Enhancing offspring quality or quantity? Different ways for using nectar amino acids in female butterflies

Butterfly-pollinated flowers offer nectar with higher amino acid concentrations than most flowers pollinated by other animals, and female butterflies of some species prefer to consume amino acid-rich nectar. However, for over 30years, there has been an ongoing discussion about whether nectar amino acids benefit butterfly fitness. A clear positive effect was only shown for the nectar-feeding Araschnia levana, and females of the fruit-feeding Bicyclus anynana also increased offspring quality when they were fed amino acids as adults. Thus, severe doubts remain about the general significance of these single positive results. We therefore tested a further species from a phylogenetically different butterfly subfamily, the small heath (Coenonympha pamphilus L., Satyrinae), taking into account feeding conditions over the whole life cycle of this species. C. pamphilus females receiving nectar amino acids as adults, irrespective of larval food quality, produced heavier larvae and also increased the hatching success of their eggs over the oviposition period. Furthermore, females raised under nitrogen-poor larval conditions tended to use nectar amino acids to increase the number of eggs laid. Thus, C. pamphilus females used nectar amino acids primarily to increase their offspring quality, and secondly tended to increase offspring quantity, if larval resources were scarce, showing a resource allocation pattern differing from both B. anynana and A. levana. Our study supports the old postulate that nectar amino acids generally enhance butterfly fitnes

Nectar amino acids enhance reproduction in male butterflies

After over 30years of research, it was recently shown that nectar amino acids increase female butterfly fecundity. However, little attention has been paid to the effect of nectar amino acids on male butterfly reproduction. Here, we show that larval food conditions (nitrogen-rich vs. nitrogen-poor host plants) and adult diet quality (nectar with or without amino acids) affected the amount of consumed nectar in Coenonympha pamphilus males. Furthermore, amino acids in the nectar diet of males increased progeny's larval hatching mass, irrespective of paternal larval reserves. Our study takes the whole reproductive cycle of male butterflies into account, and also considers the role of females in passing male nutrients to offspring, as males' realized reproduction was examined indirectly via nuptial gifts, by female performance. With this comprehensive approach, we demonstrate for the first time that nectar amino acids can improve male butterfly reproduction, supporting the old postulate that nectar amino acids generally enhance butterfly fitnes

Genotype-specific response of a lycaenid herbivore to elevated carbon dioxide and phosphorus availability in calcareous grassland

Effects of elevated CO2 and P availability on plant growth of the legume Lotus corniculatus and consequences for the butterfly larvae of Polyommatus icarus feeding on L. corniculatus were investigated in screen-aided CO2 control chambers under natural conditions on a calcareous grassland in the Swiss Jura mountains. Elevated CO2 conditions and P fertilisation increased the biomass production of L. corniculatus plants and affected the plant chemical composition. CO2 enrichment increased the C/N ratio and sugar concentration and decreased the N and P concentrations. C- and N-based allelochemicals (cyanoglycosides, total polyphenols and condensed tannins) were only marginally affected by CO2 enrichment. P fertilisation increased the specific leaf area and concentrations of water, N, sugar and P, while the C/N ratio and the concentration of total polyphenols decreased. Furthermore, P availability marginally enhanced the effect of elevated CO2 on the total dry mass and sugar concentration while the opposite occurred for the total polyphenol concentration. The changes in food-plant chemistry as a result of P fertilisation positively affected larval mass gain and accelerated the development time of P. icarus. Only a marginal negative effect on larval mass gain was found for CO2 enrichment. However, we found genotype-specific responses in the development time of P. icarus to elevated CO2 conditions. Larvae originating from different mothers developed better either under elevated CO2 or under ambient CO2 but some did not react to CO2 elevation. As far as we know this is the first finding of a genotype-specific response of an insect herbivore to elevated CO2 which suggests genetic shifts in insect life history traits in response to elevated CO

Reduced clonal reproduction indicates low potential for establishment of hybrids between wild and cultivated strawberries ( Fragaria vesca × F. × ananassa )

The genus Fragaria (Rosaceae) contains 24 species, including hybrid species such as the garden strawberry (Fragaria×ananassa Duch.). Natural hybridization between Fragaria species has repeatedly been reported, and studies on the hybridization potential between F.×ananassa and its wild relatives have become increasingly important with the outlook for genetically modified garden strawberries. In Europe, a candidate species for hybridization with garden strawberries is the common woodland strawberry (Fragaria vesca L.). Although a previous field survey indicated that the potential for hybridization between F. vesca and F.×ananassa is low, it is not clear whether the lack of natural hybrids is caused by known pre- and postzygotic barriers, or whether hybrid plants lack the fitness to establish in natural F. vesca populations. We grew different F. vesca and F. vesca×F.×ananassa hybrid clones with and without competition in a greenhouse and assessed biomass production, clonal reproduction, and sexual reproduction of plants. While some hybrid clones exceeded F. vesca in biomass production, general clonal reproduction was much lower and delayed in hybrids. Furthermore, hybrids were sterile. These results demonstrate a mechanism by which the general lack of F. vesca×F.×ananassa hybrids in natural habitats can be explained, in addition to the known low hybridization potential between garden and woodland strawberries. We conclude that hybrids have a competitive disadvantage against co-occurring F. vesca plants due to inferior and delayed clonal reproduction, and that the potential for hybrid establishment under natural conditions is lo

Elevated carbon dioxide increases nectar production in Epilobium angustifolium L

Effects of elevated CO2 and nutrient availability on nectar production and onset of flowering in five different seed families (genotypes) of Epilobium angustifolium were investigated in a greenhouse experiment. Elevated CO2 significantly increased nectar production per day (+51%, p < 0.01), total sugar per flower (+41%, p < 0.05), amino acid concentration (+65%, p < 0.05) and total amino acids per flower (+192%, p < 0.001). All other parameters tested, i.e., nectar sugar concentration, proportion of glucose/fructose and proportion of sucrose/(glucose + fructose), were not significantly affected by elevated CO2 and/or fertilization. However, elevated CO2 caused a marginally significant trend for earlier flowering in highly fertilized plants. No significant family × CO2 interaction was found in any of the tested parameters, but the response in nectar production varied considerably among seed families (+10 to +104%) and was significantly positive in two of the five seed families investigated. Our results are not consistent with earlier studies on effects of elevated CO2 on nectar production and flowering phenology in other plant species. It seems, on the other hand, that CO2 effects on nectar production are specific to species and genotype. Hence, no general conclusions about effects of elevated CO2 on these floral traits can be drawn at present, but it must be cautioned that elevated CO2 might not only increase floral rewards as in E. angustifolium, but might also lead to shifts or even disruptions in fine-tuned plant-pollinator interaction

Effects of fertilization and elevated CO2 on larval food and butterfly nectar amino acid preference in Coenonympha pamphilus L

The effects of larval diet on the nutritional preferences of butterflies has rarely been examined. This study investigates whether alterations in the larval diet result in changes in adult preferences for nectar amino acids. Larvae of Coenonympha pamphilus were raised on fertilized or unfertilized Festuca rubra, grown under ambient (350ppm) or elevated (750ppm) atmospheric CO2 environments. Fertilization led to marked increases in leaf nitrogen concentration. In plants grown under elevated CO2 conditions, leaf water and nitrogen concentrations were significantly lower, and the C/N-ratio increased significantly. Fertilization of the host plant shortened the development time of C. pamphilus larvae, and pupal weight increased. In contrast, larvae of C. pamphilus developed significantly slower on F. rubra grown under elevated CO2, but adult emergence weight was not affected by CO2 treatment of the plant. C. pamphilus females showed a clear preference for nectar mimics containing amino acids, whereas males, regardless of treatment, either preferred the nectar mimic void of amino acids or showed no preference for the different solutions. Female butterflies raised on fertilized plants showed a significant decline in their preference for nectar mimics containing amino acids. A slight, but not significant, trend towards increased nectar amino acid preference was found in females raised on plants grown under elevated CO2. We clearly demonstrate that alterations in larval host quality led to changes in butterfly nectar preferences. The ability of the butterfly to either rely less on nectar uptake or compensate for poor larval conditions represents a trade-off between larval and adult butterfly feedin

Elevated carbon dioxide increases nectar production in Epilobium angustifolium L

Effects of elevated CO2 and nutrient availability on nectar production and onset of flowering in five different seed families (genotypes) of Epilobium angustifolium were investigated in a greenhouse experiment. Elevated CO2 significantly increased nectar production per day (+51%, p > 0.01), total sugar per flower (+41%, p > 0.05), amino acid concentration (+65%, p > 0.05) and total amino acids per flower (+192%, p > 0.001). All other parameters tested, i.e., nectar sugar concentration, proportion of glucose/fructose and proportion of sucrose/(glucose + fructose), were not significantly affected by elevated CO2 and/or fertilization. However, elevated CO2 caused a marginally significant trend for earlier flowering in highly fertilized plants. No significant family x CO2 interaction was found in any of the tested parameters, but the response in nectar production varied considerably among seed families (+10 to +104%) and was significantly positive in two of the five seed families investigated. Our results are not consistent with earlier studies on effects of elevated CO2 on nectar production and flowering phenology in other plant species. It seems, on the other hand, that CO2 effects on nectar production are specific to species and genotype. Hence, no general conclusions about effects of elevated CO2 on these floral traits can be drawn at present, but it must be cautioned that elevated CO2 might not only increase floral rewards as in E. angustifolium, but might also lead to shifts or even disruptions in fine-tuned plant-pollinator interactions

Searching for gene flow from cultivated to wild strawberries in Central Europe

Background and Aims Experimental crosses between the diploid woodland strawberry (Fragaria vesca L.) and the octoploid garden strawberry (F. × ananassa Duch.) can lead to the formation of viable hybrids. However, the extent of such hybrid formation under natural conditions is unknown, but is of fundamental interest and importance in the light of the potential future cultivation of transgenic strawberries. A hybrid survey was therefore conducted in the surroundings of ten farms in Switzerland and southern Germany, where strawberries have been cultivated for at least 10 years and where wild strawberries occur in the close vicinity. Methods In 2007 and 2008, 370 wild F. vesca plants were sampled at natural populations around farms and analysed with microsatellite markers. In 2010, natural populations were revisited and morphological traits of 3050 F. vesca plants were inspected. DNA contents of cell nuclei of morphologically deviating plants were estimated by flow cytometry to identify hybrids. As controls, 50 hybrid plants from interspecific hand-crosses were analysed using microsatellite analysis and DNA contents of cell nuclei were estimated by flow cytometry. Key Results None of the wild samples collected in 2007 and 2008 contained F. × ananassa microsatellite markers, while all hybrids from hand-crosses clearly contained markers of both parent species. Morphological inspection of wild populations carried out in 2010 and subsequent flow cytometry of ten morphologically deviating plants revealed no hybrids. Conclusions Hybrid formation or hybrid establishment in natural populations in the survey area is at best a rare even

The relative importance of sexual and clonal reproduction for population growth in the perennial herb Fragaria vesca

The relative importance of sexual and clonal reproduction for population growth in clonal plants is highly variable. Clonal reproduction is often more important than sexual reproduction but there is considerable interspecific variation and the importance of the two reproductive modes can change with environmental conditions. We carried out a demographic study on the woodland strawberry (Fragaria vesca), a widespread clonal herb, at 12 sites in Switzerland during 2years. Study sites were selected in two different habitats, i.e., forest and forest edge. We used periodic matrix models to estimate annual population growth rates and carried out prospective analyses to identify life cycle components that influence population growth rates most. Retrospective analyses were applied to study how the two different habitats affected population dynamics. Furthermore, we tested whether trade-offs between sexual and clonal reproduction occurred. There were large differences in annual population growth rates between sites and large within-site differences between years. Results of the prospective analyses clearly indicate that clonal reproduction is the dominant reproductive pathway whereas sexual reproduction is rather insignificant for population growth. Compared to forest habitats, forest edge habitats had higher population growth rates in the first year but smaller growth rates in the second year. We attribute these differing habitat effects to different water availabilities during consecutive years. No trade-offs between sexual and clonal reproduction were found. In conclusion, population growth of F. vesca relies heavily on clonal reproduction. Furthermore, reproduction and survival rates of F. vesca depend highly on spatio-temporal variation of environmental condition

Positive effects of cyanogenic glycosides in food plants on larval development of the common blue butterfly

Cyanogenesis is a widespread chemical defence mechanism in plants against herbivory. However, some specialised herbivores overcome this protection by different behavioural or metabolic mechanisms. In the present study, we investigated the effect of presence or absence of cyanogenic glycosides in birdsfoot trefoil (Lotus corniculatus, Fabaceae) on oviposition behaviour, larval preference, larval development, adult weight and nectar preference of the common blue butterfly (Polyommatus icarus, Lycaenidae). For oviposition behaviour there was a female-specific reaction to cyanogenic glycoside content; i.e. some females preferred to oviposit on cyanogenic over acyanogenic plants, while other females behaved in the opposite way. Freshly hatched larvae did not discriminate between the two plant morphs. Since the two plant morphs differed not only in their content of cyanogenic glycoside, but also in N and water content, we expected these differences to affect larval growth. Contrary to our expectations, larvae feeding on cyanogenic plants showed a faster development and stronger weight gain than larvae feeding on acyanogenic plants. Furthermore, female genotype affected development time, larval and pupal weight of the common blue butterfly. However, most effects detected in the larval phase disappeared for adult weight, indicating compensatory feeding of larvae. Adult butterflies reared on the two cyanogenic glycoside plant morphs did not differ in their nectar preference. But a gender-specific effect was found, where females preferred amino acid-rich nectar while males did not discriminate between the two nectar mimics. The presented results indicate that larvae of the common blue butterfly can metabolise the surplus of N in cyanogenic plants for growth. Additionally, the female-specific behaviour to oviposit preferably on cyanogenic or acyanogenic plant morphs and the female-genotype-specific responses in life history traits indicate the genetic flexibility of this butterfly species and its potential for local adaptatio