Outcrossing breeding system does not compromise invasiveness in Buddleja davidii

Introductions of alien plant species are often likely to consist of a few individuals. Thus, invasion success may strongly depend on their reproductive biology. A high number of self-compatible plants species are known to be successful colonizers of new habitats, even able to establish populations from single propagules. However, many other invasive species require pollen vectors. Here, we investigated the mating system of Buddleja davidii, a fast growing shrub native to China that colonizes quickly in disturbed habitats such as quarries, river banks, along railways and roads, both in its native and invasive regions. It was intentionally introduced to Europe as an ornamental plant because of its fragrant and showy flowers. We additionally studied its vulnerability to biparental inbreeding depression by performing a controlled crossing experiment using pollen from the same population or from geographically close and distant populations, respectively. As a measure for pollination success, we used capsule weight, seed number per capsule and seed weight for each treatment.     

The Stalk Domain and the Glycosylation Status of the Activating Natural Killer Cell Receptor NKp30 Are Important for Ligand Binding

The natural cytotoxicity receptors are a unique set of activating proteins expressed mainly on the surface of natural killer (NK) cells. The human natural cytotoxicity receptor family comprises the three type I membrane proteins NKp30, NKp44, and NKp46. Especially NKp30 is critical for the cytotoxicity of NK cells against different targets including tumor, virus-infected, and immature dendritic cells. Although the crystal structure of NKp30 was recently solved (Li, Y., Wang, Q., and Mariuzza, R. A. (2011) J. Exp. Med. 208, 703-714; Joyce, M. G., Tran, P., Zhuravleva, M. A., Jaw, J., Colonna, M., and Sun, P. D. (2011) Proc. Natl. Acad. Sci. U.S.A. 108, 6223-6228), a key question, how NKp30 recognizes several non-related ligands, remains unclear. Therefore, we investigated the parameters that impact ligand recognition of NKp30. Based on various NKp30-hIgG1-Fc fusion proteins, which were optimized for minimal background binding to cellular Fcγ receptors, we identified the flexible stalk region of NKp30 as an important but so far neglected module for ligand recognition and related signaling of the corresponding full-length receptor proteins. Moreover, we found that the ectodomain of NKp30 is N-linked glycosylated at three different sites. Mutational analyses revealed differential binding affinities and signaling capacities of mono-, di-, or triglycosylated NKp30, suggesting that the degree of glycosylation could provide a switch to modulate the ligand binding properties of NKp30 and NK cell cytotoxicity.     

Studies of binding of tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) to fibroblast growth factor inducible 14 (Fn14)

To perform highly sensitive cellular binding studies with TNF-like weak inducer of apoptosis (TWEAK), we developed a bioluminescent variant of soluble TWEAK (GpL-FLAG-TNC-TWEAK) by fusing it genetically to the C terminus of the luciferase of Gaussia princeps (GpL). Equilibrium binding studies on human (HT1080 and HT29) and murine (Renca and B16) cell lines at 37 °C revealed high affinities of human TWEAK from 53 to 112 pm. The dissociation rate constant of the TWEAK-Fn14 interaction was between 0.48×10(-3) s(-1) (HT29) and 0.58×10(-3) s(-1) (HT1080) for the human molecules, and the association rate constant obtained was 3.3×10(6) m(-1) s(-1) for both cell lines. It has been shown previously that oligomerization of soluble TWEAK trimers results in enhanced Fn14-mediated activation of the classical NFκB pathway. Binding studies with GpL-FLAG-TNC-TWEAK trimers oligomerized by help of a FLAG tag-specific antibody gave no evidence for a major increase in Fn14 occupancy by oligomerized ligand despite strongly enhanced induction of the NFκB target IL8. Thus, aggregated complexes of soluble TWEAK and Fn14 have a higher intrinsic activity to stimulate the classical NFκB pathway and qualitatively differ from isolated trimeric TWEAK-Fn14 complexes. Furthermore, determination of IL8 induction as a function of occupied activated receptors revealed that the intrinsic capability of TNFR1 to stimulate the classical NFκB pathway and IL8 production was ～100-fold higher than Fn14. Thus, although ～25 activated TNFR1 trimers were sufficient to trigger half-maximal IL8 production, more than 2500 cell-bound oligomerized TWEAK trimers were required to elicit a similar response.     

Mixed reproduction strategy and polyploidy facilitate dominance of Kobresia pygmaea on the Tibetan Plateau

Aims The sedge Kobresia pygmaea is the dominant species of high-altitude pastures in Tibet, and it is the most important source of forage in animal husbandry. We present the first comprehensive reproduction study for this perennial key species that adopts a molecular approach and tests how sexual and vegetative reproduction, as well as ploidy, relate to survival and dominance under harsh conditions.Methods We assessed inflorescence numbers of K. pygmaea across two Tibetan alpine pastures with differing grazing regimes. Germination was tested in untreated diaspores and then following mechanical and chemical scarification. In a 4-year experiment, we assessed diaspore viability and seed bank formation. Using eight microsatellite markers, we recorded multilocus genotypes in hierarchical grids and measured their ploidy using flow cytometry. Adjusted analysis of variance models were used to analyse data on sexual reproduction, while the complement of the Simpson index and the Shannon diversity index were used to characterize the spatial distribution of multilocus genotypes and clonal richness.Important findings Inflorescence production was high and differed significantly between years (2010: 617±460 SD; 2012: 2015±1213 SD) but not between grazing regimes. Diaspore viability was high (94%) and gradually decreased after 3 and 4 years of storage in the soil. Diaspores not exposed to further scarification failed to germinate, while mechanical and chemical (H 2 SO 4) scarification increased germination to 9 and 44%, respectively. Clonal diversity was high, although in situ germination was rarely observed. Multilocus genotypes intersected and covered a mean area of 0.74 m 2. Most individuals were found to be tetraploid, with only 0.8% of all ramets being triploid. We conclude that K. pygmaea survives on the Tibetan Plateau by employing a mixed reproduction strategy involving both sexual and clonal propagation. The species' adaptability and dominance is further facilitated by its polyploidy. As pasture restoration using diaspores would be difficult, existing Kobresia pasture should be managed more sustainably.     

The invasive shrub Buddleja davidii performs better in its introduced range

It is commonly assumed that invasive plants grow more vigorously in their introduced than in their native range, which is then attributed to release from natural enemies or to microevolutionary changes, or both. However, few studies have tested this assumption by comparing the performance of invasive species in their native vs. introduced ranges. Here, we studied abundance, growth, reproduction, and herbivory in 10 native Chinese and 10 invasive German populations of the invasive shrub Buddleja davidii (Scrophulariaceae; butterfly bush). We found strong evidence for increased plant vigour in the introduced range: plants in invasive populations were significantly taller and had thicker stems, larger inflorescences, and heavier seeds than plants in native populations. These differences in plant performance could not be explained by a more benign climate in the introduced range. Since leaf herbivory was substantially reduced in invasive populations, our data rather suggest that escape from natural enemies, associated with increased plant growth and reproduction, contributes to the invasion success of B. davidii in Central Europe.     

Effects of altitude and livestock on the regeneration of two tree line forming <Emphasis Type="Italic">Polylepis</Emphasis> species in Ecuador

Regeneration is known to be limited at many temperate tree lines, but very little data is available on the impacts of altitude and anthropogenic disturbance on regeneration patterns along tropical tree lines. The study focused on the reproductive traits of two Polylepis species in the Páramo de Papallacta in Ecuador along an altitudinal gradient, and involved different intensities of cattle trampling within subsequent altitudinal ranges. We analyzed flowering, fruit set, seed viability, germination, and seedling establishment as well as stand structure of Polylepis incana and P. pauta. The numbers of P. incana inflorescences and seedlings per m2 showed a marginally significant decrease with increasing altitude. Mean tree height was significantly lower at higher altitudes, while stem number increased. The number of P. pauta inflorescences also decreased significantly upslope. In both forest types, trampling was found to have a positive impact on seedling abundance, presumably due to the removal of the litter layer. Thus, there was no evidence of negative effects of moderate cattle grazing on both tree line species. However, sapling establishment was minimal inside the forest stands at all altitudes and grazing levels, and we consequently observed a low proportion of narrow stems within all investigated forests. Our results show that, along with vegetative growth limitations of adult trees, important regeneration traits such as seedling and inflorescence numbers are also influenced by altitude, which might contribute to the formation of the upper tree line. Nevertheless, recruitment in the forest interior was low overall indicating that further factors, such as light conditions, affect regeneration of the studied species.     

Why get big in the cold? Towards a solution to a life-history puzzle

The temperature–size rule (TSR), which states that body size increases at lower developmental temperatures, appears to be a near-universal law for ectotherms. Although recent studies seem to suggest that the TSR might be adaptive, the underlying developmental mechanisms are thus far largely unknown. Here, we investigate temperature effects on life-history traits, behaviour and physiology in the copper butterfly Lycaena tityrus in order to disentangle the mechanistic basis for the above rule. In L. tityrus the larger body size produced at a lower temperature was proximately due to a greater increase in mass, which was caused by both behavioural and physiological mechanisms: a much-increased food intake and a higher efficiency in converting ingested food into body matter. These mechanisms, combined with temperature-induced changes at the cellular level, may provide general explanations for the TSR. Body fat and protein content increased in butterflies reared at the higher temperature, indicating favourable growth conditions. As predicted from protandry theory, males showed reduced development times, caused by higher growth rates compared to females. The latter was itself related to a higher daily food consumption, while the total food consumption (due to the females’ longer developmental period) and assimilation was higher in females and may underly the sexual body size dimorphism.     

Adaptive and non‐adaptive evolution of trait means and genetic trait correlations for herbivory resistance and performance in an invasive plant

The EICA‐hypothesis predicts that invading plants adapt to their novel environment by evolving increased performance and reduced resistance in response to the release from natural enemies, and assumes a resource allocation tradeoff among both trait groups as mechanistic basis of this evolutionary change. Using the plant Silene latifolia as a study system, we tested these predictions by investigating whether 1) invasive populations evolved lower resistance and higher performance, 2) this evolutionary change is indeed adaptive, and 3) there is a negative genetic correlation between performance and resistance (i.e. a tradeoff) in native and introduced individuals. Moreover, we sampled eight native and eight invasive populations and determined their population co‐ancestry based on neutral SSR‐markers. We performed controlled crossings to produce five sib‐groups per population and exposed them to increased and reduced levels of enemy attack in a full‐factorial experiment to estimate performance and resistance. With these data, we performed trait‐by‐trait comparisons between ranges with ‘animal models’ that account for population co‐ancestry to quantify the amount of variance in traits explained by non‐adaptive versus adaptive evolution. Moreover, we tested for genetic correlations among performance and resistance traits within sib‐groups. We found significant reductions in resistance and increases in performance in invasive versus native populations, which could largely be attributed to adaptive evolution. While we detected a non‐significant trend towards negative genetic performance × resistance correlations in native populations, invasive populations exhibited both significant and non‐significant positive correlations. In summary, these results do not support a shift of performance and resistance trait values along a tradeoff line in response to enemy release, as predicted by EICA. They rather suggest that the independent evolution of both traits is not constrained by a tradeoff, and that various selective agents (including resource availability) interact in shaping both traits and in weakening negative genetic correlations in the invaded habitat.     

Intra- and interspecific hybridization in invasive Siberian elm

Hybridization creates unique allele combinations which can facilitate the evolution of invasiveness. Frequent interspecific hybridization between the Siberian elm, Ulmus pumila, and native elm species has been detected in the Midwestern United States, Italy and Spain. However, Ulmus pumila also occurs in the western United States and Argentina, regions where no native elm species capable of hybridizing with it occurs. We examined whether inter- or intraspecific hybridization could be detected in these regions. Nuclear markers and the program STRUCTURE helped detect interspecific hybridization and determine the population genetic structure in both the native and the two non-native ranges. Chloroplast markers identified sources of introduction into these two non-native ranges. No significant interspecific hybridization was detected between U. pumila and U. rubra in the western United States or between U. pumila and U. minor in Argentina and vice versa. However, the genetic findings supported the presence of intraspecific hybridization and high levels of genetic diversity in both non-native ranges. The evidence presented for intraspecific hybridization in the current study, combined with reports of interspecific hybridization from previous studies, identifies elm as a genus where both inter- and intraspecific hybridization may occur and help maintain high levels of genetic diversity potentially associated with invasiveness.