Is ovule packaging strategy in animal-pollinated plants correlated to the level of specialization of their pollination systems?

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Katarzyna Roguz, Justyna Ryniewicz, Anna Szaciłło, Robert Junker, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8038291/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract The number of ovules per flower varies over several orders of magnitude among angiosperms, with some families exhibiting remarkable conservatism and others displaying extensive variation among closely related species. Numerous hypotheses have been advanced to explain plant strategies in ovule packaging, often linking this trait to the statistical dispersion of pollen receipt. Given that, in animal-pollinated plants, the pollen load received depends on flower visitor identity, abundance, and efficiency, we hypothesized that ovule packaging should be directly related to the pollination system. Specifically, we predicted that (1) specialist plants, serviced by a few efficient floral visitors, would bet-hedge on rare visits by producing more ovules per flower and thereby maximizing seed output when pollination is successful, (2) compared to generalists that operate in more predictable pollinator environments and consequently produce fewer ovules per flower. To test our hypothesis, we analyzed literature-derived data encompassing 192 zoogamous plant species representing 61 taxonomic families. We observed fewer ovules with greater visitor diversity in trees compared to perennials, but these effects disappeared under phylogenetic models, which showed that ovule number has a strong phylogenetic signal. These findings indicate that while pollination system characteristics may influence ovule packaging strategies, they do not fully account for the observed variation. Instead, ovule number appears subject to strong and conflicting selective pressures, suggesting that the generalization/specialization of pollination systems does not singularly explain its evolution. Angiospermae ovule bet-hedging evolution flower generalization meta-analysis sexual reproduction plant-pollinator network Evolutionary ecology Figures Figure 1 Introduction The number of ovules, one of the most important floral traits determining plants' reproductive success, exhibits significant variability among angiosperms. Despite the importance of ovule production for species reproduction and evolution, this trait is variable both among the basal lineages (Doyle and Endress 2000 ) and among the derived plant families (e.g., Matthews et al. 1999 ; Potter et al. 2007 ). It can vary over several orders of magnitude, being conservative in some plant families (e.g., Apiaceae, Boraginaceae, or Lamiaceae) or very flexible across closely related species (e.g., in Rosaceae; Stebbins 1974 ; Burd 1995 ; Herrera 1999 ; Burd et al. 2009 ; Banks et al. 2010 ; Arroyo et al. 2019 ), and even among populations of the same species (Herrera 1999 ; Chalcoff and Aizen 2016 ; Arena et al. 2018 ). Strategies in ovule packaging, i.e., the number of ovules produced by a single flower, are a long-discussed topic in plant evolutionary biology, resulting in numerous hypotheses explaining the evolution of the phenomenon. Possible explanations include, among all, features related to plant physiology, e.g., resource allocation (Lloyd 1980 ; Campbell et al. 2022 ), quantity of the pollen tube transmitting tissue (Matthews et al. 1999 ), flower size (Arroyo et al. 2019 ), and regulation of offspring quality via seed abortion (Kozłowski and Stearns 1989 ). Also, the life form may potentially shape ovule number through a complex interplay of resource allocation strategies, reproductive assurance mechanisms, and/or environmental adaptations (Cruden and Lyon 1989 ; Plitmann and Levin 1990 ; Jürgens et al. 2002 ). There are studies suggesting the presence of a correlation between life form and breeding system (e.g., Wyatt et al. 2000 ; Lord and Westoby 2012 ). The available data indicate that long-lived plants like trees often produce flowers with a limited number of ovules, optimizing reproductive success over extended lifespans. While annuals, which have shorter life cycles, tend to produce numerous flowers with many ovules, to maximize seed output within a single growing season (Cruden and Lyon 1989 ). The number of ovules also depends on several ecological factors acting on the species level, like sibling- (Kress 1981 ) and parent-offspring competition (Shaanker et al. 1988 ; Greenway and Harder 2007 ), but also on species interactions, e.g., with pollinators (Friedman and Barrett 2011 ; Arroyo et al. 2019 ). Available data show that in the case of plant-pollinator interaction, several factors can play an important role in shaping the number of ovules, e.g., pollinators efficiency, time of flowering, or mode of pollination (Friedman and Barrett 2011 ; Arroyo et al. 2019 ; Bochynek and Burd 2024 ; Harder and Johnson 2025 ). Finally, the number of ovules may be explained by a way of exploiting unpredictably favorable conditions for seed ripening (Lee 1988 ) or seed dispersal (fleshy fruits vs. wind-dispersed propagules (Burd 1995 ). Burd ( 1995 ) proposed that the evolution of ovule number may be driven by the probability distribution of pollination success (measured, e.g., as the mean receipt of pollen tubes), suggesting that producing higher ovule numbers is a bet-hedging strategy to deal with unpredictable pollen receipt (Knight et al. 2005 ) or uncertain environmental factors (Philippi and Seger 1989 ). Indeed, a large meta-analysis by Burd et al. ( 2009 ), confirmed a clear positive relationship between ovule number per flower and probability of pollen receipt (assessed by counting stigmatic pollen loads). Also, a recent analysis from the Andean communities showed a positive correlation between the number of ovules and the probability of efficient pollination. This tendency may be, however, true only for habitats with especially low and variable pollination rates - greater variability in stigmatic pollen loads is likely in species pollinated by efficient pollinators because relatively more pollen will be deposited on the stigmas of flowers that have any visitors in comparison with those that fail to be visited (Arroyo et al. 2019 ). The overall explanation, however, most likely reflects selection acting simultaneously on many of the above-mentioned factors. In the case of animal-pollinated plants, stigmatic pollen load is a function of flower visitor identity, abundance and efficiency on flowers (e.g., Wilson and Thomson 1991 ; Thøstesen and Olesen 1996 ; Adler and Irwin 2006 ; Sahli and Conner 2007 ; Fumero-Cabán and Meléndez-Ackerman 2007 ; Junker et al. 2010 ; Sánchez-Lafuente et al. 2012 ; Zych et al. 2013a ; Kulbaba et al. 2014 ; Parker et al. 2016 ). Since pollinators vary greatly in their dependence on a particular plant species, a successful transfer of pollen grains to the conspecific stigma can result from both highly generalist or very specialist interactions (or anything in between; Ollerton 1996 ; Waser et al. 1996 ; Johnson and Steiner 2000 ; Willmer 2011 ). In the first instance, flowers are visited relatively frequently by taxonomically wide pollinator assemblages which, however, mainly include highly generalist foragers (e.g., Apis mellifera ) and/or largely opportunistic species such as hoverflies and beetles (Willmer 2011 ; Zych et al. 2014 ; Inouye et al. 2015 ; Hung et al. 2018 ). In such cases, the size of conspecific pollen loads carried by insects may vary greatly (see e.g. Zych et al. 2019 ). Furthermore, mass flowering plants serviced by super generalist pollinators can be composed of low-quality self-pollen (Diller et al. 2022 ). In more specialized interactions, such as those observed in the Orchidaceae family, the overall visitation frequency tends to be lower and extremely variable, compared to generalist interactions, however, the proportion of conspecific pollen loads increases, so the natural selection is expected to favor strategies allowing the production of a higher number of seeds after a single visit (Schreiber et al. 2015 ). This compensates for the unpredictable nature of pollination events in specialized interactions. A meta-analysis of orchids found that while pollination success is highly variable and often low, when pollination does occur, orchids tend to maximize reproductive output by producing a large number of seeds per pollination event, partly because relying on pollinia, which transfer large pollen loads efficiently (Tremblay et al. 2005 ). According to the bet-hedging hypothesis, ovule packaging in animal-pollinated plants should reflect the pollination system, specifically, the number and diversity of potential pollinators. Specialist systems, which channel pollen more directly among conspecifics, tend to deliver more efficient pollen transfer per visit. In such cases, plants serviced by one or a few efficient visitors may produce many seeds from a single visit, but successful visits are relatively rare. The optimal strategy, therefore, is to produce more ovules per flower to capitalize on infrequent yet efficient visits and to buffer against the likely failure of other flowers on the same plant to receive pollen (Burd 1995 ; Burd et al. 2009 ; Rosenheim et al. 2016 ). By contrast, generalists should produce fewer ovules per flower: each visit yields fewer seeds, but a large pool of potential pollinators increases visitation rates and compensates for per-visit inefficiency. Although direct tests linking visitor diversity to variance in pollen receipt remain scarce, large-scale analyses indicate that greater wild insect diversity stabilizes pollinator abundance across years, implying more consistent service and supporting this expectation (Senapathi et al. 2021 ). Recent evidence from an elevational gradient in the South American Andes is consistent with bet-hedging theory: ovule number increased with elevation, peaking where pollination is most unpredictable, with early-flowering species also having higher ovule number (Arroyo et al. 2019 ). Arroyo et al. further proposed that higher ovule number should be associated with more specialized pollination, but their alpine dataset did not confirm this, likely because in that specific environment, generalization is the more advantageous strategy. In the present paper, we sought evidence for the more universal application of this assumption. To do so, we check if the (1) generalization/specialization in the pollination system, (2) life-form, and (3) phylogenetic relationships can explain some of the observed variation in ovule numbers. We tested these assumptions on our pollination network studies and literature-derived data for 192 zoogamous plant species representing 61 taxonomic families. Since detailed analyses of pollination effectiveness of animal visitors are rarely reported,d and comparing the performance of pollinators in pollination is problematic (see e.g., Ne’Eman et al. 2010 )), as a measure of generalization/specialization we used the number of different species of floral visitors (the alpha diversity), treating all recorded animal visitors as legitimate and equally efficient pollinators. Materials and methods We performed a literature survey for data on the number of ovules per flower and pollinators of angiosperm plant species. In our investigation, we used Google Scholar, employing keyword combinations such as "number of ovules", "ovule packaging", along with terms "floral visitors", "pollination", and "pollinators". In cases where we encountered incomplete data (lacking information on number of ovules or insect visitors), we strived to fill in gaps by obtaining data from other studies. Our search encompassed available literature published until 2016. In cases where various sources reported different values of the ovule number per flower, we used means calculated for all available data. For data on floral visitors, we used α-diversity, and if several papers reported different numbers of taxa we chose the highest available result noting also, if available, taxonomic diversity of animals (in the case of insects at the rank of order and for vertebrates at the rank of class). This initial dataset was accompanied by data on floral visitors from additional two datasets: (1) a survey of the pollination network 26 plant species and their floral visitors) from a lowland meadow in NE Poland described in Goldstein and Zych (Goldstein and Zych 2016 ), and (2) fallow land pollination network (7 plant species and their floral visitors) reported by Junker et al. 2013 (Junker et al. 2013 ). For 27 plant species from the above networks lacking literature records of ovule numbers, in the summer of 2016, we collected flowers on-site and preserved them in 70% EtOH for further analysis of their ovaries’ content in the lab. For each species enlisted in Appendix 1, this was done for 20 fully developed flowers from at least 10 individual plants. Ovules from dissected flowers were counted under the stereoscopic microscope. Plants used in our database were classified according to the life form as annuals, biennials, perennials, trees, or shrubs. Our survey resulted in data for 192 zoogamous plant species representing 61 taxonomic families of the global flora (source papers from which the final data were extracted are listed in Appendix 2). As our dataset includes three plant families with distinctive pollination systems or floral architecture: Orchidaceae, Asclepiadaceae, and Asteraceae, the final analysis was done in several variants (1) database with all species, (2) database with Orchidaceae and Asclepiadaceae excluded, (3) database with Asteraceae excluded, and (4) database with Orchidaceae, Asclepiadaceae, and Asteraceae excluded. Orchids often have specialized interactions, low visitation frequency, high proportion of conspecific pollen loads (Faegri and Van Der Pijl 1966 ; Proctor et al. 1996 ), and are known to produce very numerous ovules in a single flower. However, pollination in the Orchidaceae family occurs generally via transfer of the whole pollen sacks (pollinia; packaged pollen), which is largely different from other flowering plant families (Dahlgren et al. 1985 ) but similar to Asclepiadaceae(Ollerton and Liede 1997 )). The Asteraceae, in turn, are characterized by very distinct inflorescences: numerous flowers are arranged in a compound capitulum (pseudanthium), which is a functional equivalent of a flower. Often, e.g., in coneflowers ( Echinacea purpurea ) or chamomile ( Chamomilla recutita ), the capitulum is composed of specialized rewardless ray (ligulate) florets, responsible for insect attraction, and nectariferous seed-producing disc florets (Wist and Davis 2006 ; Sulborska 2011 ). Although members of Asteraceae are very conservative in the number of ovules per flower (always one), we assumed that for legitimate comparisons, the whole inflorescence should be treated as a single unit of pollination. Because the literature data largely lacked information on the number of individual flowers forming capitula of various species, and this trait showed to have some correlation with the altitude in the study of Arroyo et al. (Arroyo et al. 2019 ), we also decided to exclude Asteraceae from our analysis. Full results of these exclusions are provided in Appendix 1. Statistical analyses All analyses were conducted in R (Posit Software, PBC 2025). To construct a phylogenetic tree as a framework for the Phylogenetic Generalized Least Squares (PGLS) analysis, we used a previously available phylogenetic tree from (Zanne et al. 2014 ; Supplementary Fig. 1). To assess whether ovule number variation significantly covaries with pollinator richness and life form, we performed both non-phylogenetically corrected and phylogenetically corrected analyses using Ordinary Least Squares (OLS) regression and PGLS regression (function gls from the nlm package; Pinheiro et al. 2025 ), respectively. To evaluate the extent to which trait variation is influenced by phylogenetic relationships, we estimated the phylogenetic signal for ovule number, pollinator richness, and life form. Specifically, we calculated Pagel’s λ and Blomberg’s K, which capture different aspects of phylogenetic signal: Pagel’s λ is widely used in community ecology, while Blomberg’s K is more suited for closely related species. Using both metrics allowed for a more robust interpretation of phylogenetic patterns within our dataset. Phylogenetic signal estimates were obtained using the phytools package in R (Revell 2024 ). Results Apart from the excluded Asteraceae and Orchidaceae, the most abundant plant families in our database were Fabaceae (14 species), Apiaceae, Caryophyllaceae, and Ranunculaceae (10 species each), and Lamiaceae (6 species). Most of the recorded herbaceous species were perennials (123 species) and annuals (13 species), while 10 species were assigned as biennials. The rest of the species were classified as shrubs (19 species) or trees (16 species). Some of the studied species were assigned to two categories, e.g., annuals and biennials. For example, Silene latifolia is mostly annual; however, it also occasionally occurs as a biennial plant (Appendix 2, Table 1 ). The average number of ovules in the studied species was 1132 ± 2978, with a median of 15, a minimum value of 1, and a maximum value of 10000 ovules. Across all species, the number of ovules per flower and visitor richness was negatively correlated with flower-visitor richness (Pearson’s r = − 0.26, p < 0.001; Fig. 1 . The analyses excluding Asteraceae, Orchidaceae, and/or Asclepiadaceae yielded qualitatively similar patterns (Appendix 1): non-phylogenetic effect sizes ranged from r = − 0.23 to − 0.31. Plant life form affected ovule number (ANOVA: F₄,₁₈₆ = 3.55, p = 0.01): trees had fewer ovules than perennial herbs (Tukey HSD p = 0.007). Visitor richness did not differ among life forms (ANOVA: F₄, 186 = 2.03, p = 0.09). However, we accounted for shared ancestry using PGLS (and, for comparison, OLS with the same model), we found no significant relationship between ovules per flower and visitor richness; this result was unchanged after excluding Asteraceae, Orchidaceae, Asclepiadaceae, or all three (Table 1 ). Based on AIC, PGLS consistently fit better than OLS, highlighting the importance of phylogenetic non-independence even though the slope did not differ from zero. Quantitative traits showed strong phylogenetic signal (ovules per ovary: Pagel’s λ = 0.92, p < 0.01; Blomberg’s K = 0.72, p = 0.001), and neither PGLS nor OLS detected an effect of life form on visitor richness (PGLS had the lower AIC; Table 2 ). Table 1 Ovule number vs. pollinator richness (OLS and PGLS). Relationships between ovule number and pollinator richness across taxonomic subsets. Linear models showed no significant association between pollinator richness and ovule number (all p > 0.09p > 0.09p > 0.09 in OLS and PGLS). Dataset Model Intercept (± SE) visitor_ric (± SE) t (slope) p (slope) Adj. R² AIC RSE (df); n All species OLS 1333 ± 251 -8.61 ± 5.48 -1.57 0.12 0.01 3596 2936 (189); 191 No Asteraceae OLS 1537 ± 284 -9.72 ± 5.97 -1.63 0.11 0.01 3145 3112 (164); 166 No Asclepiadaceae & Orchidaceae OLS 150 ± 62 * -0.74 ± 1.43 -0.51 0.61 0.00 2583 659 (161); 163 No Asteraceae, Asclepiadaceae & Orchidaceae OLS 1542 ± 291 -9.89 ± 6.87 -1.44 0.15 0.01 3074 3150 (160); 162 All species PGLS 1333 ± 251 -8.61 ± 5.48 -1.57 0.12 — 3223 2921 (189); 191 No Asteraceae PGLS 1537 ± 284 -9.72 ± 5.97 -1.63 0.11 — 2868 3093 (164); 166 No Asclepiadaceae & Orchidaceae PGLS 1338 ± 256 -8.83 ± 6.26 -1.41 0.16 — 3168 2951 (185); 187 No Asteraceae, Asclepiadaceae & Orchidaceae PGLS 1542 ± 291 -9.88 ± 6.87 -1.44 0.15 — 2813 3131 (160); 162 Table 2 Ovule number vs. life form (OLS and PGLS). Relationships between ovule number and life form across taxonomic subsets. Linear models showed no significant association between ovule number and life form (all p > 0.09p > 0.09p > 0.09 in OLS and PGLS). Dataset Model Intercept (± SE) life_formb (± SE) life_formp (± SE) life_forms (± SE) life_formt (± SE) Adj. R² / note AIC RSE (df); n All species OLS 64.8 ± 771 -6.29 ± 1195 1551 ± 811 † -11.14 ± 1006 -48.54 ± 1056 0.04; model p = 0.02 3593 2886 (186); 191 No Asteraceae OLS 75.4 ± 876 -2.54 ± 1386 1849 ± 923 -21.77 ± 1109 -59.17 ± 1159 0.06; model p = 0.01 3140 3036 (161); 166 No Asclepiadaceae & Orchidaceae OLS 64.8 ± 177 -6.29 ± 274 118 ± 189 -11.14 ± 231 -48.54 ± 242 -0.01; model p = 0.80 2587 662 (158); 163 No Asteraceae, Asclepiadaceae & Orchidaceae OLS 75.4 ± 883 -2.54 ± 1397 1918 ± 932 -21.77 ± 1117 -59.17 ± 1169 Adj. R² = 0.06; model p = 0.01 3067 3060 (157); 162 All species PGLS 64.8 ± 771 -6.29 ± 1195 1551 ± 811 † -11.14 ± 1006 -48.54 ± 1056 — 3219 2848 (186); 191 No Asteraceae PGLS 75.4 ± 876 -2.54 ± 1386 1849 ± 923 -21.77 ± 1109 -59.17 ± 1159 — 2863 2990 (161); 166 No Asclepiadaceae & Orchidaceae PGLS 64.8 ± 177 -6.29 ± 274 118 ± 189 -11.14 ± 231 -48.54 ± 242 — 2272 652 (158); 163 No Asteraceae, Asclepiadaceae & Orchidaceae PGLS 75.4 ± 883 -2.54 ± 1397 1918 ± 932 -21.77 ± 1117 -59.17 ± 1169 — 2806 3012 (157); 162 Discussion Our results show that phylogenetic constraints play an important role in shaping the ovule number per flower. Across a taxonomically diverse sample of species in our database, ovule number was not related to the degree of specialization or generalization in a plant’s pollination system. Although we initially found that flowers with a higher number of ovules per flower received visits from a less diverse set of pollinators than flowers with fewer ovules, this relationship was not supported when we accounted for phylogenetic non-independence in the analysis. Our results are therefore in contrast with the ovule bet-hedging hypothesis by Arroyo et al. 2019 (Arroyo et al. 2019 ), which, according to Burd et al. 1995 (Burd 1995 ), predicts that the unpredictability of the pollinator environment may select for an increase in ovule number. According to this hypothesis, plants that produce more ovules per flower might take advantage of rare pollination visits through higher stigmatic pollen loads, enabling them to produce more seeds. Our results revealed a correlation between ovule number and the life form of the studied species, which was also previously detected in other systems (Raven and Ra Yen 1979 ; Plitmann and Levin 1990 ). For example, in Caryophylloideae ( Agrostemma , Dianthus , Saponaria , Silene , and Vaccaria ), where ovule number was significantly higher in perennials compared to annual species (Jürgens et al. 2002 ). This may suggest that ovule number, floral visitor diversity, and life form evolved independently in the species studied. These traits may be influenced by different ecological or evolutionary factors that are not directly related. Specialized pollinators have been proposed as a prerequisite for the evolution of large ovule numbers because they promote more direct conspecific pollen transfer and reduce losses during transport or grooming, thereby increasing the odds of successful outcrossing (Willson 1979 ; Rademaker et al. 1997 ). Yet recent work suggests that pollinator dependence and pollination efficiency more strongly shape pollen production than ovule production: higher pollen output, and thus higher P/O ratios, may compensate for reduced or inefficient interactions, whereas ovule number is comparatively insensitive to variation in pollination efficiency or syndrome (Cunha et al. 2022 ; Nepal et al. 2023 ; Harder and Johnson 2025 ). Accordingly, environments with greater pollination uncertainty (e.g., generalist-dominated systems) are expected to favor increased pollen production rather than larger ovule numbers (Cunha et al. 2022 ; Nepal et al. 2023 ; Harder and Johnson 2025 ). Consistent with the role of mating structure, Burd ( 2025 ) further proposes that high pollination efficiency often deposits related pollen cohorts or repeatedly targets nearby recipients, generating local mating competition; this framework predicts low P/O ratios coupled with high correlated paternity. By contrast, in generalist Apiaceae the ovule number per ovary is developmentally fixed and low, and is decoupled from pollen-load quantity or quality. Visitor taxa vary widely in pollen-carrying capacity and often deliver mixed loads; despite high visitation (Zych 2007 ; Zych et al. 2014 , 2019 ), conspecific deposition can be limited because generalist flowers are serviced by equally generalist visitors. For example, in the supergeneralist Angelica sylvestris (Apiaceae), 66% of visitors carried mixed pollen (Zych et al. 2019 ), and in Ostericum palustre (Apiaceae), all guilds carried similar amounts of conspecific and heterospecific pollen (Zych et al. 2014 ); a similar pattern occurs in Polemonium caeruleum (Polemoniaceae; Zych et al. 2013b ). Even so, the low, fixed ovule numbers are typically sufficient to achieve a full seed set (e.g., Apiaceae: Zych et al. 2019 ). Consequently, visitor taxa are functionally equivalent in pollination service (Zamora 2000 ) and exert limited selective pressure toward specialization, aligning with the expectation that, under diffuse and variable pollination, selection acts more on pollen output than on ovule number (Gómez and Zamora 2006 ). Our results suggest that ovule production is primarily constrained by phylogeny, consistent with earlier work. For example, both simple-flowered species and species with pseudanthium in the central Chilean Andes show a strong phylogenetic signal in ovule number (Arroyo et al. 2019 ), and broad phylogenetic structuring of ovule development is evident in orchids (Mayer et al. 2021 ) By contrast, pollination systems appear more labile: closely related species can rely on different pollinators, indicating substantial evolutionary plasticity in pollination strategies (e.g., Niet et al. 2014 ; Smith and Kriebel 2018 ; Roguz et al. 2021 ). This asymmetry raises a key question: does ovule number constrain the evolution of pollination systems? Clades with a fixed, low ovule number, as in many Lamiaceae (Mabberley 2017 ), may have limited scope to adopt highly specialized pollination and instead tend toward generalized systems. When ovules are few, reproductive success hinges on efficient pollen transfer, favoring high visitation rates or highly effective pollinators. Species with few ovules may also evolve mitigating traits, such as self-compatibility or clonal reproduction, to ensure reproductive success. Therefore, ovule number may play a fundamental role in shaping the evolution of pollination systems by influencing pollinator dependence, interactions, and reproductive strategies under varying ecological conditions. It is important to note that a limitation of our dataset is assessing the variability in pollen receipt indirectly through the diversity of floral visitors rather than direct measurements of pollen deposition or seed set. While this approach provides insight into long-term pollination environments, it does not quantitatively capture variation in pollen delivery. Direct methods, such as measuring stigmatic pollen loads or seed-set (Burd et al. 2009 ), offer a more precise assessment but rely on the assumption that short-term observations represent long-term trends. In contrast, visitor diversity may better reflect the evolutionary pollination environment but does not measure variance in pollen receipt explicitly (Arroyo et al. 2019 ). This trade-off should be considered when interpreting our results. Overall, our results show an important role of phylogenetic constraints in the context of ovule production in a larger geographic context. Our results also follow a theory pointing at pollen production as an important response to pollinators' efficiency and reproductive strategy. It is still important, however, to call for a more in-depth study of the ovule oversupply in flowers also taking into account other evolutionary, ecological, and, perhaps, geographical explanations for this phenomenon. To shed more light on these questions further studies on a broader list of species are needed. Declarations The authors declare that they have no financial or non-financial interests that are directly or indirectly related to the work submitted for publication. Author Contribution MZ and RRJ conceived the idea; JR, AS, KR and MZ performed the literature survey, JR, AS and MZ made ovule counting, RRJ and KR performed statistical analyses, MZ, KR and RRJ drafted the first version of the manuscript, all authors contributed to the final version of the text. Acknowledgement We thank the organizers and participants of the 29th Annual Meeting of the Scandinavian Association of the Pollination Ecologists in Silkeborg, Denmark, for stimulating discussions that led to formulating the idea of this paper. Literature surveys and ovule counting was financed via the Statutory Research Programme of the Faculty of Biology, University of Warsaw. A short sabbatical of RRJ was financially supported by the University of Warsaw IDUB Program “Mentor”. References Adler LS, Irwin RE (2006) Comparison of Pollen Transfer Dynamics by Multiple Floral Visitors: Experiments with Pollen and Fluorescent Dye. Ann Bot 97:141–150. https://doi.org/10.1093/aob/mcj012 Arena ME, Lencinas MV, Radice S (2018) Variability in floral traits and reproductive success among and within populations of Berberis microphylla G. Forst., an underutilized fruit species. 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Plant Syst Evol 263:159–179. https://doi.org/10.1007/s00606-006-0493-y Zych M, Goldstein J, Roguz K, Stpiczyńska M (2013a) The most effective pollinator revisited: Pollen dynamics in a spring-flowering herb. Arthropod-Plant Interact 7:315–322. https://doi.org/10.1007/s11829-013-9246-3 Zych M, Junker RR, Nepi M et al (2019) Spatiotemporal variation in the pollination systems of a supergeneralist plant: Is Angelica sylvestris (Apiaceae) locally adapted to its most effective pollinators? Ann Bot 123:415–428. https://doi.org/10.1093/aob/mcy140 Zych M, Michalska B, Krasicka-Korczyńska E (2014) Myophily in the critically endangered umbelliferous plant Ostericum palustre Besser (Apiaceae). Plant Syst Evol 300:187–196. https://doi.org/10.1007/S00606-013-0870-2/TABLES/2 Zych M, Stpiczyńska M, Roguz K (2013b) Reproductive biology of the Red List species Polemonium caeruleum (Polemoniaceae). Bot J Linn Soc 173:92–107. https://doi.org/10.1111/boj.12071 Additional Declarations No competing interests reported. Supplementary Files Appendix1.docx Appendix 1. Results of studies testing ovule number vs pollination system in studied plant species with exclusion of selected plant families. Appendix2.xlsx Appendix 2. Table 1: The list of source papers from which the final data was extracted; Table 2: The final matrix of analyzed species contains data for 192 species representing 62 taxonomic families of the global flora. Supplementaryfigure1.jpeg Supplementary figures Supplementary figure 1. Phylogenetic relationship of the studied species and log ovule number. Relative values for each respective trait are reflected by the size of the symbol; the larger the symbol, the higher the value. 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1","display":"","copyAsset":false,"role":"figure","size":93349,"visible":true,"origin":"","legend":"\u003cp\u003eOvule number vs pollination system in studied plant species. The number of ovules per flower exhibited a negative correlation with the diversity of flower visitors (p = 0.001, r = -0.263).\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8038291/v1/1a55bbc05d9b232f5c754ad9.jpeg"},{"id":96603043,"identity":"47c383a3-9865-4212-a53a-1fbae0a0bb0c","added_by":"auto","created_at":"2025-11-24 09:06:19","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":712077,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8038291/v1/b139386a-70b2-4f19-98d2-7a2adba2b143.pdf"},{"id":96453153,"identity":"4710affc-7673-434e-accf-c4cc5e3f9606","added_by":"auto","created_at":"2025-11-21 09:58:32","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":144685,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAppendix 1. \u003c/strong\u003eResults of studies testing ovule number vs pollination system in studied plant species with exclusion of selected plant families\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Appendix1.docx","url":"https://assets-eu.researchsquare.com/files/rs-8038291/v1/423e668372c0ef90bac00468.docx"},{"id":96375656,"identity":"f47603cc-33bc-4264-9d0c-323d344575f6","added_by":"auto","created_at":"2025-11-20 11:13:07","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":98473,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAppendix 2.\u003c/strong\u003e Table 1: The list of source papers from which the final data was extracted; Table 2: The final matrix of analyzed species contains data for 192 species representing 62 taxonomic families of the global flora.\u003c/p\u003e","description":"","filename":"Appendix2.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-8038291/v1/d6dcf0773fcd31c216c39d48.xlsx"},{"id":96375648,"identity":"45b3c225-7619-4844-ac6e-56f4287f641a","added_by":"auto","created_at":"2025-11-20 11:13:06","extension":"jpeg","order_by":3,"title":"","display":"","copyAsset":false,"role":"supplement","size":529708,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary figures\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSupplementary figure 1. \u003c/strong\u003ePhylogenetic relationship of the studied species and log ovule number. Relative values for each respective trait are reflected by the size of the symbol; the larger the symbol, the higher the value.\u003c/p\u003e","description":"","filename":"Supplementaryfigure1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-8038291/v1/90f321993436bb3b594c12eb.jpeg"}],"financialInterests":"No competing interests reported.","formattedTitle":"Is ovule packaging strategy in animal-pollinated plants correlated to the level of specialization of their pollination systems?","fulltext":[{"header":"Introduction","content":"\u003cp\u003eThe number of ovules, one of the most important floral traits determining plants' reproductive success, exhibits significant variability among angiosperms. Despite the importance of ovule production for species reproduction and evolution, this trait is variable both among the basal lineages (Doyle and Endress \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) and among the derived plant families (e.g., Matthews et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Potter et al. \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). It can vary over several orders of magnitude, being conservative in some plant families (e.g., Apiaceae, Boraginaceae, or Lamiaceae) or very flexible across closely related species (e.g., in Rosaceae; Stebbins \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e1974\u003c/span\u003e; Burd \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Herrera \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Burd et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Banks et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and even among populations of the same species (Herrera \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Chalcoff and Aizen \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Arena et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). Strategies in ovule packaging, i.e., the number of ovules produced by a single flower, are a long-discussed topic in plant evolutionary biology, resulting in numerous hypotheses explaining the evolution of the phenomenon. Possible explanations include, among all, features related to plant physiology, e.g., resource allocation (Lloyd \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e1980\u003c/span\u003e; Campbell et al. \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), quantity of the pollen tube transmitting tissue (Matthews et al. \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e1999\u003c/span\u003e), flower size (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and regulation of offspring quality via seed abortion (Kozłowski and Stearns \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). Also, the life form may potentially shape ovule number through a complex interplay of resource allocation strategies, reproductive assurance mechanisms, and/or environmental adaptations (Cruden and Lyon \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Plitmann and Levin \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; J\u0026uuml;rgens et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). There are studies suggesting the presence of a correlation between life form and breeding system (e.g., Wyatt et al. \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Lord and Westoby \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The available data indicate that long-lived plants like trees often produce flowers with a limited number of ovules, optimizing reproductive success over extended lifespans. While annuals, which have shorter life cycles, tend to produce numerous flowers with many ovules, to maximize seed output within a single growing season (Cruden and Lyon \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1989\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe number of ovules also depends on several ecological factors acting on the species level, like sibling- (Kress \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1981\u003c/span\u003e) and parent-offspring competition (Shaanker et al. \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e1988\u003c/span\u003e; Greenway and Harder \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2007\u003c/span\u003e), but also on species interactions, e.g., with pollinators (Friedman and Barrett \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Available data show that in the case of plant-pollinator interaction, several factors can play an important role in shaping the number of ovules, e.g., pollinators efficiency, time of flowering, or mode of pollination (Friedman and Barrett \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Bochynek and Burd \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Harder and Johnson \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Finally, the number of ovules may be explained by a way of exploiting unpredictably favorable conditions for seed ripening (Lee \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e1988\u003c/span\u003e) or seed dispersal (fleshy fruits vs. wind-dispersed propagules (Burd \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1995\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eBurd (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1995\u003c/span\u003e) proposed that the evolution of ovule number may be driven by the probability distribution of pollination success (measured, e.g., as the mean receipt of pollen tubes), suggesting that producing higher ovule numbers is a bet-hedging strategy to deal with unpredictable pollen receipt (Knight et al. \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) or uncertain environmental factors (Philippi and Seger \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). Indeed, a large meta-analysis by Burd et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), confirmed a clear positive relationship between ovule number per flower and probability of pollen receipt (assessed by counting stigmatic pollen loads). Also, a recent analysis from the Andean communities showed a positive correlation between the number of ovules and the probability of efficient pollination. This tendency may be, however, true only for habitats with especially low and variable pollination rates - greater variability in stigmatic pollen loads is likely in species pollinated by efficient pollinators because relatively more pollen will be deposited on the stigmas of flowers that have any visitors in comparison with those that fail to be visited (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). The overall explanation, however, most likely reflects selection acting simultaneously on many of the above-mentioned factors.\u003c/p\u003e\u003cp\u003eIn the case of animal-pollinated plants, stigmatic pollen load is a function of flower visitor identity, abundance and efficiency on flowers (e.g., Wilson and Thomson \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Th\u0026oslash;stesen and Olesen \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Adler and Irwin \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Sahli and Conner \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Fumero-Cab\u0026aacute;n and Mel\u0026eacute;ndez-Ackerman \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Junker et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; S\u0026aacute;nchez-Lafuente et al. \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Zych et al. \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e2013a\u003c/span\u003e; Kulbaba et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Parker et al. \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Since pollinators vary greatly in their dependence on a particular plant species, a successful transfer of pollen grains to the conspecific stigma can result from both highly generalist or very specialist interactions (or anything in between; Ollerton \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Waser et al. \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e1996\u003c/span\u003e; Johnson and Steiner \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Willmer \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). In the first instance, flowers are visited relatively frequently by taxonomically wide pollinator assemblages which, however, mainly include highly generalist foragers (e.g., \u003cem\u003eApis mellifera\u003c/em\u003e) and/or largely opportunistic species such as hoverflies and beetles (Willmer \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Zych et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Inouye et al. \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Hung et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). In such cases, the size of conspecific pollen loads carried by insects may vary greatly (see e.g. Zych et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Furthermore, mass flowering plants serviced by super generalist pollinators can be composed of low-quality self-pollen (Diller et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eIn more specialized interactions, such as those observed in the Orchidaceae family, the overall visitation frequency tends to be lower and extremely variable, compared to generalist interactions, however, the proportion of conspecific pollen loads increases, so the natural selection is expected to favor strategies allowing the production of a higher number of seeds after a single visit (Schreiber et al. \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). This compensates for the unpredictable nature of pollination events in specialized interactions. A meta-analysis of orchids found that while pollination success is highly variable and often low, when pollination does occur, orchids tend to maximize reproductive output by producing a large number of seeds per pollination event, partly because relying on pollinia, which transfer large pollen loads efficiently (Tremblay et al. \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2005\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e According to the bet-hedging hypothesis, ovule packaging in animal-pollinated plants should reflect the pollination system, specifically, the number and diversity of potential pollinators. Specialist systems, which channel pollen more directly among conspecifics, tend to deliver more efficient pollen transfer per visit. In such cases, plants serviced by one or a few efficient visitors may produce many seeds from a single visit, but successful visits are relatively rare. The optimal strategy, therefore, is to produce more ovules per flower to capitalize on infrequent yet efficient visits and to buffer against the likely failure of other flowers on the same plant to receive pollen (Burd \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Burd et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Rosenheim et al. \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). By contrast, generalists should produce fewer ovules per flower: each visit yields fewer seeds, but a large pool of potential pollinators increases visitation rates and compensates for per-visit inefficiency. Although direct tests linking visitor diversity to variance in pollen receipt remain scarce, large-scale analyses indicate that greater wild insect diversity stabilizes pollinator abundance across years, implying more consistent service and supporting this expectation (Senapathi et al. \u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eRecent evidence from an elevational gradient in the South American Andes is consistent with bet-hedging theory: ovule number increased with elevation, peaking where pollination is most unpredictable, with early-flowering species also having higher ovule number (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Arroyo et al. further proposed that higher ovule number should be associated with more specialized pollination, but their alpine dataset did not confirm this, likely because in that specific environment, generalization is the more advantageous strategy.\u003c/p\u003e\u003cp\u003eIn the present paper, we sought evidence for the more universal application of this assumption. To do so, we check if the (1) generalization/specialization in the pollination system, (2) life-form, and (3) phylogenetic relationships can explain some of the observed variation in ovule numbers. We tested these assumptions on our pollination network studies and literature-derived data for 192 zoogamous plant species representing 61 taxonomic families. Since detailed analyses of pollination effectiveness of animal visitors are rarely reported,d and comparing the performance of pollinators in pollination is problematic (see e.g., Ne\u0026rsquo;Eman et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2010\u003c/span\u003e)), as a measure of generalization/specialization we used the number of different species of floral visitors (the alpha diversity), treating all recorded animal visitors as legitimate and equally efficient pollinators.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003eWe performed a literature survey for data on the number of ovules per flower and pollinators of angiosperm plant species. In our investigation, we used Google Scholar, employing keyword combinations such as \"number of ovules\", \"ovule packaging\", along with terms \"floral visitors\", \"pollination\", and \"pollinators\". In cases where we encountered incomplete data (lacking information on number of ovules or insect visitors), we strived to fill in gaps by obtaining data from other studies. Our search encompassed available literature published until 2016. In cases where various sources reported different values of the ovule number per flower, we used means calculated for all available data. For data on floral visitors, we used α-diversity, and if several papers reported different numbers of taxa we chose the highest available result noting also, if available, taxonomic diversity of animals (in the case of insects at the rank of order and for vertebrates at the rank of class). This initial dataset was accompanied by data on floral visitors from additional two datasets: (1) a survey of the pollination network 26 plant species and their floral visitors) from a lowland meadow in NE Poland described in Goldstein and Zych (Goldstein and Zych \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), and (2) fallow land pollination network (7 plant species and their floral visitors) reported by Junker et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2013\u003c/span\u003e (Junker et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). For 27 plant species from the above networks lacking literature records of ovule numbers, in the summer of 2016, we collected flowers on-site and preserved them in 70% EtOH for further analysis of their ovaries\u0026rsquo; content in the lab. For each species enlisted in Appendix 1, this was done for 20 fully developed flowers from at least 10 individual plants. Ovules from dissected flowers were counted under the stereoscopic microscope.\u003c/p\u003e\u003cp\u003e Plants used in our database were classified according to the life form as annuals, biennials, perennials, trees, or shrubs. Our survey resulted in data for 192 zoogamous plant species representing 61 taxonomic families of the global flora (source papers from which the final data were extracted are listed in Appendix 2). As our dataset includes three plant families with distinctive pollination systems or floral architecture: Orchidaceae, Asclepiadaceae, and Asteraceae, the final analysis was done in several variants (1) database with all species, (2) database with Orchidaceae and Asclepiadaceae excluded, (3) database with Asteraceae excluded, and (4) database with Orchidaceae, Asclepiadaceae, and Asteraceae excluded. Orchids often have specialized interactions, low visitation frequency, high proportion of conspecific pollen loads (Faegri and Van Der Pijl \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1966\u003c/span\u003e; Proctor et al. \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e1996\u003c/span\u003e), and are known to produce very numerous ovules in a single flower. However, pollination in the Orchidaceae family occurs generally via transfer of the whole pollen sacks (pollinia; packaged pollen), which is largely different from other flowering plant families (Dahlgren et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1985\u003c/span\u003e) but similar to Asclepiadaceae(Ollerton and Liede \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1997\u003c/span\u003e)). The Asteraceae, in turn, are characterized by very distinct inflorescences: numerous flowers are arranged in a compound capitulum (pseudanthium), which is a functional equivalent of a flower. Often, e.g., in coneflowers (\u003cem\u003eEchinacea purpurea\u003c/em\u003e) or chamomile (\u003cem\u003eChamomilla recutita\u003c/em\u003e), the capitulum is composed of specialized rewardless ray (ligulate) florets, responsible for insect attraction, and nectariferous seed-producing disc florets (Wist and Davis \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Sulborska \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). Although members of Asteraceae are very conservative in the number of ovules per flower (always one), we assumed that for legitimate comparisons, the whole inflorescence should be treated as a single unit of pollination. Because the literature data largely lacked information on the number of individual flowers forming capitula of various species, and this trait showed to have some correlation with the altitude in the study of Arroyo et al. (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), we also decided to exclude Asteraceae from our analysis. Full results of these exclusions are provided in Appendix 1.\u003c/p\u003e\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analyses\u003c/h2\u003e\u003cp\u003eAll analyses were conducted in R (Posit Software, PBC 2025). To construct a phylogenetic tree as a framework for the Phylogenetic Generalized Least Squares (PGLS) analysis, we used a previously available phylogenetic tree from (Zanne et al. \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Supplementary Fig.\u0026nbsp;1). To assess whether ovule number variation significantly covaries with pollinator richness and life form, we performed both non-phylogenetically corrected and phylogenetically corrected analyses using Ordinary Least Squares (OLS) regression and PGLS regression (function gls from the nlm package; Pinheiro et al. \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e2025\u003c/span\u003e), respectively.\u003c/p\u003e\u003cp\u003eTo evaluate the extent to which trait variation is influenced by phylogenetic relationships, we estimated the phylogenetic signal for ovule number, pollinator richness, and life form. Specifically, we calculated Pagel\u0026rsquo;s λ and Blomberg\u0026rsquo;s K, which capture different aspects of phylogenetic signal: Pagel\u0026rsquo;s λ is widely used in community ecology, while Blomberg\u0026rsquo;s K is more suited for closely related species. Using both metrics allowed for a more robust interpretation of phylogenetic patterns within our dataset. Phylogenetic signal estimates were obtained using the phytools package in R (Revell \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e2024\u003c/span\u003e).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eApart from the excluded Asteraceae and Orchidaceae, the most abundant plant families in our database were Fabaceae (14 species), Apiaceae, Caryophyllaceae, and Ranunculaceae (10 species each), and Lamiaceae (6 species). Most of the recorded herbaceous species were perennials (123 species) and annuals (13 species), while 10 species were assigned as biennials. The rest of the species were classified as shrubs (19 species) or trees (16 species). Some of the studied species were assigned to two categories, e.g., annuals and biennials. For example, \u003cem\u003eSilene latifolia\u003c/em\u003e is mostly annual; however, it also occasionally occurs as a biennial plant (Appendix 2, Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThe average number of ovules in the studied species was 1132\u0026thinsp;\u0026plusmn;\u0026thinsp;2978, with a median of 15, a minimum value of 1, and a maximum value of 10000 ovules.\u003c/p\u003e\u003cp\u003eAcross all species, the number of ovules per flower and visitor richness was negatively correlated with flower-visitor richness (Pearson\u0026rsquo;s r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.26, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001; Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The analyses excluding Asteraceae, Orchidaceae, and/or Asclepiadaceae yielded qualitatively similar patterns (Appendix 1): non-phylogenetic effect sizes ranged from r\u0026thinsp;=\u0026thinsp;\u0026minus;\u0026thinsp;0.23 to \u0026minus;\u0026thinsp;0.31. Plant life form affected ovule number (ANOVA: F₄,₁₈₆ = 3.55, p\u0026thinsp;=\u0026thinsp;0.01): trees had fewer ovules than perennial herbs (Tukey HSD p\u0026thinsp;=\u0026thinsp;0.007). Visitor richness did not differ among life forms (ANOVA: F₄,\u003csub\u003e186\u003c/sub\u003e = 2.03, p\u0026thinsp;=\u0026thinsp;0.09).\u003c/p\u003e\u003cp\u003eHowever, we accounted for shared ancestry using PGLS (and, for comparison, OLS with the same model), we found no significant relationship between ovules per flower and visitor richness; this result was unchanged after excluding Asteraceae, Orchidaceae, Asclepiadaceae, or all three (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Based on AIC, PGLS consistently fit better than OLS, highlighting the importance of phylogenetic non-independence even though the slope did not differ from zero. Quantitative traits showed strong phylogenetic signal (ovules per ovary: Pagel\u0026rsquo;s λ\u0026thinsp;=\u0026thinsp;0.92, p\u0026thinsp;\u0026lt;\u0026thinsp;0.01; Blomberg\u0026rsquo;s K\u0026thinsp;=\u0026thinsp;0.72, p\u0026thinsp;=\u0026thinsp;0.001), and neither PGLS nor OLS detected an effect of life form on visitor richness (PGLS had the lower AIC; Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eOvule number vs. pollinator richness (OLS and PGLS). Relationships between ovule number and pollinator richness across taxonomic subsets. Linear models showed no significant association between pollinator richness and ovule number (all p\u0026thinsp;\u0026gt;\u0026thinsp;0.09p\u0026thinsp;\u0026gt;\u0026thinsp;0.09p\u0026thinsp;\u0026gt;\u0026thinsp;0.09 in OLS and PGLS).\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"9\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDataset\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModel\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eIntercept (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003evisitor_ric (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003et (slope)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003ep (slope)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003eAdj. R\u0026sup2;\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eAIC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eRSE (df); n\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAll species\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1333\u0026thinsp;\u0026plusmn;\u0026thinsp;251\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-8.61\u0026thinsp;\u0026plusmn;\u0026thinsp;5.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3596\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e2936 (189); 191\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1537\u0026thinsp;\u0026plusmn;\u0026thinsp;284\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-9.72\u0026thinsp;\u0026plusmn;\u0026thinsp;5.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3145\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3112 (164); 166\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e150\u0026thinsp;\u0026plusmn;\u0026thinsp;62 *\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-0.74\u0026thinsp;\u0026plusmn;\u0026thinsp;1.43\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-0.51\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.61\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e2583\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e659 (161); 163\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae, Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1542\u0026thinsp;\u0026plusmn;\u0026thinsp;291\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-9.89\u0026thinsp;\u0026plusmn;\u0026thinsp;6.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3074\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3150 (160); 162\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAll species\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1333\u0026thinsp;\u0026plusmn;\u0026thinsp;251\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-8.61\u0026thinsp;\u0026plusmn;\u0026thinsp;5.48\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.57\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.12\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3223\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e2921 (189); 191\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1537\u0026thinsp;\u0026plusmn;\u0026thinsp;284\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-9.72\u0026thinsp;\u0026plusmn;\u0026thinsp;5.97\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.11\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e2868\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3093 (164); 166\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1338\u0026thinsp;\u0026plusmn;\u0026thinsp;256\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-8.83\u0026thinsp;\u0026plusmn;\u0026thinsp;6.26\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.41\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.16\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e3168\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e2951 (185); 187\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae, Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e1542\u0026thinsp;\u0026plusmn;\u0026thinsp;291\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-9.88\u0026thinsp;\u0026plusmn;\u0026thinsp;6.87\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e\u003cp\u003e-1.44\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e\u003cp\u003e0.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c7\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e\u003cp\u003e2813\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c9\"\u003e\u003cp\u003e3131 (160); 162\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eOvule number vs. life form (OLS and PGLS). Relationships between ovule number and life form across taxonomic subsets. Linear models showed no significant association between ovule number and life form (all p\u0026thinsp;\u0026gt;\u0026thinsp;0.09p\u0026thinsp;\u0026gt;\u0026thinsp;0.09p\u0026thinsp;\u0026gt;\u0026thinsp;0.09 in OLS and PGLS).\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"10\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDataset\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModel\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eIntercept (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003elife_formb (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c5\"\u003e\u003cp\u003elife_formp (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c6\"\u003e\u003cp\u003elife_forms (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c7\"\u003e\u003cp\u003elife_formt (\u0026plusmn;\u0026thinsp;SE)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c8\"\u003e\u003cp\u003eAdj. R\u0026sup2; / note\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c9\"\u003e\u003cp\u003eAIC\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c10\"\u003e\u003cp\u003eRSE (df); n\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAll species\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e64.8\u0026thinsp;\u0026plusmn;\u0026thinsp;771\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-6.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1195\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1551\u0026thinsp;\u0026plusmn;\u0026thinsp;811 \u0026dagger;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-11.14\u0026thinsp;\u0026plusmn;\u0026thinsp;1006\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-48.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1056\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.04; model p\u0026thinsp;=\u0026thinsp;0.02\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3593\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2886 (186); 191\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;876\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1386\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1849\u0026thinsp;\u0026plusmn;\u0026thinsp;923\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-21.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-59.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1159\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e0.06; model p\u0026thinsp;=\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3140\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3036 (161); 166\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e64.8\u0026thinsp;\u0026plusmn;\u0026thinsp;177\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-6.29\u0026thinsp;\u0026plusmn;\u0026thinsp;274\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e118\u0026thinsp;\u0026plusmn;\u0026thinsp;189\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-11.14\u0026thinsp;\u0026plusmn;\u0026thinsp;231\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-48.54\u0026thinsp;\u0026plusmn;\u0026thinsp;242\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e-0.01; model p\u0026thinsp;=\u0026thinsp;0.80\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2587\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e662 (158); 163\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae, Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eOLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;883\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1397\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1918\u0026thinsp;\u0026plusmn;\u0026thinsp;932\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-21.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1117\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-59.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1169\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003eAdj. R\u0026sup2; = 0.06; model p\u0026thinsp;=\u0026thinsp;0.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3067\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3060 (157); 162\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAll species\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e64.8\u0026thinsp;\u0026plusmn;\u0026thinsp;771\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-6.29\u0026thinsp;\u0026plusmn;\u0026thinsp;1195\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1551\u0026thinsp;\u0026plusmn;\u0026thinsp;811 \u0026dagger;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-11.14\u0026thinsp;\u0026plusmn;\u0026thinsp;1006\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-48.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1056\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e3219\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2848 (186); 191\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;876\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1386\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1849\u0026thinsp;\u0026plusmn;\u0026thinsp;923\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-21.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1109\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-59.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1159\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2863\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e2990 (161); 166\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e64.8\u0026thinsp;\u0026plusmn;\u0026thinsp;177\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-6.29\u0026thinsp;\u0026plusmn;\u0026thinsp;274\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e118\u0026thinsp;\u0026plusmn;\u0026thinsp;189\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-11.14\u0026thinsp;\u0026plusmn;\u0026thinsp;231\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-48.54\u0026thinsp;\u0026plusmn;\u0026thinsp;242\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2272\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e652 (158); 163\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eNo Asteraceae, Asclepiadaceae \u0026amp; Orchidaceae\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePGLS\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e75.4\u0026thinsp;\u0026plusmn;\u0026thinsp;883\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e-2.54\u0026thinsp;\u0026plusmn;\u0026thinsp;1397\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e\u003cp\u003e1918\u0026thinsp;\u0026plusmn;\u0026thinsp;932\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c6\"\u003e\u003cp\u003e-21.77\u0026thinsp;\u0026plusmn;\u0026thinsp;1117\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c7\"\u003e\u003cp\u003e-59.17\u0026thinsp;\u0026plusmn;\u0026thinsp;1169\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c8\"\u003e\u003cp\u003e\u0026mdash;\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e\u003cp\u003e2806\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c10\"\u003e\u003cp\u003e3012 (157); 162\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eOur results show that phylogenetic constraints play an important role in shaping the ovule number per flower. Across a taxonomically diverse sample of species in our database, ovule number was not related to the degree of specialization or generalization in a plant\u0026rsquo;s pollination system. Although we initially found that flowers with a higher number of ovules per flower received visits from a less diverse set of pollinators than flowers with fewer ovules, this relationship was not supported when we accounted for phylogenetic non-independence in the analysis. Our results are therefore in contrast with the ovule bet-hedging hypothesis by Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), which, according to Burd et al. 1995 (Burd \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1995\u003c/span\u003e), predicts that the unpredictability of the pollinator environment may select for an increase in ovule number. According to this hypothesis, plants that produce more ovules per flower might take advantage of rare pollination visits through higher stigmatic pollen loads, enabling them to produce more seeds.\u003c/p\u003e\u003cp\u003eOur results revealed a correlation between ovule number and the life form of the studied species, which was also previously detected in other systems (Raven and Ra Yen \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1979\u003c/span\u003e; Plitmann and Levin \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). For example, in Caryophylloideae (\u003cem\u003eAgrostemma\u003c/em\u003e, \u003cem\u003eDianthus\u003c/em\u003e, \u003cem\u003eSaponaria\u003c/em\u003e, \u003cem\u003eSilene\u003c/em\u003e, and \u003cem\u003eVaccaria\u003c/em\u003e), where ovule number was significantly higher in perennials compared to annual species (J\u0026uuml;rgens et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). This may suggest that ovule number, floral visitor diversity, and life form evolved independently in the species studied. These traits may be influenced by different ecological or evolutionary factors that are not directly related.\u003c/p\u003e\u003cp\u003eSpecialized pollinators have been proposed as a prerequisite for the evolution of large ovule numbers because they promote more direct conspecific pollen transfer and reduce losses during transport or grooming, thereby increasing the odds of successful outcrossing (Willson \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e1979\u003c/span\u003e; Rademaker et al. \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). Yet recent work suggests that pollinator dependence and pollination efficiency more strongly shape pollen production than ovule production: higher pollen output, and thus higher P/O ratios, may compensate for reduced or inefficient interactions, whereas ovule number is comparatively insensitive to variation in pollination efficiency or syndrome (Cunha et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Nepal et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Harder and Johnson \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Accordingly, environments with greater pollination uncertainty (e.g., generalist-dominated systems) are expected to favor increased pollen production rather than larger ovule numbers (Cunha et al. \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Nepal et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Harder and Johnson \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Consistent with the role of mating structure, Burd (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2025\u003c/span\u003e) further proposes that high pollination efficiency often deposits related pollen cohorts or repeatedly targets nearby recipients, generating local mating competition; this framework predicts low P/O ratios coupled with high correlated paternity.\u003c/p\u003e\u003cp\u003eBy contrast, in generalist Apiaceae the ovule number per ovary is developmentally fixed and low, and is decoupled from pollen-load quantity or quality. Visitor taxa vary widely in pollen-carrying capacity and often deliver mixed loads; despite high visitation (Zych \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Zych et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), conspecific deposition can be limited because generalist flowers are serviced by equally generalist visitors. For example, in the supergeneralist \u003cem\u003eAngelica sylvestris\u003c/em\u003e (Apiaceae), 66% of visitors carried mixed pollen (Zych et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and in \u003cem\u003eOstericum palustre\u003c/em\u003e (Apiaceae), all guilds carried similar amounts of conspecific and heterospecific pollen (Zych et al. \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2014\u003c/span\u003e); a similar pattern occurs in \u003cem\u003ePolemonium caeruleum\u003c/em\u003e (Polemoniaceae; Zych et al. \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2013b\u003c/span\u003e). Even so, the low, fixed ovule numbers are typically sufficient to achieve a full seed set (e.g., Apiaceae: Zych et al. \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Consequently, visitor taxa are functionally equivalent in pollination service (Zamora \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) and exert limited selective pressure toward specialization, aligning with the expectation that, under diffuse and variable pollination, selection acts more on pollen output than on ovule number (G\u0026oacute;mez and Zamora \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2006\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eOur results suggest that ovule production is primarily constrained by phylogeny, consistent with earlier work. For example, both simple-flowered species and species with pseudanthium in the central Chilean Andes show a strong phylogenetic signal in ovule number (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), and broad phylogenetic structuring of ovule development is evident in orchids (Mayer et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) By contrast, pollination systems appear more labile: closely related species can rely on different pollinators, indicating substantial evolutionary plasticity in pollination strategies (e.g., Niet et al. \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Smith and Kriebel \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Roguz et al. \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e\u003cp\u003eThis asymmetry raises a key question: does ovule number constrain the evolution of pollination systems? Clades with a fixed, low ovule number, as in many Lamiaceae (Mabberley \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2017\u003c/span\u003e), may have limited scope to adopt highly specialized pollination and instead tend toward generalized systems. When ovules are few, reproductive success hinges on efficient pollen transfer, favoring high visitation rates or highly effective pollinators. Species with few ovules may also evolve mitigating traits, such as self-compatibility or clonal reproduction, to ensure reproductive success. Therefore, ovule number may play a fundamental role in shaping the evolution of pollination systems by influencing pollinator dependence, interactions, and reproductive strategies under varying ecological conditions.\u003c/p\u003e\u003cp\u003eIt is important to note that a limitation of our dataset is assessing the variability in pollen receipt indirectly through the diversity of floral visitors rather than direct measurements of pollen deposition or seed set. While this approach provides insight into long-term pollination environments, it does not quantitatively capture variation in pollen delivery. Direct methods, such as measuring stigmatic pollen loads or seed-set (Burd et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), offer a more precise assessment but rely on the assumption that short-term observations represent long-term trends. In contrast, visitor diversity may better reflect the evolutionary pollination environment but does not measure variance in pollen receipt explicitly (Arroyo et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). This trade-off should be considered when interpreting our results.\u003c/p\u003e\u003cp\u003eOverall, our results show an important role of phylogenetic constraints in the context of ovule production in a larger geographic context. Our results also follow a theory pointing at pollen production as an important response to pollinators' efficiency and reproductive strategy. It is still important, however, to call for a more in-depth study of the ovule oversupply in flowers also taking into account other evolutionary, ecological, and, perhaps, geographical explanations for this phenomenon. To shed more light on these questions further studies on a broader list of species are needed.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eThe authors declare that they have no financial or non-financial interests that are directly or indirectly related to the work submitted for publication.\u003c/p\u003e\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eMZ and RRJ conceived the idea; JR, AS, KR and MZ performed the literature survey, JR, AS and MZ made ovule counting, RRJ and KR performed statistical analyses, MZ, KR and RRJ drafted the first version of the manuscript, all authors contributed to the final version of the text.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eWe thank the organizers and participants of the 29th Annual Meeting of the Scandinavian Association of the Pollination Ecologists in Silkeborg, Denmark, for stimulating discussions that led to formulating the idea of this paper. Literature surveys and ovule counting was financed via the Statutory Research Programme of the Faculty of Biology, University of Warsaw. A short sabbatical of RRJ was financially supported by the University of Warsaw IDUB Program \u0026ldquo;Mentor\u0026rdquo;.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAdler LS, Irwin RE (2006) Comparison of Pollen Transfer Dynamics by Multiple Floral Visitors: Experiments with Pollen and Fluorescent Dye. 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Bot J Linn Soc 173:92\u0026ndash;107. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1111/boj.12071\u003c/span\u003e\u003cspan address=\"10.1111/boj.12071\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Angiospermae, ovule bet-hedging, evolution, flower, generalization, meta-analysis, sexual reproduction, plant-pollinator network, Evolutionary ecology","lastPublishedDoi":"10.21203/rs.3.rs-8038291/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8038291/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe number of ovules per flower varies over several orders of magnitude among angiosperms, with some families exhibiting remarkable conservatism and others displaying extensive variation among closely related species. Numerous hypotheses have been advanced to explain plant strategies in ovule packaging, often linking this trait to the statistical dispersion of pollen receipt. Given that, in animal-pollinated plants, the pollen load received depends on flower visitor identity, abundance, and efficiency, we hypothesized that ovule packaging should be directly related to the pollination system. Specifically, we predicted that (1) specialist plants, serviced by a few efficient floral visitors, would bet-hedge on rare visits by producing more ovules per flower and thereby maximizing seed output when pollination is successful, (2) compared to generalists that operate in more predictable pollinator environments and consequently produce fewer ovules per flower. To test our hypothesis, we analyzed literature-derived data encompassing 192 zoogamous plant species representing 61 taxonomic families. We observed fewer ovules with greater visitor diversity in trees compared to perennials, but these effects disappeared under phylogenetic models, which showed that ovule number has a strong phylogenetic signal. These findings indicate that while pollination system characteristics may influence ovule packaging strategies, they do not fully account for the observed variation. Instead, ovule number appears subject to strong and conflicting selective pressures, suggesting that the generalization/specialization of pollination systems does not singularly explain its evolution.\u003c/p\u003e","manuscriptTitle":"Is ovule packaging strategy in animal-pollinated plants correlated to the level of specialization of their pollination systems?","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-20 11:13:01","doi":"10.21203/rs.3.rs-8038291/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7d81f1c7-fac4-4f5c-8c22-e11592668763","owner":[],"postedDate":"November 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-04-04T18:53:47+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-20 11:13:01","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8038291","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8038291","identity":"rs-8038291","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}