Investigating the reproductive strategies of Deuterocohnia meziana (Bromeliaceae), an endangered and restricted species from South American rocky outcrops.

Studies of reproductive biology and resources availability to floral visitors by plant species are important to understand the plant-pollinator interactions that drive species adaptation. We aim to understand the relationship between reproduction mechanisms of Deuterocohnia meziana (Bromeliaceae) and pollinators. The species occurs in Bolivia and Paraguay, and it is the only species of the genus found in Brazil, where it is restricted to ironstone outcrops. These areas are currently threatened by the iron mining industry. Additionally, they face risks from fire occurrence and grazing by cattle. We analyzed the floral biology, reproductive system, phenology, and pollination ecology of a natural population of Deuterocohnia meziana, from ironstone outcrops in Brazil. The species exhibits diurnal anthesis, with stigma receptive throughout anthesis, and 77% of pollen viability. Deuterocohnia meziana produces relatively large amounts of nectar, especially early in the morning (32.8 ± 9.4 µl), with a mean sugar concentration of 23.5 (± 3.2) ºBrix. It is self-incompatible with a peak flowering occurring in August (dry season), although flowers are observed continuously throughout the year. The species exhibits two types of inflorescences, young and mature, among which an average of 13.1 and 3.6 flowers open per day, respectively. Hummingbirds and bees are the effective pollinators, although butterflies and ants also visit D. meziana flowers. The species is reliant on exogenous pollen and pollinators for fruit set. The continuous conservation of D. meziana populations and their communities is essential for preserving plant-pollinator mutualism and the floral community adapted to ironstone outcrops.

Development, characterization, and transferability of SSR markers for Vriesea carinata (Bromeliaceae) based on RNA sequencing.

PREMISE OF THE STUDY: Expressed sequence tag-simple sequence repeat (EST-SSR) markers were isolated for Vriesea carinata, an endemic bromeliad from the Brazilian Atlantic Forest. These SSR loci may be used to investigate the genetic diversity and population structure of this species and related bromeliads. METHODS AND RESULTS: Based on the transcriptome data of V. carinata, 30 primer pairs were designed and selected for initial validation. Of these primer pairs, 16 generated suitable SSR loci in 69 individuals. The number of alleles per locus ranged from one to 13; the levels of observed and expected heterozygosity per locus ranged from 0.000 to 1.000 and from 0.000 to 0.935, respectively. All loci produced heterologous amplification. Transferability of the loci was tested in 15 species belonging to three Bromeliaceae subfamilies. CONCLUSIONS: The developed EST-SSR markers revealed polymorphism in the four studied populations and could be useful to investigate the genetic diversity of V. carinata and related species. The markers may also be suitable for novel gene annotation and discovery.

Microsatellites in the endangered species Dyckia distachya (Bromeliaceae) and cross-amplification in other bromeliads.

Microsatellite markers were isolated in Dyckia distachya, an endangered bromeliad from southern Brazil, which will be useful to assess the population genetic structure and reproductive success in introduced and natural populations of this species. Twenty microsatellite loci were developed from an enriched genomic library, and nine of these were amplified. The loci were characterized in 43 individuals from introduced and wild D. distachya populations. All nine loci were polymorphic, with four to ten alleles per locus. In an introduced population the observed and expected heterozygosities ranged from 0.136-0.667 and 0.543-0.877, respectively, while in a wild population it ranged from 0.000 to 0.895 and from 0.050 to 0.811, respectively. The development of these microsatellite markers will contribute to investigations of the reproductive potential and viability of introduced populations of D. distachya as well as the single known wild population. Cross-amplification in other Bromeliaceae species was successful, with high rates in four loci, demonstrating the applicability of these microsatellite markers in other taxa.

Genetics, evolution and conservation of Bromeliaceae.

Bromeliaceae is a morphologically distinctive and ecologically diverse family originating in the New World. Three centers of diversity, 58 genera, and about 3,140 bromeliad species are currently recognized. We compiled all of the studies related to the reproductive biology, genetic diversity, and population structure of the Bromeliaceae, and discuss the evolution and conservation of this family. Bromeliads are preferentially pollinated by vertebrates and show marked variation in breeding systems, from predominant inbreeding to predominant outcrossing, as well as constancy in chromosome number (2n = 2x = 50). Autogamous or mixed mating system bromeliads have a high inbreeding coefficient (F(IS)), while outcrossing species show low F(IS). The degree of differentiation among populations (F(ST))of species ranges from 0.043 to 0.961, which can be influenced by pollen and seed dispersal effects, clonal growth, gene flow rates, and connectivity among populations. The evolutionary history of the Bromeliaceae is poorly known, although some studies have indicated that the family arose in the Guayana Shield roughly 100 Mya. We believe that genetic, cytogenetic, and reproductive data will be essential for diagnosing species status and for assisting conservation programs.

Cross-species transfer of nuclear microsatellite markers: potential and limitations.

Molecular ecologists increasingly require 'universal' genetic markers that can easily be transferred between species. The distribution of cross-species transferability of nuclear microsatellite loci is highly uneven across taxa, being greater in animals and highly variable in flowering plants. The potential for successful cross-species transfer appears highest in species with long generation times, mixed or outcrossing breeding systems, and where genome size in the target species is small compared to the source. We discuss the implications of these findings and close with an outlook on potential alternative sources of cross-species transferable markers.

Fertility of Vriesea gigantea Gaud. (Bromeliaceae) in southern Brazil.

Plant fertility is a central subject of many questions in plant evolutionary and conservation biology. Pollen availability, abiotic resources, and flowering pattern can limit fruit and seed production. Open pollination and pollen supplementation studies are used to estimate any pollen limitation in natural populations. To study the impact of these factors on the reproductive success of Vriesea gigantea, an epiphytic bromeliad in the Atlantic Rainforest in Brazil, its fertility in four natural populations in Itapuã State Park was assessed by considering plant and inflorescence size, flower production, fruit and seed set, flower and fruit set pattern, and seed viability and germination rate. Supplemental pollination in adult plants was used to determine whether fruit production in V. gigantea is limited by reception of pollen. The results showed that V. gigantea has a high production of flowers, fruits, and seeds. Seeds are highly viable in all populations, presenting an average germination rate of 94% (SE ± 3.5). Plants of V. gigantea from Itapuã State Park are highly fertile. The high proportion of fruit and seed set after manual hand pollination indicates that the species is self-compatible. Pollination treatments showed evidence of pollinator limitation in the Itapuã State Park population.