% less water content than lower layers. Upper layers of HD trees showed the highest phenol content, whereas lower layers of MD trees showed the lowest phenol content (36% less than the upper layers of HD). HD trees showed the largest differences in fruit maturation, water and fat content between upper and lower canopy positions, increasing quality and oil yield variability at harvest. &lsquo, showed more stable oils with a 28% higher MUFA/PUFA ratio and 12% higher phenol content than &lsquo, oils. This study provides further evidence of the fact that cultivar, planting density, and canopy architecture may be strong determinants of olive oil yield and composition in hedgerow planting systems.