ABA、NAA诱导水稻胚性愈伤组织的研究

ABA和NAA联合使用能有效地诱导水稻原生质体再生的愈伤组织向胚性发展。通过液体浅层培养由原生质体得到的愈伤组织,在含ABA和NAA的N_6培养基上培养一段时间,可以诱导原来呈非胚性状态的愈伤组织形成胚性愈伤组织,并在含ZT的N_6分化陪养基上产生绿点。通过对这两种愈伤组织的生化分析,表明二者在游离氨基酸、DNA、RNA、核酸及蛋白质含量等方面,特别是SDS-PAGE谱带存在明显的差异,其细胞的形态与结构也有显著差别,其中经ABA NAA诱导后的愈伤组织其细胞形态与结构特征与来源于种胚的胚性愈伤组织基本类似,所分析的生化指标也大多数相近。结果表明,ABA和NAA联合使用得当,能促进形成胚性愈伤组织。

STUDIES ON THE INDUCTION OF EMBRYOGENIC CALLUS IN RICE BY ABA, NAA

The embryogenic callus can be induced effectively from rice protoplast regenerated non-embryogenic callus by ABA(0.5mg/L) and NAA (2.0mg/L). When these embryogenic calli were transferred on differentiation medium(N8 ZT 2.0 mg/L); the green bud points were observed on the calli. Observations on cell ultrastruc-ture in two calli (treated and untreated with ABA, NAA) showed that there were many globular embryoids in the treated callus, the treated cells were smaller than the untreated and full of cytoplasm, these characters of ultrastructure are similar to those of embryogenic callus induced directly from mature seed embryos. Total nucleic acid, DNA, RNA, free amino acid and protein contents in cells among treated and untreated calli with ABA, NAA and embryogenic callus were compared. The results showed: (1) The contents of total nucleic acid, DNA, RNA in the treated are higher than those in the untreated. (2) The free amino acid content of the treated is more than four times that of the untreated. The proportion of Ala is increased apparently in the treated. (3) The protein content in the treated is twice as much as that of untreated. Meantime, SDS-PAGE showed that the protein patterns between the treated and the untreated were different, but the pattern of the treated was similar to that of embryogenic callus induced directly from mature seeds. These results indicate that the embryogenic process of callus is accompany with the strengthen of metabolism in nucleic acid and protein, and perhaps this character also is a character of the embryogenic callus.