The use of isoenzymes in the study of germination, development and breeding of legumes

Abstract

Amylase activities and patterns were compared in extracts from mature seeds and from different parts of seedlings at various stages of seed germination of various varieties of different legume species. Apart from soyabean, all varieties had low levels of amylase activities in extracts from mature seeds, while the former had a remarkably high level. Amylase activities and the number of bands increased during seed germination and showed time and tissue specifity. The extracts from cotyledons had the highest activities and the largest number of bands as compared to that of the various parts of embryonic axes. Amylase activities and patterns had time specific changes during the various stages of seed development of the 7 varieties of different legume species studied. All the different varieties showed changes in zymogram patterns and decreased amylase activity during seed development, except soyabean where amylase activity remained high at all stages. The different components of amylase were characterised in pea (var. Feltham First). One band of a -amylase activity, restricted to the cotyledons, was present in the middle stages of seed development only, then reappeared at the third day of seed germination. The embryonic axes β -amylase activity decreased as the seed developed, to reach zero level in mature seeds, and reappearred during seed germination. The reverse sequence occurred for the cotyledonary β -amylase. The pea a -amylase was partially purified using ethanol precipitation, glycogen complex and hydroxyapatite column chromatography. The purified protein had three major bands with few faint bands on the SDS polyacrylamide gel. The embryonic axes β -amylase of pea was partially purified using the conventional method, ammonium sulphate precipitation, ion exchange chromatography and gel filtration. The purified protein contained two thick bands and many faint bands on the SDS polyacrylamide gel. The zymogram patterns of phosphorylase, EST, GDH, GOT, LDH, ADH and MDH were investigated during seed development of various varieties of different legume species. The most noticeable changes as the seeds developed were the decrease in activities and number of bands of EST in pea and soyabean, shift in GDH activities between isoenzyme forms in pea (var. English Wonder), increase in GDH activities and number of bands in dwarf French bean and soyabean and increase in activities and number of bands of MDH in pea (var. Feltham First).