Transcriptional regulation and functional properties of <Emphasis Type="Italic">Arabidopsis</Emphasis> Pht1;4, a high affinity transporter contributing greatly to phosphate uptake in phosphate deprived plants

Phosphate mobilization into the plant is a complex process requiring numerous transporters for absorption and translocation of this major nutrient. In the genome of Arabidopsis thaliana, nine closely related high affinity phosphate transporters have been identified but their specific roles remain unclear. Here we report the molecular, histological and physiological characterization of Arabidopsis pht1;4 high affinity phosphate transporter mutants. Using GUS-gene trap and in situ hybridization, Pht1;4 was found mainly expressed in inorganic phosphate (Pi) limiting medium in roots, primarily in the epidermis, the cortex and the root cap. In addition to this, expression was also observed at the lateral root branch points on the primary root and in the stele of lateral roots, suggesting a role of Pht1;4 in phosphate absorption and translocation from the growth medium to the different parts of the plant. Pi-starved pht1;4 plantlets exhibited a strong reduction of phosphate uptake capacity (40). This phenotype appears only related to the pht1;4 mutation as there were no obvious changes in the expression of other Pht1 family members in the mutants background. However, after 10 days of growth on phosphate deficient or sufficient medium, the Pi content in the mutants was not significantly different from that of the corresponding wild type controls. Furthermore, the mutants did not display any obvious growth defects or visible phenotypes when grown on a low phosphate containing medium. The work described here offers a first step in the complex genetic dissection of the phosphate transport system in planta.     

Direct assessment of queen quality and lack of worker suppression in a paper wasp

Assessing a conspecific's potential is often crucial to increase one's fitness, e.g. in female choice, contests with rivals or reproductive conflicts in animal societies. In the latter, helpers benefit from accurately assessing the fertility of the breeder as an indication of inclusive fitness. There is evidence that this can be achieved using chemical correlates of reproductive activity. Here, we show that queen quality can be assessed by directly monitoring her reproductive output. In the paper wasp Polistes dominulus, we mimicked a decrease in queen fertility by regularly removing brood. This triggered ovarian development and egg-laying by many workers, which strongly suggests that brood abundance is a reliable cue of queen quality. Brood abundance can be monitored when workers perform regular brood care in small size societies where each brood element is kept in a separate cell. Our results also show that although the queen was not manipulated, and thus remained healthy and fully fertile, she did not control worker egg-laying. Nevertheless, when workers laid eggs, the queen secured a near reproductive monopoly by selectively destroying these eggs, a mechanism known as 'queen policing'. By contrast, workers destroyed comparatively few queen-laid eggs, but did destroy each other's eggs.     

Factors controlling soil macrofauna spatial pattern in a pure beech and a mixed beech-hornbeam forest

The soil macrofauna of a pure beech (PS) and a mixed beech-hornbeam (MS) stand was recorded using a sample design spatially explicit at stand level. Humic epipedon morphological and chemical properties, relative irradiance, soil bulk density, and the specific composition of the litter in MS were also investigated. The taxonomic diversity is nearly similar on both sites, but the average by sample is greater under PS. The main factors controlling soil macrofauna spatial variability were: litter quality under MS and humus form activity under PS. These results suggest a distal control (i.e. external to humic epipedon) of macrofauna spatial variability when the litter quality is variable and a proximal control (i.e. internal to humic epipedon) when it is uniform at the spatial scale of the study.     

Modulation of carbon and nitrogen metabolism, and of nitrate reductase, in untransformed and transformed Nicotiana plumbaginifolia during CO2 enrichment of plants grown in pots and in hydroponic culture

Transformed plants of Nicotiana plumbaginifolia Viv. constitutively expressing nitrate reductase (35S-NR) or β-glucuronidase (35S-GUS) and untransformed controls were grown for two weeks in a CO₂-enriched atmosphere. Whereas CO₂ enrichment (1000 μl · l⁻¹) resulted in an increase in the carbon (C) to nitrogen (N) ratio of both the tobacco lines grown in pots with vermiculite, the C/N ratio was only slightly modified when plants were grown in hydroponic culture in high CO₂ compared to those grown in air. Constitutive nitrate reductase (NR) expression per se did not change the C/N ratio of the shoots or roots. Biomass accumulation was similar in both types of plant when hydroponic or pot-grown material, grown in air or high CO₂, were compared. Shoot dry matter accumulation was primarily related to the presence of stored carbohydrate (starch and sucrose) in the leaves. In the pot-grown tobacco, growth at elevated CO₂ levels caused a concomitant decrease in the N content of the leaves involving losses in NO⁻ ₃ and amino acid levels. In contrast, the N content and composition were similar in all plants grown in hydroponic culture. The 35S-NR plants grown in air had higher foliar maximum extractable NR activities and increased glutamine levels (on a chlorophyll or protein basis) than the untransformed controls. These increases were maintained following CO₂ enrichment when the plants were grown in hydroponic culture, suggesting that an increased flux through nitrogen assimilation was possible in the 35S-NR plants. Under CO₂ enrichment the NR activation state in the leaves was similar in all plants. When the 35S-NR plants were grown in pots, however, foliar NR activity and glutamine content fell in the 35S-NR transformants to levels similar to those of the untransformed controls. The differences in NR activity between untransformed and 35S-NR leaves were much less pronounced in the hydroponic than in the pot-grown material but the difference in total extractable NR activity was more marked following CO₂ enrichment. Foliar NR message levels were decreased by CO₂ enrichment in all growth conditions but this was much more pronounced in pot-grown material than in that grown hydroponically. Since β-glucuronidase (GUS) activity and message levels in 35S-GUS plants grown under the same conditions of CO₂ enrichment (to test the effects of CO₂ enrichment on the activity of the 35S promoter) were found to be constant, we conclude that NR message turnover was specifically accelerated in the 35S-NR plants as well as in the untransformed controls as a result of CO₂ enrichment. The molecular and metabolic signals involved in increased NR message and protein turnover are not known but possible effectors include NO₃ ⁻, glutamine and asparagine. We conclude that plants grown in hydroponic culture have greater access to N than those grown in pots. Regardless of the culture method, CO₂ enrichment has a direct effect on NR mRNA stability. Received: 17 October 1996 / Accepted: 11 February 1997     

The immunoglobulin repertoire of the rainbow trout (Oncorhynchus mykiss): definition of nine Igh-V families

An Igh-V library was constructed from the head kidney cytoplasmic RNA of an 8.5-month-old non-immunized rainbow trout, Oncorhynchus mykiss, using the 5 RACE polymerase chain reaction. Six new Igh-V segments were identified, bringing to nine the number of Igh-V families actually defined in that species. A phylogenetic analysis shows that these nine Igh-V families can be classified into three major groups. The first includes the Igh-V1, Igh-V3, Igh-V4, and Igh-V7 families, and is homologous to the human and mouse Group III Igh-V families. The second includes the Igh-V5, Igh-V8, and Igh-V9 families and is more closely related to the Group I and Group II human and mouse Igh-V families. The third group includes the Igh-V2 and Igh-V6 families, which are not closely related to any other vertebrate Igh-V gene. Six Igh-J segments were characterized. They can recombine with Igh-V segments belonging to different families and there is a high level of junctional diversity between the Igh-V and Igh-J segments.The nucleotide sequence data reported in this paper have been submitted to the GenBank nucleotide sequence database and have been assigned the accession numbers L28741 (Igh-V4), L28742 (Igh-V5.1), L28744 (Igh-V6), L28745 (Igh-V7), L28746 (Igh-V8), L28747 (Igh-V9), and L28805 (Igh-V5.2)     

Herbaceous Angiosperms Are Not More Vulnerable to Drought-Induced Embolism Than Angiosperm Trees

The water transport pipeline in herbs is assumed to be more vulnerable to drought than in trees due to the formation of frequent embolisms (gas bubbles), which could be removed by the occurrence of root pressure, especially in grasses. Here, we studied hydraulic failure in herbaceous angiosperms by measuring the pressure inducing 50% loss of hydraulic conductance (P₅₀) in stems of 26 species, mainly European grasses (Poaceae). Our measurements show a large range in P₅₀ from −0.5 to −7.5 MPa, which overlaps with 94% of the woody angiosperm species in a worldwide, published data set and which strongly correlates with an aridity index. Moreover, the P₅₀ values obtained were substantially more negative than the midday water potentials for five grass species monitored throughout the entire growing season, suggesting that embolism formation and repair are not routine and mainly occur under water deficits. These results show that both herbs and trees share the ability to withstand very negative water potentials without considerable embolism formation in their xylem conduits during drought stress. In addition, structure-function trade-offs in grass stems reveal that more resistant species are more lignified, which was confirmed for herbaceous and closely related woody species of the daisy group (Asteraceae). Our findings could imply that herbs with more lignified stems will become more abundant in future grasslands under more frequent and severe droughts, potentially resulting in lower forage digestibility.Terrestrial biomes provide numerous ecosystem services to humans, such as biodiversity refuges, forage supply, carbon sequestration, and associated atmospheric feedback (Bonan, 2008). Drought frequency and severity are predicted to increase across various ecosystems (Dai, 2013), and its impact on the fate of terrestrial biomes has aroused great concern for stakeholders over the past decade. For instance, worldwide forest declines have been associated with drought events (Allen et al., 2010), and the sustainability of grasslands, one of the most important agro-ecosystems representing 26% of the world land area, is threatened due to increasing aridity in the light of climate change (Tubiello et al., 2007; Brookshire and Weaver, 2015). Since the maintenance of grasslands is of prime importance for livestock, and several of the most valuable crops are grasses, herbaceous species deserve more attention from a hydraulic point of view to understand how they will cope with shifts in precipitation and temperature patterns.During water deficit, hydraulic failure in trees has been put forward as one of the primary causes of forest decline (Anderegg et al., 2015, 2016). Drought exacerbates the negative pressure inside the water conducting cells, making the liquid xylem sap more metastable, and thus more vulnerable, to air entry (i.e. gas embolism; Lens et al., 2013a). Extensive levels of embolisms may lead to desiccation, leaf mortality, branch sacrifice, and ultimately plant death (Barigah et al., 2013; Urli et al., 2013). Plant resistance to embolism is therefore assumed to represent a key parameter in determining the drought tolerance of trees and is estimated using so-called vulnerability curves (VCs), from which the P₅₀, i.e. the sap pressure inducing 50% loss of hydraulic conductivity, can be estimated (Cochard et al., 2013). P₅₀ values are therefore good proxies for drought stress tolerance in woody plants and have been published for hundreds of angiosperm and gymnosperm tree species (Delzon et al., 2010; Choat et al., 2012), illustrating a wide range from −0.5 to −19 MPa (Larter et al., 2015).Studies focusing on P₅₀ values of herbs are limited to stems of ∼14 angiosperm species (see Supplemental Table S1 and references cited therein). Half of the herbaceous angiosperms studied so far (Supplemental Table S1) have a stem P₅₀ between 0 and −2 MPa, indicating that many herbs are highly vulnerable to embolism. Moreover, positive root pressure has been reported in various herbs, including many grasses (Poaceae) with hydathodes in their leaves (Evert, 2006), and root pressure is hypothesized to refill embolized conduits overnight when transpiration is low (Miller, 1985; Neufeld et al., 1992; Cochard et al., 1994; Macduff and Bakken, 2003; Saha et al., 2009; Cao et al., 2012). This could suggest that embolism formation and repair follow a daily cycle in herbs. In other words, the midday water potential that herbs experience in the field may often be more negative than P₅₀, which would result in an extremely vulnerable hydraulic pipeline characterized by a negative hydraulic safety margin (expressed as the minimum midday water potential minus P₅₀). In contrast to herbs, most trees operate at a slightly positive hydraulic safety margin (Choat et al., 2012), and woody plants are often too tall to allow refilling by positive root and/or stem pressure in the upper stems (Ewers et al., 1997; Fisher et al., 1997). Therefore, it could be postulated that herbaceous species possess a hydraulic system that is more vulnerable to embolism than that of woody species. In this study, we want to underpin possible differences in embolism resistance between stems of herbaceous and woody angiosperms.The scarcity of P₅₀ measures in herbaceous angiosperms, including grasses and herbaceous eudicots, is mainly due to their fragile stems and low hydraulic conductivity, making VCs technically more challenging. Using minor adaptations to existing centrifuge techniques (Supplemental Text S1), we obtained a P₅₀ stem data set of 26 herbaceous angiosperm species (mainly grasses) from various collection sites in France and Switzerland. In addition, we compared our data set with published data from woody (gymnosperm and angiosperm) species, confronted some of our herbaceous eudicot measurements with original P₅₀ data from derived, woody relatives, and performed anatomical observations in grasses to investigate a possible link between stem anatomical characters and differences in P₅₀ among the species studied. Three main research questions are central in our article: (1) Are stems of herbaceous angiosperms more vulnerable to embolism than those of woody angiosperms? (2) Do grasses operate with highly vulnerable, negative hydraulic safety margins? (3) Do grasses show structure-function trade-offs in their stems with respect to embolism resistance?     

Wild-Born Orangutans (Pongo abelii) Engage in Triadic Interactions During Play

It has long been held that triadic interactions, or interactions between individuals that include shared perception and goals concerning an outside entity, require elaborate cognitive processes such as joint attention. With their connection to shared intentionality, triadic interactions have been a key topic of interest for developmental and evolutionary psychologists, notably when making comparisons between humans and other ape species. There is good evidence that chimpanzees and bonobos engage in triadic interactions; however, convincing evidence for orangutans are more limited and so far have been found only in the context of feeding. I engaged 11 wild-born sanctuary orangutans through the medium of a stick, allowing them to decide how to use the object and how to interact with me. The participating orangutans developed idiosyncratic ways of using the stick and engaging with me during the activity, and six of them alternated their gaze between the stick and me. When I interrupted the activity, the participating orangutans displayed more numerous and different behaviors than before the interruption to actively reengage me in the game. Much like human infants, they appeared more interested in the social interaction than in the stick. These findings confirm that triadic interactions occur in nonenculturated orangutans and are consistent with studies of other nonhuman great ape species, which also show triadic interactions, suggesting that joint attention and potentially shared intentionality may have an early origin in our evolutionary history.     

B Cell Lymphoma-2 (BCL-2) Homology Domain 3 (BH3) Mimetics Demonstrate Differential Activities Dependent upon the Functional Repertoire of Pro- and Anti-apoptotic BCL-2 Family Proteins

The B cell lymphoma-2 (BCL-2) family is the key mediator of cellular sensitivity to apoptosis during pharmacological interventions for numerous human pathologies, including cancer. There is tremendous interest to understand how the proapoptotic BCL-2 effector members (e.g. BCL-2-associated X protein, BAX) cooperate with the BCL-2 homology domain only (BH3-only) subclass (e.g. BCL-2 interacting mediator of death, BIM; BCL-2 interacting-domain death agonist, BID) to induce mitochondrial outer membrane permeabilization (MOMP) and apoptosis and whether these mechanisms may be pharmacologically exploited to enhance the killing of cancer cells. Indeed, small molecule inhibitors of the anti-apoptotic BCL-2 family members have been designed rationally. However, the success of these “BH3 mimetics” in the clinic has been limited, likely due to an incomplete understanding of how these drugs function in the presence of multiple BCL-2 family members. To increase our mechanistic understanding of how BH3 mimetics cooperate with multiple BCL-2 family members in vitro, we directly compared the activity of several BH3-mimetic compounds (i.e. ABT-263, ABT-737, GX15-070, HA14.1, TW-37) in biochemically defined large unilamellar vesicle model systems that faithfully recapitulate BAX-dependent mitochondrial outer membrane permeabilization. Our investigations revealed that the presence of BAX, BID, and BIM differentially regulated the ability of BH3 mimetics to derepress proapoptotic molecules from anti-apoptotic proteins. Using mitochondria loaded with fluorescent BH3 peptides and cells treated with inducers of cell death, these differences were supported. Together, these data suggest that although the presence of anti-apoptotic BCL-2 proteins primarily dictates cellular sensitivity to BH3 mimetics, additional specificity is conferred by proapoptotic BCL-2 proteins.     

Social Network Analysis Shows Direct Evidence for Social Transmission of Tool Use in Wild Chimpanzees

Social network analysis methods have made it possible to test whether novel behaviors in animals spread through individual or social learning. To date, however, social network analysis of wild populations has been limited to static models that cannot precisely reflect the dynamics of learning, for instance, the impact of multiple observations across time. Here, we present a novel dynamic version of network analysis that is capable of capturing temporal aspects of acquisition—that is, how successive observations by an individual influence its acquisition of the novel behavior. We apply this model to studying the spread of two novel tool-use variants, “moss-sponging” and “leaf-sponge re-use,” in the Sonso chimpanzee community of Budongo Forest, Uganda. Chimpanzees are widely considered the most “cultural” of all animal species, with 39 behaviors suspected as socially acquired, most of them in the domain of tool-use. The cultural hypothesis is supported by experimental data from captive chimpanzees and a range of observational data. However, for wild groups, there is still no direct experimental evidence for social learning, nor has there been any direct observation of social diffusion of behavioral innovations. Here, we tested both a static and a dynamic network model and found strong evidence that diffusion patterns of moss-sponging, but not leaf-sponge re-use, were significantly better explained by social than individual learning. The most conservative estimate of social transmission accounted for 85% of observed events, with an estimated 15-fold increase in learning rate for each time a novice observed an informed individual moss-sponging. We conclude that group-specific behavioral variants in wild chimpanzees can be socially learned, adding to the evidence that this prerequisite for culture originated in a common ancestor of great apes and humans, long before the advent of modern humans.