How can structure and nectar composition explain the secretory process in super productive nuptial nectaries of Mabea fistulifera Mart. (Euphorbiaceae)?

Mabea fistulifera, a species pollinated mainly by diurnal and nocturnal vertebrates, presents pendulous inflorescences with approximately 70 pairs of nuptial nectaries (NNs). These NNs exude voluminous nectar drops that defy gravity, remaining exposed at the inflorescence for more than a day. We aimed to investigate the NN secretory process and the unique nectar presentation of M. fistulifera. NNs and their exudate were collected at different secretory stages and submitted to structural studies and chemical analysis. The epidermis is devoid of stomata and constitutes the main site of synthesis for non-sugar metabolites found on nectar and nectar-coating film. Nectary parenchyma presents few small starch grains, and vascular strands are distributed until the nectary parenchyma cells close to the epidermis. Vascular tissues at the nectary parenchyma seem to provide sugar and water for the nectar. A film composed of lipids, alkaloids, and proteins covers the nectar drops. The film guarantees the nectar offering for several hours, as it minimizes water loss and prevents falls by gravitational action. The release of large nectar drops is intrinsically linked to the NN anatomical traits and the exudate composition. Low sugar concentration and predominance of hexoses in M. fistulifera nectar are essential for maintaining nectar exudation for many hours, which results in the visitation of a broad spectrum of pollinators.

SEX EXPRESSION, BREEDING SYSTEM AND POLLINATORS OF Piper caldense (PIPERACEAE) IN THE BRAZILIAN ATLANTIC FOREST / Expresión sexual, sistema reproductivo y polinizadores de Piper caldense (Piperaceae) en un bosque Atlántico

ABSTRACT Neotropical Piper species have bisexual flowers. Such reproductive trait is considered basal in this pantropical genus. However, neotropical species having unisexual (staminate) flowers along with bisexual ones have also been reported. Dichogamy is common in the genus, associated with either self-compatibility or -incompatibility, as well as with entomophily. We analyzed a natural population of Piper caldense in a Atlantic Forest area (Viçosa municipality, Minas Gerais state, southeastern Brazil). Preliminary observations indicated that the species produces two flower types. We analyzed flower sex in spikes of 50 plants. We obtained additional information through morphological and anatomical studies and scanning electron microscopy analyses. The longevity and exposure dynamics of stigmatic papillae and the pollen release sequence of all four stamens were investigated to verify the degree of dichogamy. Pollination tests were performed, pollinators were identified and their visitation frequency was recorded. Piper caldense has both bisexual and staminate flowers, each flower type being located on separate spikes. These spikes occurred on the same plant, thus indicating andromonoecy; furthermore, plants having only spikes with staminate flowers were also observed. This gender had not yet been reported to the genus. Gradual and sequential exposure of stigmatic papillae associated with asynchronous pollen release (one stamen a day) resulted in incomplete protogyny. Hand pollination tests showed that the species is self-incompatible. Social bees, mainly Apis mellifera and Melipona spp., were the major pollinator group. Our study reinforces the need to associate morphological analysis with floral biology and indicates future changes in studies addressing reproductive traits associated with the phylogeny of the Piper genus.

RESUMEN Las especies neotropicales de Piper presentan flores bisexuales, condición considerada basal al interior de este género pantropical. Sin embargo, fueron observadas especies neotropicales con flores unisexuales (estaminadas), además de las bisexuales. La dicogamia es común en el género y se encuentra asociada a la autocompatibilidad o incompatibilidad, al igual que la entomofilia. Fue analizada una población natural de Piper caldense en un área de bosque Atlántico (municipio de Viçosa, Minas Gerais, sureste de Brasil). Las observaciones preliminares indicaron que esta especie produce dos tipos florales, para lo cual fue analizada la sexualidad de las flores en 50 espigas a través de estudios morfológicos, anatómicos y análisis con microscopia electrónica de barrido. La longevidad y dinámica de exposición de las papilas estigmáticas y la secuencia de liberación de los granos de polen en los cuatro estambres fueron observados para verificar el grado de dicogamia. El sistema reproductivo se evaluó mediante el test de autopolinización espontánea y polinización abierta. Los visitantes florales fueron identificados y se registró su frecuencia de visita. Piper caldense presenta flores bisexuales y estaminadas, cada tipo floral se encuentra en espigas separadas. Estas espigas se encuentran en la misma planta, lo que indica que es andromonoica; adicionalmente, fueron observadas plantas que solo presentaban espigas con flores estaminadas, lo que se constituye en una combinación sexual inédita para el género. La exposición gradual y secuencial de las papilas estigmáticas asociada con la liberación asincrónica de los granos de polen (un estambre por día) indica una protoginia incompleta. Los test de polinización mostraron que la especie es autoincompatible. Las abejas sociales, principalmente Apis mellifera y Melipona spp., fueron los polinizadores principales. Este estudio refuerza la necesidad de asociar los analisis morfológicos con la funcionalidad de las estructuras florales, además indica los cambios futuros en estudios que abordan características reproductivas asociadas a la filogenia de las especies de Piper.

Timing of pollen release and stigma receptivity period of Piper vicosanum: New insights into sexual reproduction of the genus.

PREMISE OF THE STUDY: Dichogamy is a common characteristic among angiosperms, including Piper species. In this genus, the tiny flowers are morphologically similar and have an asynchronous stamen development. However, there is no information on the duration of stigma receptivity and whether it overlaps with pollen release. To better understand mechanisms of floral function in Piper vicosanum, we provide a detailed characterization of the timing of pollen release from the four stamens and the period of stigma receptivity and exposure mode of the receptive areas. METHODS: We investigated plants of a natural population in a semideciduous seasonal forest (Viçosa, Minas Gerais State, southeastern Brazil), based on chemical tests, light microscopy, and scanning electron microscopy analyses. KEY RESULTS: Incomplete protogyny-a mechanism that favors outcrossing-was recorded. The period of stigma receptivity was long (14 d), and the sequential exposure and senescence of stigmatic papillae occurred gradually and in a basipetal direction. Pollen release began 2-6 d after the beginning of the pistillate phase, with an average pollen viability of 87.7%, during the bisexual flower phase. Pollen was released for up to 6 d and occurred in one stamen at a time. The fruit set observed in tests of self-pollination indicated self-compatibility. CONCLUSIONS: The gradual and sequential exposure of stigmatic papillae in P. vicosanum flowers is described here as the mechanism for the long duration of receptivity. Anther development and pollen release were also sequential. These findings are yet unreported reproductive characteristics of the genus and offer new perspectives for future studies on the floral biology of other Piper species.

Trioecy in Coccoloba cereifera Schwacke (Polygonaceae), a narrow endemic and threatened tropical species

Trioecy, the co-occurrence of the males, females, and hermaphrodite morphs in natural populations, is a rare and poorly studied breeding system. It is expressed in Coccoloba cereifera, an endemic, and endangered species from the rupestrian fields of Serra do Cipó, southeastern Brazil. Male individuals produce staminate flowers but no fruits. Female individuals produce pistillate flowers and set fruits. Both staminate and pistillate flowers present non-functional organs of the opposite sex that simulate perfect flowers. Hermaphrodite individuals produce two different perfect flowers, each one belonging to distinct individuals, and set fruits. Perfect flowers differ in the amount of pollen produced, in pollen viability, and in some morphological traits. Two Hymenoptera species visited the flowers sporadically. The low natural fructification indicated a limited pollination, while the fruit set recorded in bagged pistillate flowers indicated agamospermy. Female individuals represent more than 40 percent of the population studied and must be the result of agamospermic seeds.

A trioicia, a coocorrência, em população natural, de indivíduos masculinos, femininos e hermafroditas, é um sistema reprodutivo raro e pouco estudado. Coccoloba cereifera (Polygonaceae), espécie endêmica e em perigo de extinção dos campos rupestres da Serra do Cipó, sudeste brasileiro, apresenta esse sistema. Indivíduos masculinos produzem flores estaminadas e não frutificam. Indivíduos femininos produzem flores pistiladas e frutificam. Ambas flores estaminadas e pistiladas apresentam órgãos não-funcionais do sexo oposto, tornando-as semelhantes às flores perfeitas. Indivíduos hermafroditas produzem dois tipos de flores perfeitas, cada um pertencente a indivíduos distintos, e ambos frutificam. As flores perfeitas diferem entre si na quantidade de pólen produzido, na viabilidade do pólen e em algumas características morfológicas. Duas espécies de Hymenoptera visitaram as flores esporadicamente. A baixa frutificação natural indicou uma polinização limitada, ao passo que a frutificação registrada em flores pistiladas, isoladas por sacos, indicou agamospermia. Indivíduos femininos representaram mais que 40 por cento da população estudada e devem ser resultantes de sementes agamospérmicas.

Removal and insertion of pollinia in flowers of Oxypetalum (Asclepiadaceae) in southeastern Brazil.

Pollinarium removal and pollinium insertion of seven Oxypetalum species (O. alpinum var. alpinum, O. appendiculatum, O. banksii subsp. banksii, O. jacobinae, O. mexiae, O. pachyglossum and O. subriparium) were recorded in Viçosa. Minas Gerais. They presented a tendency of one or two pollinarium removals and one pollinium insertion (single insertion), except O. appendiculatum. In this species, mainly, two pollinia of the same pollinarium were inserted per stigmatic chamber (double insertion), resulting exceptionally in 6-10 inserted pollinia in a flower, an unusual occurrence among the Asclepiadaceae. No association between removal and insertion was found, e.g., O. subriparium and O. banksii subsp. banksii had the highest pollinarium removal (1.78 and 1.45, respectively) and one of the lowest pollinium insertions (0.02 in both species), per flower. Oxypetalum mexiae showed the lowest pollinarium removal and pollinium insertion per flower (0.09 and 0.01, respectively) among the studied species and other Asclepiadaceae. Oxypetalum subriparium, O. banksii subsp. banksii and O. mexiae might be having reproductive limitations. Pollinarium removal and pollinium insertion per flower of the studied species varied from site to site, similarly to what was recorded for other Asclepiadaceae.

Removal and insertion of pollinia in flowers of Oxypetalum (Asclepiadaceae) in southeastern Brazil

Pollinarium removal and pollinium insertion of seven Oxypetalum species (O. alpinum var. alpinum, O. appendiculatum, O. banksii subsp. banksii, O. jacobinae, O. mexiae, O. pachyglossum and O. subriparium) were recorded in Viçosa. Minas Gerais. They presented a tendency of one or two pollinarium removals and one pollinium insertion (single insertion), except O. appendiculatum. In this species, mainly, two pollinia of the same pollinarium were inserted per stigmatic chamber (double insertion), resulting exceptionally in 6-10 inserted pollinia in a flower, an unusual occurrence among the Asclepiadaceae. No association between removal and insertion was found, e.g., O. subriparium and O. banksii subsp. banksii had the highest pollinarium removal (1.78 and 1.45, respectively) and one of the lowest pollinium insertions (0.02 in both species), per flower. Oxypetalum mexiae showed the lowest pollinarium removal and pollinium insertion per flower (0.09 and 0.01, respectively) among the studied species and other Asclepiadaceae. Oxypetalum subriparium, O. banksii subsp. banksii and O. mexiae might be having reproductive limitations. Pollinarium removal and pollinium insertion per flower of the studied species varied from site to site, similarly to what was recorded for other Asclepiadaceae.