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Aspects of the Reproductive Biology of Brassavola Cebolleta Rchb.F

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Aspects of the Reproductive Biology of Brassavola Cebolleta Rchb.F DOI: 10.4025/actascibiolsci.v32i4.7148 Aspects of the reproductive biology of Brassavola cebolleta Rchb.f. (Orchidaceae) André Rodrigo Rech1*, Yara Brito Chaim Jardim Rosa2 and Fatima Cristina de Lazari 3 Manente-Balestieri 1Universidade Estadual de Campinas, Instituto de Biologia, Laboratório de Biossistemática, Rua Monteiro Lobato, 255, 13083-862, Cidade Universitária Zeferino Vaz, Campinas, São Paulo, Brazil. 2Faculdade de Ciências Agrárias, Universidade Federal da Grande Dourados, Dourados, Mato Grosso do Sul, Brazil. 3Faculdade de Ciências Biológicas e Ambientais, Universidade Federal da Grande Dourados, Dourados, Mato Grosso do Sul, Brazil. *Author for correspondence. Email: [email protected] ABSTRACT. This survey assessed some aspects of the reproductive biology of Brassavola cebolleta Rchb. f. (Orchidaceae) in the Municipality of Dourados, Mato Grosso do Sul State, Brazil. Floral biology, breeding systems, floral visitors and seed germination were analyzed. Differential success and fruit production rate were calculated. Pollination system indicated the pollen vector dependence and absence of pre-zygotic barriers related to self- incompatibility. Pollination occurred at night, being the potential pollen vector a Lepidoptera-Notodontidae of the genus Hemiceras. Differential success of male was 19%, female 9%, and the rate of effective fructification was 6.3%. Fruits produced after pollinator visits were larger than those generated by manual cross pollination, and despite the lower number of potentially viable seeds produced by the first, they presented a higher germination rate in asymbiotic media. This study warns to the vulnerability of the studied species, since the habitat fragmentation associated with pollinator scarcity and seed predation may significantly decrease new recruitment into populations. Key words: floral biology, fructification, pollination, breeding system. RESUMO. Aspectos da biologia reprodutiva de Brassavola cebolleta Rchb. f. (Orchidaceae). Este trabalho teve como objetivo analisar alguns aspectos da biologia reprodutiva de Brassavola cebolleta Rchb. f. (Orchidaceae), em Dourados, Estado do Mato Grosso do Sul, Brasil. Avaliaram-se a biologia floral, o sistema reprodutivo, os visitantes florais e a germinação das sementes. Calcularam-se o sucesso diferencial e a taxa de frutificação. O sistema de polinização indicou a dependência de vetores de pólen e a inexistência de barreiras pré-zigóticas relacionadas à autopolinização. A polinização ocorreu à noite, sendo o potencial vetor de pólen um Lepidoptera-Notodontidae, do gênero Hemiceras. O sucesso diferencial masculino foi de 19%, o feminino de 9% e a taxa de frutificação efetiva foi de 6,3%. Os frutos gerados com auxílio do agente polinizador foram maiores que os produzidos por polinização cruzada manual e, embora o número de sementes potencialmente viáveis produzidas pelo primeiro tenha sido menor, elas apresentaram maior taxa de germinação em meio assimbiótico. Este estudo alerta para a vulnerabilidade da espécie estudada uma vez que a fragmentação de hábitat associado à escassez de polinizadores e à predação das sementes pode reduzir drasticamente os novos recrutamentos às populações. Palavras-chave: biologia floral, frutificação, polinização, sistema reprodutivo. Introduction little known concerning reproductive mechanisms of some genera. The floral biology of orchid flowers, their Brassavola R. Br. presents flowers whose interactions with pollinators and diversification morphology indicates a possible sphingophilous along the evolutionary time is a more fascinating pollination, once the floral structure allows the topic than Darwin had ever suspected pollinators to feed fluttering in the air, inserting their (TREMBLAY et al., 2005). Despite the fact that proboscis into the cuniculus (CINGEL, 2001). The the number of surveys about this subject has flowers, usually cream or pale green colored, exhale increased in the last few years in Brazil, the great specific fragrances in the dusk or at night, and may diversity of plants in the country, especially have extrafloral nectaries that attract ants and other represented by families like Orchidaceae, remains herbivorous (ROEBUCK; STEINHART, 1978). Acta Scientiarum. Biological Sciences Maringá, v. 32, n. 4, p. 335-341, 2010 336 Rech et al. Hummingbirds (Trochilidae) have been observed in each population totaling up 200 hours, after the two Northern Brazil visiting flowers of B. martiana years of the study. We assessed foraging behavior of (BRAGA, 1977), and in Southern Brazil, flowers of floral visitors, time and legitimacy of the visitation B. tuberculata (CINGEL, 2001). In a study on the and parts of the contact with reproductive reproductive biology of B. nodosa it was observed the structures. Twenty floral buds in five individuals great dominance of male (pollinia removal) over the were marked during pre-anthesis in order to female success (deposition of pollinia on the stigmatic evaluate the anthesis sequence, and the occurrence cavity) (SCHEMSKE, 1980). of visitations (removal and deposition of pollinia) Concerning the flower visitors of the between 9:00 and 17:00h, and between 22:00 and Orchidaceae, there is a need for specific pollinators, 4:00h, when focal observations were not conducted. mainly in species where all the pollen are For nighttime focal observation “black light” concentrated in pollinia (TREMBLAY et al., 2005). illumination was utilized. Thus, the attraction of non effective or unspecific Morphometric characterization of the floral parts pollinators leads to a negative selection due to the was done with 30 flowers from 15 individuals, using considerable loss of reproductive success a digital caliper. The presence of osmophores was (PANSARIN, 2003; WASER; OLLERTON, 2006). tested by immersing eight flowers in a solution of Morphological adaptations to specific pollinators 1% neutral red for 10 min. and then washing them ensure the predominantly cross pollination with a solution of 5% glacial acetic acid (WIEMER (COZZOLINO; WIDMER, 2005). This et al., 2009). The presence of pigments was verified characteristic and the mycorrhizal association through the setting of eight flowers in ammonium represent preponderant agents in the diversification hydroxide atmosphere for 5 min. (SCOGIN et al., and maintenance of the great number of species of 1977), in order to detect flavonoids. The stigmatic Orchidaceae (OTERO; FLANAGAN, 2006). receptivity was tested over the study period by However, Tremblay et al. (2005) argue that the verifying the peroxidase activity, using V10 driving mechanism of diversity in Orchidaceae is hydrogen peroxide (oxygenated water) directly on based on the predominance of pollination limitation, the stigmatic surface of isolated flowers (DAFNI, 1992). The receptivity was tested at 4:00, 6:00, 8:00, which significantly favors well succeeded plant- 18:00, 20:00, and 22:00h. The subjective odor pollinator interactions, since the pollination of the assessment was obtained from 12 flowers placed in number of produced seeds is usually very expressive. odorless glass bottles (four flowers per bottle) that In this way, this survey assessed aspects of the were closed for 2 hours and afterwards opened, floral biology, pollination systems, fruit set and seed following olfactory characterization. The sugar germination in Brassavola cebolleta Rchb content was measured with a refractometer after f. (Orchidaceae). obtaining nectar with a micro syringe. To quantify the differential success along the two Material and methods years, we marked 277 flowers in the 32 studied This study was carried out from June to individuals that were monitored until the end of the September from 2006 to 2007 in Dourados flowering period. These marked flowers were Municipality, 22º21’03”S and 54º47’07”W, Mato tracked over in order to assess the number of Grosso do Sul State, Brazil, in a private reserve removed pollinia from anthers, and those deposited (Estância Aurora) on the margin of the Dourados on the stigmatic cavities. Male success was obtained river, in an altitude about 458 m. The climate of the from the ratio between the removed pollinia and the region is humid with dry winter; average total number of marked flowers. Feminine success precipitation is 1,500 mm, and an annual average was obtained from the ratio of the number of temperature of 22ºC. The vegetation is seasonally deposited pollinia in a stigmatic cavity and the total semi-deciduous forest. number of marked flowers (PARRA-TABLA et al., We studied 32 individuals of Brassavola cebolleta, 2000). Fruit set rate, in percentage, which here spread over two distinct populations that occur in a represents the real fertilization and embryo 10 m belt of riparian forest, about 3 km from each development, was obtained from the ratio of other, both on the same margin of the river, and the number of existing fruits eight months after the end host trees were about 15 m far from each other. The of the flowering period and the 277 marked flowers study of floral biology and pollination ecology was for this purpose (GARCIA-CRUZ; SOSA, 2008). done through uninterrupted observations of the We also calculated the efficiency index as the clumps with flowers of B. cebolleta, from 4:00 to proportion of removed pollinia in relation to the 9:00h, and from 17:00 to 22:00h, during five days in fruit set (TREMBLAY et al., 2005). Acta Scientiarum. Biological Sciences Maringá, v. 32, n. 4, p. 335-341, 2010 Aspects of the reproductive biology of Brassavola
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