Study On Isozyme Band Of Wheat Callus During Subculture

Induction rate of immature spike and isoenzyme bands variations of peroxidase and acid phophatase during induction and subculture was studied, using wheat cultivars Yunong 9901, Yumai 18, Dali 1, Yumai 49 and Guomai 1. The results showed that:1) During the introduction of immature spike, there was significant difference of half-induction period among different genotypes. The shorter in half-induction, the more flourishing in growth of the calli, vice verse. And Calli with long-induction pierced grew slowly, and the calli had an appearance of white, transparency, and softness.2) In calli induction of immature spikes, spike sized 0.5-1.5cm was induced more easily, and grew faster. 10 days after inoculation, the induction rate almost reached to 100%, and some of the calli sized 3mm in diameter. After 20 days, the size could reach to 5-6mm in diameter, and the calli showed a light yellow color. The results of phophatase isoenzyme band pattern showed that spike calli of more than 1.5cm in size had an extra band, and the band was very thick. 3) As influence factor, Cu2+ had important effect on the culture of calli. Cu2+ concentration of 0.1mmol/L had positive selective effect to the calli. Under this condition, high-quality calli grew faster and showed granule-like tissues. The endurance ability of different calli of different genotypes showed significant difference. Of the genotypes tested, Yumai 18 and Yumai 49 showed high endurance ability to Cu2+, and more calli survived. The results of peroxidase isoenzyme band pattern showed that calli selected by 0.1mmol/L Cu2+ had 2 more extra bands than the calli treated with 0.1mmol/L or 1mmol/L Cu2+.4) There was a variation of peroxidase isoenzyme band pattern among the same generation with different explants. The peroxidase isoenzyme band pattern of the first generation of immature spike was thicker than that of immature embryos, and appeared some extra bands. Of the fourth subculture generation, peroxidase enzyme band pattern of immature embryos was thicker than that of immature spike,and also showed more extra bands. Compared with other cultivars, Yumai 18 and Yumai 49 had an extra band. With increasing of subculture times, peroxidase was enzyme bands were decreasing.5) The results of acid posphorase were rather similar to that of peroxidase, and difference of acid posphorase was observed during different subcultures. And the 6th and 7th generations showed the least enzyme bands.