Development of ten microsatellite markers from the keystone mistletoe Tristerix corymbosus (Loranthaceae) using 454 next generation sequencing and their applicability to population genetic structure studies.

Tristerix corymbosus (Loranthaceae) is a keystone mistletoe from the South American temperate rainforests. As most mistletoes, T. corymbosus relies on biotic pollination and seed dispersal, which may cause population structure. For a better understanding of its ecology, we isolated and characterized ten polymorphic microsatellite loci for this species. We used 454 Next Generation Sequencing to build a microsatellite library from a high quality DNA sample. We tested 90 sequences from which we obtained ten polymorphic markers. In order to assess their variability, the novel markers were tested in 48 individuals from three locations of the Valdivian Coastal Reserve in Chile. We also estimated genetic differences between pairs of populations using the FST statistic. The mean number of alleles per locus in the 48 individuals studied was 7.1 (2-17 alleles per locus). The observed and expected heterozygosity ranged from 0.298 to 0.634 and from 0.310 to 0.881, respectively. There were genetic differences among the three populations assessed, according to the FST values (ranging from 0.048 to 0.100, all significant) and, the number of alleles per locus ranged from 3.9 to 5.1. These are the first microsatellite markers developed for T. corymbosus, and they arise as a powerful tool for studying population structure, genetic diversity and gene flow at the landscape scale, along its distribution.

Invasion dynamics of the European bumblebee Bombus terrestris in the southern part of South America.

Invasive species are one of the main biodiversity loss drivers. Some species can establish and thrive in novel habitats, impacting local communities, as is the case of managed pollinators. In this regard, an invasive species' expansion process over time is critical for its control and management. A good example is the European bumblebee Bombus terrestris, which has rapidly invaded the southern part of South America after being repeatedly introduced in Chile for crop pollination since 1997. We assessed the temporal dynamics of B. terrestris invasion in Argentina and Chile by compiling 562 occurrence points from 2000 to 2019. We used two estimators (minimum convex polygon and 95% fixed kernel) to estimate the increase of the invaded area over time. We found that the area invaded by B. terrestris in the southern part of South America presents a linear increase over time, which was consistent for both estimators. In this scenario, species traits, environmental characteristics, and introduction dynamics facilitate a rapid invasion process that will continue to expand, reaching other South American countries in the near future. As this bumblebee is a super-generalist, it probably will expand across South America, as climate niche modelling predicts, if no actions were taken.

Does human-induced habitat transformation modify pollinator-mediated selection? A case study in Viola portalesia (Violaceae).

Pollinator-mediated selection is one of the most important factors driving adaptation in flowering plants. However, as ecological conditions change through habitat loss and fragmentation, the interactions among species may evolve in new and unexpected directions. Human-induced environmental variation is likely to affect selection regimes, but as yet no empirical examples have been reported. In the study reported here, we examined the influence of human-induced habitat transformation on the composition of pollinator assemblages and, hence, pollinator-mediated selection on the flower phenotype of Viola portalesia (Violaceae). Our results indicate that pollinator assemblages differed substantially in terms of species composition and visitation rate between nearby native and transformed habitats. Similarly, the insect species that contributed most to visitation rates differed between plant populations. While the magnitude and sign of pollinator-mediated selection on flower length and width did not differ between sites, selection for flower number lost significance in the transformed habitat, and a significant pattern of disruptive selection for flower shape, undetected in the native habitat, was present in the transformed one. Overall, the results of this study suggest that human-induced habitat change may not only modify the species composition of pollinator assemblages, relaxing the selection process on some flower characters, but they may also create new opportunities for fitness-trait covariation not present in pristine conditions.

Different Pollinators' Functional Traits Can Explain Pollen Load in Two Solitary Oil-Collecting Bees.

Functional traits have been shown to be a good predictor of pollen load for some pollinator bee species, but little is known about solitary bees. In this study, I used two solitary oil-collecting bees to explore the impact of functional traits on the pollen load of two oil-secreting Calceolaria species. I therefore measured the visitation frequency, the time spent manipulating the flower, pollinator body size and pollen load for each bee species. The results reveal that each pollinator visits different Calceolaria species (C. cavanillesii and C. filicaulis) for pollen and/or oil and at least another four herb species for pollen in different proportions. In addition, each bee species presents different functional traits that affect Calceolaria pollen load. For C. chilensis, it was only its body size that affected the Calceolaria pollen load, while in C. subcaeruleus, both body size and handling time together account for its pollen load. Overall, these results highlight the role of pollinators' functional traits in different stages of the pollination process, and even more in specialized plant-pollination systems.

Self-Compatibility Not Associated with Morphological or Genetic Diversity Reduction in Oil-Rewarding Calceolaria Species.

One of the most common evolutionary transitions in angiosperms is the reproductive change from outcrossing to selfing, commonly associated with changes in floral biology and genetic diversity. Here, we aim to test whether self-compatibility leads to a reduction of floral traits and genetic diversity. For this, we experimentally estimate levels of self-compatibility, measure three floral traits and estimate four genetic diversity parameters using nine microsatellites in nine Calceolaria species. Our analysis indicated that four of the study species were self-incompatible. In addition, we found that self-compatible species did not show a reduction in floral traits size, but rather displayed larger corolla and elaiophore areas. Our analyses of genetic diversity identified larger allele number and observed heterozygosity in selfers than in outcrossers, but did not find larger inbreeding in the self-compatible species. Even though our results contradict our expectations, in the case of Calceolaria, their high dependence on only two genera of oil-bees puts the genus in a vulnerable reproductive position, probably facilitating the evolution of reproductive assurance mechanisms in the absence of pollinators. As a result, plants maintain their pollinator attraction traits while evolving the ability to self, possibly in a delayed way.

Maintaining close canopy cover prevents the invasion of Pinus radiata: Basic ecology to manage native forest invasibility.

Pine invasion is a global threat that is occurring in native forests of diverse regions of the world. This process is arising in a scenario of rapid forest deforestation and degradation. Therefore, elucidate which forests attributes explain invasibility is a central issue in forest ecology. The Coastal Maulino forest is an endemic forest of central Chile, which has suffered a large history of disturbance, being replaced by large extensions of Pinus radiata plantations. This land transformation conveys high rates of pines invasion into native remnants. In this study we examined to what extent structural features of forest patches explains invasibility of this forest-type. Within eight forest fragments, we sampled 162 plots (10 x 10 m2 each). We quantified seedling pine density and related these estimates with tree cover, litter depth, PAR radiation, and diversity of the resident community. Our results indicate that canopy cover was the most important variable to determine seedling pine density within forest fragments. Our investigation highlights the importance to conserve the forests cover to reduce significantly their invasibility. This action can be effective even if we cannot avoid pine plantations in the region as a source of a massive seed dispersal to forests with well conserved canopy.

Correction: Maintaining close canopy cover prevents the invasion of Pinus radiata: Basic ecology to manage native forest invasibility.

[This corrects the article DOI: 10.1371/journal.pone.0210849.].

Does Plant Origin Influence the Fitness Impact of Flower Damage? A Meta-Analysis.

Herbivory has been long considered an important component of plant-animal interactions that influences the success of invasive species in novel habitats. One of the most important hypotheses linking herbivory and invasion processes is the enemy-release hypothesis, in which exotic plants are hypothesized to suffer less herbivory and fitness-costs in their novel ranges as they leave behind their enemies in the original range. Most evidence, however, comes from studies on leaf herbivory, and the importance of flower herbivory for the invasion process remains largely unknown. Here we present the results of a meta-analysis of the impact of flower herbivory on plant reproductive success, using as moderators the type of damage caused by floral herbivores and the residence status of the plant species. We found 51 papers that fulfilled our criteria. We also included 60 records from unpublished data of the laboratory, gathering a total of 143 case studies. The effects of florivory and nectar robbing were both negative on plant fitness. The methodology employed in studies of flower herbivory influenced substantially the outcome of flower damage. Experiments using natural herbivory imposed a higher fitness cost than simulated herbivory, such as clipping and petal removal, indicating that studies using artificial herbivory as surrogates of natural herbivory underestimate the real fitness impact of flower herbivory. Although the fitness cost of floral herbivory was high both in native and exotic plant species, floral herbivores had a three-fold stronger fitness impact on exotic than native plants, contravening a critical element of the enemy-release hypothesis. Our results suggest a critical but largely unrecognized role of floral herbivores in preventing the spread of introduced species into newly colonized areas.