Floral scent diversity is differently expressed in emitted and endogenous components in Petunia axillaris lines

BACKGROUND AND AIMS: Among the subspecies of Petunia axillaris are various lines emitting sensorially different scents. Analysis of variations in floral scent among genetically close individuals is a powerful approach to understanding the mechanisms for generating scent diversity. METHODS: Emitted and endogenous components were analysed independently to gain information about evaporation and endogenous production in 13 wild lines of P. axillaris. A dynamic headspace method was used to collect emitted components. Endogenous components were extracted with solvent. Both of these sample types were subjected to quantitative and qualitative analysis by gas chromatography (GC)-flame ionization detector (FID) and GC-mass spectrometry (MS). KEY RESULTS AND CONCLUSIONS: Whereas the profiles of emitted compounds showed qualitative homogeneity, being mainly composed of methyl benzoate with quantitative variation, the profiles of endogenous compounds showed both qualitative and quantitative variation. A negative correlation was found between the evaporation ratio and boiling point of each compound examined. Lower boiling point compounds were strongly represented in the emitted component, resulting in the reduction of qualitative variation in floral scent. In conclusion, floral scent diversity results from variation in both the endogenous production and the evaporation rate of the individual volatile compounds.     

Metabolic regulation of floral scent in Petunia axillaris lines: biosynthetic relationship between dihydroconiferyl acetate and iso-eugenol

Aromatic scent-related compounds in flowers were comprehensively analyzed by high-performance liquid chromatography (HPLC) based on their absorption spectra to understand regulation of metabolism leading to floral scent diversity in Petunia axillaris lines. An unknown compound occurring at similar levels to scent compounds in some plant lines was identified to be dihydroconiferyl acetate. Based on the structure, dihydroconiferyl acetate is likely to be a biosynthetically closely related compound to aromatic scent compounds, especially iso-eugenol. Similar time-course changes of the concentrations suggest that the metabolism of dihydroconiferyl acetate is underlaid by the similar regulation to aromatic scent compounds. Dihydroconiferyl acetate and iso-eugenol occurred almost exclusively, implying that metabolism of the common precursors to each compound is selectively regulated in these plants. The branching of the biosynthetic pathway into dihydroconiferyl acetate and iso-eugenol is probably one of regulatory steps leading to scent diversity in P. axillaris lines.     

Temporal plasticity in thermal‐habitat selection of burbot Lota lota a diel‐migrating winter‐specialist

In this study, animal‐borne telemetry with temperature sensors was coupled with extensive habitat temperature monitoring in a dimictic reservoir, to test the following hypotheses: behavioural thermoregulation occurs throughout the year and temperature selection varies on a diel and seasonal basis, in a winter‐specialist diel‐migrating fish. Burbot Lota lota demonstrated nightly behavioural thermoregulation throughout the year, with a large seasonal shift between selection for very cold temperatures (<2° C) optimal for reproduction during the spawning period and selection for warmer temperatures (12–14° C) optimal for hunting and feeding during non‐reproductive periods. During daylight hours, while L. lota avoided habitats warmer than optimal for reproduction and feeding during the spawning and non‐reproductive periods, respectively, active selection was limited to selection for 4–6° C habitat during the prespawning period. Although behavioural thermoregulation explained the night‐time migration, behavioural thermoregulation only partially explained daytime behaviour, indicating that diel migration is best explained by a combination of factors. Thus, thermal‐habitat selection was a good predictor of night‐time habitat occupancy in a diel‐migrating species. Together, these results show that thermal‐habitat selection by fishes may be important throughout the year and a more seasonally plastic behaviour than previously recognized.     

A comparative analysis of characteristic floral scent compounds in Prunus mume and related species

In order to investigate the difference in their characteristic floral scents between Prunus mume Siebold & Zucc. and the related Prunus species, their headspace volatiles and endogenous extraction were analyzed by gas chromatography–mass spectrometry. The efficiency of substrate utilization of the flowers was studied by incubating them with different alcohol substrates. Our results indicated that benzyl acetate is a dominant compound influencing the characteristic floral scent of P. mume. An alcohol substrate concentration of 4?mmol?L^?1 and a reaction time of 2?h were constituted the reaction condition for catalysis of exogenous alcohol substrates by the flowers. Under these conditions, Prunus sibirica exhibited the highest utilization efficiency for benzyl alcohol substrate while the utilization efficiency of Prunus persica was the lowest. Comparative analysis of several alcohol substrates indicated that the flowers of the tested species had selective specificity for benzyl alcohol substrates.     

Pollination and floral scent differentiation in species of the Philodendron bipinnatifidum complex (Araceae)

The Philodendron bipinnatifidum complex of Philodendron subgenus Meconostigma may comprise four species, which because of only slight and not very distinct morphological differences are not all unanimously recognized as good species. To find out whether these species are reproductively isolated, we studied the flowering rhythm, thermogenesis and pollination biology of three species of this complex, namely of P. bipinnatifidum, P. aff. bipinnatifidum (provisionally named “P. form selloum”) and P. mello-barretoanum in Brazil. Of the first two mentioned taxa, floral scent was collected and scent compounds were identified by GC–MS. The results showed that the coastal forest species P. bipinnatifidum has a two-, or three-night flowering rhythm, with the pistillate stage in the first night and the staminate stage lasting the second and sometimes also the third night. Strong thermogenesis with extended heating periods of several hours during the first part of the usual two subsequent nights and the maximum temperatures of up to 40 °C absolute heating of the spadices occurred in the pistillate and staminate stages. Concomitant with the heating periods, relatively low amounts of principally (Z)-2-pentenyl acetate and (Z)-jasmone were emitted by both the pistillate and staminate stage inflorescences. The dynastid scarab beetle Cyclocephala variolosa was the only pollinator attracted. The upland forest P. form selloum always had a two-night flowering rhythm with the pistillate stage in the first and the staminate stage in the subsequent night. This world-record holder of thermogenesis can heat up to the remarkable 45 °C during a relatively short period in the evening of the pistillate stage. During the thermogenic period, enormous amounts of principally 4-methoxystyrene and 3,4-dimethoxystyrene were produced and which could attract a large number of female and male individuals of the dynastid scarab beetle Erioscelis emarginata. In the staminate stage of P. form selloum, temperature elevation is significantly lower and the scent compounds are different from the pistillate stage. The cerrado biome species P. mello-barretoanum has a flowering rhythm similar to P. form selloum, reaching a maximum heating of about 40 °C during the pistillate stage. The sole pollinator attracted was Cyclocephala atricapilla. The differences observed and analyzed among the taxa, including the flowering rhythm, thermogenic activities, scent compounds emitted, pollinating dynastid scarab beetles attracted, as well as slight morphological differences and apparent geographical exclusiveness noted in these three taxa are strong indicators that P. bipinnatifidum, P. form selloum and P. mello-barretoanum are different enough to be considered good species. The morphological affinities of these species might be a hint that speciation has been a recent event and/or also that reproductive isolation based on different, non-overlapping distribution areas, different scent compounds and different pollinators was effective enough to need further morphological differentiation.     

Floral biology and reproductive isolation by floral scent in three sympatric aroid species in French Guiana

We studied the reproductive biology of three sympatric Araceae species, Anthurium sagittatum, A. thrinax and Spathiphyllum humboldtii in French Guiana. The plants flowered simultaneously and were visited by scent‐collecting male euglossine bees, which were apparently their major pollinators. In total, each species was visited by 3–7 euglossine species, and 2–3 euglossine species accounted for at least 80% of all flower visits, with visits being plant species‐specific. Floral scent consisted of 6–10 main compounds, which made up 76–94% of the total amount of volatiles and were specific in these high amounts to each plant species. We suggest that the different floral scents lead to clear separation of the main pollinating euglossine species, providing a directed and efficient intraspecific pollen flow that results in high reproductive success. Since the simple floral (inflorescence) morphology of the studied plants does not support any morphological mechanisms to exclude visitors, as for example in euglossine‐pollinated perfume orchids, floral scent might be of major importance for the reproductive isolation and sympatric occurrence of these plants.     

Linkages between nectaring and oviposition preferences of Manduca sexta on two co‐blooming Datura species in the Sonoran Desert

1. The oviposition choices of phytophagous insects determine the environment that their offspring will experience, affecting both larval performance and host plant fitness. These choices, however, may be influenced by other activities, such as nectar foraging. 2. In the Sonoran Desert, Manduca sexta (Lepidoptera: Sphingidae) oviposits primarily on the perennial herb Datura wrightii. It has been reported to oviposit on the smaller‐flowered, co‐blooming, sympatric annual Datura discolor as well. Datura is also M. sexta's most important source of nectar in this region. Nectaring and oviposition decisions thus determine M. sexta's reproductive success, as well as the relative benefits (pollination) and costs (herbivory) that each Datura species derives from this interaction. 3. The nectaring and oviposition choices of adult M. sexta on these congeners were studied to investigate how nectar foraging influences oviposition. Larval performance on the two hosts was then assessed. 4. Nectaring and oviposition were behaviourally linked, with M. sexta preferring the perennial D. wrightii as both a nectar source and larval host when given a choice between the two species. This preference disappeared, however, when only D. discolor bore flowers. 5. In the laboratory, larvae developed at equal rates when fed D. wrightii or D. discolor leaves, but survival was higher on D. wrightii when larvae fed on intact plants. 6. These findings suggest that while female preferences match larval performance in most cases, the link between nectaring and oviposition may at times bias moths to lay eggs on inferior larval hosts.     

Sex‐specific floral attraction traits in a sequentially hermaphroditic species

• ● Many angiosperms are hermaphroditic and produce bisexual flowers in which male (pollen export) and female (stigma receptivity) functions are separated temporally. This sequential hermaphroditism may be associated with variation in flower size, color, or pattern, all of which may influence pollinator attraction. In this study, we describe variation in these traits across discrete functional sex stages within and between 225 greenhouse‐grown individuals of Clarkia unguiculata (Onagraceae). In addition, to identify the effects of floral phenotype on pollinator attraction in this species, we examine the effects of these floral traits on pollen receipt in ~180 individuals in an experimental field array.
• ● Petal area, ultraviolet (UV)‐absorbing nectar guide area, and blue and green mean petal reflectance differ significantly across the functional sex stages of C. unguiculata. Male‐ and female‐phase flowers display significantly different pollinator attraction traits. Petal and UV nectar guide area increase as flowers progress from male phase to female phase, while blue reflectance and green reflectance peak during anther maturation.
• ● In field arrays of C. unguiculata, female‐phase flowers with large UV nectar guides receive more pollen than those with small nectar guides, and female‐phase flowers with high mean blue reflectance values are more likely to receive pollen than those with low blue reflectance. Female‐phase flowers with green mean reflectance values that differ most from background foliage also receive more pollen than those that are more similar to foliage. These findings indicate that components of flower color and pattern influence pollen receipt, independent of other plant attributes that may covary with floral traits. We discuss these results in the context of hypotheses that have been proposed to explain sex‐specific floral attraction traits, and we suggest future research that could improve our understanding of sexual dimorphism in sequentially hermaphroditic species and the evolution of features that promote outcrossing.

     

The on‐again,off‐again relationship between mitochondrial genomes and species boundaries

The study of reproductive isolation and species barriers frequently focuses on mitochondrial genomes and has produced two alternative and almost diametrically opposed narratives. On one hand, mtDNA may be at the forefront of speciation events, with co‐evolved mitonuclear interactions responsible for some of the earliest genetic incompatibilities arising among isolated populations. On the other hand, there are numerous cases of introgression of mtDNA across species boundaries even when nuclear gene flow is restricted. We argue that these seemingly contradictory patterns can result from a single underlying cause. Specifically, the accumulation of deleterious mutations in mtDNA creates a problem with two alternative evolutionary solutions. In some cases, compensatory or epistatic changes in the nuclear genome may ameliorate the effects of mitochondrial mutations, thereby establishing coadapted mitonuclear genotypes within populations and forming the basis of reproductive incompatibilities between populations. Alternatively, populations with high mitochondrial mutation loads may be rescued by replacement with a more fit, foreign mitochondrial haplotype. Coupled with many nonadaptive mechanisms of introgression that can preferentially affect cytoplasmic genomes, this form of adaptive introgression may contribute to the widespread discordance between mitochondrial and nuclear genealogies. Here, we review recent advances related to mitochondrial introgression and mitonuclear incompatibilities, including the potential for cointrogression of mtDNA and interacting nuclear genes. We also address an emerging controversy over the classic assumption that selection on mitochondrial genomes is inefficient and discuss the mechanisms that lead lineages down alternative evolutionary paths in response to mitochondrial mutation accumulation.     

Relationship between floral traits and floral visitors in two coexisting Tecoma species (Bignoniaceae)

In this paper, we studied the floral biology of Tecoma fulva ssp. garrocha and T.?stans in Vaqueros (Salta, Argentina), where both species coexist. We tested the idea that floral traits are associated with the pollinator types that visit them. According to our results, T.?fulva ssp. garrocha presented traits common to bird flowers and were visited by two species of Trochilidae (Colibri serrirostris and Chlorostilbon lucidus). In addition, T.?stans exhibited traits common to hymenoptera flowers and were visited principally by Apis mellifera and Bombus atratus. This study showed that floral traits are predictive of animal visitors.     

Variation in scent emission among floral parts and inflorescence developmental stages in beetle-pollinated Protea species (Proteaceae)

Floral fragrances are an important component for pollinator attraction in beetle-pollinated flowers. Several genera in the Proteaceae contain beetle-pollinated species. However, there is no information on the floral scent chemistry of beetle-pollinated members of the family. In this paper we report on the spatial variation and differences between developmental stages in emission of inflorescence (flowerhead) volatiles of four South African Protea species (P. caffra, P. dracomontana, P. simplex, and P. welwitschii) that are pollinated by cetoniine beetles. The scents from different inflorescence parts (bracts, perianth, styles, and nectar) and from successive anthesis stages of whole inflorescences were sampled using dynamic headspace collection and identified using GC–MS. Although the four species shared many scent compounds, possibly reflecting their close phylogenetic relationships and common pollinators, they showed significant differences in overall scent composition due to various species-specific compounds, such as the unique tiglate esters found in the scent of P. welwitschii. The strongest emissions and largest number of volatiles, especially monoterpenes, were from inflorescences at full pollen dehiscence. Senescing inflorescences of two species and nectars of all species emitted proportionally high amounts of acetoin (3-hydroxy-2-butanone) and aromatic alcohols, typical fermentation products. As a consequence, the scent composition of nectar was much more similar among species than was the scent composition of other parts of the inflorescence. These results illustrate how the blends of compounds that make up the overall floral scent are a dynamic consequence of emissions from various plant parts.     

Management intensity affects the relationship between non‐native and native species in subtropical wetlands

Question: Does management intensity affect the association between non‐native and native species and between non‐native species and soil nutrients in wetlands? Location: MacArthur Agro‐Ecology Research Center, Florida, USA. Methods: We evaluated native and non‐native plant richness and relative frequency in 15 1‐m² plots in 40 wetlands across two types of pastures, highly managed (fertilized, ditched, planted, heavily grazed by cattle) and semi‐natural (unfertilized, lightly seasonally grazed). Plant biomass was collected in five 0.25‐m² plots per wetland and sorted to species. Soil cores were collected to analyse soil total nitrogen (N) and phosphorus (P). An information‐theoretic approach was used to compare mixed effects models considering the association of non‐native richness, relative frequency, and biomass with native richness, relative frequency, biomass, C₃ grass relative frequency (a dominant native group), N, P and wetland‐type. Results: Non‐native richness was negatively correlated with native richness in semi‐natural wetlands, but there was no evidence of an association between these variables in highly managed wetlands. Non‐native richness increased with increasing soil N in semi‐natural wetlands, but not in the highly managed wetlands. Soil P was positively related to non‐native frequency in semi‐natural wetlands but negatively related in highly managed wetlands. Non‐native frequency and biomass were negatively related to relative frequency of C₃ grasses in both management types. Conclusions: Our results indicate that management intensity influences relationships between native and non‐native richness. Management intensity interacts with abiotic or biotic factors, such as soil nutrients and composition, in predicting where non‐native species will most likely need control.     

Ploidy effects on the relationship between floral phenotype,reproductive investment,and fitness in an autogamous species complex

Premise

The relationships between reproductive investment, phenotype, and fitness have been broadly studied in cross-pollinated plants in contrast to selfing species, which are considered less interesting in this area because they are supposed to be a dead end in any evolutionary pathway. Still, selfing plants are unique systems to study these questions since the position of reproductive structures and traits related to flower size play an important role in female and male pollination success.

Methods

Erysimum incanum s.l. is a selfing species complex that has three levels of ploidy (diploids, tetraploids, and hexaploids) and traits that are typically associated with the selfing syndrome. Here, we used 1609 plants belonging to these three ploidies to characterize the floral phenotype and spatial configuration of reproductive structures, reproductive investment (pollen and ovule production), and plant fitness. Then, we used structural equation modelling to analyze the relationship between all these variables across ploidy levels.

Results

An increase in ploidy level leads to bigger flowers with anthers exserted farther and more pollen and ovules. In addition, hexaploid plants had higher absolute values for herkogamy, which is positively correlated with fitness. Ovule production significantly mediated the natural selection acting on different phenotypic traits and pollen production, a pattern that is maintained across ploidies.

Conclusions

The changes in floral phenotypes, reproductive investment, and fitness with ploidy level suggest that genome duplication can be a driver for transitions in reproductive strategy by modifying the investment in pollen and ovules and linking them with plant phenotype and fitness.     

Variation in floral scent composition within and between populations of Geonoma macrostachys (Arecaceae) in the western Amazon

In geonomoid palms floral scent is both an important pollinator attractant and an important factor in reproductive isolation. However, little is known about intraspecific variation in floral scent composition in these as well as in other plants. In this study the level of variation in floral scent composition found within and among five populations of Geonoma macrostachys var. macrostachys in the western Amazon is documented. Floral scent samples were collected using head-space adsorption and were analyzed by gas chromatography-mass spectrometry. Most of the 108 compounds recorded were of isoprenoid origin, but only 28 of the compounds were found in all 62 samples analyzed. No differentiation was found between the studied populations, confirming that G. macrostachys var. macrostachys is outbreeding and indicating that the individual populations are part of a metapopulation linked by sufficient gene flow to avoid local differentiation. However, a negative correlation between distance and similarity of floral scent chemistry indicates a case of clinal variation within the distribution area of G. macrostachys. Male euglossine bees are infrequent visitors to G. macrostachys, while other groups of insects are abundant. However, the level of variation and the chemical composition lend support to a suggested importance of male euglossine bees in long-distance pollen flow in G. macrostachys. Other insect groups are probably important in securing pollination of most flowers with pollen from nearby sources.     

Antagonism between herbivore‐induced plant volatiles and trichomes affects tritrophic interactions

We used tomato genotypes deficient in the jasmonic acid (JA) pathway to study the interaction between the production of herbivore‐induced plant volatiles (HIPVs) that serve as information cues for herbivores as well as natural enemies of herbivores, and the production of foliar trichomes as defence barriers. We found that jasmonic acid‐insensitive1 (jai1) mutant plants with both reduced HIPVs and trichome production received higher oviposition of adult leafminers, which were more likely to be parasitized by the leafminer parasitoids than JA biosynthesis spr2 mutant plants deficient in HIPVs but not trichomes. We also showed that the preference and acceptance of leafminers and parasitoids to trichome‐removed plants from either spr2 or wild‐type (WT) genotypes over trichome‐intact genotypes can be ascribed to the reduced trichomes on treated plants, but not to altered direct and indirect defence traits such as JA, proteinase inhibitor (PI)‐II and HIPVs levels. Although the HIPVs of WT plants were more attractive to adult insects, the insects preferred trichome‐free jai1 plants for oviposition and also had greater reproductive success on these plants. Our results provide strong evidence that antagonism between HIPV emission and trichome production affects tritrophic interactions. The interactions among defence traits are discussed.     

Association between inbreeding depression and floral traits in a generalist‐pollinated plant

Individual variation in the magnitude of inbreeding depression (ID) in plants and its association with phenotypic traits may have important consequences for mating system evolution. This association has been investigated only scarcely, and always considering traits functionally related to autogamy. Here, we explore the association between individual variation in ID and plant traits associated with pollinator attractiveness (related to plant size, corolla size and corolla shape) in two populations of Erysimum mediohispanicum (Brassicaceae). ID was calculated along the entire life cycle of the plants. In addition, we also explored the relationship between phenotypic traits and the individual levels of heterozygosity. We found significant associations between ID and corolla diameter and stalk height, being taller plants with larger corollas those undergoing a lower intensity of ID. Furthermore, we found a negative relationship between corolla diameter and heterozygosity, suggesting that plants with large flowers have purged their genetic load. Finally, we found a significant effect of corolla diameter on the intrapopulation genetic structure. All these findings suggest that plants with large flowers have secularly suffered frequent inbreeding in the study populations. Because corolla diameter is a trait frequently selected by pollinators in E. mediohispanicum, we believe that the observed relationship between this trait and ID could be mediated by pollinators, probably throughout an increasing in biparental inbreeding, geitonogamy or autogamy.     

Innate attractant for long‐tongued bumblebee,Bombus diversus diversus in floral scent of bumblebee‐pollinated epiparasitic plant,Monotropastrum humile (Ericaceae)

The epiparasitic plant Monotropastrum humile (D. Don) Hara is pollinated by workers of the long‐tongued bumblebee, Bombus diversus diversus (Smith). The interaction between the floral scents of this plant and bumblebee was investigated using electrophysiological and behavioral techniques. Three components (linalool, α‐terpineol and geraniol) elicited strong responses in antennae from B. d. diversus workers and are innate attractants, suggesting floral scents play an important role in attracting these pollinators and successful pollination.     

Flowering mechanisms,pollination strategies and floral scent analyses of syntopically co‐flowering Homalomena spp. (Araceae) on Borneo

In this study, the flowering mechanisms and pollination strategies of seven species of the highly diverse genus Homalomena (Araceae) were investigated in native populations of West Sarawak, Borneo. The floral scent compositions were also recorded for six of these species. The selected taxa belong to three out of four complexes of the section Cyrtocladon (Hanneae, Giamensis and Borneensis). The species belonging to the Hanneae complex exhibited longer anthesis (53–62 h) than those of the Giamensis and Borneensis complexes (ca. 30 h). Species belonging to the Hanneae complex underwent two floral scent emission events in consecutive days, during the pistillate and staminate phases of anthesis. In species belonging to the Giamensis and Borneensis complexes, floral scent emission was only evident to the human nose during the pistillate phase. A total of 33 volatile organic compounds (VOCs) were detected in floral scent analyses of species belonging to the Hanneae complex, whereas 26 VOCs were found in samples of those belonging to the Giamensis complex. The floral scent blends contained uncommon compounds in high concentration, which could ensure pollinator discrimination. Our observations indicate that scarab beetles (Parastasia gestroi and P. nigripennis; Scarabaeidae, Rutelinae) are the pollinators of the investigated species of Homalomena, with Chaloenus schawalleri (Chrysomelidae, Galeuricinae) acting as a secondary pollinator. The pollinators utilise the inflorescence for food, mating opportunities and safe mating arena as rewards. Flower‐breeding flies (Colocasiomyia nigricauda and C. aff. heterodonta; Diptera, Drosophilidae) and terrestrial hydrophilid beetles (Cycreon sp.; Coleoptera, Hydrophilidae) were also frequently recovered from inflorescences belonging to all studied species (except H. velutipedunculata), but they probably do not act as efficient pollinators. Future studies should investigate the post‐mating isolating barriers among syntopically co‐flowering Homalomena sharing the same visiting insects.