Cytochemical characterization and ultrastructural organization in calluses of the agarophyte Gracilariopsis tenuifrons (Gracilariales, Rhodophyta)

The culture and physiology of red macroalgae calluses are well documented. To date, however, no report has either performed a cytochemical analysis or characterized the ultrastructural organization of calluses at different stages of development and under the effect of plant growth regulators. Therefore, to undertake such analyses, this work studied the red seaweed Gracilariopsis tenuifrons (Bird et Oliveira) Fredericq et Hommersand. Morphology studies suggested three types of calluses: a) terminal callus having an irregular amorphous shape and filamentous projections originating from the cortical region of the thallus; b) apical callus growing on apical branches and having an elongated semispherical shape; and c) intercalary callus developing along the intermediary region of the thallus and having the appearance of small declivities with irregular edges. The abundance of intercalary calluses over terminal and apical calluses is most likely a result of a major cortical surface that would support the cellular growth required to generate calluses. Callus development was initially observed as a matrix of cellular disorganization with filamentous projections; then, the cellular mass seemed to become more compact with spherical uncolored aspect. The presence of starch grains in the inner part of the explant could be explained by absorption from the culture medium and by proper biosynthesis during callus development. Cell wall reaction to staining suggested cellulose and agar composition with acidic polysaccharides. Results suggest that none of the three morphological types of calluses showed any significant differences on the basis of either cytochemistry or ultrastructural organization.