Radiating diversification and niche conservatism jointly shape the inverse latitudinal diversity gradient of Potentilla L. (Rosaceae).

BACKGROUND: The latitudinal diversity gradient (LDG), characterized by an increase in species richness from the poles to the equator, is one of the most pervasive biological patterns. However, inverse LDGs, in which species richness peaks in extratropical regions, are also found in some lineages and their causes remain unclear. Here, we test the roles of evolutionary time, diversification rates, and niche conservatism in explaining the inverse LDG of Potentilla (ca. 500 species). We compiled the global distributions of ~ 90% of Potentilla species, and reconstructed a robust phylogenetic framework based on whole-plastome sequences. Next, we analyzed the divergence time, ancestral area, diversification rate, and ancestral niche to investigate the macroevolutionary history of Potentilla. RESULTS: The genus originated in the Qinghai-Tibet Plateau during the late Eocene and gradually spread to other regions of the Northern Hemisphere posterior to the late Miocene. Rapid cooling after the late Pliocene promoted the radiating diversification of Potentilla. The polyploidization, as well as some cold-adaptive morphological innovations, enhanced the adaptation of Potentilla species to the cold environment. Ancestral niche reconstruction suggests that Potentilla likely originated in a relatively cool environment. The species richness peaks at approximately 45 °N, a region characterized by high diversification rates, and the environmental conditions are similar to the ancestral climate niche. Evolutionary time was not significantly correlated with species richness in the latitudinal gradient. CONCLUSIONS: Our results suggest that the elevated diversification rates in middle latitude regions and the conservatism in thermal niches jointly determined the inverse LDG in Potentilla. This study highlights the importance of integrating evolutionary and ecological approaches to explain the diversity pattern of biological groups on a global scale.

Ecotypic divergences of the alpine herb Potentilla matsumurae adapted to fellfield-snowbed habitats across a series of mountain sky islands.

PREMISE: Divergent selection due to environmental heterogeneity can lead to local adaptation. However, the ecological and evolutionary processes of local adaptation that occurs across multiple regions are often unknown. Our previous studies reported on the ecotypic divergence within a local area of variation of Potentilla matsumurae, an alpine herb adapted to the fellfield-snowbed environment. Here we investigated large-scale geographic patterns of ecotypic differentiation in this species to infer local adaptation and selective forces across multiple regions. METHODS: We compiled information on the overall distributions of fellfield and snowbed habitats on the mountains in Japan across the distribution of the species. Next, we conducted common garden experiments to test the adaptive divergence of the fellfield-snowbed plants derived from multiple regions. Finally, we evaluated phylogeographic structures based on cpDNA and allozyme variations and inferred the evolutionary history of ecotype differentiation. RESULTS: The mosaic distribution of the fellfield-snowbed ecotypes across isolated mountaintops constitutes indirect evidence for habitat-specific natural selection. The significant difference in survivorship between the ecotypes observed in a controlled snow environment provides more substantial evidence of local selection. Phylogeographic structures support the hypothesis that ecotypic divergence events from fellfield to snowbed populations occurred independently in at least two distinct regions. CONCLUSIONS: Ecotypic divergence of P. matsumurae has occurred across a series of mountain sky islands. Local selection in snowy environments is a driving force that maintains the divergent ecotypes across multiple mountain regions and can contribute to the diversification of plants in heavy-snow regions.

Influence of spatial distribution and size of clones on the realized outcrossing rate of the marsh cinquefoil (Comarum palustre).

BACKGROUND AND AIMS: Clonal growth is a common feature in flowering plants. As clone size increases, the selfing rate in self-compatible species is likely to increase due to more frequent geitono-pollination events (i.e. pollination among flowers within the same genet). This study investigated the breeding system of the marsh cinquefoil (Comarum palustre) and assessed spatial distribution of clones, clone size and architecture, and their effects on realized outcrossing rates. In addition, pollen dispersal was investigated in two patchy populations. METHODS: The species' breeding system was investigated under controlled conditions through hand pollinations (self- vs. cross-pollination). Using microsatellite markers, an assessment was made of the realized outcrossing rates and the genetic diversity in four natural populations, the clonal structure in two populations within five 15 × 15 m sampling plots following 0.5 × 0.5 m grids, and the pollen dispersal through paternity assignment tests in those two populations. KEY RESULTS: Comarum palustre is a self-compatible species but only presents a low rate of spontaneous self-pollination. The occurrence of inbreeding depression was not detected at the seed set stage (δ(SS) = 0.04). Clones were spatially clumped (A(C) = 0.60-0.80), with intermediate to no intermingling of the ramets (D(C) = 0.40-1.00). Genet size ranged from one to 171 ramets. Patchy populations had low outcrossing rates (t(m) = 0.33-0.46). Large clones showed lower outcrossing rates than small clones. Pollen dispersal mainly occurred within patches as only 1-7 % of the pollination events occurred between patches of >25 m separation. Seedling recruitment events were detected. CONCLUSIONS: Genet size together with distances between patches, through increasing geitono-pollination events, appeared to be important factors influencing realized outcrossing rates. The study also revealed seed flow allowing seedling recruitment, which may contribute to increasing the number of new patches, and potentially further enhance gene flow within populations.

Reproductive differentiation into sexual and apomictic polyploid cytotypes in Potentilla puberula (Potentilleae, Rosaceae).

BACKGROUND AND AIMS: Intraspecific reproductive differentiation into sexual and apomictic cytotypes of differing ploidy is a common phenomenon. However, mechanisms enabling the maintenance of both reproductive modes and integrity of cytotypes in sympatry are as yet poorly understood. This study examined the association of sexual and apomictic seed formation with ploidy as well as gene flow towards sexuals within populations of purely polyploid Potentilla puberula. METHODS: The study is based on 22 populations representing various combinations of five polyploid cytotypes (tetraploid-octoploid) from East Tyrol, Austria. Embryo ploidy and the endosperm/embryo ploidy ratio obtained by a flow cytometric seed screen were used to infer reproductive modes of seed formation and to calculate the male and female genomic contributions to the embryo and endosperm. Self-incompatibility (SI) patterns were assessed and a new indirect approach was used to test for the occurrence of intercytotype matings based on the variation in the male genomic contribution to sexually derived embryos on the level of developed seed. KEY RESULTS: Tetraploids formed seeds almost exclusively via sexual reproduction, whereas penta- to octoploids were preferentially apomictic. Non-random distribution of reproductive modes within maternal plants further revealed a tendency to separate the sexual from the apomictic mode among individuals. Self-incompatibility of sexuals indicated functionality of the gametophytic SI system despite tetraploidy of the nuclear genome. We found no indication for significant cross-fertilization of tetraploids by the high polyploids. CONCLUSIONS: The study revealed a rare example of intraspecific differentiation into sexual and apomictic cytotypes at the polyploid level. The integrity of the sexual tetraploids was maintained due to reproductive isolation from the apomictic higher polyploids. Functionality of the gametophytic SI system suggested that the tetraploids are functional diploids.

Native congeners provide biotic resistance to invasive Potentilla through soil biota.

Soil biota can facilitate exotic plant invasions and these effects can be influenced by specific phylogenetic relationships among plant taxa. We measured the effects of sterilizing soils from different native plant monocultures on the growth of Potentilla recta, an exotic invasive forb in North America, and conducted plant-soil feedback experiments with P. recta, two native congeners, a close confamilial, and Festuca idahoensis, a native grass species. We also reanalyzed data comparing the ability of P. recta to invade experimentally constructed congeneric monocultures vs. monocultures of a broad suite of non-congeners. We found that monocultures as a group, other than those of the native P. arguta, were highly invasible by P. recta. In contrast, this was not the case for monocultures of P. arguta. In our first experiment, the biomass of P. recta was 50% greater when grown in soil from F. idahoensis monocultures compared to when it was grown in soils from P. arguta or P. recta monocultures. Sterilizing soil from F. idahoensis rhizospheres had no effect on the biomass of P. recta, but sterilizing soil from P. arguta and P. recta rhizospheres increased the biomass of P. recta by 108% and 90%, respectively. In a second experiment, soil trained by F. idahoensis resulted in a positive feedback for P. recta. In contrast, soils trained independently by each of the two native Potentilla species, or the closely related Dasiphora (formerly Potentilla) resulted in decreases in the total biomass of the invasive P. recta indicating strong negative feedbacks. Soil trained by P. recta also resulted in intraspecific negative feedbacks. Our results demonstrate substantial negative feedbacks for an invader in its nonnative range under certain conditions, and that native congeners can mount strong biotic resistance to an invader through the accumulation of deleterious soil biota.

Root foraging increases performance of the clonal plant Potentilla reptans in heterogeneous nutrient environments.

BACKGROUND: Plastic root-foraging responses have been widely recognized as an important strategy for plants to explore heterogeneously distributed resources. However, the benefits and costs of root foraging have received little attention. METHODOLOGY/PRINCIPAL FINDINGS: In a greenhouse experiment, we grew pairs of connected ramets of 22 genotypes of the stoloniferous plant Potentilla reptans in paired pots, between which the contrast in nutrient availability was set as null, medium and high, but with the total nutrient amount kept the same. We calculated root-foraging intensity of each individual ramet pair as the difference in root mass between paired ramets divided by the total root mass. For each genotype, we then calculated root-foraging ability as the slope of the regression of root-foraging intensity against patch contrast. For all genotypes, root-foraging intensity increased with patch contrast and the total biomass and number of offspring ramets were lowest at high patch contrast. Among genotypes, root-foraging intensity was positively related to production of offspring ramets and biomass in the high patch-contrast treatment, which indicates an evolutionary benefit of root foraging in heterogeneous environments. However, we found no significant evidence that the ability of plastic foraging imposes costs under homogeneous conditions (i.e. when foraging is not needed). CONCLUSIONS/SIGNIFICANCE: Our results show that plants of P. reptans adjust their root-foraging intensity according to patch contrast. Moreover, the results show that the root foraging has an evolutionary advantage in heterogeneous environments, while costs of having the ability of plastic root foraging were absent or very small.

Prospects and limits of the flow cytometric seed screen--insights from Potentilla sensu lato (Potentilleae, Rosaceae).

The flow cytometric seed screen allows for identification of reproductive modes of seed formation and inference of the ploidy of contributing gametes. However, the lack of a mathematical formalization to infer male/female genomic contributions, and the prerequisite of a binucleate female contribution to the endosperm limits its applicability. We evaluated this assumption combining a DNA-based progeny survey with a comparison of the cytology of reproductive pathways co-occurring within single individuals representing 14 Potentilleae species from six phylogenetic lineages. A numerical framework valid for sexual and pseudogamous taxa was developed, enabling quantification of female and male genomes contributing to embryo and endosperm independent of gametophyte origins, numbers of sperm involved and ploidy of parents. The inference strongly depended on accurate peak index estimation. The endosperm of Potentilleae species received a binucleate female contribution in five evolutionary lineages whereas endosperm formation remained uncertain in the Tormentillae. A modified flow cytometric seed screen protocol was developed to cope with low endosperm contents. Evolutionary conservation of a binucleate female contribution to the endosperm suggested wide applicability of flow cytometric seed screen - at least in the Potentilleae. However, alternative progeny surveys and precise embryo/endosperm ploidy estimates are required for a comprehensive understanding of the cytology of seed formation.

Long-term functional plasticity in plant hydraulic architecture in response to supplemental moisture.

BACKGROUND AND AIMS: Plasticity in structural and functional traits related to water balance may determine plant performance and survival in ecosystems characterized by water limitation or high levels of rainfall variability, particularly in perennial herbaceous species with long generation cycles. This paper addresses whether and the extent to which several such seasonal to long-term traits respond to changes in moisture availability. METHODS: Using a novel approach that integrates ecology, physiology and anatomy, a comparison was made of lifetime functional traits in the root xylem of a long-lived perennial herb (Potentilla diversifolia, Rosaceae) growing in dry habitats with those of nearby individuals growing where soil moisture had been supplemented for 14 years. Traditional parameters such as specific leaf area (SLA) and above-ground growth were also assessed. KEY RESULTS: Individuals from the site receiving supplemental moisture consistently showed significant responses in all considered traits related to water balance: SLA was greater by 24 %; roots developed 19 % less starch storing tissue, an indicator for drought-stress tolerance; and vessel size distributions shifted towards wider elements that collectively conducted water 54 % more efficiently - but only during the years for which moisture was supplemented. In contrast, above-ground growth parameters showed insignificant or inconsistent responses. CONCLUSIONS: The phenotypic changes documented represent consistent, dynamic responses to increased moisture availability that should increase plant competitive ability. The functional plasticity of xylem anatomy quantified in this study constitutes a mechanistic basis for anticipating the differential success of plant species in response to climate variability and change, particularly where water limitation occurs.

Do clonal plants show greater division of labour morphologically and physiologically at higher patch contrasts?

BACKGROUND: When growing in reciprocal patches in terms of availability of different resources, connected ramets of clonal plants will specialize to acquire and exchange locally abundant resources more efficiently. This has been termed division of labour. We asked whether division of labour can occur physiologically as well as morphologically and will increase with patch contrasts. METHODOLOGY/PRINCIPAL FINDINGS: We subjected connected and disconnected ramet pairs of Potentilla anserina to Control, Low, Medium and High patch contrast by manipulating light and nutrient levels for ramets in each pair. Little net benefit of inter-ramet connection in terms of biomass was detected. Shoot-root ratio did not differ significantly between paired ramets regardless of connection under Control, Low and Medium. Under High, however, disconnected shaded ramets with ample nutrients showed significantly larger shoot-root ratios (2.8∼6.5 fold) than fully-lit but nutrient-deficient ramets, and than their counterparts under any other treatment; conversely, fully-lit but nutrient-deficient ramets, when connected to shaded ramets with ample nutrients, had significantly larger shoot-root ratios (2.0∼4.9 fold) than the latter and than their counterparts under any other treatment. Only under High patch contrast, fully-lit ramets, if connected to shaded ones, had 8.9% higher chlorophyll content than the latter, and 22.4% higher chlorophyll content than their isolated counterparts; the similar pattern held for photosynthetic capacity under all heterogeneous treatments. CONCLUSIONS/SIGNIFICANCE: Division of labour in clonal plants can be realized by ramet specialization in morphology and in physiology. However, modest ramet specialization especially in morphology among patch contrasts may suggest that division of labour will occur when the connected ramets grow in reciprocal patches between which the contrast exceeds a threshold. Probably, this threshold patch contrast is the outcome of the clone-wide cost-benefit tradeoff and is significant for risk-avoidance, especially in the disturbance-prone environments.

Assessing facilitative responses to a nurse shrub at the community level: the example of Potentilla fruticosa in a sub-alpine grassland of northwest China.

Biotic interaction studies have revealed a large discrepancy among experiments in target responses to the effects of neighbours, which may in part be due to both high species-specificity of plant responses and low number of target species used in experiments. Our aim was to assess facilitative responses at the community level and the role of both functional groups and ecological attributes of target species. In a sub-alpine grassland on the eastern Tibet plateau, we assessed growth responses of all species in the community to removal of a dominant shrub. We also measured changes in the main environmental variables. Species responses were analysed by functional group and in relation to their mean regional altitudinal distribution. All significant interactions were positive and affected one-third of the total species richness of the community. All functional groups were facilitated but forbs were less strongly facilitated than in the two other groups. High-alpine species were less strongly facilitated than low-sub-alpine species, but the strength of this relationship was weaker than that reported in previous work. There was evidence of a decrease in extreme temperatures below the canopy of the shrub but no variations in soil moisture. We conclude that the highly stressful conditions induced by the dry continental climate of the eastern Tibet plateau are a main driver of the exclusive dominance of positive interactions. Assessing interactive responses at the community level is likely to provide a useful tool to better understand the role of biotic interactions in community responses to environmental changes.

Common volatiles are major attractants for neonate larvae of the specialist flea beetle Altica koreana (Coleoptera: Chrysomelidae).

Olfactory stimuli play an important role in the host searching of larval phytophagous insects. Previous studies indicate that larvae that have to find feeding sites after hatching are generally attracted to host volatiles. However, there are few studies on the olfactory responses of neonate larvae to host volatiles in cases when those larvae hatched on the host plant. In the present study, we determined the olfactory responses of neonate larvae of the specialist flea beetle, Altica koreana Ogloblin, to host and six non-host plants, using a static-air "arena." Larvae responded significantly to the host plant Potentilla chinensis Ser. and five of six non-host plants, compared to the control. Larvae did not prefer the host plant over the non-host plants (except Artemisia sp.) when offered a choice. Additionally, odours of a non-host plant, which were unattractive to neonate larvae, may have masked the attractive odour of the host plant. These results indicate that common volatiles can play a major role in attracting larvae of this specialist to plants, but attraction to such odours may not be the major mechanism of host choice.

Intraspecific variations in seedling emergence and survival of Potentilla matsumurae (Rosaceae) between alpine fellfield and snowbed habitats.

Potentilla matsumurae has a wide distribution from wind-blown fellfields to snowbeds in alpine regions of Japan. The environmental factors influencing seedling establishment differ between the fellfield and snowbed habitats; plants growing in each habitat may therefore have different germination strategies. Using a reciprocal sowing experiment, patterns of seedling emergence and survivorship were examined in both habitat types in the Taisetsu Mountains, Japan. Seeds derived from a fellfield population germinated earlier than did those derived from a snowbed population at both habitats, and the germination of fellfield seeds continued throughout the growing season. The timing of seedling emergence greatly affected subsequent survival at the fellfield. Seedlings that emerged in the first half of the growing season had low survivorship during the first year because of frost and drought damage, but the remaining seedlings had high survivorship during the winter; seedlings that emerged in the latter half of the growing season showed the opposite trend. At the snowbed, seedling survival was high throughout the growing season. Germination experiments in the laboratory highlighted a difference in the sensitivity of seeds from the fellfield and snowbed populations to fluctuating temperatures. These results indicate that intraspecific variation in emergence and survivorship may occur over a small scale in an alpine environment.