Electrophoretic analysis of soluble proteins and enzymes from selected species of Azolla and their hybrids

Differences and similarities of proteins and enzymes of selected species of azolla and their hybrids were investigated by polyacrylamide disc gel electrophoresis (PAGE). Polyvinylpyrollidone and dithiothreitol were utilized in the homogenization of Azolla. Azolla species studied were not Anabaena free and thus, Anabaena azollae was used as the control. Electrophoretic patterns of soluble proteins showed differences between selected species of Azolla although similar bands were observed. Three protein zones - zones 1, zone 2, zone 3, - were present in all the species. However, zone 2 showed variation in the number of bands. Similarly, glucose-6-phosphate dehydrogenase (G6PD) isozyme patterns showed differences between species. A slow migrating G6PD zone, designated as B, was detected. The presence and absence of three subbands-Bs, Bm, and Bf- were noted in each species. Only Azolla filiculoides 100 g showed two other unique bonds (Rf=0.197 and 0.624). On the other hand, different Azolla species exhibited different esterase phenotypes. Within each phenotype, four esterase zones were observed Est-1, Est-2, Est-3, and Est-4. Est-2 and Est-3 were present in all Azolla species but differences in the relative mobilities and intensities were observed. A. microphylla 4003 and A. filiculoides 1009 exhibited the two other esterase zones. The results indicated genetic differences between Azolla species. Moreover, it was also found out that G6PD can be used as a biochemical marker in Azolla species. Esterase, however, was not a suitable biochemical marker because of its non-specificity and variability. Comparison of electrophoretic patterns between Azolla parents and hybrids was, likewise, done. General protein patterns among the different hybrids showed the same three zones found in the parents. On the other hand, G6PD isozymes in the hybrids showed appearance and disappearance of the three B subbands. Subband Bm was seen to be present in four of the five hybrid lines. Likewise, esterase isozymes were grouped into four zones. Only one of the five hybrid lines showed the four esterase zones