Phenology, floral and reproductive biology of Dichorisandra rhizantha Aona (Commelinaceae), an endemic endangered species in an Atlantic Forest fragment.

Commelinaceae is an important component of the Atlantic Forest and its analysis can provide information on the conservation of that species. This study had the objective of analyzing the phenology and floral and reproductive biology of Dichorisandra rhizantha, growing in an Atlantic Forest fragment, to obtain data related to the form of reproduction and pollination mechanism of this species. We monitored the phenophases and reproductive biology of the D. rhizantha in the area studied using different methodologies and microscopy techniques (LM, SEM and FM). The flowering period occurs during the dry season. The species is andromonoecious and has daytime anthesis (4:30 a.m.-1:30 p.m.). It has purple zygomorphic flowers and rimose but functionally poricidal anthers. The pollen viability was 97.6% and the two floral morphs investigated contained a large quantity of pollen grains. The time of greatest stigma receptivity (anthesis) coincided with the pollen viability. Cross-pollination was the only efficient reproductive system of the species, with observation of gametophytic self-incompatibility and pollen tubes growth only as far as the middle or lower third of the pistil. We observed bee species of the genera Xylocopa and Euglossa visiting the flowers and acting as effective pollinators. The immature fruits were formed after about 3.25 ± 1 days, and the presence was noted of ants of the genus Cephalotes acting as dispersers of the seeds, with aril serving as the reward. Although the natural reproductive system is efficient, D. rhizantha is endangered, mainly due to fragmentation of habitat caused by deforestation and urban encroachment.

Seed morphoanatomy of the genus Passiflora L. (Passifloraceae) by scanning electron microscopy and light microscopy.

Morphoanatomical analysis of seeds contributes to knowledge of the development of seedlings and identification of species, as well as supporting conservation studies. The conservation of the species belonging to the Passiflora genus is crucial due to of the threats to the genetic resources of these species. Thus, the objective of this study was to morphoanatomically characterize Passiflora seeds, verify possible injuries to the tissues after cryopreservation and thus contribute to the conservation strategies of the species of this genus. Initially, seeds of Passiflora coccinea, P. edulis, P. gibertii, P. maliformis, P. morifolia, P. setacea, P. suberosa, and P. tenuifila collected from the Passion Fruit Active Germplasm Bank of the Embrapa Cassava and Fruits research unit (Embrapa Mandioca e Fruticultura) were analyzed. Then, their length, width and thickness, shape of the base and tip, and ornamentations present on the body and edge of the seeds were evaluated. The seeds of the species were placed in cryotubes and immersed in liquid nitrogen to assess possible cryoinjuries. The tegument and tissues of the seeds were examined by scanning electron microscopy. The seeds had varied biometric data, with average values of 4.63 mm for length, 3.28 mm for width, and 1.51 mm for thickness. Six ornamentation types were observed: reticulate for the species P. coccinea; finely reticulate for P. edulis; foveolate reticulate for P. gibertii and P. setacea; alveolate reticulate for P. maliformis and P. tenuifila; coarsely reticulate for P. morifolia; and falsifoveolate reticulate for P. suberosa. Some seeds suffered tegument cracks due to the freezing in liquid nitrogen, but without physiological damages to the embryo and endosperm. The cryopreservation of the seeds in the presence of the tegument significantly reduced the cryoinjuries caused to the embryo. Cryopreservation can be promising for long-term conservation of passion fruit seeds.

Leaf structure of Tillandsia species (Tillandsioideae: Bromeliaceae) by light microscopy and scanning electron microscopy.

Tillandsia L. is the largest genus of the family Bromeliaceae, containing 755 species and seven subgenera. Morphoanatomical studies of leaves provide useful characteristics to phylogenetic, taxonomic, and ecological analyses. This study aims to characterize and compare the leaves of 24 species of the four subgenera of Tillandsia that occur in Bahia and also perform adaptative inferences to environmental responses. The results of the species' morphoanatomical studies were compared through dissimilarity analysis. The species have rosulate leaves with varying lengths and widths. The peltate trichomes present variation in the indument density and the length of their wing and central disk. The stomata are longitudinally distributed in one or both sides of the limb. The mesophyll is dorsiventral and presents aquiferous and chlorophyllic parenchymas. The vascular bundles are collateral and partially covered by fibers, except for Tillandsia linearis. Based on the dissimilarity analysis, it was possible to identify the formation of five groups. Group G1 was composed of T. linearis, which diverged from the other species of the subgenus. Group G2 was formed by the remaining species of the subgenus Phytarrhiza. G3 and G4 presented the species of the subgenus Diaphoranthema and Tillandsia, respectively. Group G5 gathered 11 species of the subgenus Anoplophytum and presented higher variability than the other subgenera. Based on the results, the morphoanatomical characteristics can be used to characterize and group Tillandsia species, besides confirming the morphological variability of these species to the epiphyte habit in different environments, especially xeric ones.

Cryopreservation and low-temperature storage of seeds of Tillandsia species (Bromeliaceae) with ornamental potential.

Tillandsia (Bromeliaceae) species have high endemism, and due to their strong ornamental potential, predatory extraction is threatening the extinction or drastic population reduction of many of them. In light of this scenario, it is necessary to find strategies for the conservation of these endangered species. The objective of this study was to evaluate two seed preservation strategies (freezing at - 5 °C and cryopreservation at - 196 °C) for 20 Tillandsia species occurring in the state of Bahia. We initially evaluated the morphometry, thousand-seed weight, and water content, followed by tests of germination and desiccation. After selecting the best result of the germination test (Germitest paper and incubation at 30 °C) and desiccation (3 h on silica gel), we established conservation tests utilizing two temperatures (freezing at - 5 °C and liquid nitrogen at - 196 °C), with storage times of 1, 7, 30, 180 and 450 days. Analysis of variance indicated that the 20 species had different behaviors when submitted to the two temperatures and different storage times. After 450 days there was a reduction in the germination percentage and germination speed index (GSI) of all the species studied when the seeds were preserved in the freezer. The storage in liquid nitrogen was efficient for the preservation of Tillandsia seeds when dried to a moisture content of approximately 7%. Our results support the establishment of a cryobank for Tillandsia to conserve these endemic species.

Pollen morphology and viability of Tillandsia (Bromeliaceae) species by light microscopy and scanning electron microscopy.

Tillandsia is the bromeliad genus containing the largest number of species, with wide geographic dispersion and an important ecological role in the ecosystems. Investigations of pollen morphology are important to support taxonomic and conservation studies as well as to choose promising species for cross-pollination to obtain hybrids. The objective of this study was to evaluate the morphology, production, and viability of pollen grains of 24 Tillandsia species that naturally occur in the state of Bahia, Brazil. Pollen grains were acetolized and analyzed by scanning electron microscopy. The pollen were germinated in vitro in two culture media (BM and BKM) and collected at three floral development stages (pre-anthesis, anthesis and post-anthesis). Pollen viability also was analyzed by staining with Alexander's solution and acetocarmine in three floral development stages. Variations in the ornamentation and size of the pollen were observed among the species studied, with the majority having medium size, while T. polystachia and T. juncea had large grains and the subgenera Diaphoranthema and Phytarrhiza had small grains. The pollen of the majority of species had bilateral symmetry, with a single irregularly shaped colpus, semitectate exine and reticulated-heterobrochate surface. The pollen presented high germination percentage, tube length and viability according to the histochemistry, except for T. recurvata, T. usneoides and T. loliacea. The BKM medium and collection of pollen during anthesis produced the best results for the Tillandsia species. The results can support studies of the taxonomy and conservation of these species, which have great environmental importance and high value for ornamental purposes.

Cryopreservation of Pineapple Shoot Tips by the Droplet Vitrification Technique.

Cryopreservation is a technique that allows the conservation of many species for long periods. Among the protocols used for cryopreservation, droplet vitrification has shown efficient results in preserving shoot tips of various wild and cultivated pineapple genotypes. The method consists of extraction of shoot tips from plants grown in vitro, dehydration for a period of 48 h in a preculture medium supplemented with a high concentration of sucrose, treatment in a plant vitrification solution (PVS2), and immersion in liquid nitrogen. The method described in this chapter has produced survival and regeneration indices of around 70%, depending on the genotype and physiological conditions of the initial explants. The objective of this chapter is to describe in detail a droplet vitrification protocol for shoot tips that is easy to perform for cryopreservation of pineapple germplasm.

Cryopreservation of Pollen Grains of Pineapple and Other Bromeliads.

Cryopreservation of pollen grains is an efficient technique to overcome asynchronous flowering and to support actions for genetic improvement and conservation of important alleles. It can be used both by germplasm curators and plant breeders. In the case of Bromeliaceae, a family with wide diversity but also high vulnerability, the form of conservation can be crucial to prevent the increasing problem of genetic erosion. This chapter describes a method of cryopreservation of pollen grains of different Bromeliaceae species, including pineapple, after dehydration with silica and subsequent immersion in liquid nitrogen. The efficiency of this protocol has been demonstrated by the high pollen viability percentage and production of seeds after in vivo pollination with cryopreserved grains. The protocol can be used for cryopreserving pollen of many species of bromeliads and is easy to perform.

Volatile compounds profile of Bromeliaceae flowers.

Volatile compounds play a vital role in the life cycle of plants, possessing antimicrobial and anti-herbivore activities, and with a significant importance in the food, cosmetic, chemical, and pharmaceutical industry. This study aimed to identify the volatile compounds emitted by flowers of thirteen species belonging to four genera of Bromeliaceae, using headspace solid-phase micro-extraction and detection by gas chromatography-mass spectrometry. A total of 71 volatile compounds belonging to nine chemical groups were identified. The compounds identified represented more than 97 % of the major components in Aechmea bicolor, Ae. bromeliifolia, Ae. distichantha, Ae. fasciata, and Vriesea friburgensis. In the Ananas varieties, over 99 % of the components were identified, and around 90 % in V. simplex. V. friburgensis presented the largest diversity of volatiles with 31 compounds, while Alcantarea nahoumii presented only 14. All three Ananas varieties presented the same 28 compounds in relatively similar abundance, which has been confirmed by principal component analysis. Current taxonomy and pollination syndrome studies available can adequately explain the variation in volatile compounds among species.