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Petunia is endemic to South America grasslands; member of this genus exhibit variation in flower colour and shape, attracting bees, hawkmoths or hummingbirds. This group of plants is thus an excellent model system for evolutionary studies of diversification associated with pollinator shifts. Our aims were to identify the legitimate pollinator of Petunia secreta , a rare and endemic species, and to assess the importance of floral traits in pollinator attraction in this Petunia species. To determine the legitimate pollinator, field observations were conducted, and all floral visitors were recorded and evaluated. We also measured the nectar volume and sugar concentration. To characterize morphological cues for pollinators, we assessed the ultraviolet (UV)-light response in detached flowers, and characterized the floral pigments and pollen volatile scents for four different Petunia species that present different pollination syndromes. Petunia secreta shares the most recent ancestor with a white hawkmoth-pollinated species, P. axillaris , but presents flavonols and anthocyanin pigments responsible for the pink corolla colour and UV-light responses that are common to bee-pollinated Petunia species. Our study showed that a solitary bee in the genus Pseudagapostemon was the most frequent pollinator of P. secreta , and these bees collect only pollen as a reward. Despite being mainly bee-pollinated, different functional groups of pollinators visit P. secreta . Nectar volume, sugar concentration per flower, morphology and components of pollen scent would appear to be attractive to several different pollinator groups. Notably, the corolla includes a narrow tube with nectar at its base that cannot be reached by Pseudagapostemon , and flowers of P. secreta appear to follow an evolutionary transition, with traits attractive to several functional groups of pollinators. Additionally, the present study shows that differences in the volatiles of pollen scent are relevant for plant mutualistic and antagonist interactions in Petunia species and that pollen scent profile plays a key role in characterizing pollination syndromes.

The analysis of genetic structure and variability of isolated species is of critical importance in evaluating whether stochastic or human-caused factors are affecting rare species. Low genetic diversity compromises the ability of populations to evolve and reduces their chances of survival under environmental changes. Petunia secreta, a rare and endemic species, is an annual and heliophilous herb that is bee-pollinated and easily recognizable by its purple and salverform corolla. It was described as a new species of the Petunia genus in 2005. Few individuals of P. secreta have been observed in nature and little is known about this species. All the natural populations of P. secreta that were found were studied using 15 microsatellite loci, two intergenic plastid sequences and morphological traits. Statistical analysis was performed to describe the genetic diversity of this rare species and the results compared with those of more widespread and frequent Petunia species from the same geographic area to understand whether factors associated with population size could affect rare species of this genus. The results showed that despite its rarity, P. secreta presented high genetic diversity that was equivalent to or even higher than that of widespread Petunia species. It was shown that this species is divided into two evolutionary lineages, and the genetic differentiation indices between them and other congeneric species presented different patterns. The major risk to P. secreta maintenance is its rarity, suggesting the necessity of a preservation programme and more biological and evolutionary studies that handle the two evolutionary lineages independently., (Published by Oxford University Press on behalf of the Annals of Botany Company.)