ISOLATION AND PRESERVATION OF THE LIVING EMBRYO SAC OF CRINUM-ASIATICUM L VAR JAPONICUM BAKER

The enzymatic maceration method was used to isolate an intact embryo s ac of Crinum asiaticum and its component cells. Best results were obta ined when using enzyme solutions that contained pectinase, hemicellula se, cellulase and pectolyase. Aseptic ovules were incubated in the enz yme solution for 1.5 hr at 25 C. This allowed the isolation of embryo sacs to yield up to 20% of the amount present. An isolated embryo sac usually consists of an egg cell, synergids, antipodals and a central c ell. Some embryo sacs can be digested as gametophytic protoplast. The size, shape and position of the isolated embryo sac seemingly possesse d similarities with those of the fixed embryo sac in the ovary. An iso lated embryo sac can be in a living state when the result of the fluor ochromatic reaction (FCR) and protoplasmic streaming is positive. When cultured in proper media, 68% of the isolated gametophytic protoplast s were observed to have sustained their positive FCR for more than 1 m onth.