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Team-based learning (TBL) is an active learning pedagogy developed for in-class sessions and based on the collaborative work of small groups of students. The increasing push to online and blended learning has enhanced the need to expand this pedagogy to a virtual environment, but little evidence has been produced on how students accept online synchronous sessions of TBL. The purpose of this study, that relies on 427 responses, is to present a comparative perspective of traditional in-class versus adapted fully synchronous online TBL and across different disciplinary fields. Students of two different academic years and different programs were surveyed for their acceptance of TBL. They were invited to answer closed-ended questions focused on their engagement in all TBL learning process and the final outcomes provided. Results obtained from this unique comparative study revealed a wide approval of TBL, regardless of the environment (online or in-class TBL sessions), scientific area of courses and student gender. The acceptance of fully online TBL sessions, in a similar way as traditional in-class sessions, could be a rationale for giving more use to the `virtual' context. Other results corroborated previous researches on TBL, such the need of student awareness of TBL benefits to get more engaged in the process or the impact of student activities overload on the TBL process. Implications are informative for pedagogical practice., info:eu-repo/semantics/publishedVersion

Background Soil microbiomes are important to maintain soil processes in forests and confer protection to plants against abiotic and biotic stresses. These microbiomes can be affected by environmental changes. In this work, soil microbial communities from different cork oak Portuguese forests under different edaphoclimatic conditions were described by using a metabarcoding strategy targeting ITS2 and 16S barcodes. Results A total of 11,974 fungal and 12,010 bacterial amplicon sequence variants (ASVs) were obtained, revealing rich and diverse microbial communities associated with different cork oak forests. Bioclimate was described as the major factor influencing variability in these communities (or bioclimates/cork oak forest for fungal community), followed by boron and granulometry. Also, pH explained variation of fungal communities, while C:N ratio contributed to bacterial variation. Fungal and bacterial biomarker genera for specific bioclimates were described. Their co-occurrence network revealed the existence of a complex and delicate balance among microbial communities. Conclusions The findings revealed that bacterial communities are more likely to be affected by different edaphoclimatic conditions than fungal communities, also predicting a higher impact of climate change on bacterial communities. The integration of cork oak fungal and bacterial microbiota under different bioclimates could be further explored to provide information about useful interactions for increasing cork oak forest sustainability in a world subject to climate changes.

Plant growth promoting rhizobacteria (PGPR) are in increasing demand due to their role in promoting sustainable practices, not only in agriculture but also in forestry. Keeping in mind the future application of PGPR for increasing cork oak sustainability, the aim of this study was to find cork oak PGPR isolates with increased nutrient solubilisation traits, able to promote root morphological changes and/or antagonize cork oak bark phytopathogens. Soils from three cork oak forests with distinct bioclimates (humid, semi-humid and semi-arid) were used for isolating bacteria. From the 7634 colony-forming units, 323 bacterial isolates were biochemically assayed for PGPR traits (siderophores production, phosphate solubilizing and organic acids production), and 51 were found to display all these traits. These PGPR were able to induce root morphological changes on Arabidopsis thaliana, like suppression of primary root growth, increase of lateral roots or root hairs formation. However, the most proficient PGPR displayed specific ability in changing a single root morphological trait. This ability was related not only to bacterial genotype, but also with the environment where bacteria thrived and isolation temperature. Bacteria from semi-arid environments (mainly Bacillus megaterium isolates) could hold a promising tool to enhance plant development. Other isolates (Serratia quinivorens or B. cereus) could be further explored for biocontrol purposes., This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundação para a Ciência e a Tecnologia) in the framework of the projects SuberControl (PTDC/ASP-SIL/28635/2017), BioISI (UIDB/04046/2020) and CIMO (UIDB/00690/2020).

Team-based learning (TBL) is an active learning pedagogy developed for in-class sessions and based on the collaborative work of small groups of students. The increasing push to online and blended learning has enhanced the need to expand this pedagogy to a virtual environment, but little evidence has been produced on how students accept online synchronous sessions of TBL. The purpose of this study, that relies on 427 responses, is to present a comparative perspective of traditional in-class versus adapted fully synchronous online TBL and across different disciplinary fields. Students of two different academic years and different programs were surveyed for their acceptance of TBL. They were invited to answer closed-ended questions focused on their engagement in all TBL learning process and the final outcomes provided. Results obtained from this unique comparative study revealed a wide approval of TBL, regardless of the environment (online or in-class TBL sessions), scientific area of courses and student gender. The acceptance of fully online TBL sessions, in a similar way as traditional in-class sessions, could be a rationale for giving more use to the ‘virtual’ context. Other results corroborated previous researches on TBL, such the need of student awareness of TBL benefits to get more engaged in the process or the impact of student activities overload on the TBL process. Implications are informative for pedagogical practice.

Ectomycorrhizal fungi are crucial for forests sustainability. For Castanea sativa, ectomycorrhizal fungus Pisolithus tinctorius is an important mutualist partner. Saprotrophic fungi Hypholoma fasciculare, although used for biocontrol of Armillaria root disease, it negatively affected the interaction between the P. tinctorius and plant host roots, by compromise the formation of P. tinctorius-C. sativa mycorrhizae. In this work, fungal morphology during inhibition of H. fasciculare against P. tinctorius was elucidated. P. tinctorius growth was strongly affected by H. fasciculare, which was significantly reduced after six days of co-culture and become even more significant through time. During this period, P. tinctorius developed vesicles and calcium oxalate crystals, which were described as mechanisms to stress adaption by fungi. H. fasciculare produced different volatile organic compounds in co-cultures over time and differ between single or in dual-species. H. fasciculare highly produced sesquiterpenes (namely, alpha-muurolene) and nitrogen-containing compounds, which are recognised as having antimicrobial activity., This work was funded by FEDER through the Operational Competitiveness Program (COMPETE) and by Portuguese national funds through the Foundation for Science and Technology (FCT) within the scope of project PTDC/AGR-AAM/099556/2008, UID/MULTI/04046/2013, and the CIMO (UIDB/00690/2020) and the Applied Molecular Biosciences Unit-UCIBIO (UID/Multi/04378/2020). Nathalie Moreira acknowledge FCT for her grant, SFRH/BDP/109668/2015.

Microorganisms colonizing internal tissues of insects are known important players in the insect’s reproduction, nutrition, and fitness. Thus, a better knowledge and characterization of this microbial community can lead to their exploitation in the management of insect populations. Philaenus spumarius has been of interest in the recent years due to its confirmed ability of vectoring the plant pathogen Xylella fastidiosa. In this work the microbial community of different organs (head, abdomen, and genitalia) of 41 P. spumarius was assessed by culturing methods. Overall, 86 isolates classified in 51 operational taxonomic units (OTUs) (41 bacteria and 10 fungi) were obtained, being found 2 OTUs on average per insect. The most abundant genera were Curtobacterium (within bacteria) and Cladosporium (within fungi). The abdomen has the highest number of microbial isolates and fungal diversity with about 0.32 OTUs per sample, while genitalia harbored the highest bacterial diversity with about 0.68 OTUs per sample. The bacterial isolates able to growth in artificial medium were selected, and their influence on the insect cycle and reproduction were assessed using an insect model. The observed effects on insects were variable depending on the bacterial isolates. Some bacteria demonstrated ability to increase insect population while other bacterial isolates displayed a negative effect by decreasing insect reproduction or by causing insect premature death. Although preliminary, these results are very promising for the management of P. spumarius population and, consequently, for the transmission of X. fastidiosa., PT; PPT; ccameirao@ipb.pt

Plant-inhabiting microorganisms interact directly with each other affecting disease progression. However, the role of host plant and plant habitat in shaping pathobiome composition and their implication for host susceptibility/resistance to a particular disease are currently unknown. For the elucidation of these questions, both epiphytic and endophytic bacterial communities, present in asymptomatic and symptomatic twigs from olive cultivars displaying different susceptibilities to olive knot (OK) disease, were investigated using culturing methods. OK disease was the main driver of the bacterial community, causing changes on their diversity, abundance and composition. OK disease effect was most notorious on OK-susceptible cultivar and when considering the endophytic communities. Plant habitat (epiphytes vs. endophytes) also contributed to the bacterial community assembling, in particular on symptomatic twigs (knots) of OK-susceptible cultivar. In contrast, host cultivar had little effect on the bacterial community composition, but OK-symptomatic twigs (knots) revealed to be more affected by this driver. Overall, the pathobiome seems to result from an intricate interaction between the pathogen, the resident bacteria, and the plant host. Specific bacterial genera were associated to the presence or absence of OK disease in each cultivar. Their ability to trigger and/or suppress disease should be studied in the future., The authors are grateful to the FEDER and Foundation for Science and Technology (FCT, Portugal) under Programme PT2020 for financial support to CIMO (UIDB/00690/2020) and BioISI (UID/MULTI/04046/2013), as well as the Horizon 2020, the European Union's Framework Programme for Research and Innovation, for financial support the project PRIMA/0002/2018 INTOMED -Innovative tools to combat crop pests in the Mediterranean. D. Mina thanks FCT, POPH-QREN and FSE for SFRH-BD-105341/2014 grant.

The common bean (Phaseolus vulgaris L.) is the most consumed legume and the major protein source for poor populations, especially in developing Afro-Asian countries. Common bean has a high nutritional value, being rich in protein, carbohydrates, essential fatty acids, vitamins, minerals, and fibers. Besides these nutrients, common bean is also enriched in bioactive compounds, such as polyphenols, mainly flavonoids. These compounds reveal important antioxidant activities through their capacity to interfere with oxidative processes and consequently exhibit antiinflammatory activity. Common bean also reveals anticarcinogenic and antimutagenic activities. Accordingly the consumption of common bean has been associated with the prevention and control of several age-related problems, such as diabetes, obesity, cardiovascular and gastrointestinal diseases, and cancer. This chapter focuses on the bioactivities present in common bean grains that make this legume an excellent functional food. Therefore the increase in common bean consumption, in traditional or nutraceutical forms, should be encouraged for improving human health.

The phyllosphere comprises the aerial parts of plants and is colonized by a great diversity of microorganisms, either growing inside (as endophytes) or on the surface (as epiphytes) of plant tissues. The factors that structure the diversity of epiphytes and the importance of these microorganisms for host plant protection have been less studied when compared to the case of endophytes. In this work, the epiphytic fungal communities from fruits of the olive tree (olives) in different maturation stages (green and semi-ripened), obtained from different olive orchard managements (integrated and organic production) and from distinct cultivars displaying different susceptibilities to olive anthracnose (Cobrançosa and Madural), are compared by using a metabarcoding approach. We discuss whether such differences in host resistance against anthracnose depend on both the fungal taxa or fungal community composition. A total of 1565 amplicon sequence variants (ASVs) were obtained, mainly belonging to the Ascomycota phylum and Saccharomycetes class. Although significant differences on epiphytic fungal richness were observed among olives obtained in different production systems and maturation stages, these factors in addition to host cultivar did not influence the composition of the epiphytes. Despite these results, a co-inertia analysis showed that Aureobasidium spp. and Sporocadaceae spp. were positively associated with the green olives of the cv. Madural produced under integrated production, while Saccharomycetales spp. (Kluyveromyces, Candida, Kazachstania and Saccharomyces) were positively associated with the semi-ripened olives of the cv. Cobrançosa obtained from organic production. The discriminant power of these fungi, some of them recognized as biocontrol agents, suggest that they might be important in conferring differences on host plant susceptibility to anthracnose., This research was funded by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundação para a Ciência e a Tecnologia) in the framework of the projects PTDC/ASP-PLA/31133/2017, as well as BioISI (UIDB/04046/2020), CBMA (UIDB/04050/2020) and Mountain Research Center—CIMO (UIDB/00690/2020).

Genetic diversity is fundamental to breeding programs and consequently has an important role in obtaining new varieties. To properly use the genetic diversity present in germplasm collections, a good knowledge of the agro-morphological traits of each accession is needed. The aim of this study was to explore the production capacity of 24 cowpea landraces from southern Europe, through phenotypic characterization and evaluation in three different locations in Greece and Portugal.; Results: Most qualitative parameters tested showed a high stability among the three locations. A wide difference was observed among the three locations with respect to number of days to flowering, ranging from 55 to 99 days. Quantitative traits showed a higher genotype × environment than genetic variance component. In general, an inverse relationship between σ2ge /σ2g ratio (where σ2ge is genotype × genotype interaction and σ2g is genotype impact) and heritability value was observed. Principal component analysis was able to group accessions based on their origin. The first two principal components explained 97.52% of variation, being the number of seeds per plant, plant height and seed protein content, the traits which contributed most to variability.; Conclusion: The results show that sufficient variation exists in different traits within landraces in the studied cowpea germplasm to pursue a breeding program. However, the quantitative traits showed a higher genotype × environment component. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Cowpea is a grain legume native from Africa and is a primary source of protein for millions of people in sub-Saharan Africa and other parts of the developing world. The main important characteristics of this crop include a good protein quality with a high nutritional value, its nitrogen-fixing ability, and an ability to be more drought- and heat-tolerant than most of its legume relatives. In a research perspective, studies of cowpea are relatively scarce, despite its relevance to agriculture in the developing world and its resilience to stress. The present review provides an overview of different aspects of cowpea, with a special emphasis on the molecular markers for assessing genetic diversity, as well as on biochemical and transcriptomic data with respect to evaluating cowpea drought stress tolerance. The integration of both datasets will be useful for the improvement of cowpea because research on drought stress tolerance is of major interest for this crop in a challenging environment. © 2017 Society of Chemical Industry.; © 2017 Society of Chemical Industry.

Quercus suber L. is an evergreen tree species with high economic, ecological and social importance within the Mediterranean Basin. Cork oak forests occupy more than 2 million hectares worldwide, being mainly located in Algeria, Morocco, Portugal and Spain. As in other Mediterranean ecosystems, cork oak forests have been reported as important reservoirs of biological diversity, including endemic species that are currently under threat due to abiotic and biotic stress. Despite the adaptation of cork oak to the Mediterranean climate, which is characterised by warm, dry summers and wet winters, the growth and productivity of this species is sensitive to climatic change and variability. Extended periods of high temperature and/or low precipitation leading to low level of available water in the soil, can trigger the decline of cork oak and increase vulnerability to pathogen attack. Plant microbiomes are major factors for preserving plant health and productivity under challenging climates and their endophytic components can have dual ecological function, both as detrimental microbes or as beneficial symbionts. Endophytes can play beneficial roles for plant health and productivity but some can become opportunistic pathogens that take advantage of weakened plants that are stressed by environmental conditions. This review discusses endophytes in the context of Mediterranean bioclimates, the geographic distribution of cork oaks and the spread of opportunistic disease-causing agents. Some studies have begun to characterise and isolate endophytes from cork oak, which represents the first steps towards understanding how cork oak endophytes might help ameliorate the negative impacts of climate change for this tree species., (undefined)

Supplementary material related to this article can be found, in the online version, at doi: https://doi.org/10.1016/j.scienta.2018.11.082., Global warming has an increasing impact on the availability of water for agriculture. Crops tolerant to high temperatures and drought, such as cowpea (Vigna unguiculata L. Walp.), have an added value in the near future. The main objective of this study was to evaluate the effect of drought on seed germination and seedling emergence of cowpea genotypes, in order to screen the most tolerant genotypes. Seeds from 58 cowpea genotypes all over the world were submitted to two stress conditions, induced by PEG-6000 (corresponding to osmotic potentials of -0.75 bars and -1.5 bars). Germination and seedling growth parameters, vigor index and proline content were determined to assess drought tolerance. The results revealed significant differences of all parameters among genotypes after treatments and interaction of both. Water stress caused a general decrease in germination and seedling growth, while an increase in proline content was observed. A high variation of drought responses were detected among genotypes, being possible to select seven genotypes (C11, C18, C44, C46, C47, C50 and C54) as tolerant to drought at germination stage. These results will be useful to select the best suitable parents for insertion in future breeding programs., This study was supported by EUROLEGUME project. This project has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no 613781. European Investment Funds by FEDER/COMPETE/POCI - Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the projects UID/AGR/04033/2013 and UID/AGR/04046/2013.

Plants are able to interact with plentiful bacteria resulting in a number of positive or negative outcomes for plant health. The ecological balance between pathogens and beneficial bacteria could be strategically disturbed and manipulated for improving host plant protection. As bacterial communities present in the phyllosphere of herbaceous plants have been largely studied, a number of biocontrol agents for controlling host diseases are already identified and used with promising results. A few studies on the use of phyllosphere biocontrol agents on woody crop tree plants have revealed encouraging results toward a future where plant disease control could be attained without the application of chemical compounds. In addition to the use of biocontrol agents, disease suppression can be achieved by the manipulation of microbial communities through plant management practices. In this review, an overview of the available knowledge on phyllosphere bacterial communities of woody tree crop species is provided, giving special emphasis to the structural differences of bacterial communities living on and within important tree crop species. Studies and challenges on the application and/or manipulation of these bacteria under in planta conditions are discussed, disclosing new sustainable ways for dealing with woody crop diseases.

In nature, pathogens live and interact with other microorganisms on plant tissues. Yet, the research area exploring interactions between bacteria-fungi and microbiota-plants, within the context of a pathobiome, is still scarce. In this study, the impact of olive knot (OK) disease caused by the bacteria Pseudomonas savastanoi pv. savastanoi (Psv) on the epiphytic and endophytic fungal communities of olive tree twigs from three different cultivars, was investigated in field conditions. The ITS-DNA sequencing of cultivable fungi, showed that OK disease disturbs the resident fungal communities, which may reflect changes in the habitat caused by Psv. In particular, a reduction on epiphyte abundance and diversity, and changes on their composition were observed. Compared to epiphytes, endophytes were less sensitive to OK, but their abundance, in particular of potential pathogens, was increased in plants with OK disease. Host genotype, at cultivar level, contributed to plant fungal assembly particularly upon disease establishment. Therefore, besides fungi - Psv interactions, the combination of cultivar - Psv also appeared to be critical for the composition of fungal communities in olive knots. Specific fungal OTUs were associated to the presence and absence of disease, and their role in the promotion or suppression of OK disease should be studied in the future., This research was partially supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds through FCT (Fundacao para a Ciencia e a Tecnologia) in the framework of the project EXCL/AGR-PRO/0591/2012. This work was supported by FCT under the project UID/MULTI/04046/2013. T. Gomes thanks FCT, POPH-QREN and FSE for PhD SFRH/BD/98127/2013 grant; and also the COST Action FA1405 for a short-term scientific mission (STSM) grant.

Supplementary material related to this article can be found, in the online version, at doi: https://doi.org/10.1016/j.micres.2020.126693., A wide array of fungal endophytes is known to inhabit plant tissues and were recently recognized as essential for plant health. A better description of the scarcely known endophyte microbiota in olive tree phyllosphere is the first step for elucidating the microbial interactions that lead to olive disease establishment. In this work, the fungal endophytic community of the phyllosphere of different olive tree cultivars (Cobrancosa, Galega vulgar, Madural, Picual, Verdeal Transmontana) is revealed by using a metabarcoding strategy targeting ITS1 barcode. A total of 460 OTUs were obtained, increasing the broad view of fungal endophytes inhabiting the olive tree phyllosphere, in particular yeast endophytes. New endophytes were persistently found in all cultivar tissues. Different olive tree cultivars depicted distinct endophyte communities. Olive cultivars exhibited dissimilar amounts of fungi with distinct ecological functions, which could explain at least in part their differential susceptibility/tolerance to olive diseases., This work was supported by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundacao para a Ciencia e a Tecnologia) in the framework of the projects EXCL/AGR-PRO/0591/2012, PTDC/AGR-PRO/4354/2012, and PTDC/ASP-PLA/31133/2017, as well as BioISI (UIDB/04046/2020) and Mountain Research Center - CIMO (UIDB/00690/2020).

The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2020.02051/full#supplementary-material, Olive knot (OK) is a widespread bacterial disease, caused byPseudomonas savastanoipv.savastanoi(Pss), which currently has not effective control methods. The use of naturally occurring microbial antagonists, such as bacteria, as biocontrol agents could be a strategy to manage this disease. The objective of this work was to select bacteria from olive tree phyllosphere able to antagonizePssusingin vitroandin plantaexperiments. The elucidation of their modes of action and the potential relationship between antagonism and bacteria origin has been investigated, as well. To this end, 60 bacterial isolates obtained from the surface and inner tissues of different organs (leaves, twigs, and knots), from two olive cultivars of varying susceptibilities to OK, were screened for theirin vitroantagonistic effect againstPss. A total of 27 bacterial strains were able to significantly inhibitPssgrowth, being this effect linked to bacteria origin. Strains from OK-susceptible cultivar and colonizing the surface of plant tissues showed the strongest antagonistic potential. The antagonistic activity was potentially due to the production of volatile compounds, siderophores and lytic enzymes.Bacillus amyloliquefaciensP41 was the most effective antagonistic strain and their capacity to control OK disease was subsequently assayed usingin plantaexperiments. This strain significantly reduces OK disease severity (43.7%), knots weight (55.4%) and population size ofPss(26.8%), while increasing the shoot dry weight (55.0%) and root water content (39.6%) ofPss-infected olive plantlets. Bacterial isolates characterized in this study, in particularB. amyloliquefaciensP41, may be considered as promising biocontrol candidates for controlling OK disease., This work was funded by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade), national funds through FCT (Fundacao para a Ciencia e a Tecnologia) and by Horizon 2020, the European Union's Framework Programme for Research and Innovation, within the project PRIMA/0002/2018 (INTOMED - Innovative tools to combat crop pests in the Mediterranean), and the Mountain Research Center - CIMO (UIDB/00690/2020 and UIDB/04046/2020).

Variation on bacterial communities living in the phyllosphere as epiphytes and endophytes has been attributed to plant host effects. However, there is contradictory or inconclusive evidence regarding the effect of plant genetics (below the species' level) and of plant tissue type on phyllosphere bacterial community assembly, in particular when epiphytes and endophytes are considered simultaneously. Here, both surface and internal bacterial communities of two olive (Olea europaea) cultivars were evaluated in twigs and leaves by molecular identification of cultivable isolates, with an attempt to answer these questions. Overall,Proteobacteria,ActinobacteriaandFirmicuteswere the dominant phyla, being epiphytes more diverse and abundant than endophytes. Host genotype (at cultivar level) had a structuring effect on the composition of bacterial communities and, in a similar way, for both epiphytes and endophytes. Plant organ (leaf vs. twig) control of the bacterial communities was less evident when compared with plant genotype and with a greater influence on epiphytic than on endophytic community structure. Each olive genotype/plant organ was apparently selective towards specific bacterial operational taxonomic units (OTUs), which may lead to specific feedbacks on fitness of plant genotypes. Bacterial recruitment was observed to happen mainly within epiphytes than in endophytes and in leaves as compared with twigs. Such host specificity suggested that the benefits derived from the plant-bacteria interaction should be considered at genetic levels below the species., The authors are grateful to the FEDER and Foundation for Science and Technology (FCT, Portugal) under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019) and BioISI (UID/MULTI/04046/2013), as well as the Horizon 2020, the European Union’s Framework Programme for Research and Innovation, for financial support the project PRIMA/0002/2018 INTOMED—Innovative tools to combat crop pests in the Mediterranean. D. Mina thanks FCT, POPH-QREN and FSE for SFRH-BD-105341/2014 grant.

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., This research was funded by the EU H2020 Research Project XF-ACTORS "Xylella Fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy" (Grant Agreement 727987) and by the Mountain Research Center-CIMO (UIDB/00690/2020) and BioISI (UIDB/04046/2020). The Funding Agencies are not responsible for any use that may be made of the information it contains.

Climate changes comprise increasing global temperature and water cycle deregulation (precipitation storms and long dry seasons). Many affected ecosystems are located within the Mediterranean basin, where cork oak (Quercus suber L.) is one of the most important forest ecosystems. Despite cork oak tolerance to drought, the decrease of water availability and increase of temperature is causing a serious decline of cork oak populations. In the present work, the bacterial community of cork oak soils was assessed by metabarcoding using Illumina Miseq. Soils from seven independent cork oak forests were collected along a climate gradient. In all forest soils, Proteobacteria and Actinobacteria were the richest and more abundant bacteria. Acidobacteria also presented a high relative abundance, and Chloroflexi was a rich phylum. The soil bacterial community diversity and composition was strongly affected by the climatic region where cork oak resides and specific bacterial taxa were differently affected by precipitation and temperature. Accordingly, cork oak bacterial communities clustered into three distinct groups, related with humid, sub-humid and arid/semi-arid climates. Driest and warmer forests presented more diverse bacterial communities than humid and coolest forests. However, driest climates presented more homogenous bacterial communities among forests than humid climates. Climate (mainly precipitation) revealed to be the strongest driver leading to significant variations of bacterial community profiles. The most impacted bacterial taxa by climatic variables were Proteobacteria, in particular Gammaproteobacteria and Deltaproteobacteria, Chloroflexi, and Firmicutes. Humid forests presented mainly Acidobacteria as good indicators of climate, whereas Actinobacteria members were better indicators for arid forests (mainly Gaiellales and Frankiales). Some indicator species for different climate conditions were members of the bacterial core of cork oak stands (7% of the total bacterial community). Taken together, different, This work was supported by FEDER through the Operational Competitiveness Program (COMPETE) and by Portuguese national funds through the Foundation for Science and Technology (FCT) within the scope of the project POCI-01-0145-FEDER-028635; FCT/MCTES/PIDDAC (Portugal) under the project (PEst-OE/BIA/UI4046/2014; UID/MULTI/04046/2013) and PhD grant to F.R. (SFRH/BD/86519/2012).

Pest control service provided by natural enemies of Bactrocera oleae, the key pest of the olive tree, is nowadays recognized as fundamental. B. oleae has developed resistance to common insecticides, and negative effects both on consumers' health and non-target species arc the major drawbacks of conventional control strategics. Carabid beetles are potential B. oleae pupae predators, but their predation on field still need to be assessed. We tested adult Pseudoophonus rufipes, a species known to be active in olive orchard when pest pupae are abundant in the soil, in order to detect B. oleae pupae consumption at different post feeding times for both male and female carabids. An already existing protocol was used for detecting B. oleae mtDNA sequences of the cytochrome oxidase subunit I gene in carabids' gut, and its versatility improved. B. oleae mtDNA was detected up to 20 h after pupa ingestion with a high percentage of success, without significant differences between sexes and pair primers used. Prey DNA extraction was tested from both dissected and non-dissected carabids, obtaining comparable results. The trapping system used to collect carabids for molecular assays and the new elements introduced in the protocol represent cost-effective solutions that may be beneficial for future laboratory trials and, mostly, for the analysis of field-collected predators. Fostering the investigation of soil predators in olive orchard may increase the design of conservation control strategies against B. oleae., Scuola Superiore Sant’Anna, Pisa, Italy (PhD Programme in Agrobiodiversity).

The online version of this article (https://doi.org/10.1007/s00248-018-1161-9) contains supplementary material, which is available to authorized users., The diversity and factors influencing fimgal 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., This work is funded by FEDER funds through COMPETE (Programa Operacional Factores de Competitividade) and by national funds by FCT (Fundacao para a Ciencia e a Tecnologia) within the framework of the project EXCL/AGR-PRO/0591/2012. T. Gomes thanks FCT, POPH-QREN, and FSE for PhD SFRH/BD/98127/2013 grant.

Cowpea (Vigna unguiculata L. Walp) is an important legume crop due to its high protein content, adaptation to heat and drought and capacity to fix nitrogen. Europe has a deficit of cowpea production. Knowledge of genetic diversity among cowpea landraces is important for the preservation of local varieties and is the basis to obtain improved varieties. The aims of this study were to explore diversity and the genetic structure of a set of Iberian Peninsula cowpea accessions in comparison to a worldwide collection and to infer possible dispersion routes of cultivated cowpea., This study was supported by EUROLEGUME project. This project has received funding from the European Union’s Seventh Framework Programme for research, technological development and demonstration under grant agreement no 613781. European Investment Funds by FEDER/COMPETE/ POCI – Operational Competitiveness and Internationalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT – Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013. MMA was partially supported by the Feed the Future Innovation Lab for Climate Resilient Cowpea (USAID Cooperative Agreement AID-OAA-A-13-00070), which is directed by TJC. The funding entities had no role in the design of the study, collection, analysis and interpretation of data, or in writing the manuscript., info:eu-repo/semantics/publishedVersion

Cork oak (Quercus suber L.) forests play an important ecological and economic role. Ectomycorrhizal fungi (ECMF) are key components for the sustainability and functioning of these ecosystems. The community structure and composition of ECMF associated with Q. suber in different landscapes of distinct Mediterranean bioclimate regions have not previously been compared. In this work, soil samples from cork oak forests residing in different bioclimates (arid, semi-arid, sub-humid, and humid) were collected and surveyed for ectomycorrhizal (ECM) root tips. A global analysis performed on 3565 ECM root tips revealed that the ECMF community is highly enriched in Russula, Tomentella, and Cenoccocum, which correspond to the ECMF genera that mainly contribute to community differences. The ECMF communities from the rainiest and the driest cork oak forests were distinct, with soils from the rainiest climates being more heterogeneous than those from the driest climates. The analyses of several abiotic factors on the ECMF communities revealed that bioclimate, precipitation, soil texture, and forest management strongly influenced ECMF structure. Shifts in ECMF with different hyphal exploration types were also detected among forests, with precipitation, forest system, and soil texture being the main drivers controlling their composition. Understanding the effects of environmental factors on the structuring of ECM communities could be the first step for promoting the sustainability of this threatened ecosystem., This work was supported by Fundacao Ciencia e Tecnologia (FCT/MCTES/PIDDAC, Portugal), under the project (PEst-OE/BIA/UI4046/2014; UID/MULTI/04046/2013) and PhD grant to F.R. (SFRH/BD/86519/2012).

The community of entomopathogenic fungi associated with the olive moth Prays oleae is specific for each generation of this insect. The entomopathogen Beauveria bassiana is almost exclusive to the phyllophagous or leaf generation, whereas Paecilomyces formosa mainly occurs in the carpophagous or fruit generation. The main objective of this work was to evaluate olive plant-mediated effects on the development (germination, growth, sporulation and spore viability) of both entomopathogens. While B. bassiana was strongly inhibited by olives, P. formosa development was more restricted by leaves. These inhibitory effects were caused by both volatile and diffusible compounds produced by olives and leaves. Specifically, the effects of two leaf volatile compounds, (Z)-3-hexen-1-ol and (Z)-3-hexenyl acetate were studied. The results suggest that olive plant organs affect entomopathogens by controlling their abundance and function, which may partly explain the occurrence of a fungal species in a specific P. oleae generation., This work is funded by FEDER through the Operational Competitiveness Programme—COMPETE—and by national funds through the Foundation for Science and Technology—FCT—in the scope of project PTDC/AGR-AAM/102600/2008 “Entomopathogenic fungi associated to olive pests: isolation, characterization and selection for biological control”. The first author is grateful to the Science and Technology Foundation for the PhD grant SFRH/BD/44265/2008., info:eu-repo/semantics/publishedVersion

Common bean is a legume important in Mediterranean diet with interesting nutritional food characteristics due to its proteins, fibers and antioxidant activity. There is an increased interest among consumers in foods rich in natural antioxidants for maintaining and improving health. Beyond this, beans are a low-price food and an excellent complement for the population with low income. There are some differences in the phenolic and flavonoid compounds content and antioxidant capacity between accessions of common bean. In this study, 15 traditional Portuguese and 1 Greek accessions of Phaseolus vulgaris L. were analyzed. Total phenols, ortho-diphenols and flavonoid contents were investigated and antioxidant activity were measured by different assays (DPPH, ABTS) which may give a better characterization of this activity.

Drought impact on plants is an increasing concern under the climate change scenario. Cowpea (Vigna unguiculata L. Walp.) is considered as one of the most tolerant legume crops to drought, being the search for the best well-adapted genotypes crucial to face the future challenges. Different approaches have been used for differentiating plant responses to drought stress. Plants of four cowpea genotypes were submitted to three watering regimens (a severe and moderate drought stress, and well-watered control) during 15 days, and several physiological, biochemical and molecular parameters were evaluated. Stressed plants revealed commonly-described drought stress characteristics, but not all assayed parameters were useful for discriminating plants with different drought severities or genotypes. The analyses which have contributed most to genotype discrimination were those related with stomatal function, and biochemical markers such as proline and anthocyanin contents. Antioxidant enzymes activities and related genes expression did not differed among genotypes or upon drought stress treatments, suggesting that scavenging enzymes are not involved in the differential ability of cowpea plants to survive under drought stress. This information will be useful to evaluate and use genetic resources, as well as design strategies for breeding cowpea resistance to drought stress., This study was supported by EUROLEGUME project. This project has received funding from: the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no 613781; National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2019; and the project PEstOE/BIA/UI4046/2014; UID/MULTI/04046/2013.

The search for new antimicrobial compounds and the optimization of production methods turn the use of antimicrobial susceptibility tests a routine. The most frequently used methods are based on agar diffusion assays or on dilution in agar or broth. For filamentous fungi, the most common antimicrobial activity detection methods comprise the co-culture of two filamentous fungal strains or the use of fungal extracts to test against single-cell microorganisms. Here we report a rapid, effective and reproducible assay to detect fungal antimicrobial activity against single-cell microorganisms. This method allows an easy way of performing a fast antimicrobial screening of actively growing fungi directly against yeast. Because it makes use of an actively growing mycelium, this bioassay also provides a way for studying the production dynamics of antimicrobial compounds by filamentous fungi. The proposed assay is less time consuming and introduces the innovation of allowing the direct detection of fungal antimicrobial properties against single cell microorganisms without the prior isolation of the active substance(s). This is particularly useful when performing large screenings for fungal antimicrobial activity. With this bioassay, antimicrobial activity of Hypholoma fasciculare against yeast species was observed for the first time., Authors are grateful to Fundacao para a Ciencia e Tecnologia (FCT) for financial support (Project PTDC/AGR-AAM/099556/2008 and Pest-C/BIA/UI4050/2011, FEDER-COMPETE, FCT, Portugal)., info:eu-repo/semantics/publishedVersion

Genetic diversity is fundamental to breeding programs and consequently has an important role in obtaining new varieties. To properly use the genetic diversity present in germplasm collections, a good knowledge of the agro-morphological traits of each accession is needed. The aim of this study was to explore the production capacity of 24 cowpea landraces from southern Europe, through phenotypic characterization and evaluation in three different locations in Greece and Portugal.Most qualitative parameters tested showed a high stability among the three locations. A wide difference was observed among the three locations with respect to number of days to flowering, ranging from 55 to 99 days. Quantitative traits showed a higher genotype × environment than genetic variance component. In general, an inverse relationship between σThe results show that sufficient variation exists in different traits within landraces in the studied cowpea germplasm to pursue a breeding program. However, the quantitative traits showed a higher genotype × environment component. © 2017 Society of Chemical Industry.

Cowpea is a grain legume native from Africa and is a primary source of protein for millions of people in sub-Saharan Africa and other parts of the developing world. The main important characteristics of this crop include a good protein quality with a high nutritional value, its nitrogen-fixing ability, and an ability to be more drought- and heat-tolerant than most of its legume relatives. In a research perspective, studies of cowpea are relatively scarce, despite its relevance to agriculture in the developing world and its resilience to stress. The present review provides an overview of different aspects of cowpea, with a special emphasis on the molecular markers for assessing genetic diversity, as well as on biochemical and transcriptomic data with respect to evaluating cowpea drought stress tolerance. The integration of both datasets will be useful for the improvement of cowpea because research on drought stress tolerance is of major interest for this crop in a challenging environment. © 2017 Society of Chemical Industry.

Bactrocera oleae (Rossi) (Diptera: Tephritidae) is considered the most devastating pest of the olive tree worldwide. In an effort to develop management and biological control strategies against this pest, new molecular tools are urgently needed. In this study, we present the design of B. oleae-specific primers based on mitochondrial DNA sequences of cytochrome oxidase subunit I (COI) gene. Two pairs of B. oleae-specific primers were successfully designed and named as SBo1-F/SBo1-R and SBo2-F/SBo1-R, being able to amplify 108 and 214 bp COI fragments, respectively. The specificity of designed primers was tested by amplifying DNA from phylogenetically related (i.e. Diptera order) and other non-pest insects living in olive groves from the Mediterranean region. When using these primers on a PCR-based diagnostic assay, B. oleae DNA was detected in the gut content of a soil-living insect, Pterostichus globosus (Fabricius) (Coleoptera: Carabidae). The detection of B. oleae DNA in the guts of arthropods was further optimized by adding bovine serum albumin enhancer to the PCR reaction, in order to get a fast, reproducible and sensitive tool for detecting B. oleae remains in the guts of soil-living arthropods. This molecular tool could be useful for understanding pest-predator relationships and establishing future biological control strategies for this pest., This work was supported by FEDER Funds throughout Programa Operacional Factores de Competitividade – COMPETE and National Funds throughout FCT – Fundação para a Ciência e Tecnologia, within the project EXCL/ AGR-PRO/0591/2012: Olive crop protection in sustainable production under global climatic changes: linking ecological infrastructures to ecosystem functions. M. Rejili was supported by a fellowship on the frame of Erasmus Mundus EU MARE NOSTRUM., info:eu-repo/semantics/publishedVersion

Quercus suber (cork oak) is a West Mediterranean species of key economic interest, being extensively explored for its ability to generate cork. Like other Mediterranean plants, Q. suber is significantly threatened by climatic changes, imposing the need to quickly understand its physiological and molecular adaptability to drought stress imposition. In the present report, we uncovered the differential transcriptome of Q. suber roots exposed to long-term drought, using an RNA-Seq approach. 454-sequencing reads were used to de novo assemble a reference transcriptome, and mapping of reads allowed the identification of 546 differentially expressed unigenes. These were enriched in both effector genes (e.g., LEA, chaperones, transporters) as well as regulatory genes, including transcription factors (TFs) belonging to various different classes, and genes associated with protein turnover. To further extend functional characterization, we identified the orthologs of differentially expressed unigenes in the model species Arabidopsis thaliana, which then allowed us to perform in silico functional inference, including gene network analysis for protein function, protein subcellular localization and gene co-expression, and in silico enrichment analysis for TFs and cis-elements. Results indicated the existence of extensive transcriptional regulatory events, including activation of ABA-responsive genes and ABF-dependent signaling. We were then able to establish that a core ABA-signaling pathway involving PP2C-SnRK2-ABF components was induced in stressed Q. suber roots, identifying a key mechanism in this species' response to drought., This work was supported by FEDER through the Operational Competitiveness Program – COMPETE – and by national funds through the Foundation for Science and Technology – FCT – within the scope of projects SOBREIRO/0033/2009 “Cork oak ESTs Consortium – Abiotic stress: drought, salt and oxidative stresses” and PTDC/AGRGPL/118505/2010“An integrated approach to identify stress-related regulatory genes in cork oak (SuberStress).” PHC was supported by FEDER through COMPETE, and by national funds (FCT) within the scope of project “SUMOdulator” [Refs. FCOMP-01-0124-FEDER-028459 and PTDC/BIA-PLA/3850/2012]. HA was supported by the “Genomics and Evolutionary Biology” project, co-financed by North Portugal Regional Operational Programme 2007/2013 (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF)., info:eu-repo/semantics/publishedVersion