ABSTRACT
A method for obtaining algal chromosomal preparations is described employing the Feulgen method for DNA staining, Fe-propionocarmine as an enhancing stain, and cupra-ammonium to remove cell wall material. Fe-propionocarmine applied as a gradient to the side provides cells stained with the Feulgen stain alone or with the Feulgen Fe-propionocarmine stain, thereby facilitating useful comparison. Where dilute the Fe-propionocarmine enhances nuclear staining without staining orthe organelles; where more concentration it also stains the nucleolus, spindle, spindle polar bodies, pyrenoid and protoplast. Treatment with cupra-ammonium, to remove polysaccharide wall material, followed by neutralization with propionocarmine, enables thinner squashes and better chromosome spreads without loss of differential staining. Preparations mounted in euparal are long-lasting.
Subject(s)
Chlorophyta/ultrastructure , Chromosomes/ultrastructure , Histocytochemistry , Polysaccharides , Species Specificity , Staining and LabelingSubject(s)
Mitosporic Fungi/cytology , Amino Acids/analysis , Ammonium Sulfate , Cell Fractionation , Cell Wall/analysis , Culture Media , Cytoplasmic Granules , Galactose/analysis , Glucose/analysis , Hexosamines/analysis , Lipids/analysis , Mannose/analysis , Melanins , Methylation , Microscopy, Electron , Mitosporic Fungi/analysis , Mitosporic Fungi/growth & development , Nitrates , Polysaccharides/analysis , Spores/cytology , Spores, Fungal/analysis , Spores, Fungal/cytology , Spores, Fungal/growth & developmentABSTRACT
A chemically resistant cuticle fraction was isolated from 5 phaeophycean, 1 rhodophycean, and 11 chlorophycean marine algae using acid treatment alone, or acid treatment followed by leaching in cupra-ammonium. In Cladophora rupestris and Chaetomorpha melagonium this fraction consists of several alternate microfibrillar and amorphous layers similar in appearance to those seen in innermost carbohydrate-rich regions and amount to about 1/10 or more of the cell wall thickness. In Porphyra umbilicalis and Padina vickersiae it is a single layer less than I µ thick, accounting for 1/50-1/100 of the cell wall in Porphyra, and 1/5-1/10 of the cell wall in Padina. The cuticle fractions of all 4 algae contain surprisingly large amounts of protein (about 70% in Cladophora and 80% in Porphyra). Similarities in the behavior of cuticles obtained from the other 12 species studied suggest that they may have a similar protein-rich composition.