Evolution of the relaxin-like peptide family

Background  

The relaxin-like peptide family belongs in the insulin superfamily and consists of 7 peptides of high structural but low sequence similarity; relaxin-1, 2 and 3, and the insulin-like (INSL) peptides, INSL3, INSL4, INSL5 and INSL6. The functions of relaxin-3, INSL4, INSL5, INSL6 remain uncharacterised. The evolution of this family has been contentious; high sequence variability is seen between closely related species, while distantly related species show high similarity; an invertebrate relaxin sequence has been reported, while a relaxin gene has not been found in the avian and ruminant lineages.     

Growth irradiance effects on photosynthesis and growth in two co-occurring shade-tolerant neotropical perennials of contrasting photosynthetic pathways

Dieffenbachia longispatha (C3) and Aechmea magdalenae (Crassulacean acid metabolism, CAM) are syntopic, neotropical forest perennials in central Panama that are restricted to shaded habitats. This is of particular interest for A. magdalenae because, like other understory CAM bromeliad species, it appears functionally and structurally to be better suited to life in full sun. Growth irradiance (GI) effects on photosynthesis and growth in both species were explored in the context of sun/shade trade-off concepts largely derived from studies of C3 plants. Potted plants were grown outdoors in 1, 55, and 100% full sun for 5 mo under well-watered conditions. While both species grew faster in high compared to low light, maximum relative growth rates (RGR) in full sun were still extremely slow with A. magdalenae showing a RGR approximately half that of D. longispatha. Photosynthetic capacity increased with GI in D. longispatha but not in A. magdalenae. Aechmea magdalenae responded to GI with shifts in the activity of the different CAM phases. Both species were photoinhibited in full sun, but more so in A. magdalenae. Despite possessing many traits considered adaptive in high light, these results suggest that A. magdalenae is unlikely to attain sufficient growth rates to thrive in productive, high-light habitats.     

Seasonal changes in the response of winter wheat to elevated atmospheric CO2 concentration grown in Open-Top Chambers and field tracking enclosures

Winter wheat was grown at ambient and elevated (ambient plus 350 μL L^–1) CO₂ concentrations in open top chambers and in field-tracking sun-lit climatized enclosures (elevated is 718 μL L^–1). There was no significant effect of CO₂ concentration on sheath, leaf and root biomass and leaf area in the early spring (January to April). 24-h canopy CO₂ exchange rate (CCER) was not significantly affected either. However, elevated CO₂ concentration increased CCER at midday, decreased evapotranspiration rate and increased instantaneous water-use-efficiency during early spring. Leaf, sheath and root nitrogen concentration per unit dry weight decreased and nonstructural carbohydrate concentration increased under elevated CO₂, and N-uptake per unit ground area decreased significantly (– 22%) towards the end of this period. These results contrast with results from the final harvest, when grain yield and biomass were increased by 19% under elevated CO₂. N concentration per dry weight was reduced by 5%, but N-uptake per unit ground area was significantly higher (+ 11%) for the elevated CO₂ treatment. 24-h and midday-CCER increased significantly more in late spring (period of 21 April to 30 May) (respectively by + 40% and 53%) than in the early spring (respectively 5% and 19%) in response to elevated CO₂. Midday evapotranspiration rate was reduced less by elevated CO₂ in the late spring (– 13%) than in early spring (– 21%). The CO₂ response of midday and 24-h CCER decreased again (+ 27% and + 23% resp.) towards the end of the growing season. We conclude that the low response to CO₂ concentration during the early spring was associated with a growth-restriction, caused by low temperature and irradiance levels. The reduction of nitrogen concentration, the increase of nonstructural carbohydrate, and the lower evapotranspiration indicated that CO₂ did have an effect towards the end of early spring, but not on biomass accumulation. Regression analysis showed that both irradiance and temperature affected the response to CO₂.     

Influence of nitrogen supply and growth irradiance on photoinhibition and recovery in Heuchera americana (Saxifragaceae)

Prior work demonstrated that Heuchera americana, an evergreen herb inhabiting the deciduous forest understory in the southeastern United States, has a 3-4-fold greater photosynthetic capacity under the low-temperature, strong-light, open canopies of winter compared to the high-temperature, weak-light, closed canopies of summer. Moreover, despite the reductions in soil nitrogen, the chilling temperatures, and the increased quantum flux associated with winter, chronic photoinhibition was not observed in this species at this time of the year. We were interested in the photosynthetic acclimation and photoinhibition characteristics of this species when grown under contrasting light and nitrogen regimes. Newly expanded shade-acclimated leaves of forest-grown plants exposed to strong light varying in intensity and duration at 25°C showed a reduction in F_v/F_m (the ratio of variable to maximum room temperature chlorophyll fluorescence measured after dark adaptation), which was correlated with a decline in ø_a (the intrinsic quantum yield of CO₂-saturated O₂ evolution on an absorbed light basis). Plants grown in the glasshouse under contrasting light (high and low light; HL and LL, respectively) and nitrogen supply (high and low nitrogen; HN and LN, respectively) regimes showed that photosynthetic acclimation to HL was impaired in LN regimes. The HL-LN plants also had the lowest values of F_v/F_m and of ø on both incident and absorbed light bases and had 50% less chlorophyll (per unit area) compared to plants from other growth regimes. Controlled exposure to bright light at low temperatures (2-3°C) for 3 h resulted in a sharp decrease in F_v/F_m (and rise in F_o, the minimum fluorescence yield) in all plants. Shade-grown plants from both N regimes were highly susceptible to chronic photoinhibition, as indicated by a greater reduction in F_v/F_m and incomplete recovery after 18 h in weak light at 25°C. The HL-HN plants were the least susceptible to chronic photoinhibition, having the smallest decrease in F_v/F_m with near full recovery within 6 h. The decline in Fv/Fm in HL-LN plants was comparable to that of shade-acclimated plants, but recovered fully within 6 h. Low-N plants from both light regimes displayed greater increases in F_o which did not return to pretreatment levels after 18 h of recovery. These studies indicate that HL-LN plants were sensitive to chronic photoinhibition and, at the same time, had a high capacity for dynamic photoinhibition. Experimental garden studies showed that H. americana grown in an open field in summer were photoinhibited and did not fully recover overnight or during extended periods of weak light. These results are discussed in relation to the photosynthetic acclimation of H. americana under natural conditions.     

Ca2+, annexins, and GTP modulate exocytosis from maize root cap protoplasts

Protoplasts isolated from root cap cells of maize were shown to secrete fucose-rich polysaccharides and were used in a patch-clamp study to monitor changes in whole-cell capacitance. Ca2+ was required for exocytosis, which was measured as an increase in cell capacitance during intracellular dialysis with Ca2+ buffers via the patch pipette. Exocytosis was stimulated significantly by small increases above normal resting [Ca2+]. In the absence of Ca2+, protoplasts decreased in size. In situ hybridization showed significant expression of the maize annexin p35 in root cap cells, differ-entiating vascular tissue, and elongating cells. Dialysis of protoplasts with maize annexins stimulated exocytosis at physiological [Ca2+], and this could be blocked by dialysis with antibodies specific to maize annexins. Dialysis with milli-molar concentrations of GTP strongly inhibited exocytosis, causing protoplasts to decrease in size. GTPgammaS and GDPbetaS both caused only a slight inhibition of exocytosis at physiological Ca2+. Protoplasts were shown to internalize plasma membrane actively. The results are discussed in relation to the regulation of exocytosis in what is usually considered to be a constitutively secreting system; they provide direct evidence for a role of annexins in exocytosis in plant cells.     

Effects of climate change on productivity of cereals and legumes; model evaluation of observed year-to-year variability of the CO2 response

The effect of elevated [CO₂] on the productivity of spring wheat, winter wheat and faba bean was studied in experiments in climatized crop enclosures in the Wageningen Rhizolab in 1991–93. Simulation models for crop growth were used to explore possible causes for the observed differences in the CO₂ response. Measurements of the canopy gas exchange (CO₂ and water vapour) were made continuously from emergence until harvest. At an external [CO₂] of 700 μmol mol^?1 Maximum Canopy CO₂ Exchange Rate (CCERmax) at canopy closure was stimulated by 51% for spring wheat and by 71% for faba bean. At the end of the growing season, above ground biomass increase at 700 μmol mol^?1 was 58% (faba bean), 35% (spring wheat) and 19% (winter wheat) and the harvest index did not change. For model exploration, weather data sets for the period 1975-88 and 1991–93 were used, assuming adequate water supply and [CO₂] at 350 and 700 μmol mol^?1. For spring wheat the simulated responses (35–50%) were at the upper end of the experimental results. In agreement with experiments, simulations showed smaller responses for winter wheat and larger responses for faba bean. Further model explorations showed that this differential effect in the CO₂ response may not be primarily due to fundamental physiological differences between the crops, but may be at least partly due to differences in the daily air temperatures during comparable stages of growth of these crops. Simulations also showed that variations between years in CO₂ response can be largely explained by differences in weather conditions (especially temperature) between growing seasons.     

Evolutionarily divergent, unstable filamentous actin is essential for gliding motility in apicomplexan parasites

Apicomplexan parasites rely on a novel form of actin-based motility called gliding, which depends on parasite actin polymerization, to migrate through their hosts and invade cells. However, parasite actins are divergent both in sequence and function and only form short, unstable filaments in contrast to the stability of conventional actin filaments. The molecular basis for parasite actin filament instability and its relationship to gliding motility remain unresolved. We demonstrate that recombinant Toxoplasma (TgACTI) and Plasmodium (PfACTI and PfACTII) actins polymerized into very short filaments in vitro but were induced to form long, stable filaments by addition of equimolar levels of phalloidin. Parasite actins contain a conserved phalloidin-binding site as determined by molecular modeling and computational docking, yet vary in several residues that are predicted to impact filament stability. In particular, two residues were identified that form intermolecular contacts between different protomers in conventional actin filaments and these residues showed non-conservative differences in apicomplexan parasites. Substitution of divergent residues found in TgACTI with those from mammalian actin resulted in formation of longer, more stable filaments in vitro. Expression of these stabilized actins in T. gondii increased sensitivity to the actin-stabilizing compound jasplakinolide and disrupted normal gliding motility in the absence of treatment. These results identify the molecular basis for short, dynamic filaments in apicomplexan parasites and demonstrate that inherent instability of parasite actin filaments is a critical adaptation for gliding motility.     

Phenology of brown marmorated stink bug described using female reproductive development

Temperature‐based degree‐day models describe insect seasonality and to predict key phenological events. We expand on the use of a temperature‐based process defining timing of reproduction through the incorporation of female reproductive physiology for the invasive pentatomid species Halyomorpha halys, the brown marmorated stink bug. A five‐stage ranking system based on ovary development was able to distinguish between the reproductive statuses of field‐collected females. Application of this ranking method described aspects of H. halys’ seasonality, overwintering biology, and phenology across geographic locations. Female H. halys were collected in the US from NJ, WV, NC, OR, and two sites in PA in 2006–2008 (Allentown, PA only) and 2012–2014. Results identify that H. halys enters reproductive diapause in temperate locations in the fall and that a delay occurs in developmental maturity after diapause termination in the spring. Modification of the Snyder method to identify biofix determined 12.7‐hr photoperiod as the best fit to define initiation of reproduction in the spring. Applying the biofix, we demonstrated significant differences between locations for the rate at which the overwintering generation transition into reproductive status and the factors contributing to this difference require further study. For example, after including abiotic variables influencing development such as temperature and photoperiod (critical diapause cue), reproduction occurred earlier in OR and for an extended period in NJ. This data describe a method to investigate insect seasonality by incorporating physiological development across multiple regions that can clarify phenology for insects with overlapping generations.     

The effects of heterospecific mating frequency on the strength of cryptic reproductive barriers

Heterospecific mating frequency is critical to hybrid zone dynamics and can directly impact the strength of reproductive barriers and patterns of introgression. The effectiveness of post‐mating prezygotic (PMPZ) reproductive barriers, which include reduced fecundity via heterospecific matings and conspecific sperm precedence, may depend on the number, identity and order of mates. Studies of PMPZ barriers suggest that they may be important in many systems, but whether these barriers are effective at realistic heterospecific mating frequencies has not been tested. Here, we evaluate the strength of cryptic reproductive isolation in two leaf beetles (Chrysochus auratus and C. cobaltinus) in the context of a range of heterospecific mating frequencies observed in natural populations. We found both species benefited from multiple matings, but the benefits were greater in C. cobaltinus and extended to heterospecific matings. We found that PMPZ barriers greatly limited hybrid production by C. auratus females with moderate heterospecific mating frequencies, but that their effectiveness diminished at higher heterospecific mating frequencies. In contrast, there was no evidence for PMPZ barriers in C. cobaltinus females at any heterospecific mating frequency. We show that integrating realistic estimates of cryptic isolation with information on relative abundance and heterospecific mating frequency in the field substantially improves our understanding of the strong directional bias in F1 production previously documented in the Chrysochus hybrid zone. Our results demonstrate that heterospecific mating frequency is critical to understanding the impact of cryptic post‐copulatory barriers on hybrid zone structure and dynamics, and that future studies of such barriers should incorporate field‐relevant heterospecific mating frequencies.