Further use of fluorochromes in the cytochemical characterization of phytoplankton.

The rapid, specific effects of 25 fluorochromes at low concentration and physiological conditions of pH and temperature were investigated on live cells of five phytoplankton species (Prorocentrum micans, Amphidinium carterae, Dunaliella tertiolecta, Chlamydomonas moewusii and Fragilaria crotonensis). They allowed the identification of cellular components such as the plasma membrane, endoplasmic reticulum, Golgi apparatus, thecal plates, nucleus, mitochondria, trichocysts, vacuoles/lysosomes, polyphosphate and starch granules, lipid bodies and hydrolytic enzymes. Morphological alterations of some of these constituents were examined in cells at different metabolic states. It was found that the thickness of Prorocentrum thecal plates increases during cell development while surface pores appear to be formed in the early stages of thecal formation. The number and size of mitochondria varies among cells at different stages of growth. The number of trichocysts, the size of vacuoles and the quantity of polyphosphates, starch or lipid inclusions increases in nitrogen-depleted cells. Photodegradation and photoenhancement phenomena are described. Some important factors helping to avoid quenching and some applications of the fluorochroming technique are presented.

The use of fluorochromes in the cytochemical characterization of some phytoflagellates.

Sixteen fluorochromes were tested for the cytochemical characterization of two dinoflagellates (Amphidinuim carterae, Prorocentrum micans) and one chlorophycean flagellate (Dunaliella tertiolecta). Depending on the fluorochrome used, various cellular components (including the plasma membrane, thecal plates, pusule, trichocysts, nucleus, lipid bodies and vacuoles) were revealed. The different colours obtained from single or double fluorochrome staining enabled the differentiation and identification of most cellular components. Protoplasmic staining with Fluorescein diacetate suggested the occurrence of esterases in the three phytoflagellates. Rhodamine B, Neutral Red, FluoroBora P and Nile Blue revealed extensive occurrence of lipoid bodies in A. carterae, but Nile Blue showed considerable difference from the other stains in the inclusion size and intracellular location of these bodies. Chlortetracycline binding, and its inhibition by the Ca2+ionophore A23187, indicated that the plasma membrane, pusule system and trichocysts contain sites of Ca2+ binding. Calcofluor White ST proved superior to Congo Red and Lucifer Yellow in elucidating structural details of the thecal plates of P. micans. While Acridine Orange revealed the presence of surface-coat acidic polysaccharides, the fluoresceinated lectins established their glycoconjugate nature in all the three flagellates. Possible mechanisms of fluorochrome uptake are discussed.

Chloroplast and mitochondrial DNA in a brown alga Egregia menziesii.

Chloroplasts and mitochondria of the brown alga Egregia menziesii were studied with the electron microscope. In both organelles, 15-25-A fibrils with DNA characteristics are found within areas of electron transparency. In each chloroplast there are two DNA-containing areas, one at each tip of the chloroplast. This localization, the shape and size of each DNA-containing area, and its close association with lamellae in a nondividing chloroplast are noted. One or occasionally two DNA-containing areas are found within the mitochondrion and they are compared with a similar structure in the chloroplast.