Climate change is associated with increased allocation to potential outcrossing in a common mixed mating species.

PREMISE: Although the balance between cross- and self-fertilization is driven by the environment, no long-term study has documented whether anthropogenic climate change is affecting reproductive strategy allocation in species with mixed mating systems. Here, we test whether the common blue violet (Viola sororia; Violaceae) has altered relative allocation to the production of potentially outcrossing flowers as the climate has changed throughout the 20th century. METHODS: Using herbarium records spanning from 1875 to 2015 from the central United States, we quantified production of obligately selfing cleistogamous (CL) flowers and potentially outcrossing chasmogamous (CH) flowers by V. sororia, coupled these records with historic temperature and precipitation data, and tested whether changes to the proportion of CL flowers correlate with temporal climate trends. RESULTS: We find that V. sororia progressively produced lower proportions of CL flowers across the past century and in environments with lower mean annual temperature and higher total annual precipitation. We also find that both CL and CH flower phenology has advanced across this time period. CONCLUSIONS: Our results suggest that V. sororia has responded to lower temperatures and greater water availability by shifting reproductive strategy allocation away from selfing and toward potential outcrossing. This provides the first long-term study of how climate change may affect relative allocation to potential outcrossing in species with mixed mating systems. By revealing that CL flowering is associated with low water availability and high temperature, our results suggest the production of obligately selfing flowers is favored in water limited environments.

Metabolic conversion of ß-pinene to ß-ionone in rats.

Exposure to or ingestion of turpentine can alter the scent of urine, conferring it a flowery, violet-like scent. Turpentine's effect on urine was initially noticed after its use either as medicine or as a preservative in winemaking. Regardless of the source of exposure, the phenomenon requires metabolic conversion of turpentine component(s) to ionone, the molecule mainly responsible for the scent of violets.The purpose of this study was to identify the presence of ionone in the urine of rats that received ß-pinene, and thus to demonstrate that the postulated conversion occurs.We treated rats intraperitoneally with normal saline (negative control), ß-ionone (positive control), low-dose ß-pinene (1/3 of LD50), and high-dose ß-pinene (1/2 of LD50). Urine samples were collected up to 72 h after administration of the compounds, followed by gas chromatography/mass spectrometry identification of the presence of ionone.ß-Ionone was found in the urine of rats exposed to both low and high doses of ß-pinene. In contrast, α-ionone appears unlikely to have been formed in rats exposed to either low or high doses of ß-pinene. ß-pinene was converted to ß-ionone, followed by partial excretion in the urine of rats. ß-Ionone is a minor metabolite of ß-pinene.

Species-specific effects of production practices on genetic diversity in plant reintroduction programs.

Plant production practices can influence the genetic diversity of cultivated plant materials and, ultimately, their potential to adapt to a reintroduction site. A common step in the plant production process is the application of seed pretreatment to alleviate physiological seed dormancy and successfully germinate seeds. In production settings, the seeds that germinate more rapidly may be favored in order to fill plant quotas. In this study, we investigated how the application of cold-moist stratification treatments with different durations can lead to differences in the genetic diversity of the propagated plant materials. Specifically, we exposed seeds of three Viola species to two different cold stratification durations, and then we analyzed the genetic diversity of the resulting subpopulations through double-digestion restriction site-associated sequencing (ddRADseq). Our results show that, in two out of three species, utilizing a short stratification period will decrease the genetic diversity of neutral and expressed loci, likely due to the imposition of a genetic bottleneck and artificial selection. We conclude that, in some species, the use of minimal stratification practices in production may jeopardize the adaptive potential and long-term persistence of reintroduced populations and suggest that practitioners carefully consider the evolutionary implications of their production protocols. We highlight the need to consider the germination ecology of target species when selecting the length of dormancy-breaking pretreatments.

Flower visitors of Streptocarpus teitensis: implications for conservation of a critically endangered African violet species in Kenya.

BACKGROUND: The African violets are endangered plant species restricted mainly to the Eastern Arc Mountains biodiversity hotspots in Kenya and Tanzania. These plants grow well in shaded environments with high humidity. Given their restricted geographical range and published evidence of dependance on insect vectors to facilitate sexual reproduction, understanding their pollination biology is vital for their survival. METHODS: We conducted an empirical study using flower visitor observations, pan trapping and bagging experiments to establish the role of flower visitors in the fruit set of a locally endemic and critically endangered species of African violet in Taita Hills, Kenya, Streptocarpus teitensis. RESULTS: The study found that fruit set is increased by 47.8% in S. teitensis when flowers are visited by insects. However, it is important to note the presence of putative autogamy suggesting S. teitensis could have a mixed breeding system involving self-pollination and cross-pollination since bagged flowers produced 26.9% fruit set. CONCLUSIONS: Insects appear to be essential flower visitors necessary for increased fruit set in S. teitensis. However, there is evidence of a mixed breeding system involving putative self-pollination and cross-pollination suggesting that S. teitensis is somewhat shielded from the negative effects of pollinator losses. Consequently, S. teitensis appears to be protected to a degree from the risks such as reproduction failure associated with pollinator losses by the presence of a safety net in putative self-pollination.

Deciphering the phylogeny of violets based on multiplexed genetic and metabolomic approaches.

Molecular phylogenetics based on nucleotide sequence comparisons has profoundly influenced plant taxonomy. A comprehensive chemotaxonomical approach based on GC-MS and UHPLC-HRMS profiling was evaluated for its ability to characterize a large collection of plants all in the violet family Violaceae (n = 111) and thus decipher the taxonomy. A thorough identification of violets is challenging due to their natural hybridization and phenotypic variability. Phylogenetic inference performed on ribosomal internal transcribed spacer sequences using maximum likelihood and neighbor-joining distance methods allowed the clear identification of 58% of the collection. Metabolomic approaches with multivariate data analysis were performed on SPME/GC-MS chromatograms of volatile compounds emitted by fresh mature flowers and on UHPLC-HRMS/MS leaf extracts for non-volatile compounds. Interestingly, molecular and biochemical approaches provided separate classifications while highlighting several common clusters. The profiling of secondary metabolites was proved most suitable for the classification of hundreds of extracts. The combination of phylogenetic and chemotaxonomic approaches, allowed the classification of 96% of the entire collection. A correlation network revealed specific chemotaxonomic biomarkers, in particular flavonoids, coumarins and cyclotides. Overall, our pioneering approach could be useful to solve misclassification issues within collections of close plant species.

Speyeria (Lepidoptera: Nymphalidae) Conservation.

Speyeria (Nymphalidae) are a conspicuous component of the North American butterfly fauna. There are approximately 16 species and >100 associated subspecies (or geographical variants). Speyeria are univoltine, occupy a wide range of habitats, overwinter as first instar larvae, and feed only on native violets. Speyeria species have become a model group for studies of evolution, speciation, and conservation. Several species and subspecies are threatened or endangered. The reasons for this vary with the taxa involved, but always involve the degradation or loss of quality habitat for larvae and adults. The impacts of climate change must be considered among the causes for habitat degradation and in the establishment of conservation measures. In addition to increasing the available habitat, conservation efforts should consider maintaining habitat in a seral "disturbed" successional stage that selectively favors the growth of violets and preferred adult nectar sources. A major future challenge will be determining the most effective allocation of conservation resources to those species and subspecies that have the greatest potential to respond favorably to these efforts.

Albanian violets of the section Melanium, their morphological variability, genetic similarity and their adaptations to serpentine or chalk soils.

Violets of the section Melanium from Albanian serpentine and chalk soils were examined for their taxonomic affiliations, their ability to accumulate heavy metals and their colonization by arbuscular mycorrhizal fungi (AMF). The sequence analysis of the ITS1-5.8S rDNA-ITS2 region showed that all the sampled six Albanian violets grouped between Viola lutea and Viola arvensis, but not with Viola tricolor. The fine resolution of the ITS sequences was not sufficient for a further delimitation of the Albanian violets within the V. lutea-V. arvensis clade. Therefore, the Albanian violets were classified by a set of morphological characters. Viola albanica, Viola dukadjinica and Viola raunsiensis from serpentine soils as well as Viola aetolica from a chalk meadow were unambiguously identified, whereas the samples of Viola macedonica showed high morphological variability. All the violets, in both roots and shoots contained less than or similar levels of heavy metals as their harboring soils, indicating that they were heavy metal excluders. All the violets were strongly colonized by AMF with the remarkable exception of V. albanica. This violet lived as a scree creeper in shallow serpentine soil where the concentration of heavy metals was high but those of P, K and N were scarce.