Changes in the cell wall accompanying drying and maturation determine the ease of isolation of protoplasts from wheat aleurone layers.

Cell walls of aleurone tissue of developing wheat grains (25 days post anthesis) are degraded by cellulase to give a good yield of protoplasts. As grains become older the aleurone cell walls become increasingly resistant to cellulase, which is completely ineffective on 45-day old material. Resistance to cellulase is provoked even in aleurone cells of young grains by enforced drying of the grains to 12% water content. This suggests that the decreasing effectiveness of cellulase that occurs as grains develop and mature may be caused partly by the natural loss of water.Resistance to degradation by the aleurone cell walls of older or dried grains is to a large extent overcome by the addition of hemicellulase and xylanase to the digestion mixture but completely if acetyl esterase is also included. Changes involving hemicelluloses and xylans are therefore implicated in the effects of drying or maturation, including acetylation processes especially in grains which are almost mature. Arising from these findings a method has been developed for obtaining a high yield of protoplasts from aleurone layers of mature wheat grains, having a high viability and response to gibberellic acid.