Search

Increasing exposure to unfavorable temperatures and water deficit imposes major constraints on most crops worldwide. Despite several studies regarding coffee responses to abiotic stresses, transcriptome modulation due to simultaneous stresses remains poorly understood. This study unravels transcriptomic responses under the combined action of drought and temperature in leaves from the two most traded species: Coffea canephora cv. Conilon Clone 153 (CL153) and C. arabica cv. Icatu. Substantial transcriptomic changes were found, especially in response to the combination of stresses that cannot be explained by an additive effect. A large number of genes were involved in stress responses, with photosynthesis and other physiologically related genes usually being negatively affected. In both genotypes, genes encoding for protective proteins, such as dehydrins and heat shock proteins, were positively regulated. Transcription factors (TFs), including MADS-box genes, were down-regulated, although responses were genotype-dependent. In contrast to Icatu, only a few drought- and heat-responsive DEGs were recorded in CL153, which also reacted more significantly in terms of the number of DEGs and enriched GO terms, suggesting a high ability to cope with stresses. This research provides novel insights into the molecular mechanisms underlying leaf Coffea responses to drought and heat, revealing their influence on gene expression. [ABSTRACT FROM AUTHOR]

This document is an editorial from the International Journal of Molecular Sciences titled "Molecular Insight of Plants Response to Drought Stress: Perspectives and New Insights towards Food Security." The editorial emphasizes the importance of understanding the molecular mechanisms underlying drought tolerance in plants in order to enhance crop productivity and environmental conservation. It highlights several research contributions that investigate the molecular responses of plants to drought stress, including studies on transcription factors, epidermal patterning factors, genetic variation, and gene expression in response to cold stress. The editorial also discusses the challenges and new perspectives in the field, such as understanding dynamic regulatory networks, epigenetic regulation, environmental interactions, and translational research. The authors conclude by emphasizing the need for continued interdisciplinary research to address the remaining challenges and pave the way for sustainable agricultural practices and resilient food systems in a changing climate. [Extracted from the article]

Salinity is an increasing problem worldwide that limits crop production. The cultivation of salt-tolerant flowers is a potential sustainable strategy for the exploitation of saline soils while avoiding the use of freshwater resources. However, our understanding of how flowers can adapt to salinity is still limited. In this study, we investigated the effects of salinity on a widely-used ornamental plant (Tagetes patula) by submitting three cultivars (cv. Aurora Orange, Fireball, Safari Scarlet) to different salinity levels (0, 50, 100, 300 mM). The plants were grown under controlled conditions and followed over two weeks. We tested the effect of salinity on plant growth and flower production, as well as on the levels of total carotenoids, polyphenols, and flavonoids, and the activities of antioxidant enzymes (catalase, peroxidase activity, ascorbate peroxidase). The findings indicate a substantial decline in both plant growth and flower production under saline conditions. Overall, plant height was ¼ lower, and there were 1/3 less flowers under salinity. Additionally, there was a consistent rise in antioxidant compounds, highlighting the swift response of defense mechanisms, both enzymatic and non-enzymatic, to combat stress. The high levels of antioxidant compounds sustain the use of marigold flowers as a new source of nutritional compounds, with enriched nutritional contents. Yet, cultivation of these plants in saline conditions should carefully consider the pronounced adverse effects observed at high salinity levels (e.g., 100 and 300 mM) on both plant and flower production. [ABSTRACT FROM AUTHOR]

This document is an editorial from the International Journal of Molecular Sciences titled "Plant Responses to Climate Change." It discusses the risks that climate change poses to the natural environment and agriculture, particularly in relation to abiotic stresses such as extreme temperatures and drought. The editorial highlights the importance of scientific research in understanding the molecular mechanisms of plant responses to these stresses and improving crop tolerance. The document also provides a list of ten research contributions from various countries, covering topics such as autophagy-related genes, the circadian clock, drought and elevated CO2 effects, and salt tolerance in different plant species. The editorial concludes by emphasizing the need for further research on the interactions between abiotic stresses and their impacts on different plant components, as well as the potential of emerging technologies like genome editing and artificial intelligence in supporting sustainable agriculture. [Extracted from the article]

We studied the impacts of moderate and severe drought on different pot marigold (Calendula officinalis L.) genotypes, evaluating the antioxidant performance of leaves and flowers concerning the levels of proline and malondialdehyde, the activity of antioxidant enzymes (catalase, peroxidase, and ascorbate peroxidase), as well as impacts on flower production. Overall, we found high resilience to moderate drought. However, the severe drought significantly affected flower production, despite the high level of antioxidants, proline, and malondialdehyde. Results also indicate significant variation in drought tolerance among pot marigolds, providing an opportunity to identify valuable tolerance traits. [ABSTRACT FROM AUTHOR]

Salinization is an increasing problem worldwide, limiting crop production. Soil salinity causes ion toxicity, osmotic stress, nutrient deficiency, and oxidative stress in plants, leading to the overproduction of reactive oxygen species (ROS). To counterbalance these effects, plants activate a complex detoxification system through the action of antioxidant pigments, carotenoids, phenolics, and flavonoids, and the accumulation of minerals, that play an important role in human health against several diseases. In this study, we investigated the impacts of salinity (0, 50, 100, and 300 mM NaCl) on the flowers of three Tagetes patula cultivars harvested after 14 days, recording mineral, total phenol and protein contents. Overall, all compounds increased with an increase in salinity levels, in comparison with control conditions. Results showed that edible marigold flowers are a promising crop with enriched nutritional contents and antioxidant activity that can be a new source of source of nutraceuticals. However, further tests are needed to evaluate the implications that salinity might have on the viability and yield of flowers. [ABSTRACT FROM AUTHOR]

Drought is a major constraint to plant growth and productivity worldwide and will aggravate as water availability becomes scarcer. Although elevated air [CO2] might mitigate some of these effects in plants, the mechanisms underlying the involved responses are poorly understood in woody economically important crops such as Coffea. This study analyzed transcriptome changes in Coffea canephora cv. CL153 and C. arabica cv. Icatu exposed to moderate (MWD) or severe water deficits (SWD) and grown under ambient (aCO2) or elevated (eCO2) air [CO2]. We found that changes in expression levels and regulatory pathways were barely affected by MWD, while the SWD condition led to a down-regulation of most differentially expressed genes (DEGs). eCO2 attenuated the impacts of drought in the transcripts of both genotypes but mostly in Icatu, in agreement with physiological and metabolic studies. A predominance of protective and reactive oxygen species (ROS)-scavenging-related genes, directly or indirectly associated with ABA signaling pathways, was found in Coffea responses, including genes involved in water deprivation and desiccation, such as protein phosphatases in Icatu, and aspartic proteases and dehydrins in CL153, whose expression was validated by qRT-PCR. The existence of a complex post-transcriptional regulatory mechanism appears to occur in Coffea explaining some apparent discrepancies between transcriptomic, proteomic, and physiological data in these genotypes. [ABSTRACT FROM AUTHOR]

Terpenyl acetate, which is used for the production of flavors and fragrances, is industrially prepared from turpentine oil in a two‐step process including hydration and catalytic esterification. Such processes have several shortcomings like too‐large amounts of catalysts, non‐recyclability of catalysts, long reaction times, and serious environmental pollution. Here, a direct one‐step synthesis of terpenyl acetate from a feed mixture of turpentine oil and acetic acid with fluoroantimonic acid as superacid catalyst is described. Only 0.2 mol % of catalyst is necessary for up to 96 % conversion of starting material at room temperature. The desired product was obtained with a selectivity of up to 60 %. Thus, a clean and environmentally friendly strategy for synthesizing α‐terpenyl acetate is provided. [ABSTRACT FROM AUTHOR]

Although tef provides high-quality foods and grows under conditions unsuitable for many cereal crops, it still shows a very low productivity. Achieving crop tolerance to environmental changes via modern breeding techniques while exploiting the natural variation in underutilized crops and genotypes is an excellent way to accomplish food requirements. World food production will need to increase by at least 50% to meet our future needs [[1]]. [Extracted from the article]

In contrast, under heat stress, the gene expression of antioxidant genes and the enzyme activity of antioxidants in I S. trilobata i remained high, suggesting a stronger ROS-scavenging ability. This special edition brings recent signs of progress towards understanding the molecular mechanisms underlying the influence of cold, high temperatures, elevated CO SB 2 sb , and salinity in several tropical plants. The climate crisis is pushing the planet's tropical plants towards their limits [[1]]. [Extracted from the article]

Background and Aims It is accepted that contemporary allopolyploid species have originated recurrently, but very few cases have been documented using multiple natural formations of the same species. To extend our knowledge, we have investigated the multiple origins, genetic variation and structure of the allotetraploid grass Brachypodium hybridum with respect to its progenitor diploid species B. distachyon (D genome) and B. stacei (S genome). For this, our primary focus is the Iberian Peninsula, an evolutionary hotspot for the genus Brachypodium. Methods We analysed 342 B. hybridum individuals from 36 populations using ten nuclear SSR loci and two plastid loci. The B. hybridum genetic profiles were compared with those previously reported for B. stacei and B. distachyon. In addition, phylogenetic analysis of the plastid data was performed for a reduced subset of individuals. Key Results The nuclear simple sequence repeat (SSR) genetic analysis detected medium to high genetic diversity, with a strong south-to-north genetic structure cline, and a high selfing rate in B. hybridum. Comparative genetic analysis showed a close relatedness of current B. hybridum D allelic profiles with those of B. distachyon , but a lack of similarity with those of B. stacei , suggesting another B. stacei source for the B. hybridum S alleles. Plastid analysis detected three different bidirectional allopolyploidization events: two involved distinct B. distachyon -like ancestors and one involved a B. stacei -like ancestor. The south-eastern Iberian Peninsula B. hybridum populations were more genetically diverse and could have originated from at least two hybridization events whereas north-eastern/north-western Iberian Peninsula B. hybridum populations were less diverse and may have derived from at least one hybridization event. Conclusions The genetic and evolutionary evidence supports the plausible in situ origin of the south-eastern and northern Iberian Peninsula B. hybridum allopolyploids from their respective local B. distachyon and unknown B. stacei ancestors. The untapped multiple origins and genetic variation detected in these B. hybridum populations opens the way to future evolutionary analysis of allopolyploid formation and genomic dominance and expression in the B. hybridum – B. distachyon – B. stacei grass model complex. [ABSTRACT FROM AUTHOR]

Biomass is an alternative and renewable carbon feedstock for fossil fuel in chemical industry. An important pathway for biomass conversion is the conversion of turpentine oil into different chemical intermediates. Herein we described the hydration of turpentine assisted by ultrasound waves in the presence of an inorganic acid. The desired product α‐terpineol was obtained up to 60% of selectivity after 100% of conversion of the turpentine oil, and short reaction times were needed (15 minutes to 1 hour). [ABSTRACT FROM AUTHOR]

Rhodosporiodum toruloides NCYC 921 yeast biorefinery is able to simultaneously provide a gaseous energy carrier, lipids and high-value carotenoids, either using glucose or carob pulp syrup as carbon source in the cultivation step for biomass production. The resulting leftover materials from the yeast biorefinery (YR- de-oiled yeast biomass residue, GS- glucose supernatant, CS- carob supernatant and CR- carob residue) were valued through anaerobic digestion in different mixtures, according to the generated effluents volume. Biogas productions of 55, 31 and 29 mL and methane yield of 236, 179 and 144 mL CH 4 /g substrate volatile solids (VS) were recorded for the studied mixtures MIX I (YR + GS), MIX II (YR + CS) and MIX III (YR + CS + CR) respectively. The comparison between the biogas production and volatile solids removal obtained for the individual substrates and for the admixtures substrates digestions revealed that co-digestion favoured the conversion of the biorefinery wastes into biogas/methane. Flow cytometry analysis of cells stained with propidium iodide revealed that the proportion of cells with injured membrane was lower in the substrate mixtures digestions than in the individual waste digestions, indicating that co-digestion mitigates the negative effect of potential toxic compounds present in the wastes or produced during the process digestion on the microbial consortium. [ABSTRACT FROM AUTHOR]

Background: Brachypodium distachyon (Poaceae), an annual Mediterranean Aluminum (Al)-sensitive grass, is currently being used as a model species to provide new information on cereals and biofuel crops. The plant has a short life cycle and one of the smallest genomes in the grasses being well suited to experimental manipulation. Its genome has been fully sequenced and several genomic resources are being developed to elucidate key traits and gene functions. A reliable germplasm collection that reflects the natural diversity of this species is therefore needed for all these genomic resources. However, despite being a model plant, we still know very little about its genetic diversity. As a first step to overcome this gap, we used nuclear Simple Sequence Repeats (nSSR) to study the patterns of genetic diversity and population structure of B. distachyon in 14 populations sampled across the Iberian Peninsula (Spain), one of its best known areas. Results: We found very low levels of genetic diversity, allelic number and heterozygosity in B. distachyon, congruent with a highly selfing system. Our results indicate the existence of at least three genetic clusters providing additional evidence for the existence of a significant genetic structure in the Iberian Peninsula and supporting this geographical area as an important genetic reservoir. Several hotspots of genetic diversity were detected and populations growing on basic soils were significantly more diverse than those growing in acidic soils. A partial Mantel test confirmed a statistically significant Isolation-By-Distance (IBD) among all studied populations, as well as a statistically significant Isolation-By-Environment (IBE) revealing the presence of environmental-driven isolation as one explanation for the genetic patterns found in the Iberian Peninsula. Conclusions: The finding of higher genetic diversity in eastern Iberian populations occurring in basic soils suggests that these populations can be better adapted than those occurring in western areas of the Iberian Peninsula where the soils are more acidic and accumulate toxic Al ions. This suggests that the western Iberian acidic soils might prevent the establishment of Al-sensitive B. distachyon populations, potentially causing the existence of more genetically depauperated individuals. [ABSTRACT FROM AUTHOR]

The reasons for plant rarity have been the focus of many studies, especially because rare species are more prone to extinction than common species. Under the same habitat conditions, rare plants are expected to attract fewer flower visitors and to show some limitation in their reproductive success. Here, using one of the most emblematic Neotropical plant genus ( T ibouchina) we tested whether narrow endemic and threatened species in Ecuador have a lower reproductive success or are visited by fewer pollinators than common species, in 13 populations monitored from 2011 to 2013. We also assessed whether interspecific gene flow could be considered a threat to the rare species. However, contrary to expectations, we found that few pollinators visited the flowers, independently of species rarity. Natural outcross pollinations were always very low in all small-size populations, leading to high levels of pollen limitation. Interspecific crossing experiments also revealed weak reproductive barriers in some species. This study reveals that both narrow and common species of T ibouchina have similar reproductive and pollinator patterns in Ecuador and, therefore, other causes of the rarity of these species should be considered. [ABSTRACT FROM AUTHOR]

Hybridization can generate new species if some degree of isolation prevents gene flow between the hybrids and their progenitors. The recruitment of novel pollinators by hybrids has been hypothesized to be one way in which such reproductive isolation can be achieved., We tested whether pollinators contributed to isolation between two natural Narcissus hybrids and their progenitors using pollination experiments, observations, plus morphological and floral-volatile measurements. These hybrids share the same maternal but different paternal progenitors., We found that only the hybrids were visited by and pollinated by ants. The two hybrids exceeded their progenitors in floral-tube aperture size and nectar production. The emission of floral volatiles by hybrid plants was not only equal to or higher than the progenitor species, but also contained some new compounds not produced by the progenitors., The recruitment of ants as novel pollinators in the hybrids involved the combination of increased nectar secretion and the production of novel floral scent compounds. A breakdown of chemical defence against ants may also be involved. This study provides support for the hypothesis that the recruitment of novel pollinators can contribute to reproductive isolation between hybrids and their progenitors. [ABSTRACT FROM AUTHOR]

Relatively little is known about species-level genetic diversity in flowering plants outside the eudicots and monocots, and it is often unclear how to interpret genetic patterns in lineages with whole-genome duplications. We addressed these issues in a polyploid representative of Hydatellaceae, part of the water-lily order Nymphaeales., We examined a transcriptome of Trithuria submersa for evidence of recent whole-genome duplication, and applied transcriptome-derived microsatellite (expressed-sequence tag simple-sequence repeat ( EST- SSR)) primers to survey genetic variation in populations across its range in mainland Australia., A transcriptome-based Ks plot revealed at least one recent polyploidization event, consistent with fixed heterozygous genotypes representing underlying sets of homeologous loci. A strong genetic division coincides with a trans-Nullarbor biogeographic boundary., Patterns of 'allelic' variation (no more than two variants per EST- SSR genotype) and recently published chromosomal evidence are consistent with the predicted polyploidization event and substantial homozygosity underlying fixed heterozygote SSR genotypes, which in turn reflect a selfing mating system. The Nullarbor Plain is a barrier to gene flow between two deep lineages of T. submersa that may represent cryptic species. The markers developed here should also be useful for further disentangling species relationships, and provide a first step towards future genomic studies in Trithuria. [ABSTRACT FROM AUTHOR]

As drought threatens crop productivity it is crucial to characterize the defense mechanisms against water deficit and unveil their interaction with the expected rise in the air [CO 2 ]. For that, plants of Coffea canephora cv. Conilon Clone 153 (CL153) and C. arabica cv. Icatu grown under 380 (aCO 2) or 700 μL L−1 (eCO 2) were exposed to moderate (MWD) and severe (SWD) water deficits. Responses were characterized through the activity and/or abundance of a selected set of proteins associated with antioxidative (e.g., Violaxanthin de-epoxidase, Superoxide dismutase, Ascorbate peroxidases, Monodehydroascorbate reductase), energy/sugar (e.g., Ferredoxin-NADP reductase, NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, sucrose synthase, mannose-6-phosphate isomerase, Enolase), and lipid (Lineolate 13S-lipoxygenase) processes, as well as with other antioxidative (ascorbate) and protective (HSP70) molecules. MWD caused small changes in both genotypes regardless of [CO 2 ] level while under the single imposition to SWD, only Icatu showed a global reinforcement of most studied proteins supporting its tolerance to drought. eCO 2 alone did not promote remarkable changes but strengthened a robust multi-response under SWD, even supporting the reversion of impacts already observed by CL153 at aCO 2. In the context of climate changes where water constraints and [CO 2 ] levels are expected to increase, these results highlight why eCO 2 might have an important role in improving drought tolerance in Coffea species. [ABSTRACT FROM AUTHOR]

Extended flower longevity has been widely defended to occur as a consequence of low pollination activity. To test this hypothesis in Neotropical rainforests, we selected three species of Epidendrum that flower during rainy conditions and commonly exhibit low levels of insect visits. Experimental studies carried out in 15 populations distributed along a wide spatial gradient detected the presence of pollinator limitation in all populations. Pollinia removal without pollen deposition increased longevity, and thus extended longevity could apparently be considered as an ecological advantage for these orchids. Nevertheless, flower longevity was severely shortened by abiotic factors like precipitation and humidity. Comparison with other tropical orchids showed that flower longevity was higher even in species that flower during periods of more pollinator activity than the ones studied here, which is contrary to the hypothesis that pollinators are the only driving force for long-lived flowers in Epidendrum. Thus, although long floral longevities are apparently favorable for the reproductive success of these species, the high maintenance costs experienced during rainy conditions are also an important factor constraining the selection of long-lived flowers in Epidendrum. [ABSTRACT FROM AUTHOR]

The Aristaless-related homeobox ( ARX) gene is implicated in intellectual disability with the most frequent pathogenic mutations leading to expansions of the first two polyalanine tracts. Here, we describe analysis of the ARX gene outlining the approaches in the Australian and Portuguese setting, using an integrated clinical and molecular strategy. We report variants in the ARX gene detected in 19 patients belonging to 17 families. Seven pathogenic variants, being expansion mutations in both polyalanine tract 1 and tract 2, were identifyed, including a novel mutation in polyalanine tract 1 that expands the first tract to 20 alanines. This precise number of alanines is sufficient to cause pathogenicity when expanded in polyalanine tract 2. Five cases presented a probably non-pathogenic variant, including the novel HGVS: c.441_455del, classified as unlikely disease causing, consistent with reports that suggest that in frame deletions in polyalanine stretches of ARX rarely cause intellectual disability. In addition, we identified five cases with a variant of unclear pathogenic significance. Owing to the inconsistent ARX variants description, publications were reviewed and ARX variant classifications were standardized and detailed unambiguously according to recommendations of the Human Genome Variation Society. In the absence of a pathognomonic clinical feature, we propose that molecular analysis of the ARX gene should be included in routine diagnostic practice in individuals with either nonsyndromic or syndromic intellectual disability. A definitive diagnosis of ARX-related disorders is crucial for an adequate clinical follow-up and accurate genetic counseling of at-risk family members. [ABSTRACT FROM AUTHOR]

Background In addition to energy benefits, anaerobic digestion offers other interesting advantages. The cork industry is of great environmental, economic and social significance in the western Mediterranean region, with Portugal beingthe world-leading producer and exporter. Cork boiling wastewater (CBW) is a toxic and recalcitrant organic effluent produced by this sector, which constitutes a serious environmental hazard. However, there is no documented research on anaerobic treatment/valorization performed with this effluent. The work presented here was developed with the aim to use the anaerobic digestion process to convert the CBW polluting organic load into an energy carrier gas and valuable molecules for industry. Results No lag phases were observed and a methane yield of 0.126 to 0.142 m3 kg-1chemical oxygen demand (COD)addedwas registered in the mesophilic consortium experiments carried out in batch flasks at 37 ± 1°C. Anaerobic digestion can be advantageously connected to ultrafiltration or electrochemical processes, due to the following: 1) reduction of ellagic acid content and consequentdecrease of CBW viscosity; and 2) increase in conductivity after the anaerobic process, avoiding the electrolyte application of the electrochemical process. The improvement of several CBW biochemical features shows that anaerobic digestion may provide additionally useful molecules. The rise in concentration of some of these compounds, belonging to the benzoic acid family (gallic, protocatechuic, vanillic and syringic acids), is responsible for the increase of antiradical activity of the phenolic fraction. Additionally, some enzymatic activitywas also observed and while the laccase activity increased in the digested effluent by anaerobiosis, xylanase was formed in the process. Conclusions The multidisciplinary approach adopted allowed the valorization of CBW in terms of energy and valuable biomolecules. By exploiting the anaerobic digestion process potential, a novel methodology to toxic and recalcitrant cork processing wastewater was developed. [ABSTRACT FROM AUTHOR]

Background Hybridization and polyploidy are central processes in evolution and speciation. These mechanisms often lead to complex patterns of genetic variation and the creation of novel genotypes, which may establish if they become isolated from gene flow. However, in the absence of reproductive isolation, species boundaries might easily be disrupted. Here, we used a combination of AFLPs, chloroplast DNA markers and flow cytometry to investigate the evolutionary outcomes of hybridization between two endemic Ecuadorian species of Epidendrum (E. madsenii and E. rhopalostele) in three hybrid zones. Postmating isolation was also quantified to determine the role of this barrier in restraining gene flow between hybrids and the parental species. In addition, future ecological niche models were constructed to predict the outcomes of hybridization between these species. Results Our results confirmed the presence of hybrids in all hybrid zones, but revealed that a third parental species (E. falcisepalum) has contributed to one of the hybrid zones studied. Backcross genotypes were frequent in all hybrid zones, which was in accordance with the absence of strong reproductive barriers. The process of hybridization was highly asymmetric and followed in some cases by polyploidy. The projection of future niche models predicted a severe reduction in the area suitable for the occurrence of these species, although favorable conditions will still occur for the existence of the current hybrid zones. Conclusions The recurrent process of hybridization has compromised the genetic integrity of the parental species. Most individuals of the parental species can no longer be considered as pure-bred individuals because most were classified as backcrossed hybrids. Novel genetic lineages occur in all hybrid zones implying that hybrids are fertile and can compete with the parental species. These results, together with the prediction of suitable conditions for the future occurrence of these hybrid zones, highlight the importance of conserving these geographic areas as sources of novel taxonomic entities. [ABSTRACT FROM AUTHOR]

Small fragmented populations often exhibit reduced plant-pollinator interactions and scarce outcrossing opportunities. In this context, mixed-mating systems can be advantageous since selfing can provide reproductive assurance, but they may also carry relevant costs such as those involved in inbreeding depression. This study examines the advantages and costs of selfing in Narcissus serotinus L., a mixed-mating geophyte that currently occurs in several fragmented populations in Portugal, that resulted from the construction of the largest European dam. Observation of pollinators revealed that cross-pollination is less frequent in small than in large populations. Manual self-pollinations significantly increased fruit and seed set in small-size populations, also suggesting limited insect pollination. The existence of selfing may provide reproductive assurance in small-size populations of N. serotinus where outcrossing pollination is reduced. Although floral biology and experimental pollinations showed that N. serotinus is capable of autonomous selfing, four of the six fitness traits studied showed significant inbreeding depression in all populations. The high levels of inbreeding depression found in N. serotinus suggest that the initial reproductive assurance advantage of selfing may be counterbalanced by lower survival of the resulting individuals and a decrease in the evolutionary potential of the populations. [ABSTRACT FROM AUTHOR]

The interdisciplinary nature of bioinformatics makes it an ideal framework to develop activities enabling enquiry-based learning. We describe here the development and implementation of a pilot project to use bioinformatics-based research activities in high schools, called “Bioinformatics@school.” It includes web-based research projects that students can pursue alone or under teacher supervision and a teacher training program. The project is organized so as to enable discussion of key results between students and teachers. After successful trials in two high schools, as measured by questionnaires, interviews, and assessment of knowledge acquisition, the project is expanding by the action of the teachers involved, who are helping us develop more content and are recruiting more teachers and schools. [ABSTRACT FROM AUTHOR]

Highlights: [•] We characterized MTBC populations from Portugal and Northeast Brazil. [•] We used a SNP-based Bayesian tree for identification of the MTBC isolates. [•] Our approach allowed to define the usefulness of SNPs in strain identification. [•] MTBC populations from Portugal and Northeast Brazil presented striking similarities. [ABSTRACT FROM AUTHOR]

Flower longevity, the period of time that a flower remains open, is a crucial factor in plant reproductive fitness. Though longevity varies substantially among angiosperm species, longer flower longevities may be a selective advantage under unpredictable or scarce pollination activity. This study examined this hypothesis in Narcissus serotinus L. (Amaryllidaceae), a small Mediterranean geophyte that blooms during the autumnal rainy season. Field experiments performed in three natural populations through 3 consecutive years shows that N. serotinus has the ability to expand flower longevity when pollination has not occurred. Flowers that were open longer to pollination have higher reproductive fitness than younger flowers. As N. serotinus flowers age, there was a shift in pollination patterns since flower visits by butterflies became scarcer and are replaced by one small pollen beetle. Given that there was evidence of pollen limitation and that this species produces one single flower, we suggest that extended flower longevity is selected because it assures pollination success under scarce insect visits. [ABSTRACT FROM AUTHOR]

The unpredictable seasonality of the Mediterranean climate imposes on the development of specific life-cycle phenologies such as those seen in geophyte species that have a period of dormancy to avoid stressful abiotic conditions. To date, two life-cycle strategies are recognized in Mediterranean geophytes. In some species, growth and reproduction occur in different seasons (hysteranthous geophytes), whereas in others these two processes occur at the same time (synanthous geophytes). A study of the life cycle of the autumnal-flowering Narcissus serotinus in three populations over two consecutive years confirmed that neither the hysteranthy nor the synanthy strategy could be applied to this geophyte. Rather, reproduction and growth occur simultaneously, but in different plants. A strong correlation was found between bulb weight and the presence of reproductive or vegetative plants. [ABSTRACT FROM AUTHOR]

Background and Aims Although there is evidence that both allopolyploid and homoploid hybridization lead to rapid genomic changes, much less is known about hybrids from parents with different basic numbers without further chromosome doubling. Two natural hybrids, Narcissus × alentejanus (2n = 19) and N. × perezlarae (2n = 29), originated by one progenitor (N. cavanillesii, 2n = 28) and two others (N. serotinus, 2n = 10 and N. miniatus, 2n = 30, respectively) allow us to study how DNA content and composition varies in such hybrids. Methods Flow cytometry measurements with two staining techniques, PI and DAPI, were used to estimate 2C values and base composition (AT/GC ratio) in 390 samples from 54 wild populations of the two natural hybrids and their parental species. In addition, 20 synthetic F1 hybrid individuals were also studied for comparison. Key Results Natural hybrids presented 2C values intermediate between those found in their parental species, although intra-population variance was very high in both hybrids, particularly for PI. Genome size estimated from DAPI was higher in synthetic hybrids than in hybrids from natural populations. In addition, differences for PI 2C values were detected between synthetic reciprocal crosses, attributable to maternal effects, as well as between natural hybrids and those synthetic F1 hybrids in which N. cavanillesii acted as a mother. Conclusions Our results suggest that natural hybrid populations are composed of a mixture of markedly different hybrid genotypes produced either by structural chromosome changes, consistent with classic cytogenetic studies in Narcissus, or by transposon-mediated events. [ABSTRACT FROM AUTHOR]

Summary 1. The evolutionary fate of natural hybridization is determined by the fitness of hybrids. Specifically, the survival of newly formed hybrids is affected by intrinsic genetic incompatibilities and/or genotype-environment interactions. The influence of these factors on the evolutionary potential of two natural hybrids of Narcissus has been assessed. Fitness components were estimated in early experimental F1 hybrids as well as in natural hybrids of unknown descent. 2. To test the efficiency of reproductive isolation, fruit set, seed number and seed mass from interspecific crosses were measured, as were germination rate, radicle vigour and leaf appearance time in F1 experimental hybrids in the greenhouse. 3. Germination rate, survival and the number of leaves produced were also measured in F1 plants transplanted to eight natural sites and compared with plants growing under greenhouse conditions. In addition, to overcome the difficulty of obtaining experimental late-generation hybrids, fruit set, leaf production, bulb production and bulb size were measured in natural hybrids of unknown descent. 4. Experimental crosses indicated weak post-mating barriers. In the case of N. cavanillesii, the barriers were virtually absent and interspecific crosses produced the same level of fruit set, seed set and seed mass as conspecific crosses. Fitness of F1 experimental hybrids was similar to or greater than conspecific progeny, although some reciprocal hybrids differed on some of the traits measured. No differences were found in hybrid performance between the various natural sites and the greenhouse, indicating a lack of exogenous selection against hybrids. Mature natural hybrids of unknown descent exhibited higher levels of growth and bulb propagation than parental species but very low fertility. 5. Synthesis. Hybrid vigour in early life stages and the absence of exogenous selection against hybrids provide an initial advantage for the establishment of hybrids in a wide variety of habitats. These factors, together with higher levels of growth and bulb propagation recorded in hybrids as compared to parental species, may compensate for low fertility, thus conferring evolutionary potential to Narcissus natural hybrids. [ABSTRACT FROM AUTHOR]

Evolutionary consequences of natural hybridization between species may vary so drastically depending on spatial, genetic, and ecological factors that multiple approaches are required to uncover them. To unravel the evolutionary history of a controversial hybrid ( Narcissus× perezlarae), here we use four approaches: DNA sequences from five regions (four organellar, one nuclear), cytological studies (chromosome counts and genome size), crossing experiments, and niche modeling. We conclude that (1) it actually consists of two different hybrid taxa, N.× perezlarae s.s. ( N. cavanillesii× N. miniatus) and N.× alentejanus ( N. cavanillesii × N. serotinus); (2) both have been formed several times independently, that is, polytopically; (3) N. cavanillesii was the mother progenitor in most hybridization events. We also address the origin of orphan hybrid populations of N.× perezlarae in eastern Spain, hundreds of kilometers away from N. cavanillesii. Although long-distance dispersal of already formed hybrids cannot be completely rejected, extirpation of N. cavanillesii via demographic competition is a more likely explanation. Low-reproductive barriers to fertilization by foreign pollen in N. cavanillesii, molecular footprints of the former presence of this species in the area, active asexual propagation by bulbs in N.× perezlarae, and overlapping ecological niches are consistent with the extirpation scenario. [ABSTRACT FROM AUTHOR]

Drought is a major threat to coffee, compromising the quality and quantity of its production. We have analyzed the core proteome of 18 Coffea canephora cv. Conilon Clone 153 and C. arabica cv. Icatu plants and assessed their responses to moderate (MWD) and severe (SWD) water deficits. Label-free quantitative shotgun proteomics identified 3000 proteins in both genotypes, but less than 0.8% contributed to ca. 20% of proteome biomass. Proteomic changes were dependent on the severity of drought, being stronger under SWD and with an enrolment of different proteins, functions, and pathways than under MWD. The two genotypes displayed stress-responsive proteins under SWD, but only C. arabica showed a higher abundance of proteins involved in antioxidant detoxification activities. Overall, the impact of MWD was minor in the two genotypes, contrary to previous studies. In contrast, an extensive proteomic response was found under SWD, with C. arabica having a greater potential for acclimation/resilience than C. canephora. This is likely supported by a wider antioxidative response and an ability to repair photosynthetic structures, being crucial to develop new elite genotypes that assure coffee supply under water scarcity levels. [ABSTRACT FROM AUTHOR]

Water scarcity is the most significant factor limiting coffee production, although some cultivars can still have important drought tolerance. This study analyzed leaf transcriptomes of two coffee cultivars with contrasting physiological responses, Coffea canephora cv. CL153 and Coffea. arabica cv. Icatu, subjected to moderate (MWD) or severe water deficits (SWD). We found that MWD had a low impact compared with SWD, where 10% of all genes in Icatu and 17% in CL153 reacted to drought, being mainly down-regulated upon stress. Drought triggered a genotype-specific response involving the up-regulation of reticuline oxidase genes in CL153 and heat shock proteins in Icatu. Responsiveness to drought also included desiccation protectant genes, but primarily, aspartic proteases, especially in CL153. A total of 83 Transcription Factors were found engaged in response to drought, mainly up-regulated, especially under SWD. Together with the enrollment of 49 phosphatases and 272 protein kinases, results suggest the involvement of ABA-signaling processes in drought acclimation. The integration of these findings with complementing physiological and biochemical studies reveals that both genotypes are more resilient to moderate drought than previously thought and suggests the existence of post-transcriptional mechanisms modulating the response to drought. [ABSTRACT FROM AUTHOR]

Natural hybrids between rare and common sympatric species are commonly eradicated to avoid the potential extinction of the rare species, although there is currently no clear predictive framework to quantify this risk. As hybrids can have intrinsic value as new evolutionary pathways, further knowledge on the factors controlling hybridization is needed. In this study we evaluated the role of pollination patterns in hybridization events in two sympatric populations of Narcissus cavanillesii and N. serotinus in Portugal. Narcissus cavanillesii is a rare species, while N. serotinus is widely distributed across the Mediterranean. The hybrid, N. Xperezlarae, is quite frequent in southeastern Spain but is scarce in Portugal. Reciprocal manual crossings confirmed compatibility between the two species, although hybridization was more successful when N. cavanillesii participated as female. Narcissus cavanillesii and N. serotinus only shared one pollinator, Megachile sp. (Hymenoptera), which had low visitation rates and high flower constancy. No single isolation mechanism was fully effective in preventing hybridization. Temporal displacement of flowering peaks, strong pollinator specificity, and high flower constancy in the shared pollinator all contributed to limiting hybridization in this site. In other sympatric occurrences, different phenological windows and pollination assemblages may allow greater frequency of the hybrid. [ABSTRACT FROM AUTHOR]

Mitochondrial complex I exists as a mixture of two inter-convertible forms: active (A) and de-activated (D), the latter being sensitive to SH-modifying compounds. To investigate if the conserved cysteine-rich 11.5 kDa subunit of Neurospora crassa complex I is involved in this process, we subjected the corresponding genomic DNA to site-directed mutagenesis. The four cysteine residues of the subunit were separately substituted with serine residues and the resulting proteins were independently expressed in a null-mutant strain. All of the obtained mutant strains were able to assemble a complex I with similar kinetic properties to those observed in the wild-type enzyme, indicating that none of the cysteine residues of the 11.5 kDa protein is individually relevant for the A/D transition process. Diminished amounts of assembled complex I seem to be the major effect of these specific mutations. The cysteine residues are likely important to the acquisition and stabilization of the correct 11.5 kDa protein conformation and this is reflected in the assembly/stability of complex I. [ABSTRACT FROM PUBLISHER]

Abstract: Respiratory chain complex I of the fungus Neurospora crassa contains at least 39 polypeptide subunits, of which 35 are conserved in mammals. The 11.5 kDa and 14 kDa proteins, homologues of bovine IP15 and B16.6, respectively, are conserved among eukaryotes and belong to the membrane domain of the fungal enzyme. The corresponding genes were separately inactivated by repeat-induced point-mutations, and null-mutant strains of the fungus were isolated. The lack of either subunit leads to the accumulation of distinct intermediates of the membrane arm of complex I. In addition, the peripheral arm of the enzyme seems to be formed in mutant nuo14 but, interestingly, not in mutant nuo11.5. These results and the analysis of enzymatic activities of mutant mitochondria indicate that both polypeptides are required for complex I assembly and function. [Copyright &y& Elsevier]

A nuclear gene encoding a 9.8 kDa subunit of complex I, the homologue of mammalian MWFE protein, was identified in the genome of Neurospora crassa. The gene was cloned and inactivated in vivo by the generation of repeat-induced point mutations. Fungal mutant strains lacking the 9.8 kDa polypeptide were subsequently isolated. Analyses of mitochondrial proteins from mutant nuo9.8 indicate that the membrane and peripheral arms of complex I fail to assemble. Respiration of mutant mitochondria on matrix NADH is rotenone-insensitive, confirming that the 9.8 kDa protein is required for the assembly and activity of complex I. We found a similarity between the MWFE homologues and the C-terminal part of the nqrA subunit of bacterial Na+-translocating NADH:quinone oxidoreductases (Na+-NQR), suggesting a link between proton-pumping and sodium-pumping NADH dehydrogenases. [Copyright &y& Elsevier]

The present study assesses hotel workers' perceptions regarding their soft and hard skills, during COVID-19 pandemic. To this end, data were collected through a survey applied to 4 and 5 star hotel workers in the Historical Center of Porto (Portugal). The results show that workers have a generally positive perception on their skills, especially in the case of teamwork, interpersonal relationship and flexibility (soft skills). Digital competencies (hard skills), present the lowest level of confidence. Data also show that essential competencies are rarely provided by the employer. Moreover, the evaluation of workers with a higher education degree only differs significantly regarding hard skills (linguistic). [ABSTRACT FROM AUTHOR]

Presents a continuous-flow system for on-line water monitoring using back-side contact ISFET-based sensors. ISFET response characteristics; Reference electrode characterization; Continuous-flow system design; System evaluation.

As atmospheric [CO2] continues to rise to unprecedented levels, understanding its impact on plants is imperative to improve crop performance and sustainability under future climate conditions. In this context, transcriptional changes promoted by elevated CO2 (eCO2) were studied in genotypes from the two major traded coffee species: the allopolyploid Coffea arabica (Icatu) and its diploid parent, C. canephora (CL153). While Icatu expressed more genes than CL153, a higher number of differentially expressed genes were found in CL153 as a response to eCO2. Although many genes were found to be commonly expressed by the two genotypes under eCO2, unique genes and pathways differed between them, with CL153 showing more enriched GO terms and metabolic pathways than Icatu. Divergent functional categories and significantly enriched pathways were found in these genotypes, which altogether supports contrasting responses to eCO2. A considerable number of genes linked to coffee physiological and biochemical responses were found to be affected by eCO2 with the significant upregulation of photosynthetic, antioxidant, and lipidic genes. This supports the absence of photosynthesis down-regulation and, therefore, the maintenance of increased photosynthetic potential promoted by eCO2 in these coffee genotypes. [ABSTRACT FROM AUTHOR]

Framework: Health tourism is currently one of the fastest growing forms of tourism. In Portugal, the “Tourism Strategy 2027” highlighted thermal activity as a means of economic development in the country's regions. On March 11, 2020, WHO declared a covid-19 pandemic. In Portugal, all thermal establishments took the initiative to suspend activity. After the lockdown period the units had to adapt to restart following the guidelines of the health authorities. Goals This study aims to emphasize the spa health tourism sector in Portugal in the context of the Covid-19 pandemic, analyze consumer behavior, and suggest future research on the topic. Paper organization: We present qualitative research, through content analysis using WebQDA software and a bibliometric analysis of co-occurrence of keywords in the thermal field, using the VOSviewer software. The analysis was conducted considering public statements from representatives of 21 units in mainland Portugal, published in the media, as of March 2020. Results: The analyses point to a high appreciated value by the customers to the item safety and to the aesthetic and medicinal treatments. There were no differences in behavior between public and private entities. All regions in mainland Portugal behaved the same way in terms of compliance with the health organization's guidelines, seeking to give the customer more safety and confidence. The training and actions of the human resources were essential to adapt the spas to guarantee strict safety and hygiene rules, as well as the respect for physical distance. In terms of marketing and innovation, the biggest references from the clients are at the product level. Final considerations: The current context of the Covid-19 pandemic favors the use of spas for their curative aspect related to lung problems, post-disease. There are several opportunities for hydrotherapy and research associated with the sector. It is important to draw new lines connected to its potential role in accelerating healing and prevention of coronavirus. [ABSTRACT FROM AUTHOR]

A methodology was developed to assess the allocation of different types of endogenous waste biomass to eight technologies for producing electricity, heat, biogas and advanced biofuels. It was based on the identification of key physicochemical parameters for each conversion process and the definition of limit values for each parameter, applied to two different matrices of waste biomass. This enabled the creation of one Admissibility Grid with target values per type of waste biomass and conversion technology, applicable to a decision process in the routing to energy production. The construction of the grid was based on the evaluation of 24 types of waste biomass, corresponding to 48 sets of samples tested, for which a detailed physicochemical characterization and an admissibility assessment were made. The samples were collected from Municipal Solid Waste treatment facilities, sewage sludges, agro-industrial companies, poultry farms, and pulp and paper industries. The conversion technologies and energy products considered were (trans)esterification to fatty acid methyl esters, anaerobic digestion to methane, fermentation to bioethanol, dark fermentation to biohydrogen, combustion to electricity and heat, gasification to syngas, and pyrolysis and hydrothermal liquefaction to bio-oils. The validation of the Admissibility Grid was based on the determination of conversion rates and product yields over 23 case studies that were selected according to the best combinations of waste biomass type versus technological solution and energy product. [ABSTRACT FROM AUTHOR]

Chloroplast genome diversity in cork oak (Quercus suber) is characterised by the occurrence of haplotypes that are akin to those found in other Mediterranean oak species, particularly in Q. ilex and Q. rotundifolia, suggesting the possible presence of an introgressed chloroplast lineage. To further investigate this pattern, we reconstructed chloroplast haplotypes by sequencing four chloroplast markers (cpDNA), sampled across 181 individuals and 10 taxa. Our analyses resulted in the identification of two diversified chloroplast haplogroups in Q. suber, corresponding to a geographically widespread lineage and an Afro-Iberian lineage. Time-calibrated phylogenetic analyses of cpDNA point to a Miocene origin of the two haplogroups in Q. suber, suggesting that the Afro-Iberian lineage was present in cork oak before the onset of glaciation periods. The persistence of the two haplogroups in the western part of the species distribution range may be a consequence of either ancient introgression events or chloroplast lineage sorting, combined with different fixation in refugia through glaciation periods. Our results provide a comprehensive insight on the origins of chloroplast diversity in these ecologically and economically important Mediterranean oaks. [ABSTRACT FROM AUTHOR]

Features various dance companies from several countries throughout the world. Aims of the Caleidos dance education company in Brazil; Research on dancers' health conducted by Dance UK; Research funding acquired by Mind the Gap, a German contemporary dance company.

Ecuador has a high diversity of orchids, but little is known about levels of genetic diversity for the great majority of species. Understanding how orchids might adapt to changes is crucial as deforestation and fragmentation of forest ecosystems threaten the survival of many epiphytic orchids that depend on other species, such as fungi and their host trees, for germination, growth, and establishment. Overcollection and the illegal trade are also major concerns for the survival of wild populations of orchids. Despite increasing awareness, effective interventions are often limited by a lack of data concerning the impacts that overexploitation might have. To fill this gap, we tested the effects of overcollection in the genetic diversity and structure of Masdevallia rosea, a narrow distributed epiphytic orchid historically collected in Ecuador, in comparison with the widely distributed Pleurothallis lilijae. Genotyping based on AFLPs showed reduced levels of diversity in wild populations but most especially in the overcollected, M. rosea. Overall, genetic admixture was high in P. lilijae segregating populations by altitude levels while fewer genetic groups were found in M. rosea. Genetic differentiation was low in both species. A spatial genetic structure was found in P. lilijae depending on altitude levels, while no spatial genetic structure was found in M. rosea. These results suggest different scenarios for the two species: while gene flow over long distance is possible in P. lilijae, the same seems to be unlikely in M. rosea possibly due to the low levels of individuals in the known populations. In situ and ex situ conservation strategies should be applied to protect the genetic pool in these epiphytic orchid species, and to promote the connectivity between wild populations. Adopting measures to reduce overexploitation and to understand the impacts of harvesting in wild populations are necessary to strengthen the legal trade of orchids. [ABSTRACT FROM AUTHOR]

A letter to the editor is presented in response to the dwindling botany courses at academic institutions and the need for botanists to devise new approaches to teaching coming generations regarding the importance of plants.

This study evaluated the operation of a hybrid anaerobic reactor fed with algal biomass cultivated in effluent from the brewery industry. Three stages of operation were distinguished during the 72 days of semi-continuous functioning of the reactor: Stage 1 (S1), in which algal biomass was used as substrate; Stage 2 (S2), in which 10% (v/v) of the algal biomass was substituted by olive mill wastewater (OMW); and Stage 3 (S3), in which algal biomass was heat pre-treated. During S1, a loss of solids was observed, with an increment of organic matter in the outlet. The substitution of 10% of the volume of algal biomass by OMW tripled the methane productivity obtained in the previous stage by digestion of pure algal biomass. Heat pre-treatment was not efficient in rupturing the cell wall, and consequently did not have any effect on the increase in biogas production. The complementarity of substrates in the assessed conditions led to better results than the pre-treatment of the algal biomass. [ABSTRACT FROM AUTHOR]

Hybridization has the potential to contribute to phenotypic and genetic variation and can be a major evolutionary mechanism. However, when hybridization is extensive it can also lead to the blurring of species boundaries and the emergence of cryptic species (i.e., two or more species not distinguishable morphologically). In this study, we address this hypothesis in Epidendrum, the largest Neotropical genus of orchids where hybridization is apparently so common that it may explain the high levels of morphological diversity found. Nonetheless, this hypothesis is mostly based on the intermediacy of morphological characters and intermediacy by itself is not a proof of hybridization. Therefore, in this study, we first assessed the existence of hybrids using cpDNA and AFLP data gathered from a large-scale sampling comprising 1038 plants of three species of Epidendrum (E. calanthum, E. cochlidium and E. schistochilum). Subsequently, a Bayesian assignment of individuals into different genetic classes (pure species, F1, F2 or backcross generations) revealed that hybrid genotypes were prevalent in all sympatric populations. In most cases, parental species were not assigned as pure individuals, rather consisting in backcrossed genotypes or F1 hybrids. We also found that reproductive barriers are apparently very weak in Epidendrum because the three species largely overlapped in their flowering periods and interspecific crosses always produced viable seeds. Further, hybridization contributed to enhance floral variability, genome size and reproductive success since we found that these traits were always higher in hybrid classes (F1, F2 and backcrosses) than in pure parental species, and offer an explanation for the blurring of species boundaries in this genus of orchids. We hypothesize that these natural hybrids possess an evolutionary advantage, which may explain the high rates of cryptic species observed in this genus. [ABSTRACT FROM AUTHOR]

We present the first phylogenomic analysis of relationships among all ten families of Liliales, based on 75 plastid genes from 35 species in 29 genera, and 97 additional plastomes stratified across angiosperm lineages. We used a supermatrix approach to extend our analysis to 58 of 64 genera of Liliales, and calibrated the resulting phylogeny against 17 fossil dates to produce a new timeline for monocot evolution. Liliales diverged from other monocots 124 Mya and began splitting into separate families 113 Mya. Our data support an Australian origin for Liliales, with close relationships between three pairs of lineages (Corsiaceae/Campynemataceae, Philesiaceae/Ripogonaceae, tribes Alstroemerieae/Luzuriageae) in South America and Australia or New Zealand reflecting teleconnections of these areas via Antarctica. Long-distance dispersal ( LDD) across the Pacific and Tasman Sea led to re-invasion of New Zealand by two lineages ( Luzuriaga, Ripogonum); LDD allowed Campynemanthe to colonize New Caledonia after its submergence until 37 Mya. LDD permitted Colchicaceae to invade East Asia and Africa from Australia, and re-invade Africa from Australia. Periodic desert greening permitted Gloriosa and Iphigenia to colonize Southeast Asia overland from Africa, and Androcymbium-Colchicum to invade the Mediterranean from South Africa. Melanthiaceae and Liliaceae crossed the Bering land-bridge several times from the Miocene to the Pleistocene. [ABSTRACT FROM AUTHOR]

Dengue viruses (DENVs) are mosquito-borne flaviviruses causing millions of human infections each year and pose a challenge for public health systems worldwide. Aedes aegypti is the principal vector species transmitting DENVs to humans. Controlling Ae. aegypti is difficult due to the abundance of breeding sites and increasing insecticide resistance in the vector populations. Developing new vector control strategies is critical for decreasing the disease burden. One potential approach is genetically replacing Ae. aegypti populations with vector populations highly resistant to DENV transmission. Here, we focus on an alternative strategy for generating dengue 2 virus (DENV-2) resistance in genetically-modified Ae. aegypti in which the mosquitoes express an inactive form of Michelob_x (Mx), an antagonist of the Inhibitor of Apoptosis (IAP), to induce apoptosis in those cells in which actively replicating DENV-2 is present. The inactive form of Mx was flanked by the RRRRSAG cleavage motif, which was recognized by the NS2B/NS3 protease of the infecting DENV-2 thereby releasing and activating Mx which then induced apoptosis. Our transgenic strain exhibited a significantly higher mortality rate than the non-transgenic control when infected with DENV-2. We also transfected a DNA construct containing inactive Mx fused to eGFP into C6/36 mosquito cells and indirectly observed Mx activation on days 3 and 6 post-DENV-2 infections. There were clear signs that the viral NS2B/NS3 protease cleaved the transgene, thereby releasing Mx protein into the cytoplasm, as was confirmed by the detection of eGFP expression in infected cells. The present study represents proof of the concept that virus infection can be used to induce apoptosis in infected mosquito cells. [ABSTRACT FROM AUTHOR]

The genus Limonium Mill. (sea lavenders) includes species with sexual and apomixis reproductive strategies, although the genes involved in these processes are unknown. To explore the mechanisms beyond these reproduction modes, transcriptome profiling of sexual, male sterile, and facultative apomictic species was carried out using ovules from different developmental stages. In total, 15,166 unigenes were found to be differentially expressed with apomictic vs. sexual reproduction, of which 4275 were uniquely annotated using an Arabidopsis thaliana database, with different regulations according to each stage and/or species compared. Gene ontology (GO) enrichment analysis indicated that genes related to tubulin, actin, the ubiquitin degradation process, reactive oxygen species scavenging, hormone signaling such as the ethylene signaling pathway and gibberellic acid-dependent signal, and transcription factors were found among differentially expressed genes (DEGs) between apomictic and sexual plants. We found that 24% of uniquely annotated DEGs were likely to be implicated in flower development, male sterility, pollen formation, pollen-stigma interactions, and pollen tube formation. The present study identifies candidate genes that are highly associated with distinct reproductive modes and sheds light on the molecular mechanisms of apomixis expression in Limonium sp. [ABSTRACT FROM AUTHOR]