The anatomy of the seed-coat includes diagnostic characters in the subtribe Eugeniinae (Myrteae, Myrtaceae).

The subtribe Eugeniinae comprises of two genera, Eugenia (ca. 1,100 species) and Myrcianthes (ca. 40 species). Eugenia is the largest genus of neotropical Myrtaceae and its latest classification proposes 11 sections. This study describes the seed anatomy of forty-one species of Eugeniinae in order to provide possible diagnostic characteristics. Following standard anatomical techniques, flower buds, flowers, and fruits were processed and analyzed using microtome sections and light microscopy. The phylogeny used the regions ITS, rpl16, psbA-trnH, trnL-rpl32, and trnQ-rps16, following recent studies in the group. Ancestral character reconstruction uncovered that: (1) the ancestral ovule in Eugeniinae was campylotropous (98.9% probability), bitegmic (98.5% probability), and unitegmic ovules arose on more than one lineage independently within Eugenia; (2) the pachychalazal seed-coat appeared with a 92% probability of being the ancestral type; (3) non-lignified seed-coat (24,5% probability) and aerenchymatous mesotesta (45.8% probability) are diagnostic characters in Myrcianthes pungens (aerenchymatous mesotesta present in the developing seed-coat) and in the species of E. sect. Pseudeugenia until the species of E. sect. Schizocalomyrtus and it is the type of seed-coat that predominates in most basal sections on the tree; (4) the partial sclerification (only in the exotesta-exotestal seed-coat) is mainly observed in species of E. sect. Excelsae, E. sect. Jossinia (group X), and E. sect. Racemosae (22.2% probability); (5) and in the species of the recent lineages of Eugenia, with a probability of 27.2%, predominate the exomesotestal or testal construction of the seed-coat [character observed in almost all species analyzed of E. sect. Jossinia (group Y) and E. sect. Umbellatae]. A dehiscent fruit is considered as a plesiomorphic state in Myrtaceae; the ancestor of this family had seeds with a completely sclerified testa, and the other testa types described for the current species with dehiscent and indehiscent fruits are simplified versions of this ancestral type. Perhaps, this means that the sclerified layers in the seed-coat have remained in whole or in part as a plesiomorphic condition for taxa with a capsule and bacca. Maintaining the plesiomorphic condition may have represented a selective advantage at some point in the evolutionary history of the family and its groups.

Phylogeny of the Alismatales (Monocotyledons) and the relationship of Acorus (Acorales?).

A phylogenetic analysis of the early branching lineages of the monocotyledons is performed using data from two plastid genes (rbcL and matK), five mitochondrial genes (atp1, ccmB, cob, mttB and nad5) and morphology. The complete matrix includes 93 terminals representing Acorus, the 14 families currently recognized within Alismatales, and numerous lineages of monocotyledons and other angiosperms. Total evidence analysis results in an almost completely resolved strict consensus tree, but all data partitions, genomic as well as morphological, are incongruent. The effects of RNA editing and potentially processed paralogous sequences are explored and discussed. Despite a decrease in incongruence length differences after exclusion of edited sites, the major data partitions remain significantly incongruent. The 14 families of Alismatales are all found to be monophyletic, but Acorus is found to be included in Alismatales rather than being the sister group to all other monocotyledons. The placement is strongly supported by the mitochondrial data, atp1 in particular, but it cannot be explained as an artifact caused by patterns of editing or by sampling of processed paralogues.

Plastid phylogenomics and molecular evolution of Alismatales.

Past phylogenetic studies of the monocot order Alismatales left several higher-order relationships unresolved. We addressed these uncertainties using a nearly complete genus-level sampling of whole plastid genomes (gene sets representing 83 protein-coding and ribosomal genes) from members of the core alismatid families, Tofieldiaceae and additional taxa (Araceae and other angiosperms). Parsimony and likelihood analyses inferred generally highly congruent phylogenetic relationships within the order, and several alternative likelihood partitioning schemes had little impact on patterns of clade support. All families with multiple genera were resolved as monophyletic, and we inferred strong bootstrap support for most inter- and intrafamilial relationships. The precise placement of Tofieldiaceae in the order was not well supported. Although most analyses inferred Tofieldiaceae to be the sister-group of the rest of the order, one likelihood analysis indicated a contrasting Araceae-sister arrangement. Acorus (Acorales) was not supported as a member of the order. We also investigated the molecular evolution of plastid NADH dehydrogenase, a large enzymatic complex that may play a role in photooxidative stress responses. Ancestral-state reconstructions support four convergent losses of a functional NADH dehydrogenase complex in Alismatales, including a single loss in Tofieldiaceae.

Anatomy of fleshy fruits in the monocots.

PREMISE OF THE STUDY: An anatomical and developmental study of distantly related fleshy fruits in the monocots was undertaken to better understand the evolution of baccate fruits in the monocot clade as a whole. We studied 14 species with fleshy fruits spanning the Alismatales, Arecales, Asparagales, Commelinales, Dioscoreales, Liliales, and Poales to determine various mechanisms through which baccate fruits attain fleshiness at maturity. METHODS: Flowers and fruits of various stages were collected, sectioned, stained, and examined using light microscopy and scanning electron microscopy. KEY RESULTS: Three basic pathways for attaining fleshiness were identified within the species examined (true berries, with a uniform pericarp; typical drupes, with an endocarp differentiated by the presence of stony pyrenes; and specialized drupes, involving mesocarp and endocarp differentiated by stone pyrenes). Furthermore, developmental characters differentiating basic fruit types were identified. CONCLUSIONS: Fleshy fruits in the monocots do not develop through a single shared pathway, indicating that fleshiness has evolved multiple times within the clade.

Preparation of samples for leaf architecture studies, a method for mounting cleared leaves.

PREMISE OF THE STUDY: Several recent waves of interest in leaf architecture have shown an expanding range of approaches and applications across a number of disciplines. Despite this increased interest, examination of existing archives of cleared and mounted leaves shows that current methods for mounting, in particular, yield unsatisfactory results and deterioration of samples over relatively short periods. Although techniques for clearing and staining leaves are numerous, published techniques for mounting leaves are scarce. • METHODS AND RESULTS: Here we present a complete protocol and recommendations for clearing, staining, and imaging leaves, and, most importantly, a method to permanently mount cleared leaves. • CONCLUSIONS: The mounting protocol is faster than other methods, inexpensive, and straightforward; moreover, it yields clear and permanent samples that can easily be imaged, scanned, and stored. Specimens mounted with this method preserve well, with leaves that were mounted more than 35 years ago showing no signs of bubbling or discoloration.

Anatomical and histochemical characterization of extrafloral nectaries of Prockia crucis (Salicaceae).

Besides being vital tools in taxonomic evaluation, the anatomy of plant secretory structures and the chemical composition of their secretions may contribute to a more thorough understanding of the roles and functions of these secretory structures. Here we used standard techniques for plant anatomy and histochemistry to examine secretory structures on leaves at different stages of development of Prockia crucis, to evaluate the origin and development of the structures, and to identify the disaccharides and monosaccharides in the exudates. Fructose, glucose, and sucrose constituted up to 49.6% of the entire secretion. The glands were confirmed to be extrafloral nectaries (EFNs); this is the first report of their presence in the genus Prockia. These EFNs are globular, sessile glands, with a central concavity occurring on the basal and marginal regions of the leaf. The epidermis surrounding the concavity is secretory, forming a single-layered palisade that strongly reacts with periodic acid-Schiff's reagent (PAS) and xylidine Ponceau, indicators of total polysaccharides and total proteins, respectively, in the exudate. On the basis of the similarity of these glands to the salicoid teeth in Populus and Salix, we suggest that these three taxa are phylogenetically close.