Cytological attributes of storage tissues in nematode and eriophyid galls: pectin and hemicellulose functional insights.

Cell walls and protoplast may work together or distinctly in the establishment of the functional profiles of gall tissue compartments. This presumption is herein evaluated in three gall systems by immunocytochemical and ultrastructural analyses. The common storage tissues (CSTs) of leaf galls induced by Eriophyidae on Miconia ibaguensis leaves and by Ditylenchus gallaeformans on M. ibaguensis and M. albicans have rigid and porous cell walls due to their composition of pectins. Hemicelluloses in CST cell walls are scarcer when compared to the cell walls of the control leaves, being functionally compensated by rigid pectate gels. The typical nutritive tissues (TNTs) in galls induced by Ditylenchus gallaeformans are similar to promeristematic and secretory cells regarding their enriched cytoplasm, several mitochondria, and proplastids, as well as multivesicular and prolamellar bodies in cell membranes. The cytological features of the feeding cells of Eriophyidae galls indicate that they are not as metabolically active as the cells of the TNT in nematode galls. However, their cell wall composition suggests more plasticity and porosity than the cells of the TNT, which can compensate the less production of nutrients with more transport. The ultrastructural and immunocytochemical profiles of CST cells reveal functional similarities, which are independent of the taxa of the gall inducer or of the host plant. Despite their analogous functionalities, the protoplast and cell wall features of TNT cells of nematode galls and of the feeding cells of the Eriophyidae galls are distinct, and work out through different strategies toward keeping gall developmental site active.

How the activity of natural enemies changes the structure and metabolism of the nutritive tissue in galls? Evidence from the Palaeomystella oligophaga (Lepidoptera) -Macairea radula (Metastomataceae) system.

Insect-induced galls usually develop nutritional cells, which they induce and consume directly, and any metabolic modification of those cells may reflect changes of the insect's own metabolism. The system Palaeomystella oligophaga (Lepidoptera)-Macairea radula (Melastomataceae) presents a series of natural enemies, including parasitoids and cecidophages that can function as a natural experiment, respectively removing the specific galling feeding stimulus and providing a nonspecific one. Considering that the process of induction and maintenance of gall tissues strictly depends on the constant specific stimulus of galling, question I:What kind of metabolic changes these different groups of natural enemies can promote in chemical and structural composition of these galls? II: How the specialized tissues are metabolically dependent on the constant specific stimulus of galling in latter stages of gall development? Galls without natural enemies, with parasitoids or cecidophages in larvae or pupae stages were analyzed through histochemistry and cytological profiles and all compared to galls in natural senescence state. The analysis revealed the accumulation of proteins and lipids in typical nutritive tissue and starch in the storage tissue, as well a high integrity of cellular organelles and membrane systems on galls with gallings in the larval stage. Both parasitoids and cecidophages stop galling feeding activities, which resulted in the paralysis of the stimulus that maintain the metabolism of gall tissues, leading to generalized collapse. We demonstrate that the development and metabolic maintenance of a typical nutritive tissue in these galls are completely dependent on constant larval stimulus.

Preventing False Negatives for Histochemical Detection of Phenolics and Lignins in PEG-Embedded Plant Tissues.

Polyethylene glycol (PEG) is a low-cost and advantageous embedding medium, which maintains the majority of cell contents unaltered during the embedding process. Some hard or complex plant materials are better embedded in PEG than in other usual embedding media. However, the histochemical tests for phenolics and lignins in PEG-embedded plant tissues commonly result in false negatives. We hypothesize that these false negatives should be prevented by the use of distinct fixatives, which should avoid the bonds between PEG and phenols. Novel protocols for phenolics and flavanols detection are efficiently tested, with fixation of the samples in ferrous sulfate and formalin or in caffeine and sodium benzoate, respectively. The differentiation of lignin types is possible in safranin-stained sections observed under fluorescence. The Maule's test faultlessly distinguishes syringyl-rich from guaiacyl- and hydroxyphenyl-rich lignins in PEG-embedded material under light microscopy. Current hypothesis is corroborated, that is, the adequate fixation solves the false-negative results, and the new proposed protocols fill up some gaps on the detection of phenolics and lignins.

Additions to the mycobiota of the invasive weed Miconia calvescens (Melastomataceae).

Miconia calvescens (Melastomataceae) is a shrub or small tree native to the Neotropics that has become one of the worst invaders of forest ecosystems, particularly in Pacific islands such as Hawaii and French Polynesia. It has been a target for biological control for more than 10 y, both with arthropod and pathogen natural enemies. Until now Colletotrichum gloeosporioides f. sp. miconiae was the only organism to be used in biological control against this weed. This fungus was introduced both in Hawaii and in French Polynesia in the late 1990s/early 2000s, where it has caused some damage to the weed, but it became evident that additional agents are needed to achieve adequate control. Exploratory surveys for plant pathogens as potential biocontrol agents of M. calvescens were undertaken in Brazil, Costa Rica and Ecuador and yielded a diverse list of pathogens, including one phytoplasma, two nematodes, one oomycete and numerous fungi. A study including the description of five fungal species found attacking M. calvescens was published recently. Herein the following additional fungi also belonging to the mycobiota of M. calvescens are described: Hyalosphaera ornata sp. nov, Microsphaeropsis miconiae sp. nov., Myrothecium miconiae sp. nov., Phyllachora miconiiphila sp. nov., as well as Hyalosphaera miconiae, Lembosia melastomatum and Microsphaeropsis clidemiae, which are recorded here for the first time on this host. Although preliminary our observations of damage to M. calvescens caused by these seven fungal species did not indicate any potential for use in classical biological control.

Comparative anatomy and morphology of nectar-producing Melastomataceae.

BACKGROUND AND AIMS: Most neotropical Melastomataceae have bee-pollinated flowers with poricidal anthers. However, nectar rewards are known to be produced in about 80 species in eight genera from four different tribes. These nectar-producing species are pollinated by both vertebrates and invertebrates. METHODS: The floral morphology and anatomy of 14 species was studied in six genera of nectar-producing Melastomataceae (Blakea, Brachyotum, Charianthus, Huilaea, Meriania and Miconia). Anatomical methods included scanning electron microscopy, and serial sections of paraffin-embedded flowers. KEY RESULTS: All vertebrate-pollinated melastome flowers have petals that do not open completely at anthesis, thus forming a pseudo-tubular corolla, while closely related species that are bee pollinated have rotate or reflexed corollas. In most species, nectar secretion is related to stomatal or epidermal nectaries and not filament slits as previously reported. Moreover, the nectar is probably supplied by large vascular bundles near the release area. Blakea and Huilaea have nectary stomata located upon the dorsal anther connective appendages. Brachyotum also has nectary stomata on the anther connectives, but these are distributed lengthwise along most of the connective. Meriania may release nectar through the anther connective, but has additional nectary stomata on the inner walls of the hypanthium. Miconia has nectary stomata on the ovary apex. Charianthus nectaries were not found, but there is circumstantial evidence that nectar release occurs through the epidermis at the apex of the ovary and the lower portions of the inner wall of the hypanthium. CONCLUSIONS: Nectar release in Melastomataceae is apparently related to nectary stomata and not filament slits. The presence of nectary stomata on stamens and on ovary apices in different lineages suggests that the acquisition of nectaries is a derived condition. Nectary location also supports a derived condition, because location is strongly consistent within each genus, but differs between genera.

Leaf emergences in Microlepis oleaefolia (DC.) Triana (Melastomataceae) and their probable function: an anatomical and ultrastructural study.

Microlepis oleaefolia (DC.) Triana, an endemic species of Melastomataceae from the Brazilian cerrado, presents very complex leaf structures called as "hairs with root-like base" embedded in the mesophyll. This paper describes the ontogenesis, histochemical and structural aspects of these leaf structures as a framework for further functional studies. Samples of leaves in different developmental stages were processed according to common light and electron microscopy techniques. Fresh material was subjected to histochemical tests to examine the chemical composition of cell walls. The apoplastic transport between leaf emergences and the vascular system was verified by staining with 1% aqueous safranin. The structures are emergences of mixed protoderm and ground meristem origin. They are persistent, predominantly short-stalked and dendritic. Their arms have thick non-lignified cellulosic walls with a loose and heterogeneous aspect; on the inner face, the wall, which appears labyrinthine, presents small irregularly shaped projections directed towards the protoplast. The base of the emergences is composed of sclereids embedded in the mesophyll that reach the vascular system. Assays with aqueous safranin solution revealed it penetrates the cell walls of the arms and showed a connection between the emergence and xylem. Anatomical, chemical and ultrastructural features of leaf emergences of M. oleaefolia indicate that these structures are able to transport substances via apoplast and can absorb or exude solutions.