| II. Acid phosphatase isozymes About 100 mg of seeds of each strain were soaked in petri-dishes for 24 hours in a growth chamber (23C). Polyacrylamide gel electrofocusing of seed extracts was carried out using a minor modification of DAVIS' (1962) method of disc electrophoresis. This method was described in detail in a previous publication (NAKAI and TSUNEWAKI 1971). The procedure for staining acid phosphatase isozymes was the same as that described by ALLEN et al. (1963). The acid phosphatase of Ae. umbellulata showed a simple zymogram (Fig. 3a); the pI 6.0 band showed high activity, while three bands of pI 5.7, 5.8 and 6.4 had low activities. Separation of bands at pI 7.4-7.7 was very poor. In general, both varieties of Ae. caudata (var. typica and var. polyathera) showed similar zymograms with two highly active bands at pI 5.7 and 6.5, and three other bands at pI 6.0, 6.3 and 7.1 (Fig. 3b). However, a single strain of var. typica differed from the other strains of Ae. caudata in that the pI 5.85 band showed high activity (Fig. 3c). Zymograms of both natural and artificially synthesized strains were similar, i.e., they had bands at pI 5.7, 5.8, 6.0, 6.3, 6.4, 6.5, 6.85 and 7.5-7.7 in common (Fig. 3d, e). However, the activities of two bands at pI 6.4 and 6.5 were low in the synthesized strains as compared to those of the natural strain. The zymogram of Ae. triuncialis involved all the bands of Ae. caudata (pI 5.7, 6.3 and 6.5) and Ae. umbellulata (pI 6.0, 6.4). The pI 5.8 band (No. 2 band) of natural and synthesized Ae. triuncialis was not found in either of the parental species. Furthermore, this band did not appear in a 1 : 1 mixture by weight of the extracts of Ae. caudata and Ae. umbellulata (Fig. 3f). These results suggest that the pI 5.8 band in Ae. triuncialis is a hybrid enzyme. Literature cited ALLEN, S. L., M. S. MISCH and M. MORRISON 1963. Variations in the electrophoretically separated acid phosphatase of Tetrahymena. J. Histochem. Cytochem. 11: 706-719. BREWER, G. J., C. F. SING and E. R. SEARS 1969. Studies of isozyme patterns in nullisomic-tetrasomic combinations of hexaploid wheat. Proc. N.A.S. 64: 1224-1229. KIHARA, H. and N. KONDO 1943. Studies on amphidiploids of Aegilops caudata x Ae. umbellulata induced by colchicine. Seken Ziho 2: 24-42. KIHARA, H., K. YAMASHITA and M. TANAKA 1965. Morphological, physiological and cytological studies in Aegilops and Triticum collected from Pakistan, Afghanistan and Iran. Results of the Kyoto Univ. Scientific Expedi. to the Karakoram and Hindukush. Vol. 1: 1 -72. KONDO, N. 1941.Chromosome doubling in Secale, Haynaldia and Aegilops by colchine treatment. Jap. Jour. Genet. 17: 46-54. NAKAI, Y., T. TERAMURA and K. TSUNEWAKI 1969. Variation in the esterase of Triticum and Aegilops analyzed by the gel isoelectrofocusing technique. Jap. J. Genet. 44: 259-270. NAKAI. Y., and K. TSUNEWAKI 1971. Isozyme variations in Aegilops and Triticum. I. Esterase isozymes in Aegilops studied using the gel isoelectrofocusing method. Jap. J. Genet. 46: 321-336. SEARS, E. R. 1939. Amphidiploids in the Triticinae induced by colchicine. The Journal of Heredity 30: 38-43. SENJANINOVA-KORZAGINA, M. 1932. Karyo-systematical investigation of the genus Aegilops L. Bull. Appl. Bot., Genet. and Plant-Breed. Ser. 2: 1-90. WAINES. J. G. and B. L. JOHNSON 1971. New protein bands in an amphiploid of Aegilops caudata and Ae. umbellulata. Wheat Inf. Service 32 : 22-24. ZOHARY, O. and M. FELDMAN 1962. Hybrization between amphiploids and the evolution of polyploids in the wheat (Aegilops-Triticum) group. Evolution 16: 44-61. (Received January 21, 1972) |
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