摘要
用非酶手工显微解剖法分离小麦(TriticumaestivumL.)合子与幼胚。应用金霉素(CTC)和氟奋乃静(FPZ)荧光探针,观察了合子与胚胎发育过程中膜钙和活化钙调素的分布。合子原生质体CTC荧光呈极性分布。二细胞至十几个细胞原胚时期各细胞间的CTC和FPZ荧光均匀分布。幼小梨形胚胚体CTC和FPZ荧光稍强于胚柄。接近分化的梨形胚CTC荧光仅位于胚体与胚柄之间的几层细胞和胚体腹侧中部的几个细胞中。胚芽刚分化时,胚基部CTC荧光较强,FPZ荧光则仅出现在胚基部。讨论了合子分离技术和膜钙与钙调素(CaM)在胚胎发育中的可能作用。
The method of nonenzymatic,manual microdissection was established to isolate zygotes and young embryos in Triticum aestivum L. The distribution of membranebound calcium and activated calmodulin in the isolated zygotes and young embryos was visualized by chlorotetracycline (CTC) and fluphenanize (FPZ) fluorescence probe respectively. The CTC fluorescence was polar distributed in the zygote protoplast. The distribution of the CTC and FPZ fluorescence from twocelled embryos to multicellular embryos was observed. In the young pearshaped embryos the CTC and FPZ fluorescence of the embryos was slightly higher than that of the suspensor. In a pearshaped embryo beginning with differentiation the CTC fluorescence was restricted to severallayer of cells between embryo and suspensor and the several ventral cells of the embryo. In the embryos with newly differentiated plumule the basal part of the embryo possessed a higher CTC fluorescence,while the FPZ fluorescence was only distributed in the basal part. It indicated that the distribution of CTC and FPZ fluorescence was in coincidence with the sites that plumule and radicle were beginning to differentiate. The technique of isolated zygotes and the possible function of calcium and calmodulin during embryo development are discussed.
基金
国家自然科学基金
关键词
分离合子
胚胎发育
膜钙
钙调素
小麦
Zygote, Embryo development, Isolation, Membranebound calcium, Calmodulin, Triticum aestivum