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Genomic and chromosomal organization of Ty1- copia-like sequences in Olea europaea and evolutionary relationships of Olea retroelements.

The Ty1- copia-like retrotransposon is one of the commonest class of transposable elements in the plant kingdom, often comprising several percent of the total DNA content. We aimed to study the evolutionary relationships of Olea retroelements, using part of the reverse transcriptase domain, as well as the genomic and chromosomal organization of these sequences in Olea europaea chromosomes and their transcription activity and copy number. Fourteen clones, that were isolated from four different species, were sequenced and a phylogenetic tree was constructed based on their predicted amino acids. Five clones derived from O. europaea were clustered together with a 87% nucleotide sequence homology and two Olea oleaster clones showed 98% sequence homology. The rest of the clones showed heterogeneity among them, leading to a common ancestral transposon that existed before the genus arose. The Ty1- copia-like sequences have a dispersed genomic organization, physically distributed on all chromosomes, showing minor clustering in some cases and low copy numbers in the smallest chromosome pair. The total copy number in the O. europaea genome was estimated by dot blotting to be 40,000 in a haploid nucleus, but a number of these are non-functional since the sequenced clones contained stop codons and frame-shifts. Some Ty1- copia-like copies, present in O. europaea, were found to be methylated, while no differences in methylation were observed between DNA isolated from young leaves and callus-suspension cultures.

Journal Article↗

An allergen from Olea europaea pollen (Ole e 7) is associated with plant-derived food anaphylaxis.

BACKGROUND: Several cross-reacting proteins have been identified as responsible of the co-occurrence of pollinosis and plant-derived food allergy. This association has been mainly described in the birch-apple syndrome but other pollens such as Olea europaea and other fruits may also contain homologous proteins. OBJECTIVE: To evaluate the associations between sensitization to allergens of Olea europaea pollen and confirmed plant-derived food allergy, in addition to investigate if any pattern of clinical hypersensitivity of food allergy reaction (oral allergy syndrome (OAS) or anaphylaxis) and/or any fresh fruit or nut allergy, are associated to one or several Olea pollen allergen(s). METHODS: One-hundred and thirty-four consecutive patients diagnosed with pollinosis by Olea were studied. Of these patients only 40, reported adverse reaction to plant-derived food. Twenty-one (group A) were classified as OAS and 19 (group B) as anaphylaxis. Skin-tests with six Olea pollen allergens and several groups of fruits, were performed. Double-blind placebo-controlled food challenge (DBPCFC), confirmed the diagnostics of food allergy with the exception of patients who suffered previous anaphylactic reaction. RESULTS: All patients, showed a positive skin prick test (SPT), against one or more of Olea europaea allergens. Sensitization to Ole e 7, was more frequent (P = 0.02) in patients from group B. A total of 84 DBPCFC were performed with 44% positive results. Challenge confirmed at least the 50% of positive SPT in any case (peach: 68.42%; pear: 50%; melon: 71.42% and kiwi: 53.84%). In patients from group B, significant association with O. europaea pollen allergens were found between positive SPT to Rosaceae fruits and Ole e 3 (P = 0.045) and Ole e 7 (P = 0.03); Cucurbitaceae and Ole e 7 (P = 0.03) and Actinidiaceae with Ole e 3 (P = 0.04). CONCLUSIONS: The results of this study, establish a new spectrum of associations between pollens and plant-derived foods: sensitization to olive profilin (Ole e 2) is not more frequent in OAS patients. Patients with anaphylactic reaction after eating fruit are also sensitized to Ole e 7, a LTP present in Olea pollen, and suffer pollinic symptoms. Finally a polcalcin (Ole e 3) could be also associated to Olea pollen respiratory and food allergy.

Adult↗

[Hypersensitivity to pollen of Olea europea in patients with pollen allergy in the area of Zadar and Dubrovnik].

Olive pollen is one of the most important causes of inhalant allergy in countries around the Mediterranean sea. Due to the lack of information on hypersensitivity to the pollen of Olea europea from Croatian coast, the aim of this investigation was to establish the frequency of hypersensitivity to the pollen of Olea europea in pollen allergic patients in Zadar and Dubrovnik. Also, we compared two areas of Dalmatia regarding the number of patients and expression of allergy to the Olea europea. A total of 810 patients, children and adults, with pollen allergy were examined in both areas, Zadar and Dubrovnik. In the area of Zadar we examined 546 participants and in the area of Dubrovnik 264 participants. The patients were assessed by anamnestic data, clinical examination, measurement of pulmonary function (adults and children older than 7 years), skin prick test and enzymo-immunologic UniCAP test for measurements of specific IgE antibodies. For statistical analysis we used chi square test. Hypersensitivity to the allergy of Olea europea occurred in 66/810 (8.15%) assessed participants with pollen allergy. The comparison between the two areas didn't show any statistical difference in the number of affected participants with hypersensitivity to the Olea europea. Also, we didn't show any statistical difference in comparison of skin prick tests, immunologic measurements of specific IgE antibodies, or clinical manifestations between participants in the two investigated areas. The most prevalent clinical manifestation was rhinitis registered in 39/66 (59%) patients with hypersensitivity to the olive pollen. The majority of patients with hypersensitivity to olive pollen 51/66 (77%) live in towns. Only 3/66 (4%) participants live on an island. Rhinitis was the most prevalent clinical manifestation in our patients with hypersensitivity to allergen of Olea europea. Hypersensitivity to olive pollen was higher in participants who lived in towns. The hypersensitivity to Olea europea didn't show any difference between the two investigated areas of Dalmatia.

Adolescent↗

Olive (Olea europea) pollen allergens--II. Isolation and characterization of two major antigens.

A dialyzed extract of olive (Olea europea) pollen was fractionated by anion exchange chromatography on DEAE-Sepharose CL-6B using a discontinuous gradient of ammonium bicarbonate. The most important protein allergen was obtained from the 0.3 M fraction after gel filtration on Sephadex G-100 and separation by lentil-lectin Sepharose-4B. The major allergen of olive pollen was contained in the effluent and was designated Olea Antigen I. This material inhibited the RAST activity of 15 patients' sera that were tested. Analytical IEF demonstrated a major band at pH 5.3 and two minor ones at pH 5.6 and 5.0. When these were run into SDS-polyacrylamide gel electrophoresis in a second dimension, all were separated into two bands of mol. wt 17 and 19 K. A second protein, which is the next most important allergen, Olea Antigen II, was obtained from the 0.5 M fraction by chromatofocusing in a 4-7 pH range followed by filtration on Bio-gel P-30. Olea Antigen II had a mol. wt of 8 K as assessed by SDS-PAGE. IEF analysis displayed one main band at pH 3.6 and two minor bands at pH 3.8 and 4.0, respectively. OL-1, an anti-Olea europea monoclonal antibody (MAb) previously reported by us Lauzurica et al. (1988) reacted with the 17 and 19 K antigens from the crude extract and with Olea Antigen I but not with Olea Antigen II.

Allergens↗

Diversity of Olea genotypes and the origin of cultivated olives.

Tandem repeats belonging to three DNA sequence families ( OeTaq80, OeTaq178, and OeGEM86) were isolated from the nuclear DNA of Olea europaea cv. Carolea and dot-hybridized to the genomic DNA of 14 hypothetically different Olea species, 78 olive cultivars, and 14 wild olives. The copy number per unreplicated haploid genome of OeTaq80- and OeTaq178-related sequences was in the 10(7)-10(6) range and that of OeGEM86-related sequences was in the 10(5) range in cultivars, wild olives and some Olea species. A large variation in the frequency of repeats belonging to each sequence family was observed within each group of plants. Positive correlations existed in each genome between the frequencies of repeats belonging to each family, and their overall frequency was positively correlated to the genome size. Duncan grouping showed that the frequency variation of tandem repeats within each group of plants was not continuous. Two main groups and several subgroups of genotypes could be separated within both the olive cultivars and the wild olives. Discrete areas in the Mediterranean Basin could be delimited by the geographic distribution of cultivated olives with different genotypes and the wild plants were associated with the cultivars in these areas according to genotypic similarity. The Olea species could be divided into four genotypic groups. Three of these, comprising accessions from Asia and North Africa, showed similarity with the genotypes of cultivars and wild olives. These results suggest a polyphyletic origin of cultivated olives from different wild Olea forms distributed throughout the Mediterranean Basin.

Journal Article↗

Olea europaea airborne pollen in southern Spain.

BACKGROUND: Olea europaea pollen is one of the most abundant constituent pollens in Seville (southern Spain). It is responsible for many documented cases of pollinosis in the area. OBJECTIVE: To contribute to the useful knowledge of Olea europaea for allergists. METHODS: The number of Olea europaea pollen grains in the atmosphere was recorded during 8 consecutive years (1987-1994), using a Cour collector. RESULTS: The concentration of Olea europaea pollen was as high as 250 to 1015 grains/m3 only during 1 or 2 weeks in April and May. Overall annual production of O. europaea pollen alternated between years. The beginning of the main pollination period was related to the mean temperature of the preceding months (February and March). Pollination occurred when the mean temperature in both months was higher than 14 degrees C; conversely it was delayed when the mean temperature was lower. Main pollination period length depended upon both temperature and rainfall during this period: temperatures higher than 19.5 degrees C and absence of rainfall shortened the main pollination period, while lower temperatures (15 to 18 degrees C) together with rainfall rates above 100 mm lengthened it. CONCLUSIONS: Climatic variables such as preceding mean temperature and rainfall impact on pollen anthesis of Olea europaea affect onset and duration of pollination. A consideration of yearly cycles of pollen production as well as these variables should allow pollen forecasting.

Air Pollution↗

Tracing the history of an enzyme polymorphism: the case of alcohol dehydrogenase-2 (Adh-2) of the olive fruit fly Bactrocera oleae.

In the olive fruit fly Bactrocera oleae, previous studies have described a one-locus three-allele electrophoretic polymorphism of the enzyme alcohol dehydrogenase and provided evidence that the polymorphism is under the influence of selection. A recent study has shown that this species carries a two-locus duplication for alcohol dehydrogenase. Here, we show that the polymorphism maps at one of the duplicated loci, Adh2, and identify the nucleotide and, therefore, the inferred amino acid differences among the three allozymes. At the amino acid level, the polymorphism is of the simplest possible form: there is no intra-allozyme variation, and interallozyme differences are restricted to one amino acid for two pairs of alleles and to two amino acids for the third pair. Consideration of the amino acid residues at the sites that segregate in B. oleae in four congeneric species and the phylogenetic trees produced from the nucleotide sequences of the Adh2 gene of these species point to the same allozyme as the ancestral form of the polymorphism. Interestingly, this allozyme comprises less than 1% of the gene pool of present-day natural populations of B. oleae, where the other two allozymes appear to form a stable polymorphism. Previous studies have shown that the frequency of the rare allozyme rises rapidly in laboratory colonies maintained on artificial diet and declines again when the artificial diet is replaced with olive fruit, the natural substrate of B. oleae. The geographical distribution of several congeneric species suggests that B. oleae originated in the Indian subcontinent, where the olive tree is practically absent. The poor performance of the ancestral allele on the olive fruit suggests the possibility that the decline of this allele and the concomitant rise of the presently common alleles might be associated with the expansion of the insect's geographical distribution to areas where the olive tree has become its main and perhaps sole host. The estimated age of the polymorphism is compatible with this hypothesis, but firmer support could be difficult to obtain.

Alcohol Dehydrogenase↗

High levels of Olea europaea pollen and relation with clinical findings.

BACKGROUND: Olea europaea pollen is an important cause of seasonal allergic rhinitis and bronchial asthma in southern Spain. For patients allergic to grass pol- len the critical concentration of airborn pollen is 50 grains/m3, but in the case of Olea pollinosis no data is available. METHODS: Fifty-six seasonal allergic rhinitis patients (29 in 1994 and 27 in 1995) were included in this study, all of whom lived in Jaen. Daily symptom card were filled in and pollen counts during May and June were performed in both years. A linear regression model was used for analysis of the airborne pollen concentration and the symptom score. RESULTS: Significant correlations among daily counts of Olea pollen and rhinitis symptoms were obtained. Most of our monosensitized patients needed a high Olea pollen concentration in the atmosphere (around 400 grains/m3) to suffer at least from mild allergic rhinitis symptoms. CONCLUSION: Local conditions with a wide area dedicated to olive tree cultivars result in a high concentration of this pollen in the atmosphere. Monosensitized Olea patients in our area seem to need exceptionally high levels to suffer from allergic symptoms.

Adolescent↗

Allergy to Olea europaea pollen: relationship between skin prick tests, RAST, ELISA and bronchial provocations tests.

Olea Europaea is the most important allergenic tree in Southern Italy and in the Naples area. Its pollination period lasts from the middle of April to the end of June. In our pollinosis patients we observed frequency of sensitization of 13.05% to Olea pollen. That means that this pollen follows Parietaria (47.80%) and Gramineae (34.70%) regarding the frequency of allergic sensitization. In an attempt to evaluate the degree of allergic sensitization to Olea pollen and the concordance of various tests, we studied 24 asthmatic patients with skin positivity to only this pollen using commercially available allergen extracts and reagents. The patients were examined by SPT, RAST, ELISA and BPT with allergen. Our study has shown that there is no relationship between the results of SPT and Phadebas RAST; a slight relationship between SPT and Phadezym RAST, and a very good relationship between Phadebas and Phadezym RAST. Bronchial challenge with Olea extract was positive in all patients. These results may be interpreted considering the low degree of purification of Olea pollen extracts available commercially.

Adolescent↗

Genomic organization, sequence interrelationship, and physical localization using in situ hybridization of two tandemly repeated DNA sequences in the genus Olea.

Two tandemly repeated DNA sequences, the 81-bp family and pOS218, have been isolated from a Sau3AI Olea europaea ssp. sativa partial genomic library. Sequencing of the 81-bp element showed the monomer to be between 78 and 84 bases long and to contain 51-58% adenine and thymidine residues. Comparison between the monomers revealed heterogeneity of the sequence primary structure. The clone pOS218 is 218 bases long, and sequence comparison between the two elements revealed that an internal region of the pOS218 repeated DNA sequence had 79% homology to the 81 bp repeat sequence. A breakage-reunion mechanism, involving the CAAAA sequence, could be responsible for the derivation of pOS218 from the 81 bp family element. By using double target in situ hybridization, co-localization of the two sequences on Olea chromosomes was observed. The sequences were present at DAPI stained heterochromatic regions, as major or minor sites having a subtelomeric or interstitial location. Methylation studies using two sets of isoschizomers, Sau3AI-MboI and MspI-HpaII, demonstrated that most cytosine residues in the GATC sites and the internal cytosine in the CCGG sites of both elements were methylated in O. europaea ssp. sativa. No major difference in methylation was apparent between DNA extracted from young leaves or from callus of O. europaea ssp. sativa. Both elements are also present in Olea chrysophylla, Olea oleaster, and Olea africana, but are absent from other Oleaceae genera, including Phillyrea, Forsythia, Ligustrum, Parasyringa, and Jasminum.

Base Sequence↗

Transferability of olive microsatellite loci across the genus Olea.

The transferability of microsatellite markers developed for olive cultivars ( Olea europaea L.) has been tested and confirmed in the Olea complex. Thirty two genotypes, belonging to different taxa of the genus Olea, have been analyzed with four olive SSRs. Positive amplifications at all loci were obtained in 13 taxa (at least one accession per species). Sixty seven different alleles have been detected at the four loci analyzed. Polymorphic products have been observed at the inter- and intra-species level. Some SSR loci have shown multiple amplification products in some species. The high number of unique alleles has allowed the unambiguous discrimination of most accessions. Similarity coefficients and relationships among the Olea taxa have been calculated based on SSR amplification results. The reliability of SSRs as markers for intra-species variability evaluation has been confirmed while their use to explore relationships at the inter-species level is discussed, being dependent on the locus analyzed.

DNA, Plant↗

The dual function of ovo/shavenbaby in germline and epidermis differentiation is conserved between Drosophila melanogaster and the olive fruit fly Bactrocera oleae.

The olive fruit fly Bactrocera oleae (B. oleae) is a major olive damaging pest in the Mediterranean area. As a first molecular analysis of a developmental gene in this insect, we characterised the ovo/shavenbaby (ovo/svb) gene. In Drosophila, ovo/svb encodes a family of transcription regulators with two distinct functions: ovo is required for female germline differentiation and svb controls morphogenesis of epidermal cells. Here, we report the cloning and characterisation of ovo/svb in B. oleae, showing that the ovo genomic organisation and complex pattern of germline transcription have been conserved between distantly related Dipterae. We further show that B. oleae svb embryonic expression precisely prefigures the pattern of larval trichomes, supporting the conclusion that regulatory changes in svb transcription underlie evolutionary diversification of trichome patterns seen among Dipterae.

Amino Acid Sequence↗

Genetic and cytogenetic analysis of the olive fruit fly Bactrocera oleae (Diptera: Tephritidae).

The genetic and cytogenetic characteristics of one of the major agricultural pests, the olive fruit fly Bactmcera oleae, are presented here. The mitotic metaphase complement of this insect consists of six pairs of chromosomes including one pair of heteromorphic sex chromosomes, with the male being the heterogametic sex. The analysis of the polytene complements of three larval tissues, the fat body, the salivary glands and the Malpighian tubules of this pest has shown (a) a total number of five long chromosomes (10 polytene arms) that correspond to the five autosomes of the mitotic nuclei and a heterochromatic mass corresponding to the sex chromosomes, (b) the constancy of the banding pattern of the three somatic tissues, (c) the absence of a typical chromocenter as an accumulation of heterochromatin, (d) the existence of reverse tandem duplications, and (e) the presence of toroid tips of the chromosome arms. The in situ hybridization of genes or DNA sequences to the salivary gland polytene chromosomes of B. oleae provided molecular markers for all five autosomes and permitted the establishment of chromosomal homologies among B. olea, B. tryoni and Ceratitis capitata. The heat shock response of B. oleae, as revealed by heat-inducible puffing and protein pattern, shows a higher thermotolerance than Drosophila melanogaster.

Animals↗

Effect of acetone feeding on alcohol dehydrogenase activity in the olive fruit fly, Bactrocera oleae.

The purpose of this study is to demonstrate a clear connection between the presence of acetone in larval diet and alcohol dehydrogenase (ADH) activity in laboratory raised populations of Bactrocera oleae. ADH activity of B. oleae is depressed in acetone-impregnated diets. At the same time the change of activity is accompanied by a change in the relative proportions of the multiple forms of ADH. The bulk of activity in the most cathodally migrating form is lost, and all the activity becomes localized in the less cathodally migrating forms of the enzyme. Moreover, ADH activity, expressed in vivo, appears to drop after exposure to acetone, as shown by the fact that larvae become less sensitive to pentenol poisoning. Our results show clear selective differences imposed by acetone on three homozygous genotypes involving the ADH alleles F, S and I in B. oleae. The directions of these differences were found to vary with the fitness component under test. Acetone treatment seems to affect developmental time and larva's viability as well as allele frequencies of ADH under artificial rearing. The effect of acetone on the maintenance of ADH polymorphism in artificially reared populations of B. oleae is further discussed.

Acetone↗

Amount and organization of the heterochromatin in Olea europaea and related species.

The amount and spatial organization of the heterochromatin in nuclei of the shoot meristem and the frequency in the nuclear DNA of sequences belonging to a family of tandem repeats were investigated in cultivars of Olea europaea and related species. Significant differences between Olea species and between cultivars of O. europaea were observed: (i) in the spatial organization of the heterochromatin in interphase nuclei as determined by the number and surface area of the chromocentres; (ii) in genome size; and (iii) in the amount of condensed chromatin as measured by cytophotometry carried out at different thresholds of optical density. DNA elements belonging to a family of tandem repeats about 80 bp in length (OeTaq80 repeats) were isolated from the genomic DNA of an olive cultivar. It was shown: (i) by nucleotide sequence comparisons, that these repeats display variability in structure even within the same array, where different elements may share no more than 74% homology; (ii) by in situ hybridization, that OeTaq80-related DNA sequences are mainly localized in the heterochromatin at the chromosome ends; (iii) by dot-blot hybridization experiments, that these sequences are highly represented in the genome of all the olive cultivars and the majority of Olea species studied, and that their frequency may differ significantly even between olive cultivars; and (iv) by calculating the copy number of OeTaq80-related sequences per haploid (1C) genome, that the redundancy of these DNA elements may differ significantly between the genomes tested. It is suggested that the inter- and intraspecific changes in the nuclear and genomic traits observed can contribute to the understanding of the phylogenetic relationships between Olea species and in defining parameters to be exploited in varietal identification within cultivated olives.

Base Sequence↗

Partial purification and insecticidal activity of toxic metabolites secreted by a Mucor hiemalis strain (SMU-21) against adults of Bactrocera oleae and Ceratitis capitata (Diptera: Tephritidae).

The secondary metabolites present in the methanol extract of a Mucor hiemalis strain (SMU-21) mycelia, cultured in liquid medium, were evaluated for toxicity to Bactrocera oleae (Gmelin) and Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) adults. Feeding and contact bioassays revealed that the methanol fraction of the crude supernatant was strongly toxic to both species. Symptoms of toxicity developed quickly; lethargy occurred 1-2 h posttreatment, and mortality reached 82-97% after 24 h. Both feeding and contact bioassays showed that B. oleae was more susceptible than C. capitata. Concentrations producing 50% mortality (LC50) to B. oleae and C. capitata after 24 h in feeding bioassays were 0.52 and 1.28 mg/ 0.1 ml diet, respectively, but 34.8 and 64.0 microg/ cm2, respectively, after 4 h in contact bioassays. Solvent partition, precipitation, and chromatographic procedures were used to isolate the active principles from the crude supernatant. These resulted in the isolation of one high-pressure liquid chromatography fraction with insecticidal activity on B. oleae flies equal to the initial crude supernatant.

Animals↗

Immunotherapy with a depigmented, polymerized vaccine of Olea europaea pollen allergens. Significantly reduces specific bronchial and skin test reactivity in sensitized patients after one year of treatment.

Clinical manifestations after the inhalation of Olea europaea pollen are very frequent in Spain. Forty-five patients with a clinical history of asthma and sensitivity to O. europaea pollen were included in a randomized trial to evaluate the safety and the clinical efficacy of immunotherapy with a new chemically modified extract (depigmented and glutaraldehyde-polymerized) of Olea europaea. The study was conducted following good clinical practices and appropriate consent forms were signed. Patients were divided into three groups of 15 individuals: Group A received a maximum concentration of 44 micrograms/ml of the depigmented, polymerized allergen extract (equivalent to 100 HEPL of the native unmodified extract). Group B received 10 times less; Group C did not receive any specific immunotherapy. Any adverse event was recorded to assess safety. Efficacy was evaluated by measuring the amount of allergen needed to elicit a positive response in specific bronchial challenges before and after 12 months of immunotherapy. The treatment schedule consisted of an incremental phase of five injections and a maintenance dosage of 0.5 ml per month. Each patient received a total of 14 injections during this period. No moderate or serious adverse events related to immunotherapy were recorded. At the beginning of the study, no significant differences were observed between the three groups in specific bronchial hyperreactivity (p > 0.05). A significant difference (p < 0.05) was obtained after 12 months. Patients in Group A needed four times more native unmodified allergen than Group C to elicit the same degree of bronchoconstriction. The analysis of the individual groups before and after 12 months of treatment showed that patients in Groups B and C did not improve. Patients in Group A showed a significant improvement (p < 0.05) in specific bronchial hyperreactivity, and at the end of the study needed 5.5 times more native unmodified allergen to obtain the same degree of bronchial response as in the beginning. Depigmented and glutaraldehyde-polymerized vaccines of Olea europaea pollen are very safe for treating patients with asthma and clinical sensitivity to allergens of this pollen. The clinical efficacy of this new allergen vaccine seems to be dose-dependent as shown by specific bronchial challenges as well as by symptom and medication scores. These modified extracts induce protection against unmodified native allergens.

Allergens↗

Airborne pollen of Olea in five regions of Portugal.

The aim of this work was to study spatial and temporal distribution of Olea europeae airborne pollen in different Portuguese regions: Reguengos de Monsaraz (south); Bairrada (west); Braga (northwest); Valença do Douro and Foz Côa (north-east). Airborne pollen sampling was conducted from 1998-2003 using "Cour" type samplers located in each region. The main pollen season (MPS) of Olea lasted on average 36 days and occurred from late April until middle-to-end of June. During the studied period, inter-annual variations among and within regions, concerning the total annual pollen counts and the beginning, peak and ending dates of the MPS, were reported. Reguengos de Monsaraz and Bairrada registered the earliest MPS starting date, followed by Valença do Douro and Foz-Côa, and the latest date was verified in Braga that also had the shortest MPS. Reguengos de Monsaraz presented the longest MPS with the highest differences in the beginning and ending dates, but minimum differences in the dates of the maximum pollen peak. Our results showed an increase in the Olea annual pollen index, from north to south, and from the west to the east regions of the country.

Air↗