Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation.
Karyotype analysis of Lilium longiflorum and Lilium rubellum by chromosome banding and fluorescence in situ hybridisation.
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DOI：
10.1139/g01-066
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发表时间：
2001-10
期刊：
影响因子：
3.1
通讯作者：
Ki-Byung Lim;Jannie Wennekes;J. Jong;Evert Jacobsen;Jaap M. van Tuyl
中科院分区：
文献类型：
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作者：
Ki-Byung Lim;Jannie Wennekes;J. Jong;Evert Jacobsen;Jaap M. van Tuyl
以5S和45S rDNA序列为探针，通过染色体臂长、C-带、AgNO_3染色和PI-DAPI显带，结合荧光原位杂交(FISH)技术，构建了长花百合和橡胶百合的详细核型。用标准的BSG技术获得的C-带型在长花乳杆菌和红花乳杆菌染色体的异源位置上只显示了很少的少量条带。长花乳杆菌中期复合体的5S和45S rDNA探针FISH显示，5S和45S rDNA探针在第4、7号染色体短臂的近端有重叠信号，在第3染色体的次级收缩处有单一的5S rDNA信号，在第3染色体长臂的亚远端有1个45S rDNA信号。在长花乳杆菌中，我们观察到5S和45S rDNA序列在第2和4染色体的短臂以及第2和3染色体的长臂上共定位。核仁和NORs的银染(Ag-NOR)显示，间期核中信号数目变化很大，而有丝分裂中期染色体的NORs上只有少量微弱的银染色质。在PI和DAPI染色的标本中，我们观察到长花乳杆菌和红花乳杆菌染色体上不同位置的红色和蓝色荧光带。红色荧光或所谓的反向PI-DAPI带总是与rDNA位点重合，而蓝色荧光DAPI带对应于C-带。基于这些技术，我们可以鉴定出长花乳杆菌和红花乳杆菌的大部分染色体。
Detailed karyotypes of Lilium longiflorum and L. rubellum were constructed on the basis of chromosome arm lengths, C-banding, AgNO3 staining, and PI-DAPI banding, together with fluorescence in situ hybridisation (FISH) with the 5S and 45S rDNA sequences as probes. The C-banding patterns that were obtained with the standard BSG technique revealed only few minor bands on heterologous positions of the L. longiflorum and L. rubellum chromosomes. FISH of the 5S and 45S rDNA probes on L. longiflorum metaphase complements showed overlapping signals at proximal positions of the short arms of chromosomes 4 and 7, a single 5S rDNA signal on the secondary constriction of chromosome 3, and one 45S rDNA signal adjacent to the 5S rDNA signal on the subdistal part of the long arm of chromosome 3. In L. rubellum, we observed co-localisation of the 5S and 45S rDNA sequences on the short arm of chromosomes 2 and 4 and on the long arms of chromosomes 2 and 3, and two adjacent bands on chromosome 12. Silver staining (Ag-NOR) of the nucleoli and NORs in L. longiflorum and L. rubellum yielded a highly variable number of signals in interphase nuclei and only a few faint silver deposits on the NORs of mitotic metaphase chromosomes. In preparations stained with PI and DAPI, we observed both red- and blue-fluorescing bands at different positions on the L. longiflorum and L. rubellum chromosomes. The red-fluorescing or so-called reverse PI-DAPI bands always coincided with rDNA sites, whereas the blue-fluorescing DAPI bands corresponded to C-bands. Based on these techniques, we could identify most of chromosomes of the L. longiflorum and L. rubellum karyotypes.