水仙荧光原位杂交体系的建立

Establishment of Fluorescence in situ Hybridization in Narcissus

建立以水仙染色体为靶、rDNA为探针的荧光原位杂交实验技术,为进一步利用荧光原位杂交技术分析水仙的亲缘和进化关系奠定了基础。水仙的染色体制片以去壁低渗-火焰干燥法较好,容易获得大量清晰、分散的有丝分裂中期相;切口平移法地高辛标记探针、染色体和探针90℃共变性5min能有效的进行水仙rDNA的染色体荧光原位杂交定位。

Fluorescent in situ hybridization （FISH） technique, 45S rDNA as probe, was established in narcissus in the study. It showed that the modified procedures could be further used to identify narcissus phylogenetic process and relationship. Flame-drying protocol after enzyme digestion of cell wall was a good method for getting clear and well dispersed narcissus chromosome preparation. With probe DNA labeled by nick translation, and co-denaturation of chromosome DNA and probe for 5 min at 90℃, the rDNA was successfully localizated by FISH method.