Do plants adjust their sex allocation and secondary sexual morphology in response to their neighbours?

Background and aims Changes in the sex allocation (i.e. in pollen versus seed production) of hermaphroditic plants often occur in response to the environment. In some homosporous ferns, gametophytes choose their gender in response to chemical cues sent by neighbours, such that spores develop as male gametophytes if they perceive a female or hermaphrodite nearby. Here it is considered whether a similar process might occur in the androdioecious angiosperm species Mercurialis annua, in which males co-occur with hermaphrodites; previous work on a Spanish population of M. annua found that individuals were more likely to develop as males at high density. Methods Using a novel approach to treat plants with leachate from pots containing males or hermaphrodites of M. annua, the hypothesis that individuals assess their mating opportunities, and adjust their sex expression accordingly, was tested through an exchange of chemical cues through the soil. Key Results For the population under study, from Morocco, no evidence was found for soil-signal-dependent sex expression: neither sex ratios nor sex allocation differed among experimental treatments. Conclusions The results imply either that the Moroccan population under study behaves differently from that previously studied in Spain (pointing to potential geographical variation in plasticity for sex expression), or that our method failed to capture the signals used by M. annua for adjustment of sex expressio

Environmental variation in sex ratios and sexual dimorphism in three wind‐pollinated dioecious plant species

Variation in plant sex ratios is often attributable to sex-specific mortality in heterogeneous environments that differentially limit male and female plant reproduction. Yet sexual dimorphism and plastic responses to environmental heterogeneity are common and may co-vary with variation in sex ratios. Here, we show that the sex ratio and the degree of sexual dimorphism for a number of plant traits varied along climatic and elevation gradients in three wind-pollinated dioecious species, Rumex lunaria, Urtica dioica and Salix helvetica. Some of the observed sex-specific responses to climatic variation are consistent with greater sensitivity of females to water scarcity, but most responses rather point to the greater sensitivity of males to ecological stress, consistent with larger male reproductive effort, as has been commonly reported for wind-pollinated plants. In contrast, we found no evidence for variation in either sex ratios or sexual dimorphism expected under sexual selection. Interestingly, sex ratios and sexual dimorphism varied both along distinct and the same ecological axes of variation, suggesting that the evolution of sexual dimorphism in the measured traits was not sufficient to prevent sex-specific mortality

Simulated herbivory enhances leaky sex expression in the dioecious herb Mercurialis annua

BACKGROUND AND AIMS: Plant reproductive traits are widely understood to be responsive to the selective pressures exerted by pollinators, but there is also increasing evidence for an important role for antagonists such as herbivores in shaping these traits. Many dioecious species show leaky sex expression, with males and females occasionally producing flowers of the opposite sex. Here, we asked to what extent leakiness in sex expression in Mercurialis annua (Euphorbiaceae) might also be plastically responsive to simulated herbivory. This is important because enhanced leakiness in dioecious populations could lead to a shift in both the mating system and in the conditions for transitions between combined and separate sexes. METHODS: We examined the effect of simulated herbivory on the sexual expression of males and females of M. annua in two experiments in which different levels of simulated herbivory led to enhanced leakiness in both sexes. KEY RESULTS: We showed that leaky sex expression in both males and females of the wind-pollinated dioecious herb M. annua is enhanced in response to simulated herbivory, increasing the probability for and the degree of leakiness in both sexes. We also found that leakiness was greater in larger females but not in larger males. CONCLUSIONS: We discuss hypotheses for a possible functional link between herbivory and leaky sex expression, and consider what simulated herbivory-induced leakiness might imply for the evolutionary ecology of plant reproductive systems, especially the breakdown of dioecy and the evolution of hermaphroditism

Exogenous selection shapes germination behaviour and seedling traits of populations at different altitudes in a Senecio hybrid zone

Background and Aims The Senecio hybrid zone on Mt Etna, Sicily, is characterized by steep altitudinal clines in quantitative traits and genetic variation. Such clines are thought to be maintained by a combination of ‘endogenous' selection arising from genetic incompatibilities and environment-dependent ‘exogenous' selection leading to local adaptation. Here, the hypothesis was tested that local adaptation to the altitudinal temperature gradient contributes to maintaining divergence between the parental species, S. chrysanthemifolius and S. aethnensis. Methods Intra- and inter-population crosses were performed between five populations from across the hybrid zone and the germination and early seedling growth of the progeny were assessed. Key Results Seedlings from higher-altitude populations germinated better under low temperatures (9-13 °C) than those from lower altitude populations. Seedlings from higher-altitude populations had lower survival rates under warm conditions (25/15 °C) than those from lower altitude populations, but also attained greater biomass. There was no altitudinal variation in growth or survival under cold conditions (15/5 °C). Population-level plasticity increased with altitude. Germination, growth and survival of natural hybrids and experimentally generated F1s generally exceeded the worse-performing parent. Conclusions Limited evidence was found for endogenous selection against hybrids but relatively clear evidence was found for divergence in seed and seedling traits, which is probably adaptive. The combination of low-temperature germination and faster growth in warm conditions might enable high-altitude S. aethnensis to maximize its growth during a shorter growing season, while the slower growth of S. chrysanthemifolius may be an adaptation to drought stress at low altitudes. This study indicates that temperature gradients are likely to be an important environmental factor generating and maintaining adaptive divergence across the Senecio hybrid zone on Mt Etn

Small-scale and regional spatial dynamics of an annual plant with contrasting sexual systems

Plant demography is known to depend on both spatial dynamics and life history, but how these two factors interact is poorly understood. We conducted a longitudinal study of the wind-pollinated annual plant Mercurialis annua that varies geographically in its sexual system to investigate this interaction. Metapopulation demographic models predict that regular population turnover should be a more common feature of monomorphic than dimorphic populations because males and females cannot found new populations by selfing but hermaphrodites can. We tested the prediction that rates of population turnover would be higher in monomorphic compared to dimorphic regions. We surveyed 356 populations of M. annua along five regional transects in Morocco and the Iberian Peninsula over a 3-year period to examine their demography and persistence. Each transect crossed a transition in the sexual system, from a monomorphic region where almost all populations were hermaphroditic to a dimorphic one in which most populations had separate sexes (males with females or hermaphrodites). As predicted, rates of local apparent extinctions (i.e., the disappearance of adult plants) were nearly 50% higher in monomorphic compared to dimorphic regions. Local extinctions appeared to be driven by changes in vegetation cover, with extinctions tending to occur in sites in which perennial cover also declined. This suggests that disturbance is a primary agent of local extinctions. We further examined the influence of regional dynamics on local demographic properties by investigating patterns of spatial autocorrelation in population density across years. We found positive spatial autocorrelations in plant densities within regions for both sexual systems. However, these positive autocorrelations extended over shorter distances in monomorphic regions, perhaps as a result of greater population flux in these regions. Synthesis. Our study shows that population dynamics may be influenced by processes acting at a range of spatial scales: within patches, across patches within sites, and across sites within regions, as well as by life-history variation. In Mercurialis annua, regional variation in apparent extinction rates is affected by life history and implicated in regulating the geographical distribution of populations with different sexual systems

Heritabilities of lateral and vertical herkogamy in Lysimachia arvensis

Herkogamy, spatial separation between stigma and anthers within a flower, is important in regulating plant-mating systems. We studied phenotypic variation and heritability of herkogamy traits in Lysimachia arvensis (=Anagallis arvensis) that show both lateral and vertical herkogamy in the same flower, a rare strategy in flowering plants. Both lateral and vertical herkogamy showed continuous variation in 15 natural populations. Lateral herkogamy, measured as the angle between style and stamens, ranged from 5.6 to 66.5°; vertical herkogamy ranged from reverse to approach herkogamy. Herkogamy traits were constant within plants but variable among plants and populations. Flowers with marked lateral herkogamy showed mainly reverse herkogamy, whereas flowers with low lateral herkogamy showed mainly approach herkogamy. Both herkogamy traits showed a high degree of narrow sense heritability (h2 = 0.843 for lateral and h2 = 0.635 for vertical herkogamy). We discuss the possibility that variation in both herkogamy traits among populations of L. arvensis is a consequence of differential selective pressures under different pollination environments.Ministerio de Economía y Competitividad BES-2013-062859, CGL2015-63827, CGL2012-3327

Two's Company, Three's a Crowd: Experimental Evaluation of the Evolutionary Maintenance of Trioecy in Mercurialis annua (Euphorbiaceae)

Trioecy is an uncommon sexual system in which males, females, and hermaphrodites co-occur as three clearly different gender classes. The evolutionary stability of trioecy is unclear, but would depend on factors such as hermaphroditic sex allocation and rates of outcrossing vs. selfing. Here, trioecious populations of Mercurialis annua are described for the first time. We examined the frequencies of females, males and hermaphrodites across ten natural populations and evaluated the association between the frequency of females and plant densities. Previous studies have shown that selfing rates in this species are density-dependent and are reduced in the presence of males, which produce substantially more pollen than hermaphrodites. Accordingly, we examined the evolutionary stability of trioecy using an experiment in which we (a) indirectly manipulated selfing rates by altering plant densities and the frequency of males in a fully factorial manner across 20 experimental plots and (b) examined the effect of these manipulations on the frequency of the three sex phenotypes in the next generation of plants. In the parental generation, we measured the seed and pollen allocations of hermaphrodites and compared them with allocations by unisexual plants. In natural populations, females occurred at higher frequencies in denser patches, a finding consistent with our expectations. Under our experimental conditions, however, no combination of plant densities and male frequencies was associated with increased frequencies of females. Our results suggest that the factors that regulate female frequencies in trioecious populations of M. annua are independent of those regulating male frequencies (density), and that the stable co-existence of all three sex phenotypes within populations is unlikely

Revisiting a pollen-transmitted ilarvirus previously associated with angular mosaic of grapevine

We report the characterization of a novel tri-segmented RNA virus infecting Mercurialis annua, a common crop weed and model species in plant science. The virus, named "Mercurialis latent virus" (MeLaV) was first identified in a mixed infection with the recently described Mercurialis orthotospovirus 1 (MerV1) on symptomatic plants grown in glasshouses in Lausanne (Switzerland). Both viruses were found to be transmitted by Thrips tabaci, which presumably help the inoculation of infected pollen in the case of MeLaV. Complete genome sequencing of the latter revealed a typical ilarviral architecture and close phylogenetic relationship with members of the Ilarvirus subgroup 1. Surprisingly, a short portion of MeLaV replicase was found to be identical to the partial sequence of grapevine angular mosaic virus (GAMV) reported in Greece in the early 1990s. However, we have compiled data that challenge the involvement of GAMV in angular mosaic of grapevine, and we propose alternative causal agents for this disorder. In parallel, three highly-conserved MeLaV isolates were identified in symptomatic leaf samples in The Netherlands, including a herbarium sample collected in 1991. The virus was also traced in diverse RNA sequencing datasets from 2013-2020, corresponding to transcriptomic analyses of M. annua and other plant species from five European countries, as well as metaviromics analyses of bees in Belgium. Additional hosts are thus expected for MeLaV, yet we argue that infected pollen grains have likely contaminated several sequencing datasets and may have caused the initial characterization of MeLaV as GAMV

Female sterility associated with increased clonal propagation suggests a unique combination of androdioecy and asexual reproduction in populations of Cardamine amara (Brassicaceae)

Background and Aims The coexistence of hermaphrodites and female-sterile individuals, or androdioecy, has been documented in only a handful of plants and animals. This study reports its existence in the plant species Cardamine amara (Brassicaceae), in which female-sterile individuals have shorter pistils than seed-producing hermaphrodites. Methods Morphological analysis, in situ manual pollination, microsatellite genotyping and differential gene expression analysis using Arabidopsis microarrays were used to delimit variation between female-sterile individuals and hermaphrodites. Key Results Female sterility in C. amara appears to be caused by disrupted ovule development. It was associated with a 2.4- to 2.9-fold increase in clonal propagation. This made the pollen number of female-sterile genets more than double that of hermaphrodite genets, which fulfils a condition of co-existence predicted by simple androdioecy theories. When female-sterile individuals were observed in wild androdioecious populations, their ramet frequencies ranged from 5 to 54%; however, their genet frequencies ranged from 11 to 29%, which is consistent with the theoretically predicted upper limit of 50%. Conclusions The results suggest that a combination of sexual reproduction and increased asexual proliferation by female-sterile individuals probably explains the invasion and maintenance of female sterility in otherwise hermaphroditic populations. To our knowledge, this is the first report of the coexistence of female sterility and hermaphrodites in the Brassicacea