Pollination success across an elevation and sex ratio gradient in gynodioecious Daphne laureola

Gynodioecy is a dimorphic breeding system in which hermaphrodite and female individuals coexist in populations. Sex ratio and gender-relative lifetime seed production determine the stability of gynodioecy, and both genetic and ecological factors may influence these parameters. I analyzed the consequences of variation in population sex ratio and site elevation for the relative pollination success of female and hermaphrodite individuals of Daphne laureola in southern Spain, where previous studies failed to detect female fecundity advantages at two mid-elevation sites. Pollination success, estimated as stigmatic pollen loads, number of pollen tubes per style, and percentage of fertilized flowers, was higher for hermaphrodites than females in populations with 20-56% females. Furthermore, female quantitative disadvantage in pollination success increased with elevation, suggesting that the higher availability of pollen due to the increased proportion of hermaphrodites could not mitigate the negative effect that other factors associated with elevation apparently had on pollination. Supplemental hand pollinations showed that female seed production was pollen limited in populations with a proportion of females >50%, although both pollination success and natural fruit set of females in these sites were the highest recorded.Peer Reviewe

Variation in herbivory by Yponomeuta mahalebella on its only host plant Prunus mahaleb along an elevational gradient

1. The effect of natural variation in abiotic conditions on the herbivory interaction between Prunus mahaleb (Rosaceae) and its monophagous folivore, larvae of Yponomeuta mahalebella (Lepidoptera, Yponomeutidae), was analysed for 2 consecutive years along an elevational gradient in Sierra de Cazorla, south-east Spain. 2. There was a negative correlation between site elevation and mean population herbivory level measured at the end of the growing period of Y. mahalebella. Mortality during larval development was higher at higher elevation sites, and mean adult body mass was higher in lower elevation populations. 3. Variation in temperatures recorded during the larval growth period at different altitudes was the only study factor related to abundance of Y. mahalebella larvae, neither differences in parasitisation rates nor plant features covaried significantly with herbivore abundance. 4. These results support the existence of geographical variation in plant-animal interactions in relation to environmental heterogeneity.Peer Reviewe

Epigenetic contribution to successful polyploidizations: variation in global cytosine methylation along an extensive ploidy series in Dianthus broteri (Caryophyllaceae)

Polyploidization is a significant evolutionary force in plants which involves major genomic and genetic changes, frequently regulated by epigenetic factors. We explored whether natural polyploidization in Dianthus broteri complex resulted in substantial changes in global DNA cytosine methylation associated to ploidy. Global cytosine methylation was estimated by high-performance liquid chromatography (HPLC) in 12 monocytotypic populations with different ploidies (2×, 4×, 6×, 12×) broadly distributed within D. broteri distribution range. The effects of ploidy level and local variation on methylation were assessed by generalized linear mixed models (GLMMs). Dianthus broteri exhibited a higher methylation percent (˜33%) than expected by its monoploid genome size and a large variation among study populations (range: 29.3–35.3%). Global methylation tended to increase with ploidy but did not significantly differ across levels due to increased variation within the highest-order polyploidy categories. Methylation varied more among hexaploid and dodecaploid populations, despite such cytotypes showing more restricted geographic location and increased genetic relatedness than diploids and tetraploids. In this study, we demonstrate the usefulness of an HPLC method in providing precise and genome reference-free global measure of DNA cytosine methylation, suitable to advance current knowledge of the roles of this epigenetic mechanism in polyploidization processes.Peer Reviewe

To be or not to be better pollinated: Differences between sex morphs in marginal gynodioecious populations

PREMISE OF THE STUDY: Changes in the pollinator communities of marginal plant populations can affect their pollination quantity or quality. Geographic variation in pollination success can alter the reproductive advantage that female plants require to persist within gynodioecious populations. Particularly valuable is determining the pollination success at the prezygotic stage in self-compatible gynodioecious species whose females do not exhibit enhanced seed production. METHODS: In core and marginal populations of Daphne laureola, we analyzed the differences between hermaphrodites and females in the proportion of flowers visited, the stigma pollen loads, and the quantity of pollen tubes in styles. We also examined the relationship between the number of pollen tubes in styles vs. the number of pollen grains on stigmas using piecewise regression and binomial generalized linear mixed models. KEY RESULTS: Pollinators deposited larger pollen loads on flowers in marginal populations. In marginal populations, female flowers received more pollinator visits and more pollen grains on their stigmas, and they had more pollen tubes in their styles than did female flowers in core populations. Both piecewise regression and binomial GLMM analyses showed that females in marginal populations had a lower proportion of grains that developed tubes than females in the core populations, which suggests decreased pollination quality. CONCLUSIONS: More efficient pollination services in marginal populations decreased the overall differences in the prezygotic pollination success between the sex morphs. Our results also suggest that pollination quality is lower in females of marginal populations, which could be counteracting the increased pollination in females in marginal populations.Peer Reviewe

MSAP markers and global cytosine methylation in plants: a literature survey and comparative analysis for a wild-growing species

Methylation of DNA cytosines affects whether transposons are silenced and genes are expressed, and is a major epigenetic mechanism whereby plants respond to environmental change. Analyses of methylation-sensitive amplification polymorphism (MS-AFLP or MSAP) have been often used to assess methyl-cytosine changes in response to stress treatments and, more recently, in ecological studies of wild plant populations. MSAP technique does not require a sequenced reference genome and provides many anonymous loci randomly distributed over the genome for which the methylation status can be ascertained. Scoring of MSAP data, however, is not straightforward, and efforts are still required to standardize this step to make use of the potential to distinguish between methylation at different nucleotide contexts. Furthermore, it is not known how accurately MSAP infers genome-wide cytosine methylation levels in plants. Here, we analyse the relationship between MSAP results and the percentage of global cytosine methylation in genomic DNA obtained by HPLC analysis. A screening of literature revealed that methylation of cytosines at cleavage sites assayed by MSAP was greater than genome-wide estimates obtained by HPLC, and percentages of methylation at different nucleotide contexts varied within and across species. Concurrent HPLC and MSAP analyses of DNA from 200 individuals of the perennial herb Helleborus foetidus confirmed that methyl-cytosine was more frequent in CCGG contexts than in the genome as a whole. In this species, global methylation was unrelated to methylation at the inner CG site. We suggest that global HPLC and context-specific MSAP methylation estimates provide complementary information whose combination can improve our current understanding of methylation-based epigenetic processes in nonmodel plants.Peer reviewe

Effect of antibiotic prophylaxis for preventing infectious complications following impacted mandibular third molar surgery. A randomized controlled trial

The objective of this study was to determine the effect of antibiotic prophylaxis in preventing postoperative infections after extraction of impacted mandibular third molars. A Parallel-group, randomized, blind, placebo-controlled trial was performed. 154 patients were randomly allocated to 2 groups; experimental (n=77) receiving 2g amoxicillin 1 hour prior to surgery and control (n=77) receiving placebo. Primary outcome was postoperative infections and secondary outcome was the need for rescue analgesia. 4.5% of patients developed postoperative infections, five patients of the control group (4 alveolar osteitis, 1 surgical site infection) and two of the experimental group (1 alveolar osteitis, 1 surgical site infection). Difference between groups was not statistically significant, RR=0.4 (95%CI 0.08-1.99, ?=0.41) NNTB=26. Rescue analgesia intake was significantly higher in the control group (41 vs 18 patients of experimental group) RR=0.49 (95%CI 0.32-0.75, ?<0.05) NNTB=3. The use of 2g amoxicillin 1 hour before surgery was not effective in significantly reducing the risk of postoperative infections from impacted mandibular third molars extraction, when compared to placebo. Nevertheless, antibiotic prophylaxis was associated with a reduced need for rescue analgesia

The Role of Alien Species on Plant-Floral Visitor Network Structure in Invaded Communities

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. The interactions between pairs of native and alien plants via shared use of pollinators have been widely studied. Community level studies however, are necessary in order to fully understand the factors and mechanisms that facilitate successful plant invasion, but these are still scarce. Specifically, few community level studies have considered how differences in invasion level (alien flower abundance), and degree of floral trait similarity between native and invasive species, mediate effects on native plant-pollinator communities. Here, we evaluated the role of alien species on overall plant-floral visitor network structure, and on species-level network parameters, across nine invaded coastal communities distributed along 205 km in Yucatán, México that vary in alien species richness and flower abundance. We further assessed the potential the role of alien plant species on plant-floral visitor network structure and robustness via computational simulation of native and invasive plant extinction scenarios. We did not find significant differences between native and alien species in their functional floral phenotypes or in their visitation rate and pollinator community composition in these invaded sites. Variation in the proportion of alien plant species and flower abundance across sites did not influence plant-pollinator network structure. Species-level network parameters (i.e., normalized degree and nestedness contribution) did not differ between native and alien species. Furthermore, our simulation analyses revealed that alien species are functionally equivalent to native species and contribute equally to network structure and robustness. Overall, our results suggest that high levels of floral trait similarity and pollinator use overlap may help facilitate the integration of alien species into native plant-pollinator networks. As a result, alien species may also play a similar role than that of natives in the structure and stability of native plant and pollinator communities in the studied coastal sand dune ecosystem

Herbivores do not discriminate between leaves of female and hermaphrodite individuals of gynodioecious Daphne laureola (Thymelaeaceae)

The outcome of plant-animal interactions in dioecious plant species frequently depends on the gender of the plant individuals. It has even been proposed that these interactions could mediate the evolution of plant reproductive systems from hermaphroditism to dioecy. Gynodioecy is the most frequent intermediate stage in this evolutionary process, however, little is known about the relevance of gender dimorphism in plant-animal relationships others than pollination for gynodioecious species. In this study herbivores (Noctuid larvae) were used as subjective referees to detect differences between leaves of female and hermaphrodite Daphne laureola individuals. Larvae collected in the field were allowed to choose between leaves of female and hermaphrodite individuals at both flowering and fruiting period. There was no preference for either of the genders suggesting that the absence of male reproductive function in D. laureola shrubs does not cause dissimilarities in leaf characteristics that can affect herbivores.Peer Reviewe