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Sugar concentration in floral nectars is an assessment required in several diverse fields of application. The widely used analysis, consisting of nectar extraction with a microcapillary and sugar concentration measurement with a light refractometer, is not reliable when the nectar is secreted in small quantities, unextractable with a microcapillary. Ancillary methods adopted in such cases are destructive, rather complicated and often provide much less precise and accurate results. The microscopic-size, low cost and biocompatibility of optical fibers were exploited to deliver light directly inside the flower with minimal invasiveness and measure instantaneously the refractometric properties of the nectar without extracting it. After comparing the new and old methods using two known nectariferous species, the new approach was validated on Primula palinuri, whose nectar is unextractable with microcapillaries. The fiber-optic probe was able to measure the nectar refractive index in P. palinuri flowers making it possible to highlight a previously undetected significant trend of the sugar concentration throughout the long anthesis of the single flowers. Changes in nectar concentrations are similar in both longistylous and brevistylous flowers. The fiber-optic refractometer is an advancement of light refractometer analysis. Further customization of the laboratory set-up into portable equipment will boost applications., (© 2019 The Authors. New Phytologist © 2019 New Phytologist Trust.)

The trait-fitness relationship influences the strength and direction of floral evolution. To fully understand and predict the evolutionary trajectories of floral traits, it is critical to disentangle the direct and indirect effects of floral traits on plant fitness in natural populations. We experimentally quantified phenotypic selection on floral traits through female fitness and estimated the casual effects of nectar robbing with different nectar robbing intensities on trait-fitness relationships in both the L- (long-style and short-anther phenotype) and S-morph (short-style and long-anther phenotype) flowers among Primula secundiflora populations. A larger number of flowers and wider corolla tubes had both direct and indirect positive effects on female fitness in the P. secundiflora populations. The indirect effects of these two traits on female fitness were mediated by nectar robbers. The indirect effect of the number of flowers on female fitness increased with increasing nectar robbing intensity. In most populations, the direct and/or indirect effects of floral traits on female fitness were stronger in the S-morph flowers than in the L-morph flowers. In addition, nectar robbers had a direct positive effect on female fitness, but this effect varied between the L- and S-morph flowers. These results show the potential role of nectar robbers in influencing the trait-fitness relationships in this primrose species., (© 2019 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

Heterostyly is a floral polymorphism characterized by reciprocal herkogamy maintained through high levels of mating between morphs, serviced by appropriate pollinators. We studied how differential efficiency and abundance of distinct pollinators affect plant female reproduction in self- and intra-morph incompatible distylous Primula secundiflora. Bumblebees and syrphid flies were found to be the most abundant floral visitors. Bumblebees frequently exhibited nectar-robbing behavior. Because the robbing holes were always situated between the high- and low-level organs on both morphs, nectar-robbing bumblebees only pollinated S-styled flowers. L-styled flowers set four times as many seeds as did S-styled flowers after being visited by pollen-collecting syrphid flies. The natural female fecundity and the magnitude of pollen limitation varied between the morphs within populations because of the mosaic distribution of nectar-robbing bumblebees and syrphid flies. L-styled flowers and S-styled flowers set the same number of seeds after supplemental hand pollination, indicating equivalent female reproductive potential. We suggest that bumblebee nectar robbers and syrphid flies play an important role in sustaining the floral dimorphism of heterostyly in P. secundiflora because of their complementary roles in the pollination system.