Are leaf glandular trichomes of oregano hospitable habitats for bacterial growth?

Phyllospheric bacteria were isolated from microsites around essential-oil-containing glands of two oregano (Origanum vulgare subsp. hirtum) lines. These bacteria, 20 isolates in total, were subjected to bioassays to examine their growth potential in the presence of essential oils at different concentrations. Although there were qualitative and quantitative differences in the essential oil composition between the two oregano lines, no differences were recorded in their antibacterial activity. In disk diffusion bioassays, four of the isolated strains could grow almost unrestrained in the presence of oregano oil, another five proved very sensitive, and the remaining 11 showed intermediate sensitivity. The strain least inhibited by oregano essential oil was further identified by complete16s rRNA gene sequencing as Pseudomonas putida. It was capable of forming biofilms even in the presence of oregano oil at high concentrations. Resistance of P. putida to oregano oil was further elaborated by microwell dilution bioassays, and its topology on oregano leaves was studied by electron microscopy. When inoculated on intact oregano plants, P. putida was able not only to colonize sites adjacent to essential oil-containing glands, but even to grow intracellularly. This is the first time that such prolific bacterial growth inside the glands has been visually observed. Results of this study further revealed that several bacteria can be established on oregano leaves, suggesting that these bacteria have attributes that allow them to tolerate or benefit from oregano secondary metabolites.

Contemporary seasonal and altitudinal variations of leaf structural features in oregano (Origanum vulgare L.).

The effects of elevation (200, 950 and 1760 m) and season (April-October) on leaf morphological, anatomical, ultrastructural, morphometrical and photosynthetic parameters were studied in Origanum vulgare plants. Observations aimed at the determination of the alterations in leaf structure and function associated with differential growth and adaptation of plants. Raising elevation results in a progressive decrease of plant height. During the growing period, summer plants are taller than spring and autumn plants at all elevations examined. In high-altitude populations (O. vulgare ssp. vulgare), the blade size becomes reduced in June leaves as compared with October leaves, while it does not change remarkably in low-altitude populations (O. vulgare ssp. hirtum). Leaf thickness remains more or less stable during the growing period. Expanded leaves in June and October at 200 m elevation contain dark phenolics only in their epidermis, whereas leaves of August are densely filled with phenolics in all of their tissues. In June at 1760 m elevation, leaves are devoid of phenolics, which, however, occur in the epidermis of the leaves in August and October. At higher altitudes, larger mesophyll chloroplasts with more starch grains are present in June leaves, whereas in August and October leaves chloroplasts are smaller with fewer starch grains. Leaf stomata and non-glandular hairs increase in number from the lowland to the upland habitats, whereas glandular hairs decrease in number. During the growing season, the density of stomata and of glandular and non-glandular hairs progressively increases. In the low- and mid-altitude oregano populations, leaf chlorophyll a content and PSII activity significantly increase in October, whereas they simultaneously decrease in the high-altitude population, suggesting a phenomenon of chilling-induced photoinhibition. The highest photochemical efficiency of PSII appears in the mid-altitude population (having characteristics intermediate between those of O. vulgare ssp. hirtum and ssp. vulgare) where environmental conditions are more favourable. This conclusion is also confirmed by the observation that the 950 m O. vulgare population has larger and thicker leaves with highly developed palisade and spongy parenchymas.

A possible new mode of dictyosome duplication in plant cells.

Dictyosomes are found in a large number in the glandular scales of Origanum dictamnus during the early developmental stages. Later they significantly diminish when essential oil secretion starts. Phases of dictyosome duplication are frequently observed at the stage of growth of the Golgi apparatus. The process of dictyosome division starts in the middle region of the stack where a Golgi cisterna undergoes a central dilation. An analogous dilation is progressively formed in the adjacent cisternae. Finally, by membrane fusion the stack separates into two daughter stacks which organize into normal dictyosomes.

Ultrastructure, organization and cytochemistry of cytoplasmic crystalline inclusions in Origanum dictamnus L. leaf chlorenchyma cells.

In the cytoplasm of chlorenchyma cells in primary Origanum dictamnus leaves, rectangular prism-like crystals occur, which, as shown by pepsin digestion, are proteinaceous. In sections they usually show either a square lattice or striations running parallel to the short or long axis. In both cases the interspacings are estimated to be about 100 A, suggesting that the arrangement of the crystallographic planes possesses tetragonal symmetry. High magnifications of the crystalline inclusions together with analysis of their diffraction patterns showed that the striations are composed of double helices of protein macromolecules. In leaves 3-4 mm long, eroded crystals are often observed. When leaves are larger than 5 mm, no crystalline bodies can be identified in chlorenchyma cells. It is possible that they represent a storage form of protein, which is used later by the developing cells.

Cell-type-specific gene expression and acatalasemic peroxisomes in a null Cat2 catalase mutant of maize.

Cell separation studies in conjunction with immunocytochemical studies indicate that mesophyll cells and bundle-sheath cells, the dimorphic photosynthetic cell types in the leaves of the C(4) plant Zea mays, differ in their catalase composition. In particular, catalase-2, the product of the Cat2 gene, is found primarily in the bundle-sheath cells, whereas catalase-3, the product of the Cat3 gene, is found primarily in the mesophyll cells. Electron microscopic observations reveal that bundle-sheath cells of A16, a mutant line lacking expression of the Cat2 gene in all tissues examined, contain numerous peroxisomes, but they are acatalasemic as determined by staining with 3,3'-diaminobenzidine. The significance of this mutant in physiological studies is discussed.