Histochemical study of the development of the phytomelan layer in the seed coat ofGasteria verrucosa (Mill.) H. Duval

Summary In this study we document the development of the phytomelan layer in the outer epidermis of the outer integument ofGasteria verrucosa. Phytomelan has been described as a black, melanin-like substance which is chemically very inert. Using histochemical techniques we show that phytomelan development in the cell wall can be divided into three stages. The first stage is deposition of a callosic layer against the tangential wall, with simultaneous thickening of the adjacent parts of the radial walls. The second stage is the conversion of this callosic wall, which we call a tertiary wall, into a noncallosic inner and outer layer. The inner layer stains predominantly for cellulose and a little for pectin. The outer layer is of unknown composition, since it did not react with the stains that were used. In the third stage the outer tertiary layer becomes black, the phytomelan. The callosic wall deposited in the first developmental stage seems to function as a carbohydrate source and as a mould for the tertiary cell wall. The conversion of the callose in the second stage might be the result of penetration of substances which react with callose. All the components for phytomelan seem to be present in the outer layer before the conversion. Phenolics might be involved in this second conversion.Abbreviations DAP days after pollination - PAS periodic acid Schiff's reagent - PEG polyethylene glycol