The epidermal cell structure of the secondary pollen presenter in Vangueria infausta (Rubiaceae: Vanguerieae) suggests a functional association with protruding onci in pollen grains.

Secondary pollen presentation is a well-known phenomenon in the Rubiaceae with particularly conspicuous pollen presenters occurring in the tribe Vanguerieae. These knob-like structures are formed by a modification of the upper portion of the style and stigma, together known as the stylar head complex. In the flower bud and shortly before anthesis, the anthers surrounding the stylar head complex dehisce and release pollen grains which adhere to the pollen presenter. The epidermal cells of the pollen presenter facing the anthers are radially elongated with a characteristic wall thickening encircling the anticlinal walls of each cell towards the distal end. These cells were studied in the pollen presenter of Vangueria infausta using electron and light microscopy in conjunction with histochemical tests and immunohistochemical methods. Other prominent thickenings of the cell wall were also observed on the distal and proximal walls. All these thickenings were found to be rich in pectin and possibly xyloglucan. The terms "thickenings of Igersheim" and "bands of Igersheim" are proposed to refer, respectively, to these wall structures in general and those encircling the anticlinal walls of each cell near the distal end. The epidermal cells have an intricate ultrastructure with an abundance of organelles, including smooth and rough endoplasmic reticulum, Golgi apparatus, mitochondria and secretory vesicles. This indicates that these cells are likely to have an active physiological role. The pollen grains possess prominent protruding onci and observations were made on their structure and development. Walls of the protruding onci are also rich in pectin. Pectins are hydrophilic and known to be involved in the dehydration and rehydration of pollen grains. We hypothesise that the thickenings of Igersheim, as well as the protruding onci of the pollen grains, are functionally associated and part of the adaptive syndrome of secondary pollen presentation, at least in the Vanguerieae.

Immunohistochemical localization of caffeine in young Camellia sinensis (L.) O. Kuntze (tea) leaves.

The anatomical localization of caffeine within young Camellia sinensis leaves was investigated using immunohistochemical methods and confocal scanning laser microscopy. Preliminary fixation experiments were conducted with young C. sinensis leaves to determine which fixation procedure retained caffeine the best as determined by high-performance liquid chromatography analysis. High pressure freezing, freeze substitution, and embedding in resin was deemed the best protocol as it retained most of the caffeine and allowed for the samples to be sectioned with ease. Immunohistochemical localization with primary anti-caffeine antibodies and conjugated secondary antibodies on leaf sections proved at the tissue level that caffeine was localized and accumulated within vascular bundles, mainly the precursor phloem. With the use of a pressure bomb, xylem sap was collected using a micro syringe. The xylem sap was analyzed by thin-layer chromatography and the presence of caffeine was determined. We hypothesize that caffeine is synthesized in the chloroplasts of photosynthetic cells and transported to vascular bundles where it acts as a chemical defense against various pathogens and predators. Complex formation of caffeine with chlorogenic acid is also discussed as this may also help explain caffeine's localization.

Structural evidence in Plectroniella armata (Rubiaceae) for possible material exchange between domatia and mites.

Domatia are small structures on the lower surface of a leaf, usually taking the form of cavities, pouches, domes with an opening, or hairs (or a combination of these), and located in the axils between the main veins. They are found in many dicotyledons including certain members of the Rubiaceae. As part of an ongoing study of selected southern African members of the tribe Vanguerieae of this family, their structure in transverse section was investigated. In some taxa, such as Plectroniella armata, light microscopic (LM) observations revealed large numbers of stomata in the domatia as well as a number of channel-like structures extending across the cuticle toward the cavity of the domatia. The cuticle of the epidermis lining the domatia also appeared thicker than in other parts of the leaves. The epidermis in P. armata was also examined using transmission electron microscopy (TEM). Domatia have been shown to house mainly mites, many of which are predatory or fungivorous, in a symbiotic (mutualistic) relationship with the plant. To date, much research has focussed on the role of domatia in providing shelter for various organisms, their eggs and their young. However, the TEM study revealed the apparent "channels" and thick cuticle seen under LM to be electron dense non-cellulosic branching fibrils within pronounced, often closely spaced cuticular folds. The functional significance of these fibrils and folds requires further investigation. Folding of cell walls and membranes at ultrastructural level is usually functionally associated with an increased surface area to facilitate active exchange of compounds/metabolites. This may indicate that translocation of substances and/or other forms of communication is possible between the domatium and its inhabitants. This therefore suggests a far more active role for the leaf in the symbiotic relationship than was previously thought. More work is required to test such a possibility.