During the evolution of angiosperm flowers, some floral traits may undergo certain changes in order to participate in screening. The stamens and pistils of Delphinium caeruleum are covered by two “door-like” staminodes, the evolutionary function of which, however, is quite unknown. In this study, we investigated whether D. caeruleum staminodes acted as visitor filters by assessing the respective strengths of staminodes and visitor insects (six bee species). We measured the operative strength required to open the staminodes and the strength that insects were capable of exerting using a biological tension sensor. Furthermore, we compared the strength required to open staminodes at different phases of the flowering period (male and female phases) and the strength of different visitors (visitors and non-visitors of D. caeruleum). The results showed that the strength needed to open staminodes in the male phase was significantly higher than that in the female phase. There was no significant difference between the strength exerted by visitors and required by staminodes of D. caeruleum in the male phase, but the visitor strength was significantly higher than that required to open staminodes in the female phase flowers. The strength of non-visitors was significantly lower than that required to open staminodes in the male phase. Furthermore, there was a significant positive association between the strength and the body weight of the bees. These results highlighted the observation that only strong visitors could press the two staminodes to access the sex organs and achieve successful pollination. Furthermore, these results revealed the function of pollinator screening by the staminodes of D. caeruleum. The biomechanical approach to the study of flowers allowed us to address relevant ecological and evolutionary questions of the plant–pollinator interaction and explore the functional modules within the flower structure in other plant species.

The screening mechanism is an important component of the screening game between plants and pollinators, and also as a theoretical framework for understanding the maintenance and drivers of co-evolution by animals and plants. However, the composite screening mechanism in plants from biomechanical and morphological perspectives was seldomly studied. In this study, we measured the insects and flower traits in different populations of Delphinium caeruleum, and compared the operative strength of staminodes with the strength that insects were capable of exerting, as well as the length of the nectar spur and proboscis in insects. The results showed that the flower traits, insect species, visitor frequency, and efficiency differed among three population. The strength and proboscis length of the insect visitors matched with the operative strength of staminodes and the nectar spur length, whereas the opposite was found for non-visitors. Our results demonstrate that Delphinium caeruleum can screen for efficient pollinator insects through a complex mechanism based on a combination of length screening and biomechanical screening. Pollinators contribute significantly to promoting both the convergence and divergence of flower characteristics, which may be a consequence of plants adapting to local pollinators.

Continuous variation in herkogamy has been well reported, however, less attention has been paid to the phenomena that the consecutive expression of two types of herkogamy in the same flower. Euphorbia fischeriana, which have both vertical and lateral herkogamy, show vertical herkogamy during the female phase . However, their gynophores bend to one side with the male phase and show lateral herkogamy . In this study, we observed the effect of successive sexual organs movement on variation in herkogamy traits. By artificially manipulating the flower to present gynophore straightened in the floral center or bend to one side, we attempted to investigate whether herkogamy movement affects pollinator access efficiency, pollen removal and deposition, and seed set ratio. Furthermore, we conducted artificial pollination in the female phase to evaluate the effect of changes in pollination environment on the variations in herkogamy traits. The results showed that gynophore straightened in female phase favors pollen deposition, whereas gynophore bending in male phase was conducive to the removal of pollen. Visitation frequency, pollen deposition and removal, and seed set ratio decreased significantly when the gynophore movement was manipulated. Finally, the bending of gynophore was obviously promoted by pollination. Therefore, the continuous variation of herkogamy in the same flower of E. fischeriana caused by the bending of the gynophore could improve the accuracy of pollination and avoid the interference of the ovary with access efficiency. That may be an adaptive strategy when pollinators are scarce. Furthermore, our study also provides good support for the hypothesis that variations in herkogamy traits are strongly selected by differences in pollination environments.

Nectar-robbing insects, which are frequently described as cheaters in plant–pollinator mutualisms, may affect plant reproductive fitness by obtaining nectar rewards without providing pollination services. The negative effects of nectar robbing on plant reproductive success have been widely reported, but the reasons for possible positive effects demand further investigation. The goal of the study was to evaluate the effects of nectar robbing on the reproductive success of Delphinium caeruleum. Two staminodes cover the stamens and pistils in the flowers of D. caeruleum, forming a “double door” type of structure that compels pollinators to physically manipulate the staminodes to access the sex organs. In order to explore whether the operative strength required to open the staminodes is affected by actions associated with nectar robbing, we set up five different treatment groups: no nectar robbing, natural nectar robbing, artificial nectar robbing, hole making, and nectar removal. A biological tension sensor was used to measure the operative strength required to open the staminodes in the flowers. We also assessed the effect of nectar robbing on the flower-visiting behavior of pollinators and the effect of nectar robbing on reproductive fitness by the flower. The results showed that the operative strength needed to open staminodes was reduced by nectar robbers but not by artificial nectar robbing, hole making, or nectar removal. The flowers’ continuous visitation rate and visitation frequency by pollinators decreased significantly in robbed flowers. Both the pollen export and pollen deposition in naturally robbed flowers were significantly higher than those in nonrobbed flowers. Our results demonstrate that nectar robbers play an indirect positive role in the reproductive fitness of D. caeruleum flowers by reducing the operative strength of staminodes to promote pollen transfer. The reduction in operative strength of staminodes might be an adaptive mechanism that responds to nectar robbing.

The screening mechanism is an important component of the screening game between plants and pollinators, and also as a theoretical framework for understanding the maintenance and drivers of co-evolution by animals and plants. However, no previous studies have investigated the composite screening mechanism in plants from biomechanical and morphological perspectives. Thus, we observed and measured the insects and flower traits in different populations of Delphinium caeruleum J., and compared the operative strength of staminodes with the strength that insects were capable of exerting, as well as the length of the nectar spur and proboscis in insects. The flower traits, insect species, visitor frequency, and efficiency differed among three experimental sites. The strength and proboscis length of the insect visitors matched with the operative strength of staminodes and the nectar spur length, whereas the opposite was found for non-visitors. Our results demonstrate that Delphinium caeruleum J. can screen for efficient pollinator insects through a complex mechanism based on a combination of length screening and biomechanical screening. Pollinators contribute significantly to promoting both the convergence and divergence of flower characteristics, which may be a consequence of plants adapting to local pollinators.