The diversity and factors influencing fungal assemblages in phyllosphere of Mediterranean tree species have been barely studied, especially when endophytic and epiphytic communities are simultaneously considered. In this work, the endophytic and epiphytic fungal communities from olive tree phyllosphere were studied. This tree species is natural from the Mediterranean region and adapted to grow under adverse climatic conditions. The main objectives were to determine whether there are differences between both fungal communities and to examine whether different abiotic (climate-related) and biotic (plant organs) factors play a pivotal role in structuring these communities. Both communities differed in size and composition, with epiphytic community being richer and more abundant, displaying also a dominance of melanized fungi. Season was the major driver of community composition, especially of epiphytes. Other drivers shaping epiphytes were wind speed and temperature, while plant organ, rainfall, and temperature were the major drivers for endophytic composition. In contrast, canopy orientation caused slight variations in community composition of fungi, but with distinct effects in spring and autumn seasons. In conclusion, epiphytic and endophytic communities are not driven by the same factors. Several sources of variation undergo complex interactions to form and maintain phyllosphere fungal community in Mediterranean climates. Climatic parameters have influence on these fungal communities, suggesting that they are likely to be affected by climate changes in a near future.

Phenotypic characterization of Arabidopsis thaliana gain- and loss-of-function mutants is a delicate and meticulous task that often involves the analysis of multiple parameters. Arabidopsis heat tolerance has been evaluated based on direct assessments that include seed germination, seedling survival, hypocotyl and root elongation, or indirect measurements such as chlorophyll content or ion leakage. In an attempt to simplify the detection of heat stress-associated phenotypes, a collection of protocols for analysis of seed germination and seedling survival to heat treatment is proposed. Temperatures and lengths of heat treatments were combined into several heat tolerance assays, to be used as a primary approach for the search and characterization of basal and acquired heat tolerance-associated phenotypes at early developmental stages. The usefulness of this methodology was illustrated through the characterization of heat-related phenotypes in different Arabidopsis ecotypes as well as in gain- and loss-of-function mutants. The use of standardized experimental protocols designed to detect temperature-related phenotypes is proposed. The suggested plate-based assays provide an appropriate framework of experimental conditions for detection of variability amongst natural accessions or mutants lines. Functional studies could be facilitated by using this inexpensive and undemanding approach.

The dynamics of Xylella fastidiosa infections in the context of the endophytic microbiome was studied in field-grown plants of the susceptible and resistant olive cultivars Kalamata and FS17. Whole metagenome shotgun sequencing (WMSS) coupled with 16S/ITS rRNA gene sequencing was carried out on the same trees at two different stages of the infections: In Spring 2017 when plants were almost symptomless and in Autumn 2018 when the trees of the susceptible cultivar clearly showed desiccations. The progression of the infections detected in both cultivars clearly unraveled that Xylella tends to occupy the whole ecological niche and suppresses the diversity of the endophytic microbiome. However, this trend was mitigated in the resistant cultivar FS17, harboring lower population sizes and therefore lower Xylella average abundance ratio over total bacteria, and a higher α-diversity. Host cultivar had a negligible effect on the community composition and no clear associations of a single taxon or microbial consortia with the resistance cultivar were found with both sequencing approaches, suggesting that the mechanisms of resistance likely reside on factors that are independent of the microbiome structure. Overall, Proteobacteria, Actinobacteria, Firmicutes, and Bacteriodetes dominated the bacterial microbiome while Ascomycota and Basidiomycota those of Fungi.