Floral Scents of a Deceptive Plant Are Hyperdiverse and Under Population-Specific Phenotypic Selection

Floral scent is a key mediator in plant-pollinator interactions. However, little is known to what extent intraspecific scent variation is shaped by phenotypic selection, with no information yet in deceptive plants. In this study, we collected inflorescence scent and fruit set of the deceptive moth fly-pollinated Arum maculatum L. (Araceae) from six populations north vs. five populations south of the Alps, accumulating to 233 samples in total, and tested for differences in scent, fruit set, and phenotypic selection on scent across this geographic barrier. We recorded 289 scent compounds, the highest number so far reported in a single plant species. Most of the compounds occurred both north and south of the Alps; however, plants of the different regions emitted different absolute and relative amounts of scent. Fruit set was higher north than south of the Alps, and some, but not all differences in scent could be explained by differential phenotypic selection in northern vs. southern populations. This study is the first to provide evidence that floral scents of a deceptive plant are under phenotypic selection and that phenotypic selection is involved in shaping geographic patterns of floral scent in such plants. The hyperdiverse scent of A. maculatum might result from the imitation of various brood substrates of its pollinators.

Floral scents of a deceptive plant are hyperdiverse and under population-specific phenotypic selection

ABSTRACTFloral scent is a key mediator in plant-pollinator interactions; however, little is known to what extent intraspecific scent variation is shaped by phenotypic selection, with no information yet in deceptive plants. We recorded 291 scent compounds in deceptive moth fly-pollinated Arum maculatum from various populations north vs. south of the Alps, the highest number so far reported in a single plant species. Scent and fruit set differed between regions, and some, but not all differences in scent could be explained by differential phenotypic selection in northern vs. southern populations. Our study is the first to provide evidence that phenotypic selection is involved in shaping geographic patterns of floral scent in deceptive plants. The hyperdiverse scent of A. maculatum might result from the plant’s imitation of various brood substrates of its pollinators.

Within-species floral odor variation is maintained by spatial and temporal heterogeneity in pollinator communities

ABSTRACTFloral odor is a complex trait that mediates many biotic interactions, including pollination. While high intraspecific floral odor variation appears to be common, the ecological and evolutionary drivers of this variation are often unclear. Here, we investigated the influence of spatially and temporally heterogeneous pollinator communities on floral odor variation in Arum maculatum (Araceae). Through Europe-wide field surveys, we identified high floral odor diversity and shifts in the dominant pollinator species within several A. maculatum populations compared to pollinator data from the same sites ten years ago. Using common-garden experiments, we further confirmed that inflorescences from native and foreign pollinator backgrounds were equally efficient at attracting local pollinators. The substantial within-population floral odor variation we observed may therefore be advantageous when facing temporally heterogeneous pollinator communities. We propose spatio-temporal heterogeneity in pollinators as one potential mechanism maintaining diverse floral odor bouquets in angiosperms.

Degradation of mitochondrial alternative oxidase in the appendices of Arum maculatum

Cyanide-resistant alternative oxidase (AOX) is a nuclear-encoded quinol oxidase located in the inner mitochondrial membrane. Although the quality control of AOX proteins is expected to have a role in elevated respiration in mitochondria, it remains unclear whether thermogenic plants possess molecular mechanisms for the mitochondrial degradation of AOX. To better understand the mechanism of AOX turnover in mitochondria, we performed a series of in organello AOX degradation assays using mitochondria from various stages of the appendices of Arum maculatum. Our analyses clearly indicated that AOX proteins at certain stages in the appendices are degraded at 30°C, which is close to the maximum appendix temperature observed during thermogenesis. Interestingly, such temperature-dependent protease activities were specifically inhibited by E-64, a cysteine protease inhibitor. Moreover, purification and subsequent nano LC–MS/MS analyses of E-64-sensitive and DCG-04-labeled active mitochondrial protease revealed an ∼30 kDa protein with an identical partial peptide sequence to the cysteine protease 1-like protein from Phoenix dactylifera. Our data collectively suggest that AOX is a potential target for temperature-dependent E-64-sensitive cysteine protease in the appendices of A. maculatum. A possible retrograde signalling cascade mediated by specific degradation of AOX proteins and its physiological significance are discussed.

Phytochemical analysis, cytotoxicity and antioxidant activity of cuckoo pint (Arum maculatum) leaf extract

Arum maculatum is traditionally used for the control of many diseases and illnesses such as kidney pain, liver injury, hemorrhoids. However, the detailed biomedical knowledge about this species is still lacking. This study reports on the bioactive components and the possible mechanisms underlying the antioxidant, anti-inflammatory and cytotoxic activity of A. maculatum leaf extract. Gas chromatography-mass spectrometry (GC-MS) was used for phytochemical analysis. Assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide ) (MTT) was used to determine the cytotoxicity in the murine cell line L20B upon exposure to different extract concentrations for 24 h. Enzyme-linked immunosorbent assay (ELISA) was used to detect pro-inflammatory cytokines and tumor necrosis factor-α (TNF-α). GC-MS analysis identified the presence of important phytochemical components, e.g., 9-octadecenoic acid, methyl ester, (E), hexadecanoic acid, methyl ester, followed by benzenepropanoic acid, 3,5-bis(1,1-dimethylethyl)-4-hydroxy-, methyl ester (17.74%), heptadecanoic acid, 16-methyl-, methyl ester and dibutyl phthalate. The results indicated a significant dose-dependent decrease in L20B cell growth at a dose of 400 μg/ml (IC50) that is associated with a significant 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. The results suggested that the aqueous extract of A. maculatum leaves have potent antioxidant activity and cytotoxicity against L20B cell line with potential pro-inflammatory activity.