豌豆间期细胞核内核仁组织者区排列的非放射性原位杂交定位  

LOCALIZATION OF NUCLEOLAR ORGANIZER REGION ARRANGEMENT IN INTERPHASE NUCLEI OF PISUM SAT1VUM BY NON-RADIOACTIVE IN SITU HYBRIDIZATION

作　　者：王新宇[1] M.I.Highett D.J.Rawlins P.J.Shaw 

机构地区：[1]兰州大学生物系,兰州730000 [2]John Innes Institute of Plant Sciences, Norwich, NR4 7UH, UK

出　　处：《实验生物学报》1995年第2期191-201,共11页Acta Biologiae Experimentalis Sinica

摘　　要：以异羟基洋地黄毒甙(digoxigenin)标记的rRNA为探针与多聚甲醛固定的植物根尖振动切片原位杂交。结合免疫荧光检测分析了豌豆(Pisum sativum)间期细胞核内核仁组织者区的数目、分布和排列。研究结果发现豌豆根尖分生组织间期细胞核内有1—6个被探针分子强烈标记的位点。这些位点呈大小不等(0.24-1.98μm^3)的斑点状,分布在核仁的周边,并且与核仁相连。探针标记位点在核仁周边的排列方式随标记位点的数目不同而异。在具有四个标记位点的细胞核内,它们呈四面体,梯形或平行四边形排列。在显示原位杂交阳性的细胞核中,59.5%显示4个标记位点,少于或多于4个标记位点的细胞分别占35.7%和4.76%。探针标记位点的数目与标记位点本身的大小无明显联系,而与核仁的体积大体上呈负相关趋势。文章讨论了这些被rRNA探针标记的位点与核仁组织区及核仁形成的关系。The number, distribution and arrangement of nucleolar organizer regions (NORs) in interphase nuclei of Pisum sa-tivum L. were studied by the non-radioactive in situ hybridization and immuno-fluorescence staining of the vibration sections of paraformaldehyde fixed root tips using a digoxigenin-conjugated rRNA probe. It was found that there was (were) 1-6 sites intensely labelled by the probe in the interphase nuclei of the plant, which were in the shape of spots with variable sizes (0.24-1.98μm3), located around and connected with a nucleolus. The arrangement of the labelled perinuc- leolar sites were polymorphic because of their different numbers. In the interphase nuclei with four labelled sites they tended to be in a tetrahedral, trapezoid or parallelogram arrangement. Of the nuclei exhibited positive in situ hybridization, 59.5% had 4 labelled sites, 35.7% less than 4, and 4.76% more than 4 labelled sites. The numbers of the probe labelled sites were not singnificantly related to the sizes of themselves, but roughly in negative relation to the sizes of nucleoli. The relationships between the labelled sites and both NORs and nucleolar formation are discussed.

关 键 词：非放射性 原位杂交 豌豆 核仁组织者 细胞核 

分 类 号：S643.301[农业科学—蔬菜学]