The evolution of ephemeral flora in Xinjiang, China: insights from plastid phylogenomic analyses of Brassicaceae.

BACKGROUND: The ephemeral flora of northern Xinjiang, China, plays an important role in the desert ecosystems. However, the evolutionary history of this flora remains unclear. To gain new insights into its origin and evolutionary dynamics, we comprehensively sampled ephemeral plants of Brassicaceae, one of the essential plant groups of the ephemeral flora. RESULTS: We reconstructed a phylogenetic tree using plastid genomes and estimated their divergence times. Our results indicate that ephemeral species began to colonize the arid areas in north Xinjiang during the Early Miocene and there was a greater dispersal of ephemeral species from the surrounding areas into the ephemeral community of north Xinjiang during the Middle and Late Miocene, in contrast to the Early Miocene or Pliocene periods. CONCLUSIONS: Our findings, together with previous studies, suggest that the ephemeral flora originated in the Early Miocene, and species assembly became rapid from the Middle Miocene onwards, possibly attributable to global climate changes and regional geological events.

Inferring the evolution of reproductive isolation in a lineage of fossil threespine stickleback, Gasterosteus doryssus.

Darwin attributed the absence of species transitions in the fossil record to his hypothesis that speciation occurs within isolated habitat patches too geographically restricted to be captured by fossil sequences. Mayr's peripatric speciation model added that such speciation would be rapid, further explaining missing evidence of diversification. Indeed, Eldredge and Gould's original punctuated equilibrium model combined Darwin's conjecture, Mayr's model and 124 years of unsuccessfully sampling the fossil record for transitions. Observing such divergence, however, could illustrate the tempo and mode of evolution during early speciation. Here, we investigate peripatric divergence in a Miocene stickleback fish, Gasterosteus doryssus. This lineage appeared and, over approximately 8000 generations, evolved significant reduction of 12 of 16 traits related to armour, swimming and diet, relative to its ancestral population. This was greater morphological divergence than we observed between reproductively isolated, benthic-limnetic ecotypes of extant Gasterosteus aculeatus. Therefore, we infer that reproductive isolation was evolving. However, local extinction of G. doryssus lineages shows how young, isolated, speciating populations often disappear, supporting Darwin's explanation for missing evidence and revealing a mechanism behind morphological stasis. Extinction may also account for limited sustained divergence within the stickleback species complex and help reconcile speciation rate variation observed across time scales.

Impacts of stream drying depend on stream network size and location of drying.

Stream drying is increasing globally, with widespread impacts on stream ecosystems. Here, we investigated how the impacts of drying on stream ecosystem connectivity might depend on stream network size and the location of drying within the stream network. Using 11 stream networks from across the United States, we simulated drying scenarios in which we varied the location and spatial extent of drying. We found that the rate of connectivity loss varied with stream network size, such that larger stream networks lost connectivity more rapidly than smaller stream networks. We also found that the rate of connectivity loss varied with the location of drying. When drying occurred in the mainstem, even small amounts of drying resulted in rapid losses in ecosystem connectivity. When drying occurred in headwater reaches, small amounts of drying had little impact on connectivity. Beyond a certain threshold, however, connectivity declined rapidly with further increases in drying. Given the increasing stream drying worldwide, our findings underscore the need for managers to be particularly vigilant about fragmentation when managing at large spatial scales and when stream drying occurs in mainstem reaches.

Inconsistent Regulatory Mapping Quietly Threatens Rivers and Streams.

Even the most stringent environmental law cannot protect a river if its tributaries remain exposed to pollution and other threats upstream. Excluding a subset of watercourses from legal protection therefore threatens to alter freshwater ecosystems across entire river networks and the services they provide, such as drinking water and flood regulation. Considerable attention has been devoted to defining the scope of environmental laws protecting watercourses. Yet how these definitions are implemented through regulatory mapping, the cartography of waterbodies that legally qualify as watercourses and are thus protected, has not been examined outside of the United States. Here, we demonstrate the consequences of regulatory mapping on the extent of river networks that are protected, using France as a case study. By assembling the first map of France's watercourses protected under the Water Law, we estimate that a quarter of previously mapped hydrographic segments were excluded from protection and found stark geographical variations in the extent of protected ecosystems. Headwater and nonperennial segments are disproportionately excluded by 28% compared to their prevalence (67%) in the overall hydrographic network, with potentially far-reaching implications for biodiversity and people. We expect regulatory frameworks in most countries to be equally susceptible to local interpretation of legal definitions.

Forgotten fish habitats: Developing a conceptual framework for evaluating intermittent waterways and flood channels as native fish habitats: Insights from two freshwater catchments in Australia.

The ability to correctly identify Potential Key Fish Habitats (PKFH) before undertaking development assessments or stream restoration projects is a critical step for determining appropriate management interventions and offset strategies to minimize damage to all life stages of fish. However, there are no rapid, low-cost tools that can be universally utilised to identify PKFH in perennial streams and ephemeral streams in particular. We propose a conceptual framework for developing a rapid field appraisal based on a range of physical fish habitat-supporting features. Four steps are involved in developing the framework: 1) understand fish habitat features using published literature and sub-set model selection using actual data, 2) collect field data, 3) develop the conceptual framework, and 4) classify sites. We tested eight fish habitat-supporting features in the assessment and proposed a decision tree that guided the rapid appraisals at 104 ephemeral sites in southeast Australia. Out of 104 sites, 86% were identified as PKFH, 10% as likely key fish habitats and 4% as unlikely key fish habitats. The rapid field appraisal is cost-effective, simple to implement and does not require expert knowledge of fish habitats. Despite some limitations in the conceptual framework, this appraisal has great potential as a screening technique for PKFH in freshwater streams, particularly ephemeral streams in Australia and other overseas jurisdictions.

Loss of functionally important and regionally endemic species from streams forced into intermittency by global warming.

Climate change is altering hydrological cycles globally, and in Mediterranean (med-) climate regions it is causing the drying of river flow regimes, including the loss of perennial flows. Water regime exerts a strong influence over stream assemblages, which have developed over geological timeframes with the extant flow regime. Consequently, sudden drying in formerly perennial streams is expected to have large, negative impacts on stream fauna. We compared contemporary (2016/17) macroinvertebrate assemblages of formerly perennial streams that became intermittently flowing (since the early 2000s) to assemblages recorded in the same streams by a study conducted pre-drying (1981/82) in the med-climate region of southwestern Australia (the Wungong Brook catchment, SWA), using a multiple before-after, control-impact design. Assemblage composition in the stream reaches that remained perennial changed very little between the studies. In contrast, recent intermittency had a profound effect on species composition in streams impacted by drying, including the extirpation of nearly all Gondwanan relictual insect species. New species arriving at intermittent streams tended to be widespread, resilient species including desert-adapted taxa. Intermittent streams also had distinct species assemblages, due in part to differences in their hydroperiods, allowing the establishment of distinct winter and summer assemblages in streams with longer-lived pools. The remaining perennial stream is the only refuge for ancient Gondwanan relict species and the only place in the Wungong Brook catchment where many of these species still persist. The fauna of SWA upland streams is becoming homogenised with that of the wider Western Australian landscape, as drought-tolerant, widespread species replace local endemics. Flow regime drying caused large, in situ alterations to stream assemblage composition and demonstrates the threat posed to relictual stream faunas in regions where climates are drying.

Archaeal and Bacterial Diversity and Distribution Patterns in Mediterranean-Climate Vernal Pools of Mexico and the Western USA.

Biogeographic patterns in microorganisms are poorly understood, despite the importance of microbial communities for a range of ecosystem processes. Our knowledge of microbial ecology and biogeography is particularly deficient in rare and threatened ecosystems. We tested for three ecological patterns in microbial community composition within ephemeral wetlands-vernal pools-located across Baja California (Mexico) and California (USA): (1) habitat filtering; (2) a latitudinal diversity gradient; and (3) distance decay in community composition. Paired water and soil samples were collected along a latitudinal transect of vernal pools, and bacterial and archaeal communities were characterized using 16S rDNA sequencing. We identified two main microbial communities, with one community present in the soil matrix that included archaeal and bacterial soil taxa, and another community present in the overlying water that was dominated by common freshwater bacterial taxa. Aquatic microbial communities were more diverse in the north, and displayed a significant but inverted latitudinal diversity pattern. Aquatic communities also exhibited a significant distance-decay pattern, with geographic proximity, and precipitation explaining part of the community variation. Collectively these results indicate greater sensitivity to spatial and environmental variation in vernal pool aquatic microbial communities than in soil microbial communities. We conclude that vernal pool aquatic microbial communities can display distribution patterns similar to those exhibited by larger organisms, but differ in some key aspects, such as the latitudinal gradient in diversity.

Effects of the Desiccation Duration on the Dynamic Responses of Biofilm Metabolic Activities to Rewetting.

Global climate changes have increased the duration and frequency of river flow interruption, affecting the physical and community structure of benthic biofilms. However, the dynamic responses of biofilm metabolism during the dry-wet transition remain poorly understood. Herein, the dynamic changes in biofilm metabolic activities were investigated through mesocosm experiments under short-term (25 day) and long-term drought (90 day), followed by a 20 day rewetting. The biofilm ecosystem metabolism, as measured by gross primary production and community respiration, was significantly inhibited and turned heterotrophic during the desiccation phase and then recovered, becoming autotrophic during the rewetting period regardless of the desiccation periods due to the high resilience of the autotrophic community. However, long-term drought decreased the recovery rate of the ecosystem metabolism and also caused irreparable damage to the biofilm carbon metabolism, measured using Biolog Eco Plates. Specifically, the recovery of the total carbon metabolic activity is related to the specific carbon source utilized by biofilm microorganisms, such as polymers, carbohydrates, and carboxylic acids. However, the divergent changes of amino acids caused the failure of the total carbon metabolism in long-term drought treatments to recover to the control level even after 20 days of rewetting. This research provides direct evidence that the increased duration of non-flow periods affects biofilm-mediated carbon biogeochemical processes.

Comparison of network connectivity and environmental driving factors of root-associated fungal communities of desert ephemeral plants in two habitat soils.

Root-associated microorganisms regulate plant growth and development, and their distribution is likely influenced by habitat conditions. In this study, the responses of rhizosphere and root-endophytic fungi of dominant ephemeral plants to aeolian soil (AS) and grey desert soil (DS) in the Gurbantünggüt Desert were analyzed using high-throughput sequencing. This was done to understand the adaptation strategies of this vegetation in typical habitat soils from a microbial perspective. We found that the diversity of root-associated fungi of ephemeral plants differed in the two habitat soils. The diversity of rhizosphere fungi was relatively low in AS compared to DS, whereas the diversity of root-endophytic fungi was higher in AS. The community structure of root-associated fungi and relative abundances of some dominant taxa differed between the two soils. A co-occurrence network showed that the degree of coupling and interaction between root-associated fungal taxa were closer in AS than in DS and that most of the fungal taxa were cooperative in the two habitat soils. Additionally, the network properties of the root-endophytic fungi were apparent different between the two soils. Environmental factors, including electrical conductivity, soil organic carbon, carbon/nitrogen, and carbon/phosphorus ratios, were found to be key factors affecting rhizosphere fungi in DS, whereas soil available phosphorus was the main factor in AS. Several factors affect the root-endophytic fungal community and are more influential in DS than in AS. Overall, the root-associated fungal communities of ephemeral plants had different adaptation strategies to the two soils: increasing the diversity of rhizosphere fungi and their relationship with environmental factors in DS, and increasing the diversity and network relationships of root-endophytic fungi in AS. These findings provide insight into the assemblage of ephemeral plant root-associated microbial communities and the underlying environmental factors, which allows for a deeper understanding of how to construct an artificial core root microbiota to promote plant growth and resistance.

An experimental investigation of the erosion process and morphological characteristics of the tile-back type slope with non-uniform slopes based on a 3D reconstruction method.

Tile-back type slopes comprise ephemeral gullies (EGs) and hillslopes; they are a unique and widely distributed micro-landform in the Loess Plateau region of China. Gully erosion from these landforms is a serious issue, but the micro-landform makes the erosion process and its estimation complex. Quantifying soil erosion processes and their distribution characteristics at different positions on tile-back type slopes will provide a clearer picture for ecological restoration to control further soil degradation. This study investigated the erosion process of tile-back type slope with non-uniform slopes using a 3D photo-reconstruction method during eight successive simulated rainfall events. The results showed that EG erosion began with a chain of intermittent headcuts. When the accumulated rainfall reached 76 mm, serious collapses dramatically increased the amount of sediment by 216% after the first rainfall (cumulative rainfall was about 15 mm). We quantified the sediment contribution of EG erosion (46.20%), rill erosion (35.62%), and inter-rill erosion (18.18%) to total soil loss. The erosion area of the steep slope section and extremely steep slope section accounted for 33.26% and 66.74% of the total erosion area, respectively. Moreover, sediment amounts significantly correlated with morphological parameters, particularly the amount of EG erosion and maximum gully depth, with a correlation coefficient of 0.98. Cumulative gully length and erosion area had the greatest effect on rill erosion, with a correlation coefficient of 0.97. These results provide insight into the qualitative and quantitative understanding of EG erosion process on Loess Plateau of China and an important reference for the rational arrangement of EG control measures.

Changes in species diversity along the ephemeral streams in eastern Mediterranean Mountainous Ecosystem, a case study for Ahir Mountain (Kahramanmaras-Türkiye).

Ephemeral streams are important habitats that support biodiversity, especially in mountainous ecosystems in arid and semi-arid regions, and are among those that will be most affected by global climate change. This study aimed to examine the trends in plant species diversity of ephemeral stream beds and their relationship with environmental variables (aspect, elevation, soil) in the eastern Mediterranean region.For this purpose, 40 sample plots were applied in the valley cross sections of the ephemeral stream beds (valley bottom, side and ridge) on two main slopes (north and south) in 2019. The abundance values of plant species were determined and the number of individuals in each sample plot was counted. In addition, soil samples were taken at a depth of 0-30 cm and analyzed. With the data obtained, the importance value index of all species was calculated, and the diversity and species richness of the sample plots were determined.A total of 130 plant taxa were identified in the study area. The most dominant species were Astragalus kurdicus (IVI value: 25.97), Helichrysum plicatum (21.94), Taeniatherum caput-medusae subsp. crinitum (15.51), Hordeum bulbosum (15.33), Bromus erectus (15.32), and Minuartia juniperina (14.14). Both plant richness and endemism rate showed a significant relationship with increasing elevation. Plant diversity values of south-facing slopes were higher (2.23) than those of north-facing slopes (2.21). In addition, we found that phosphorus is an important factor in the distribution of plant species along the ephemeral stream beds, and as the amount of phosphorus increases, species richness and diversity also increase.

Molecular characterization and phylogenetic analysis of bovine ephemeral fever viruses in Khuzestan province of Iran in 2018 and 2020.

BACKGROUND: Bovine ephemeral fever (BEF) is an arthropod-borne viral disease caused by the BEF virus (BEFV). This single-stranded RNA virus that affects cattle and water buffalo is endemic in tropical and subtropical regions including Iran. While BEF is a major disease of cattle in Iran, information regarding its agent, molecular characterization, and circulating viruses are highly limited. The current study aimed to, firstly, determine the genetic and antigenic characteristics of BEFV strains in Khuzestan province in Southwest of Iran in 2018 and 2020 and, secondly, to compare them with strains obtained from other areas. RESULTS: By phylogenetic analysis based on the Glycoprotein gene, BEFV strains were divided into four clusters of Middle East, East Asia, South Africa, and Australia; in which the 2018 and 2020 Iranian BEFV strains were grouped in the Middle East cluster with the Turkish, Indian, and Israeli strains. Depending on the chronology and geographical area, the outbreaks of Turkey (2020), Iran (2018 and 2020), and India (2018 and 2019) are proposed to be related. These BEFVs had the highest identity matrix and the lowest evolutionary distance among the studied strains. Multiple sequence alignment of G1, G2, and G3 antigenic sites showed that these neutralizing epitopes are highly conserved among the strains of the Middle East cluster; however, the strains previously identified in Iran differed in three amino acids placed in G1 and G2 epitopes. CONCLUSION: The findings revealed that BEFVs circulating in the Middle East are closely related phylogenetically and geographically. They also have similar antigenic structures; therefore, developing a vaccine based on these strains can be effective for controlling BEF in the Middle East.

Isolation and Functional Analysis of EPHEMERAL1-LIKE (EPH1L) Genes Involved in Flower Senescence in Cultivated Japanese Gentians.

The elongation of flower longevity increases the commercial value of ornamental plants, and various genes have been identified as influencing flower senescence. Recently, EPHEMERAL1 (EPH1), encoding a NAC-type transcription factor, was identified in Japanese morning glory as a gene that promotes flower senescence. Here we attempted to identify an EPH1 homolog gene from cultivated Japanese gentians and characterized the same with regard to its flower senescence. Two EPH1-LIKE genes (EPH1La and EPH1Lb), considered as alleles, were isolated from a gentian cultivar (Gentiana scabra × G. triflora). Phylogenetic analyses revealed that EPH1L belongs to the NAM subfamily. The transcript levels of EPH1L increased along with its senescence in the field-grown flowers. Under dark-induced senescence conditions, the gentian-detached flowers showed the peak transcription level of EPH1L earlier than that of SAG12, a senescence marker gene, suggesting the involvement of EPH1L in flower senescence. To reveal the EPH1L function, we produced eph1l-knockout mutant lines using the CRISPR/Cas9 system. When the flower longevity was evaluated using the detached flowers as described above, improved longevity was recorded in all genome-edited lines, with delayed induction of SAG12 transcription. The degradation analysis of genomic DNA matched the elongation of flower longevity, cumulatively indicating the involvement of EPH1L in the regulation of flower senescence in gentians.

Responses and comprehensive evaluation of growth characteristics of ephemeral plants in the desert-oasis ecotone to soil types.

The ecological environment of the Gurbantünggüt desert-oasis ecotone is extremely fragile. Ephemeral plants are an important part of the ecosystem and play an essential role in maintaining the ecological stability of the ecotone. However, few studies have focused on the growth, soil quality and system sustainability of ephemeral plants in different soils. This study was based on two typical soil types (grey desert soil, GS; aeolian soil, AS) in the aforementioned ecotone, considered four ephemeral plants (Tetracme recurvata, TR; Tetracme contorta, TC; Malcolmia scorpioides, MS; Isatis violascens, IV) as the research object, analysed plant characteristics and soil properties, and comprehensively evaluated the ephemeral plant system by analysing the soil quality index (SQI) and sustainability index (SI). The results showed that there were significant differences in biomass and nutrient accumulation between different ephemeral plants, which were significantly affected by soil types. In the two examined soils (GS and AS), the contents of nutrients and microbial carbon (MBC) and nitrogen (MBN) in the rhizosphere soil were higher than those in the bare soil (BS), and there were significant differences among different species. The key soil factors related to total biomass in GS and AS were also different. The SQI of ephemeral plants was significantly higher than that of the BS, and varied with soil types and plant species. The species with the highest SQI of the key factor data set in GS and AS were IV and TR, respectively. The SI analysis indicated that IV in GS and MS and IV in AS were sustainable, and the plant properties can be better used to assess the sustainability of ephemeral plant systems. In conclusion, ephemeral plants improved the soil quality and system sustainability of the study ecotone. Further, the growth of ephemeral plant and rhizosphere soil properties vary with plant species and soil types; thus, selecting suitable species for large-scale planting in different soil types is of great significance for improving the ecological stability of the ecotone.

Genome-wide identification, phylogenetic and expression of MAPKKK gene family in Arabidopsis pumila.

Mitogen-activated protein kinase kinase kinases (MAPKKKs) are important components of the MAPK cascade and play crucial roles in development and stress responses. Arabidopsis pumila is an ephemeral Brassicaceae plant growing in Xinjiang desert regions, which possesses salt tolerance. To explore the evolution and function of the MAPKKK gene family in A. pumila, 143 ApMAPKKK genes were identified from A. pumila genome by genome-wide analysis, which were categorized into three subfamilies: ZIK (20), MEKK (36) and RAF (87). There existed 74 and 72 colinear genes between A. thaliana, A. lyrata and A. pumila, respectively, indicating that this gene family expanded obviously in A. pumila genome. Evolutionary analysis revealed that there were 64 duplicated gene pairs with Ka/Ks less than 1, and purifying selection was dominant. RNA-seq data were used to analyze the expression characteristics of ApMAPKKK genes in response to salt stress and in different tissues. The results showed that most ApMAPKKK genes were up-regulated under 250 mmol/L NaCl stress. For example, ApMAPKKK18-1/2 and ApMAPKKK17-1/2 were substantially up-regulated. Tissue expression profiles showed that ApMAPKKK mainly presented six expression patterns. Some duplicated genes were differentially expressed in response to salt stress and in different tissues. These results lay a foundation for further understanding the complex mechanism of MAPKKK gene family transduction pathway in response to abiotic stresses in A. pumila.

Genomic analysis of novel Yarrowia-like yeast symbionts associated with the carrion-feeding burying beetle Nicrophorus vespilloides.

BACKGROUND: Mutualistic interactions with microbes can help insects adapt to extreme environments and unusual diets. An intriguing example is the burying beetle Nicrophorus vespilloides, which feeds and reproduces on small vertebrate carcasses. Its fungal microbiome is dominated by yeasts that potentially facilitate carcass utilization by producing digestive enzymes, eliminating cadaver-associated toxic volatiles (that would otherwise attract competitors), and releasing antimicrobials to sanitize the microenvironment. Some of these yeasts are closely related to the biotechnologically important species Yarrowia lipolytica. RESULTS: To investigate the roles of these Yarrowia-like yeast (YLY) strains in more detail, we selected five strains from two different phylogenetic clades for third-generation sequencing and genome analysis. The first clade, represented by strain B02, has a 20-Mb genome containing ~ 6400 predicted protein-coding genes. The second clade, represented by strain C11, has a 25-Mb genome containing ~ 6300 predicted protein-coding genes, and extensive intraspecific variability within the ITS-D1/D2 rDNA region commonly used for species assignments. Phenotypic microarray analysis revealed that both YLY strains were able to utilize a diverse range of carbon and nitrogen sources (including microbial metabolites associated with putrefaction), and can grow in environments with extreme pH and salt concentrations. CONCLUSIONS: The genomic characterization of five yeast strains isolated from N. vespilloides resulted in the identification of strains potentially representing new YLY species. Given their abundance in the beetle hindgut, and dominant growth on beetle-prepared carcasses, the analysis of these strains has revealed the genetic basis of a potential symbiotic relationship between yeasts and burying beetles that facilitates carcass digestion and preservation.

Life-history traits are poor predictors of species responses to flow regime change in headwater streams.

Recent climate change is altering the timing, duration and volume of river and stream flows globally, and in many regions, perennially flowing rivers and streams are drying and switching to intermittent flows. Profound impacts on aquatic biota are becoming apparent, due in part to the strong influence of flow regime on the evolution of life history. We made predictions of life-history responses for 13 common aquatic invertebrate species (four caddisflies, five mayflies, two stoneflies, a dragonfly and an amphipod), to recent flow regime change in Australian mediterranean climate streams, based on historic studies in the same streams. Size distributions, phenology, voltinism and synchrony were compared, revealing five main responses. More than half of the species were restricted to perennially flowing streams and were absent from those that had switched to intermittent flows (including all four caddisfly species). These formerly common species are at risk of extinction as climate change progresses. Two mayfly species had divergent responses in voltinism and synchrony, and one relied on drought micro-refuges to persist. One stonefly species changed development timing to suit the new flow regime, and the amphipod species retreated to subterranean refuges. Two formerly common species were not detected at all during 2016-2017. In addition, a new mayfly species and a caddisfly species proliferated under new flow regimes, because they had life histories suited to brief hydroperiods. Importantly, previous life history rarely predicted species' actual responses to climate-driven flow regime change, raising doubts about the veracity of predictions based on species traits. This is because a species' potential for flexible phenology or growth rate is not necessarily indicated by life-history traits.

Expression and immunogenicity of secreted forms of bovine ephemeral fever virus glycoproteins applied to subunit vaccine development.

AIMS: Vaccines for bovine ephemeral fever virus (BEFV) are available but are difficult to produce, expensive or suffer from genetic instability. Therefore, we designed constructs encoding C-terminally truncated forms (transmembrane anchoring region deleted) of glycoproteins G and GNS such that they were secreted from the cell into the media to achieve high-level antigen expression, correct glycosylation pattern and enable further simple purification with the V5 epitope tag. METHODS AND RESULTS: In this study, synthetic biology was employed to create membrane-bound and secreted forms of G and GNS glycoprotein. Mammalian cell culture was employed as an antigen expression platform, and the secreted forms of G and GNS protein were easily purified from media using a highly effective, single-step method. The V5 epitope tag was genetically fused to the C-termini of the proteins, enabling detection of the antigen through immunoblotting and immunomicroscopy. Our data demonstrated that the C-terminally truncated form of the G glycoprotein was efficiently secreted from cells into the cell media. Moreover the immunogenicity was confirmed in mice test. CONCLUSIONS: The immuno-dot blots showed that the truncated G glycoprotein was present in the total cell extract, and was clearly secreted into the media, consistent with the western blotting data and live-cell images. Our strategy presented the expression of secreted, epitope-tagged, forms of the BEFV glycoproteins such that appropriately glycosylated forms of BEFV G protein was secreted from the BHK-21 cells. This indicates that high-level expression of secreted G glycoprotein is a feasible strategy for large-scale production of vaccines and improving vaccine efficacy. SIGNIFICANCE AND IMPACT OF THE STUDY: The antigen expression strategy designed in this study can produce high-quality recombinant protein and reduce the amount of antigen used in the vaccine.

Succession of Dung-Inhabiting Beetles and Flies Reflects the Succession of Dung-Emitted Volatile Compounds.

Chemical cues, such as volatile organic compounds (VOCs), are often essential for insects to locate food. Relative to the volume of studies on the role of VOCs in insect-plant relationships, the role of VOCs emitted by dung and carrion in mediating the behavior of insect decomposers is understudied. Such relationships may provide a mechanistic understanding of the temporal axis of community assembly processes in decomposing insect communities. We focused on the temporal succession of volatiles released by cow dung pats and the potential influence on dung-inhabiting insects. Using gas chromatography/mass spectrometry we identified and quantified VOCs released from dung 1-h, and 1, 2 3, 5, and 7 d-old. We then related changes in VOCs to successional patterns of dung-inhabiting beetles and flies. We detected 54 VOCs which could be assigned to two successional groups, with chemical turnover in dung changing around day 2. The early successional group consisted primarily of aliphatic alcohols and phenols, and the late one of aliphatic esters, nitrogen- and sulfur-bearing compounds. Flies were predominately associated with the early successional group, mainly with 1-butanol. Beetles were associated predominately with the late-successional group, mainly with dimethyl trisulfide. This association between insect and chemical successional patterns supports the idea that habitat filtering drives the community assembly of dung-inhabiting insects on an aging resource. Moreover, the affinity of both insect groups to specific VOC groups provides a mechanistic explanation for the predictability of successional patterns found in dung-inhabiting insect communities.

The paradoxical performance by different species on the ephemeral reward task.

The ephemeral reward task consists of giving an animal a choice between two distinctive stimuli, A and B (e.g., black and white), on each of which is placed a bit of food. If the animal chooses the food on A, it gets that reinforcer, but the other stimulus, B, is removed, and the trial is over. If it chooses the food on B, however, it gets that food and the stimulus A remains, so it can have that food as well. Thus, choice of stimulus B gives the animal two reinforcers rather than one. Wrasse (cleaner fish) easily learn to choose optimally, whereas surprisingly, most non-human primates do not. Parrots, however, appear to learn this task as easily as the fish. To test the hypothesis that animals that choose with their mouth can learn it, we tested pigeons and found that they show no evidence of optimal learning with this task (with either the manual presentation of the stimuli or the operant presentation of the stimuli). Similarly, rats show no evidence of optimal learning. However, if a 20-s delay (fixed-interval schedule) is inserted between stimulus choice and reinforcement, both pigeons and rats learn to perform optimally. The ephemeral reward task appears to promote impulsive choice in several species, but with this task (and others), inserting a delay between choice and reinforcement promotes more careful choice, leading to optimal performance.