Recent development of environmental DNA (eDNA) analysis allows us to survey underwater macro-organisms easily and cost effectively; however, there have been no reports on eDNA detection or quantification for jellyfish. Here we present the first report on an eDNA analysis of marine jellyfish using Japanese sea nettle (Chrysaora pacifica) as a model species by combining a tank experiment with spatial and temporal distribution surveys. We performed a tank experiment monitoring eDNA concentrations over a range of time intervals after the introduction of jellyfish, and quantified the eDNA concentrations by quantitative real-time PCR. The eDNA concentrations peaked twice, at 1 and 8 h after the beginning of the experiment, and became stable within 48 h. The estimated release rates of the eDNA in jellyfish were higher than the rates previously reported in fishes. A spatial survey was conducted in June 2014 in Maizuru Bay, Kyoto, in which eDNA was collected from surface water and sea floor water samples at 47 sites while jellyfish near surface water were counted on board by eye. The distribution of eDNA in the bay corresponded with the distribution of jellyfish inferred by visual observation, and the eDNA concentration in the bay was ~13 times higher on the sea floor than on the surface. The temporal survey was conducted from March to November 2014, in which jellyfish were counted by eye every morning while eDNA was collected from surface and sea floor water at three sampling points along a pier once a month. The temporal fluctuation pattern of the eDNA concentrations and the numbers of observed individuals were well correlated. We conclude that an eDNA approach is applicable for jellyfish species in the ocean.

When more than one closely related plant species share the same pollination niche, reproductive interference via interspecific pollen transfer should limit their coexistence. However, some studies have reported the sympatric coexistence of two native close relatives pollinated by the same pollinators under reproductive interference, even without niche partitioning. We examined the frequency dependency of reproductive interference between close relatives in natural conditions and the potential roles of autonomous selfing in mitigating the negative reproductive interference effects between congeneric species. We investigated sympatrically growing Commelina communis (Cc) and C. c. f. ciliata (Ccfc) populations. These species exhibit very large overlaps in habitat preference, flowering phenology and pollination niche, but seldom produce hybrids. First, we conducted a hand‐pollination experiment to examine the negative effects of heterospecific pollen deposition on seed production and the potential of self‐pollination to mitigate the effects in both species. Then, we examined the effects of reproductive interference on reproductive success and the potential for autonomous selfing in the field. We found significant negative effects of heterospecific pollen deposition on seed production and the mitigation effects of prior and competing self‐pollination, in both Cc and Ccfc. For both species in the field, intraspecific pollinator movements and reproductive success significantly decreased with an increase in the relative floral abundance of competing species, although the negative reproductive interference effect on reproductive success was lower in Cc than in Ccfc. We also found greater potential for prior autonomous selfing in Cc than in Ccfc. Our findings suggest that Cc flowers were less affected by reproductive interference from competing species, which was likely due to a higher prior selfing ability compared to Ccfc flowers. The asymmetry in susceptibility to reproductive interference may explain the Cc‐biased distribution in the study area. The study improves our understanding of how prior autonomous selfing can reduce the negative reproductive interference effect from competing species in mixed‐mating species with frequent pollinator visits. A plain language summary is available for this article.

Seminatural grasslands are ecosystems rich in biodiversity. However, their decline has been reported worldwide, and identification of grasslands with high conservation priority is urgently required. Recently, an increasing number of studies have reported that past vegetation history affects current biological communities. To evaluate whether the temporal continuity of grasslands promotes biodiversity, and thus can be an indicator of conservation priority, we studied vascular plant communities in old (160-1000s years) and new (52-70 years after deforestation) grasslands, as well as in forests, of Sugadaira Highland in central Japan. The number of plant species was highest in old grasslands, followed by new grasslands and forests. This pattern was much clearer in the number of grassland-dependent native and grassland-dependent endangered species, indicating the role of old grasslands as refugia for those species. The species composition differed between old and new grasslands. New grasslands had species compositions in between those of old grasslands and forests, suggesting that the plant community in new grasslands retains the influence of past forestation for more than 52 years after deforestation. Eleven indicator species were detected in old grasslands, but none in new grasslands, suggesting the uniqueness of the plant community in old grasslands. We conclude that the temporal continuity of grasslands increases plant diversity and can be an indicator of grasslands with high conservation priority.

Summary Despite the well‐known visual attraction function of angiosperm petals, additional roles of these floral organs (e.g. the provision of landing‐site platforms for pollinators) have rarely been examined. This is likely because most petals perform multiple functions, making it difficult to isolate the importance of landing sites in pollination success. We investigated the landing‐site function of dull‐coloured pinnately branched petals in Mitella pauciflora flowers, which are predominantly pollinated by fungus gnats. We conducted a field experiment, in which the effects of experimental petal removal on pollinators’ approach, landing and visit duration and floral reproductive success were examined in naturally pollinated flowers. According to direct and time‐lapse camera observations, petal removal did not influence pollinators’ approach frequency or visit duration, but did significantly decrease their landings. Fruit set and pollen dispatch both significantly decreased with petal removal, indicating that petals promote female and male reproductive success in M. pauciflora by facilitating pollinator landing. This demonstrates that inconspicuous petals primarily have a landing‐site function rather than a visual attraction function in M. pauciflora. Discriminating between diverse petal functions is a challenging problem, and new approaches are required to elucidate the functional features of angiosperm flowers. A http://onlinelibrary.wiley.com/doi/10.1111/1365-2435.12842/suppinfo is available for this article.

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The pollination and reproductive success of flowering plants can be negatively influenced in various ways by neighbouring heterospecific plants, such as resource competition and reproductive interference. We hypothesized that covering together with shading by neighbouring plants may reduce pollinator visits to and reproductive success of plants by reducing floral attractiveness and pollinator activity and by interrupting flower access, respectively. To test this hypothesis, we examined whether shaded and covered flowers suffered from pollinator limitation and low reproductive success in a population of the dwarf herb Lithospermum zollingeri, which co‐exists with woody and herbaceous plants in anthropogenically maintained forest edge meadows. Here, shaded and covered flowers were defined as those beneath the shade of the woods and those whose front portion was covered by any vegetative part of neighbouring plants, respectively. The shaded and covered flowers were visited by significantly fewer pollinators than sunlit and open flowers in the field. However, three major pollinator species responded differently to shading and covering. Significant pollen limitation reduced seed set in covered flowers, and shaded flowers produced fewer seeds. Pollen removal from the anthers was not influenced by shading or covering. Our study demonstrates the negative effects of covering on pollinator visits and seed production. It also elucidates the negative effects of shading on reproductive success in L. zollingeri, which depends on managed semi‐natural conditions. Land management abandonment, which has increased shaded and covered conditions in artificial forest edge meadows and open forest floors, might promote a rapid reduction in the populations of such dwarf plants.

Pollinator‐mediated reproductive interference can occur when two or more plant species share the same pollinators. Recent studies have suggested that prior autonomous selfing mitigates reproductive interference, potentially facilitating coexistence even in the absence of pollination niche partitioning (i.e. the pre‐emptive selfing hypothesis). However, whether the evolution of prior selfing promotes coexistence, in the context of the eco‐evolutionary dynamics of population size, selfing rates and inbreeding depression, remains poorly understood. We constructed an individual‐based model to examine the conditions under which the evolution of prior selfing promotes coexistence in the context of mutual reproductive interference. In the model, two plant species compete by way of mutual reproductive interference, and both have the potential to evolve the capacity for prior autonomous selfing. We expected that purging of deleterious mutations might result in evolutionary rescue, assuming that the strength of inbreeding depression declines as the population selfing rate increases; this would enable inferior competitors to maintain population density through the evolution of prior selfing. Our simulation demonstrated that evolution of prior selfing may promote coexistence, whereas reproductive interference in the absence of such evolution results in competitive exclusion. We found that lower pollinator availability is likely to favour rapid evolutionary shifts to higher prior selfing rates, thereby neutralising the negative effects of reproductive interference in both species. When the strength of inbreeding depression decreased with an increase in the population‐level selfing rate, moderate pollinator availability resulted in long‐term coexistence in which relative abundance‐dependent selection on the prior selfing rate served to intermittently maintain the population density of the inferior competitor. Synthesis. We demonstrate that the evolution of prior selfing may increase population growth rates of inferior competitors and may consequently promote long‐term coexistence via an evolutionary rescue. This constitutes a novel mechanism explaining the co‐evolutionary coexistence of closely related plant species without niche partitioning, and is consistent with recent studies reporting that closely related species with mixed mating systems can co‐occur sympatrically, even under conditions of mutual reproductive interference.

When the two or more plants species share the same pollinators, pollinator-mediated reproductive interference make coexistence difficult. Recent studies suggested prior autonomous selfing mitigate reproductive interference, could enabling coexistence without pollination niche partitioning (pre-emptive selfing hypothesis). However, there are no studies to test whether evolution of prior selfing promote the coexistence, considering eco-evolutionary dynamics of population size, selfing rate and inbreeding depression.To examine conditions that the evolution of prior selfing promote coexistence under mutual reproductive interference especially in the point of view for pollinator availability and dynamics of inbreeding depression, we constructed individual-based model in which two plant species compete against each other in the form of mutual reproductive interference and can evolve prior autonomous selfing rate. We expected that purging of deleterious mutations could cause evolutionary rescue because inferior species could rescue population density through the evolution of prior selfing if the strength of inbreeding depression decreases with an increase of population’s selfing rate.Our simulation demonstrated that the evolution of prior selfing could promote the coexistence while reproductive interference caused competitive exclusion without evolution. We found that lower pollinator availability tended to prefer rapid evolutionary shift to higher prior selfing rate, it neutralizes the negative effect of reproductive interference, and population dynamics exhibit neutral random walk in both species. When the strength of inbreeding depression decreased with an increase in population’s selfing rate, moderate pollinator availability resulted in long-term coexistence in which relative-abundance-dependent selection on the prior selfing rate rescue population density of inferior species intermittently.Synthesis. We showed that the evolution of prior selfing could increase population growth rate of inferior species and consequently enable the long-term coexistence with evolutionary rescue. This is the new mechanisms explaining co-evolutionary coexistence of closely related plant species without niche partitioning and consistent with recent studies reported that closely related mixed-mating species are sympatrically growing even under the mutual reproductive interference.