Metabolomic and Transcriptomic Analysis Reveal the Role of Metabolites and Genes in Modulating Flower Color of Paphiopedilum micranthum.

Food-deceptive flowers primarily use visual signals (such as color) to mimic model plants and deceive insects into achieving pollination. Paphiopedilum micranthum is a food-deceptive orchid that has a pink labellum and two purple petals with a yellow base and has been proven to be pollinated by bumblebees. However, the chemical and molecular bases of the floral color are not well understood. We conducted targeted metabolite profiling and transcriptomic analysis to determine the color signal and its genetic basis in P. micranthum. We found that both anthocyanins and carotenoids contribute significantly to the formation of floral color that determines the color signal. Higher concentrations of anthocyanins (cyanidin and peonidin) and carotenoids (primarily lutein and zeaxanthin) were detected in the petal compared to the labellum. The upregulation of structural genes of CHS, F3'H, DFR and ANS on the anthocyanin biosynthesis pathway in petals was identified, as well as three genes of LCYE, BCH, and CCD4 on the carotenoid biosynthesis pathway. Furthermore, we discovered that three R2R3-MYBs and one bHLH transcription factors were co-expressed with the expression of different genes. These genes and transcription factors may be responsible for the spatial color difference of P. micranthum. Our study emphasizes that the color of this food-deceptive orchids is achieved through specific genes and transcription factors associated with the pigment biosynthesis pathway.

Color preference and spatial distribution of glaphyrid beetles suggest a key role in the maintenance of the color polymorphism in the peacock anemone (Anemone pavonina, Ranunculaceae) in Northern Greece.

In the Mediterranean region, a group of unrelated plant species share an unusual deep-red flower color and are pollinated by glaphyrid beetles. Some of these species possess different color morphs, but the mechanisms maintaining this color polymorphism are unknown. The peacock anemone, Anemone pavonina, is a color polymorphic species with red or purple flowers. We investigated the spatial distribution of its color morphs and its potential glaphyrid pollinators, Pygopleurus spp., along an elevational gradient on the southern slopes of Mount Olympus, Greece. We found a correlation between relative proportions of the two color morphs with both elevation and beetle abundance. At low elevations (< 1000 m a.s.l.), beetles were abundant and anemone populations comprised only red flowers. Above a steep transition zone with mixed-colored populations (c. 1000-1300 m) most flowers were purple and beetles were rare. Color-trapping experiments revealed a strong preference for red over other colors in beetles and colorimetric modeling suggests that a simple chromatic mechanism is sufficient to explain their color choices. We thus hypothesize that beetles select for red flowers and that with increasing elevation and decreasing beetle density, other flower visitors (e.g., bees) gain importance as pollinators and select for a different color.

Crosstalk between Photoreceptor and Sugar Signaling Modulates Floral Signal Transduction.

Over the past decade, integrated genetic, cellular, proteomic and genomic approaches have begun to unravel the surprisingly crosstalk between photoreceptors and sugar signaling in regulation of floral signal transduction. Although a number of physiological factors in the pathway have been identified, the molecular genetic interactions of some components are less well understood. The further elucidation of the crosstalk mechanisms between photoreceptors and sugar signaling will certainly contribute to our better understanding of the developmental circuitry that controls floral signal transduction. This article summarizes our current knowledge of this crosstalk, which has not received much attention, and suggests possible directions for future research.

Tissue-Specific Emission of (E)-α-Bergamotene Helps Resolve the Dilemma When Pollinators Are Also Herbivores.

More than 87% of flowering plant species are animal-pollinated [1] and produce floral scents and other signals to attract pollinators. These floral cues may however also attract antagonistic visitors, including herbivores [2]. The dilemma is exacerbated when adult insects pollinate the same plant that their larvae consume. It remains largely unclear how plants maximize their fitness under these circumstances. Here we show that in the night-flowering wild tobacco Nicotiana attenuata, the emission of a sesquiterpene, (E)-α-bergamotene, in flowers increases adult Manduca sexta moth-mediated pollination success, while the same compound in leaves is known to mediate indirect defense against M. sexta larvae [3, 4]. Forward and reverse genetic analyses demonstrated that both herbivory-induced and floral (E)-α-bergamotene are regulated by the expression of a monoterpene-synthase-derived sesquiterpene synthase (NaTPS38). The expression pattern of NaTPS38 also accounts for variation in (E)-α-bergamotene emission among natural accessions. These results highlight that differential expression of a single gene that results in tissue-specific emission of one compound contributes to resolving the dilemma for plants when their pollinators are also herbivores. Furthermore, this study provides genetic evidence that pollinators and herbivores interactively shape the evolution of floral signals and plant defense.

"Hummingbird" floral traits interact synergistically to discourage visitation by bumble bee foragers.

Pollination syndromes are suites of floral traits presumed to reflect adaptations to attract and utilize a "primary" type of animal pollinator. However, syndrome traits may also function to deter "secondary" flower visitors that reduce plant fitness through their foraging activities. Here we use the hummingbird-pollinated plant species Mimulus cardinalis as a model to investigate the potential deterrent effects of classic bird syndrome traits on bumble bee foragers. To establish that M. cardinalis flowers elicit an avoidance response in bees, we assessed the choice behavior of individual foragers on a mixed experimental array of M. cardinalis and its bee-pollinated sister species M. lewisii. As expected, bees showed a strong preference against M. cardinalis flowers (only 22% of total bee visits were to M. cardinalis), but surprisingly also showed a high degree of individual specialization (95.2% of total plant transitions were between conspecifics). To determine M. cardinalis floral traits that discourage bee visitation, we then assessed foraging responses of individuals to M. cardinalis-like and M. lewisii-like floral models differing in color, orientation, reward, and combinations thereof. Across experiments, M. cardinalis-like trait combinations consistently produced a higher degree of flower avoidance behavior and individual specialization than expected based on bee responses to each trait in isolation. We then conducted a series of flower discrimination experiments to assess the ability of bees to utilize traits and trait combinations associated with each species. Relative to M. lewisii-like alternatives, M. cardinalis-like traits alone had a minimal effect on bee foraging proficiency but together increased the time bees spent searching for rewarding flowers from 1.49 to 2.65 s per visit. Collectively, our results show that M. cardinalis flowers impose foraging costs on bumble bees sufficient to discourage visitation and remarkably, generate such costs through synergistic color-orientation and color-reward trait interactions. Floral syndromes therefore represent complex adaptations to multiple pollinator groups, rather than simply the primary pollinator.

The effects of becoming taller: direct and pleiotropic effects of artificial selection on plant height in Brassica rapa.

Plant height is an important trait for plant reproductive success. Plant height is often under pollinator-mediated selection, and has been shown to be correlated with various other traits. However, few studies have examined the evolutionary trajectory of plant height under selection and the pleiotropic effects of plant height evolution. We conducted a bi-directional artificial selection experiment on plant height with fast cycling Brassica rapa plants to estimate its heritability and genetic correlations, and to reveal evolutionary responses to artificial selection on height and various correlated traits. With the divergent lines obtained through artificial selection, we subsequently conducted pollinator-choice assays and investigated resource limitation of fruit production. We found that plant height variation is strongly genetically controlled (with a realized heritability of 41-59%). Thus, plant height can evolve rapidly under phenotypic selection. In addition, we found remarkable pleiotropic effects in phenology, morphology, floral scent, color, nectar and leaf glucosinolates. Most traits were increased in tall-line plants, but flower size, UV reflection and glucosinolates were decreased, indicating potential trade-offs. Pollinators preferred plants of the tall selection lines over the short selection lines in both greenhouse experiments with bumblebees and field experiment with natural pollinators. We did not detect any differences in resource limitation between plants of the different selection lines. Overall, our study predicts that increased height should evolve under positive pollinator-mediated directional selection with potential trade-offs in floral signals and herbivore defense.