Anatomical characterization and technological properties of Pterogyne nitens wood, a very interesting species of the Brazilian Caatinga biome.

Pterogyne nitens is commonly known in northeastern Brazil as a lesser-known fast-growing species in the Caatinga biome, which is a difficult place for tree development due to the low natural fertility soils and low availability of water. Due to the importance of expanding information about the anatomical wood properties of Caatinga native species, the aim of this work was to characterize the anatomical elements, to macroscopically describe the wood and make inferences about its possible end-uses. Maceration was performed which enabled measuring fiber dimensions, pore frequency and the following technological indexes: cell wall fraction, slenderness ratio, Runkel index and flexibility coefficient. Histological sections enabled describing the arrangements of the cellular elements in different observation sections and to determine the pore diameter. P. nitens wood has anatomical arrangements characterized by confluent axial parenchyma, being diffuse-porous with the presence of tylosis and heterogeneous/stratified rays (biseriate). The fibers were classified as very short (length 0.81 mm), not flexible and Runkel index 0.82. The pores were few in number with a frequency of 32.9 pores/mm2, distributed in a diffuse format and many were obstructed by tylosis. Based on the anatomical results and considering other technological studies, P. nitens wood is most suitable for charcoal production.

Histology, histochemistry and ultrastructure of pre-embryogenic cells determined for direct somatic embryogenesis in the palm tree Syagrus oleracea.

Somatic embryogenesis in palm trees is, in general, a slow and highly complex process, with a predominance of the indirect route and, consequently, a lack of knowledge about the direct route. We present new knowledge related to the morphological, histochemical and ultrastructural aspects of the transition from somatic to embryogenic cells and direct formation of somatic embryos from mature zygotic embryos of Syagrus oleracea, a palm tree. The results support the general concept that 2,4-dichlorophenoxyacetic acid plays a critical role for the formation of somatic embryos of direct and multicellular origin. Seven days in medium with auxin were enough for the identification of embryogenic cells. These cells had a set of characteristics corresponding to totipotent stem cells. At 14 days on induction medium, nodular formations were observed in the distal region of inoculated embryos, which evolved into globular somatic embryos. At 120 days on induction medium, the quality of the somatic embryos was compromised. The dynamics of the mobilization of reserve compounds was also demonstrated, with emphasis on starch and protein as energy sources required for the embryogenic process. This study shows for the first time the anatomical and ultrastructural events involved in direct somatic embryogenesis in a palm tree and incites the scientific community to return to the discussion of classical concepts related to direct somatic embryogenesis, especially regarding the characteristics and location of determined pre-embryogenic cells.

Cellulases and pectinases act together on the development of articulated laticifers in Ficus montana and Maclura tinctoria (Moraceae).

The presence of articulated laticifers in the Moraceae family was recently discovered, which means that the location of pectinase and cellulase activities must be of great importance for their growth. Thus, the present study aimed to determine the role of these enzymes in the laticifer growth in Ficus montana and Maclura tinctoria. Reproductive meristems were collected and fixed in Karnovsky. Pectinase and cellulase labeling was performed in part of the samples, while another part was processed for usual TEM analyses. Pectinase and cellulase activities were detected in the vacuole and close to the middle lamella in both species. The presence of cellulases in the laticifers supports their articulated origin. Therefore, the occurrence of pectinase and cellulase activity in the laticifers points out that these enzymes could act in the dissolution of the transverse walls and in the processes of intrusive growth (through the dissolution of the middle lamella) and cell elongation (through the partial disassembly of components of the wall making it more plastic). Both enzymes are synthesized in the endoplasmic reticulum and transported to the cell wall by exocytosis or stored in the vacuole. The species studied showed a diverse subcellular composition, which is probably related to the species and not to the laticifer type (they present the same type) and to the composition of the latex (they show similar latex composition). We conclude that the presence of pectinases and cellulases can be used as a diagnostic condition for the laticifer types (articulated vs. non-articulated).

Osmophores of Caryocar brasiliense (Caryocaraceae): a particular structure of the androecium that releases an unusual scent.

Caryocar brasiliense is a flagship species of the Brazilian Cerrado. It produces flowers with a strong peculiar scent, which are pollinated by bats and occasionally moths with short mouthparts. However, the cues responsible for attracting these nocturnal pollinators remain unknown. We aimed to identify osmophores of C. brasiliense, describe the ultrastructure of the cells involved in the synthesis and release of floral odour, and identify the constituents of the floral bouquet. We performed field observations and histochemical and ultrastructural analyses of flowers focusing on the androecium. Gas chromatography-mass spectrometry was used to analyse the scents emitted. Filament epidermal cells were found to possess an unusual shape and be responsible for the main production and release of odour. These cells, called foraminous cells, are elongate and possess pores where their cell walls are abruptly thin. The cuticle is practically absent over the pores, which facilitates odour emission. The foraminous cells have conspicuous nuclei and organelle-rich cytoplasm where oil droplets can be seen prior to anthesis. The features of these cells remain similar during anthesis, but many vesicles fuse with the plasma membrane and the number of oil droplets in the cytosol decreases. Twenty-nine components were found in the scent, especially fatty acid derivatives and N- and S-bearing compounds. Our analyses revealed that the androecium of C. brasiliense has a particular structure that acts as an osmophore. The scent from the androecium resembles that of the entire flower, which is an unprecedented finding for a plant with single flowers as the pollination unit.

Cellular and Morpho-histological Foundations of In Vitro Plant Regeneration.

In vitro plant regeneration systems have turned into invaluable tools to plant biotechnology. Despite being poorly understood, the molecular mechanisms underlying the control of both morphogenetic pathways, de novo organogenesis and somatic embryogenesis, have been supported by recent findings involving proteome-, metabolome-, and transcriptome-based profiles. Notwithstanding, the integration of molecular data with structural aspects has been an important strategy of study attempting to elucidate the basis of the cell competence acquisition to further follow commitment and determination to specific a particular in vitro regeneration pathway. In that sense, morpho-histological tools have allowed to recognize cellular markers and patterns of gene expression at cellular level and this way have collaborated in the identification of the cell types with high regenerative capacity. This chapter ties together up those fundamental and important microscopy techniques that help to elucidate that regeneration occurs, most of the time, from epidermis or subepidermal cells and from the procambial cells (pericycle and vascular parenchyma). Important findings are discussed toward ultrastructural differences observed in the nuclear organization among pluripotent and totipotent cells, implying that regeneration occurs from two cellular mechanisms based on cellular reprogramming or reactivation.

Functional anatomy reveals secretory activity in papillose anthers of a buzz-pollinated Solanum species (Cyphomandra clade - Solanaceae).

Pollination in Solanum (Solanaceae) species is commonly performed by female bees, which vibrate anthers to extract pollen. Another pollen removal type is by male euglossine bees, milking the anthers when searching for floral scents produced by secretory tissues (osmophorous) at the swollen connective of the anthers of species in the Cyphomandra clade. Some species of this clade, however, are buzz-pollinated and present papillate anthers that should also have secretory activity, a hypothesis here tested. The anthers of Solanum luridifuscescens were fixed at different stages of development and analysed under light microscopy, SEM and TEM. Histochemical tests for the detection of starch and lipids were done. Epidermal cells of the abaxial surface of the anthers were visibly papillose, had large nuclei and dense cytoplasm rich in organelles such as mitochondria and plastids, typical features of secretory tissues. In this site, lipid droplets were detected, concomitantly with starch consumption, compatible with the secretory process in osmophores. No exudate or accumulation of substances was seen on the surface; in agreement with a previous pollination study performed in field conditions, where no pollinators were observed collecting floral scents, only pollen. The histochemical and structural analyses have evidenced the lipidic composition of the secretion, strongly pointing to terpenes as the secreted compounds. Ours findings show that papillae of the anthers have secretory activities that produce lipophilic compounds. This does not result in resources for bees, but could be an evolutionary step to the development of more specialised anthers in the Cyphomandra clade.

Natural history of the narrow endemics Ipomoea cavalcantei and I. marabaensis from Amazon Canga savannahs.

Amazon comprises a vast variety of ecosystems, including savannah-like Canga barrens that evolved on iron-lateritic rock plateaus of the Carajás Mountain range. Individual Cangas are enclosed by the rain forest, indicating insular isolation that enables speciation and plant community differentiation. To establish a framework for the research on natural history and conservation management of endemic Canga species, seven chloroplast DNA loci and an ITS2 nuclear DNA locus were used to study natural molecular variation of the red flowered Ipomoea cavalcantei and the lilac flowered I. marabaensis. Partitioning of the nuclear and chloroplast gene alleles strongly suggested that the species share the most recent common ancestor, pointing a new independent event of the red flower origin in the genus. Chloroplast gene allele analysis showed strong genetic differentiation between Canga populations, implying a limited role of seed dispersal in exchange of individuals between Cangas. Closed haplotype network topology indicated a requirement for the paternal inheritance in generation of cytoplasmic genetic variation. Tenfold higher nucleotide diversity in the nuclear ITS2 sequences distinguished I. cavalcantei from I. marabaensis, implying a different pace of evolutionary changes. Thus, Canga ecosystems offer powerful venues for the study of speciation, multitrait adaptation and the origins of genetic variation.

In silico characterization of 1,2-diacylglycerol cholinephosphotransferase and lysophospha-tidylcholine acyltransferase genes in Glycine max L. Merrill.

The enzymes 1,2-diacylglycerol cholinephosphotrans-ferase (CPT) and lysophosphatidylcholine acyltransferase (LPCAT) are important in lipid metabolism in soybean seeds. Thus, understand-ing the genes that encode these enzymes may enable their modification and aid the improvement of soybean oil quality. In soybean, the genes encoding these enzymes have not been completely described; there-fore, this study aimed to identify, characterize, and analyze the in silico expression of these genes in soybean. We identified two gene models encoding CPT and two gene models encoding LPCAT, one of which presented an alternative transcript. The sequences were positioned on the physical map of soybean and the promoter regions were analyzed. Cis-elements responsible for seed-specific expression and responses to biotic and abiotic stresses were identified. Virtual expression analysis of the gene models for CPT and LPCAT indicated that these genes are expressed under different stress conditions, in somatic embryos during differentiation, in immature seeds, root tissues, and calli. Putative ami-no acid sequences revealed the presence of transmembrane domains, and analysis of the cellular localization of these enzymes revealed they are located in the endoplasmic reticulum.

Development of carotenoid storage cells in Bixa orellana L. seed arils.

The arils of Bixa orellana L. seeds contain carotenoid storage cells (CSCs). The main compounds in these cells include bixin and norbixin, which are important pigments in the food and pharmaceutical industries. Although many studies have been conducted on these chemical constituents, the cellular events that occur during the development of the carotenoid-accumulating cells in the arils and their relationship with the final carotenoid accumulation in the vacuoles remain unknown. In this study, the development of the CSCs in B. orellana arils was analyzed by light and transmission electron microscopy. Carotenoids formed in specialized cells, whose number and size increased during aril development. At various stages of development, the cytoplasm of the CSCs contained chromoplasts that held an extensive network of tubules and plastoglobules. Next to the chromoplasts, lipid droplets may fuse one another to form osmiophilic bodies. In addition, vesicles were observed next to the tonoplast. At the final stages of development, both the osmiophilic bodies and vesicles, which became quadrangular or rectangular, were stored in the vacuoles of the CSCs. This study reported for the first time the occurrence of different storage unit types within the vacuole of carotenoid storage cells.

Ultrastructural studies on the sporogenous tissue and anther wall of Leucojum aestivum (amaryllidaceae) in different developmental stages.

In this study, ultrastructures of anther wall and sporogenous tissue of Leucojum aestivum were investigated during different developmental stages. Cytomictic channels were seen between pollen mother cells during prophase I. Polar distribution was described in the organelle content of pollen mother cells and microspores in early phases of microsporogenesis and also in pollen mitosis. Active secretion was observed in tapetal cells. Previous reports about developmental stages of male gametophyte were compared with the results of this study.