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 Extended Observation Descriptions 

Extended Observation Description on Micrasterias fimbriata

The meshwork of cellulose microfibrils in the primary cell wall of Micrasterias fimbriata

Critical point drying (CPT) enables the preparation of single-celled organisms for scanning electron microscopy (SEM) with minimal artifacts. Following CPT processing of a sample from the “Schwemm” peat bog near Walchsee (Tyrol, Austria) in Jürgen Stampfl’s lab, the network of cellulose microfibrils in the primary wall of Micrasterias fimbriata and the swellings of the pore apparatuses in the secondary wall could be precisely visualized (Fig. 1).

Micrasterias fimbriata
Micrasterias fimbriata

Fig. 1: Micrasterias fimbriata from the “Schwemm” moor near Walchsee in Tyrol (Austria). A net-like covering is already visible in the overview image.

During cell division in desmids, as well as in vascular plants, the primary cell wall is initially formed in each newly created cell. It is thin (50–100 nm) and soft, yet flexible, and protects the developing cell. The primary cell wall consists of cell wall matrix (pectin, hemicelluloses) but also already possesses a thin network of cellulose microfibrils. Once the nucleus, chloroplasts, and mitochondria have divided and completely migrated into the new cell, the cell begins to build the secondary cell wall, which acquires the sculpture and ornamentation typical of the species and provides the necessary stability. Only in Cosmarium species is it known that the primary cell wall is shed after cell division is complete; in Micrasterias, it envelops the gelatinous layer of mucopolysaccharides that rests on the resistant secondary cell wall. This gelatinous layer, produced in the cell’s dictyosomes, is transported to the outside via the pore apparatuses and provides additional protection to the cell (Fig. 2).

Cell wall of the Cosmarium type

Fig. 2: Schematic drawing of a cross-section through a Desmidium cell wall of the Cosmarium type, which also includes the cell walls of Micrasterias. Pw = primary wall, Sw = secondary wall, Pa = pore apparatus. Adapted from Brook modified.

Micrasterias fimbriata

Fig. 3: Detail of the upper cell half (sections of the apical lobe and lateral lobes). The meshwork of the cellulose microfibrils of the primary wall and the pore apparatuses (arrow) in the secondary wall are clearly visible.

Micrasterias fimbriata
Micrasterias fimbriata
Micrasterias fimbriata

Figs. 4–6: A series of magnifications, at the end of which even the thickness of individual network strands can be measured. The arrows point to pore structures as examples.

The magnification series (Figs. 4–6) impressively shows the network of cellulose microfibrils in the primary wall. The protuberances marked with arrows are examples of cellular structures originating from the pore apparatuses in the underlying secondary wall. In the last image, the diameters of the microfibrils become measurable at the highest magnification. Within the limits of measurement accuracy, the finest visible strands have diameters of approximately 10 nm. The order of magnitude of these measurements corresponds well with the findings of Giddings et al. (1980). They had determined that during the formation of the primary wall, individual production apparatuses of cellulose microfibrils (the so-called rosettes) produce microfibrils with a diameter of 5 nm. They were further able to show that during the formation of the secondary wall, hexagonally arranged arrays of rosettes produce complex fibrils with a diameter of 28 nm, which form the basis of the secondary wall.

Such instructive depictions of the primary cell wall in desmids are rare even in the life sciences literature. This is likely due to the fact that the ultrastructural details of the cellulose walls, as well as the processes involved in their formation, were investigated using transmission electron microscopes in the 1980s and 1990s. SEMs of the quality required to capture images like the one shown above were simply not available at that time.

References

Brook, A. J. (1981). The biology of desmids (Vol. 16). Univ of California Press.

Giddings Jr, T. H., Brower, D. L., & Staehelin, L. A. (1980). Visualization of particle complexes in the plasma membrane of Micrasterias denticulata associated with the formation of cellulose fibrils in primary and secondary cell walls. The Journal of cell biology84(2), 327-339.

Kleinig, H., & Maier, U. (1999). Zellbiologie: Begründet von Hans Kleinig und Peter Sitte.

All images:
Tescan Clara SEM, © Wolfgang Bettighofer/Jürgen Stampfl

© Wolfgang Bettighofer,
images under Creative Commons License V 3.0 (CC BY-NC-SA).
For permission to use of (high resolution) images please contact postmaster@protisten.de.

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