Reproductive biology of the herkogamous vine Chiococca alba (L.) Hitchc. (Rubiaceae) in the Atlantic Rain Forest, SE Brazil

The family Rubiaceae comprises a wide spectrum of floral mechanisms and two of them seem to be common in certain groups, e.g., distyly in Rubioidae and styllar pollen in Ixoroidae. These mechanisms include herkogamy, which is interpreted as a strategy that avoids self-pollination. This is the first report on the reproductive biology of Chiococca alba, a species that is widely distributed in America. We studied floral biology and the mating system, which were evaluated through fruit set comparisons after controlled crosses (self- and cross-pollinations and test for apomixis), as well as through the evaluation of pollen tube growth resulting from these controlled crosses. Flowers of C. alba are herkogamous, cream, protandrous and lasted for two days. No measurable nectar was found, despite the presence of a nectary-like structure at the base of the corolla tube. Chiococca alba is a preferentially self-incompatible species, but self-pollination and apomixis also contribute to the natural fruit-set. Its reproductive strategy (herkogamy associated with protandry) is different from that expected for members of Chiococceae tribe (i.e., styllar pollen presentation).

Synchrony is more than overlap: measuring phenological synchronization considering time length and intensity

The degree of flowering and fruiting synchronization is believed to have ecological and evolutionary relevance at several scales. Here we discuss some measures that have been used to estimate synchrony and propose an index that incorporates both the entire length of an individual phenophase and variation in the number of flowers or fruits over that time period. This new index describes more accurately the phenological synchrony among individuals and populations.

Are there evidences of a complex mimicry system among Asclepias curassavica (Apocynaceae), Epidendrum fulgens (Orchidaceae), and Lantana camara (Verbenaceae) in Southern Brazil?

The goal of this paper was to test the presence of mimicry in Asclepias curassavica L., Epidendrum fulgens Brong., and Lantana camara L. The study was carried out at the Parque Estadual de Itapeva, RS, southern Brazil, from 2004 to 2006. Flowering period of each of the three species was followed up; focal observations of butterflies visiting flowers, from fixed point and during random walks were carried out. We also estimated the frequency of pollinaria removal in the orchid, as well as its mode of reproduction. All these variables were important for testing the mimicry hypothesis. Despite some temporal coincidences in the flowering period of two plants in the system, there was no statistical association among the three plants as to flowering period. Twenty-nine species of butterflies, as potential pollinators, were recorded, particularly Agraulis vanillae maculosa, Dryas iulia alcionea, Urbanus simplicius, Tegosa claudina, and Heliconius erato phyllis, which were the more frequent visitors of the three plants. There was association between the number of visits to L. camara and E. fulgens, based on Pearson correlation (r = 0.4603; n = 19; P = 0.0473). Pollinaria removal of E. fulgens was low, as measured by the percentage of removal (range: 0 - 10%). The analysis of the mode of reproduction of this orchid showed its pollinator-dependence, since no fruits were formed by spontaneous self-pollination. In contrast, the percentage of fruit set that resulted from geitonogamy and xenogamy was, in average, 86%. The results here shown are not conclusive as to the occurrence of a mimicry system among the three plants.

Reproductive biology of Protium spruceanum (Burseraceae), a dominant dioecious tree in vegetation corridors in Southeastern Brazil

We investigated the reproductive biology of Protium spruceanum (Benth.) Engler in vegetation corridors of secondary Atlantic forest in Lavras, southern Minas Gerais State, Brazil. The reproductive phenology was investigated fortnightly over a one year period. Floral biology studies involved pollen viability analysis, nectar production, stigmatic receptivity, pollen tube growth, visiting insect species and visit rates. The small, pale yellowish flowers (0.3-0.4 cm diameter) are functionally unisexual and organized in dense inflorescences (ca. 45 flowers). P. spruceanum presented annual flowering between September and November. Staminate flowers supplied a high percentage of viable pollen (90.6%) and relatively abundant nectar (x = 4.5 μL). Pistillate flowers produced only nectar to flower visitors (x = 4.0 μL). The effective pollinators were Apis mellifera and Trigona sp. (Hymenoptera, Apidae). Pollen tubes of cross-pollinated flowers were observed entering the ovaries 48 h after pollination. The fruiting season is from October to March, with a peak in November, coinciding with the rainfall peak. Ecological implications of these findings, and alternative arguments to explain the high genetic diversity at regional landscape are discussed.

Flowering and fruiting of Aulonemia aristulata: a gynomonoecious woody bamboo species from Atlantic Forest in Brazil

Aulonemia aristulata (Döll) McClure is a lignified bamboo species endemic to Brazil. This species occurs in southeastern forests and can reach high density at forest edges, dominating the understory of canopy-disturbed forest patches. The goal of this study was to describe the flowering period, floral biology, fruiting and seedling recruitment of A. aristulata in natural conditions in two areas located in a segment of the Atlantic Forest. Data on the morphology of the synflorescences and florets, timing and sequence of the anthesis events and floral visitors were recorded. Natural pollinators (open pollination or control) as well as spontaneous self-pollination were also checked. Pollen viability was estimated using the acetocarmine technique. Aulonemia aristulata is monocarpic (semelparous) with gregarious flowering. All culms in both studied areas blossomed and fruited between August and November 2007, dying subsequently between December 2007 and April 2008. Two types of synflorescences and flowers were observed: terminal with bisexual and protandric florets, with the anthesis lasting for 3-4 days; and axillary, with morphologically bisexual, but functionally female, florets and anthesis lasting for 3-4 days. The latter were also observed in the rhizome of plants whose aerial portion had been removed. The presence of axillary synflorescences with pistillate flowers is described here for the first time in Aulonemia species. Moreover, this is the first report of gynomonoecy in woody bamboo. Fruiting from bisexual florets under natural conditions (35%) was superior to that obtained from bagged synflorescences (11.5%). Fruiting from functional female florets was around 20%. Pollen viability was on the average of 90%. The results suggest that Aulonemia aristulata is anemophilous. The massive bamboo seedling recruitment observed after dieback with the ability to colonize open areas could promote the regeneration of Aulonemia aristulata.

Self-, cross- and interspecific pollinations in Passiflora capsularis and P. rubra

This study aimed to characterize the reproductive system of Passiflora capsularis L. and P. rubra L. In vivo controlled pollinations, in vitro pollen germination and pollen-ovule (P:O) ratio evaluation were conducted. In self-pollination, intraspecific and interspecific pollination, P. capsularis showed means of 62.5, 68.7 and 48.4% of fertilized flowers, while in P. rubra, the averages were 67.2, 62.5 and 46.9%, respectively. For in vitro germination, 52.2% of P. capsularis pollen grains germinated while in P. rubra, the percentage was 64.4. The P:O ratio was 22.4 for P. capsularis, and 27.4 for P. rubra, which included them in the category of obligatory autogamous. Passiflora capsularis and P. rubra can reproduce both by self-pollination and cross-pollination, and crossings between the two species succeeded though the success rate was lower than 50%. The characteristics of the reproductive system of both species allow the use of greater range of options on breeding methods for production of ornamental Passiflora plants.

Mixed reproduction systems in Opuntia monacantha (Cactaceae) in Southern Brazil

The Cactaceae have morphological and physiological adaptations associated with their life histories that are reflected in different modes of reproduction and multiplication. The reproductive phenology, preferential reproductive mode, and the sexual and asexual multiplication of Opuntia monacantha were investigated between 2003 and 2006 in two restinga (sandy coastal) environment in Southern Brazil to determine the adaptive values of its reproduction modes. Flowering was annual and occurred continuous for approximately 100 days between the months of September and January, with a few flowers opening per day per individual (but many in the whole population). Facultative xenogamy was identified as the preferential sexual system, with the highest levels of fruit and seed formation following natural pollination. The seeds are recalcitrant and have high germinative capacities under laboratory conditions. Seedling recruitment and establishment in the field was extremely reduced. The skins of the fruits and the cladodes have high capacities for regeneration and clonal multiplication because of their areolas.

A protein with amino acid sequence homology to bovine insulin is present in the legume Vigna unguiculata (cowpea)

Since the discovery of bovine insulin in plants, much effort has been devoted to the characterization of these proteins and elucidation of their functions. We report here the isolation of a protein with similar molecular mass and same amino acid sequence to bovine insulin from developing fruits of cowpea (Vigna unguiculata) genotype Epace 10. Insulin was measured by ELISA using an anti-human insulin antibody and was detected both in empty pods and seed coats but not in the embryo. The highest concentrations (about 0.5 ng/µg of protein) of the protein were detected in seed coats at 16 and 18 days after pollination, and the values were 1.6 to 4.0 times higher than those found for isolated pods tested on any day. N-terminal amino acid sequencing of insulin was performed on the protein purified by C4-HPLC. The significance of the presence of insulin in these plant tissues is not fully understood but we speculate that it may be involved in the transport of carbohydrate to the fruit.

Genetic purity certificate in seeds of hybrid maize using molecular markers

One of the main features that confer high quality to the seed is its genetic purity, in which one of the major causes of contamination is the self-pollination of the female parent. Up to date, there is no accurate and fast methods for detecting such contamination. Thus, this work was carried out to certify the genetic purity in seeds of hybrid maize using different biochemical and DNA-based markers. Two single-cross hybrids and their parental lines derived from the maize breeding program at UFLA were evaluated by isoenzymatic pattern of alcohol dehydrogenase (ADH), esterase (EST), acid phosphatase (ACP), glutamate-oxaloacetate transaminase (GOT), malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphoglucomate dehydrogenase (PGDH), catalase (CAT) and ß-glucosidade (ßGLU) and by microsatellites markers. The enzymatic systems that were able to distinguish the hybrids from their parental line were the catalase, the isocitrate dehydrogenase and the esterase. The esterase showed a Mendelian segregation pattern for UFLA 8/3 hybrid, that enables a safer genetic purity certificate. Microsatellites were able to differentiate the hybrid lines and the respective parental lines. Moreover, this technique was fast, precise and without environment effects. For microsatellites, the amplification pattern was identical when young leaves or seeds were used as DNA source. The possibility of using seeds as DNA source would accelerate and facilitate the role process of the genetic purity analysis.

Physiological maturity of eggplant seeds

Determination of seed physiological maturity and ideal moment for harvesting fruits to extract their seeds are important aspects to produce seeds with high quality. To identify the best period for harvesting eggplant fruits, associated with ideal resting period of the fruit for extracting seeds, an eggplant production field was installed in municipality of Ijaci, in the State of Minas Gerais, Southwestern Brazil. The fruits were harvested at periods of 49, 56, 63, 70, and 77 days after pollination (DAP). The seeds of fruits harvested in each period were manually extracted immediately after harvest or after a post-harvest resting period of seven days, under a shed. The physiological quality of seeds was assessed by tests of: germination percentage; germination and emergence speed indexes; and electrical conductivity; which were carried out in the Central Seed Laboratory, Federal University of Lavras. Electrophoretic analyses of isoenzymes: catalase (CAT); esterase (EST); superoxide dismutase (SOD); and peroxidase (PO), were also therein performed. Results of germination and vigor of seeds have showed that the best period for harvesting the fruit is around 70 DAP; and that seeds should be extracted immediately after harvest. Electrophoretic analysis of enzymes has showed immaturity for eggplant seeds, harvested after 49 DAP.

Comics carnet : the graphic novelist as global nomad

An interdisciplinary approach is used to identify a new graphic novel genre, 'comics camet', and its key features. The study situates comics camet in a historical context and shows it to be the result of a cross-pollination between the American and French comics traditions. Comics camet incorporates features from other literary genres: journalism, autobiography, ethnography and travel writing. Its creators, primarily European rriales, document their experiences visiting countries that Europe has traditionally defined as belonging to the 'East'. A visual and narrative analysis, using theoretical perspectives derived from cultural and postcolonial studies, examines how comics camet represents the non-European other and identifies the genre's ideological assumptions. Four representative texts are examined: Joe Sacco's Palestine (2001), Craig Thompson's, Camet de Voyage (2004), Guy Delisle's Pyongyang (2005) and Mrujane Satrpi's Persespolis 2 (2004). The study concludes that the comics camet genre simultaneously reinforces and challenges stereotypical assumptions about non-European people and places.

Rôle de la protéine ScFRK1 dans le développement du sac embryonnaire et son impact sur le guidage des tubes polliniques

Le gène Solanum chacoense Fertilization-Related Kinase 1 (ScFRK1) code pour une protéine de la famille des MAPKK kinases exprimée spécifiquement dans les ovules. Son transcrit s’accumule principalement dans la zone micropylaire du sac embryonnaire à l’anthèse et diminue rapidement après pollinisation. Ces résultats suggèrent un rôle possible avant ou pendant la fécondation. Bien qu'aucune expression ne soit détectée dans le pollen à maturité, la protéine est cependant présente dans les cellules mères de microspores. Des plantes transgéniques sous-exprimant ScFRK1 ne montrent aucun phénotype au niveau des tissus végétatifs, mais présentent de petits fruits dépourvus de graines. L’étude microscopique du gamétophyte femelle révèle que son développement ne progresse pas au-delà du stade de la mégaspore fonctionnelle et une grande proportion de sacs embryonnaires anormaux est corrélée avec une faible expression de ScFRK1. De plus, la production de pollen viable diminue en fonction de la baisse des niveaux d’expression du gène, ce qui pourrait s’expliquer par un problème au cours de la mitose I. Puisque l’intégrité du sac embryonnaire est essentielle au guidage des tubes polliniques, nous avons conçu un système de guidage semi-in vivo permettant d’évaluer la capacité des ovules du mutant ScFRK1 à les attirer. L’attraction est sévèrement affectée dans de telles conditions, ce qui confirme l'implication des cellules de la zone micropylaire comme source attractive. Notre système nous a également permis de démontrer que le guidage est très spécifique à l’espèce et que cette attraction constitue un mécanisme important favorisant la spéciation et la maintenance des barrières interspécifiques dans la reproduction sexuée des végétaux.

Influence d’un supplément alimentaire sur le développement des colonies d’abeilles domestiques (Apis mellifera, Linnaeus 1758) au Québec

La malnutrition est identifiée comme l’un des facteurs potentiellement responsables des mortalités élevées de colonies d’abeilles des dernières années au Québec. Pour contrer cela, les apiculteurs donnent des suppléments de pollen à leurs colonies, mais les impacts d’une telle pratique à diverses périodes sont méconnus. Les effets de la disponibilité du pollen sur le développement de colonies d’abeilles ont été mesurés pendant 3 différentes périodes : au printemps, durant la pollinisation de la canneberge et à la fin de l’été. À chacune des périodes correspondait une expérience distincte utilisant 40 colonies. Pour chaque expérience, des conditions d’abondance de supplément de pollen et de restriction de pollen naturel étaient créées chez les colonies pendant un mois selon un plan d’expérience factorielle 2x2. L’élevage du couvain et la récolte de miel ont été mesurés jusqu’à la fin de l’été (début de l’été suivant pour l’expérience de fin d’été). Au printemps, les colonies restreintes en pollen naturel ont élevé 18% moins de couvain (p<0.05) pendant la période de restriction et 11% de moins à la fin de l’été alors que l’utilisation du supplément n’a eu aucun effet (p>0.05). Les colonies supplémentées durant la pollinisation des canneberges ont élevé moins de couvain (p<0.05) à la fin de l’été. Pour l’expérience de fin d’été, les colonies supplémentées ont eut une meilleure reprise printanière (p<0.05) de l’élevage du couvain (60% de plus) alors qu’une restriction en pollen naturel avait un effet négatif (p>0.05). Les récoltes de miel ont été augmentées (p<0.05) de 1,3 kg pendant la pollinisation de la canneberge alors qu’elles ont été diminuées (p<0.05)par une restriction en pollen naturel de 4,2 kg à la fin de l’été et de 15 kg au printemps.

La régulation génique chez Solanum chacoense : de la pollinisation jusqu’à l’embryogenèse

Chez les végétaux supérieurs, l’embryogenèse est une phase clé du développement au cours de laquelle l’embryon établit les principales structures qui formeront la future plante et synthétise et accumule des réserves définissant le rendement et la qualité nutritionnelle des graines. Ainsi, la compréhension des évènements moléculaires et physiologiques menant à la formation de la graine représente un intérêt agronomique majeur. Toutefois, l'analyse des premiers stades de développement est souvent difficile parce que l'embryon est petit et intégré à l'intérieur du tissu maternel. Solanum chacoense qui présente des fleurs relativement grande facilitant l’isolation des ovules, a été utilisée pour l’étude de la biologie de la reproduction plus précisément la formation des gamètes femelles, la pollinisation, la fécondation et le développement des embryons. Afin d'analyser le programme transcriptionnel induit au cours de la structuration de ces étapes de la reproduction sexuée, nous avons mis à profit un projet de séquençage de 7741 ESTs (6700 unigènes) exprimés dans l’ovule à différents stades du développement embryonnaire. L’ADN de ces ESTs a été utilisé pour la fabrication de biopuces d’ADN. Dans un premier temps, ces biopuces ont été utilisé pour comparer des ADNc issus des ovules de chaque stade de développement embryonnaire (depuis le zygote jusqu’au embryon mature) versus un ovule non fécondé. Trois profils d’expression correspondant au stade précoce, intermédiaire et tardive ont été trouvés. Une analyse plus approfondie entre chaque point étudié (de 0 à 22 jours après pollinisation), a permis d'identifier des gènes spécifiques caractérisant des phases de transition spécifiques. Les annotations Fonctionnelles des gènes differentiellement exprimés nous ont permis d'identifier les principales fonctions cellulaires impliquées à chaque stade de développement, révélant que les embryons sont engagés dans des actifs processus de différenciation. Ces biopuces d’ADN ont été par la suite utilisé pour comparer différent types de pollinisation (compatible, incompatible, semi-compatible et inter-espèce) afin d’identifier les gènes répondants à plusieurs stimuli avant l'arrivé du tube pollinique aux ovules (activation à distance). Nous avons pu démontrer que le signal perçu par l’ovaire était différent et dépend de plusieurs facteurs, incluant le type de pollen et la distance parcourue par le pollen dans le style. Une autre analyse permettant la comparaison des différentes pollinisations et la blessure du style nous a permis d’identifier que les programmes génétiques de la pollinisation chevauchent en partie avec ceux du stress. Cela était confirmé en traitant les fleurs par une hormone de stress, méthyle jasmonate. Dans le dernier chapitre, nous avons utilisé ces biopuces pour étudier le changement transcriptionnel d’un mutant sur exprimant une protéine kinase FRK2 impliqué dans l’identité des ovules. Nous avons pu sélectionner plusieurs gènes candidat touchés par la surexpression de cette kinase pour mieux comprendre la voie se signalisation. Ces biopuces ont ainsi servi à déterminer la variation au niveau transcriptionnelle des gènes impliqués lors de différents stades de la reproduction sexuée chez les plantes et nous a permis de mieux comprendre ces étapes.