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为了筛选着丝粒探针,在棉花染色体荧光原位杂交中标记染色体着丝粒区域,以便于构建棉花粗线期染色体细胞遗传学图谱.在棉花遗传连锁图上选择尽可能接近着丝粒区的单拷贝的分子标记,以海岛棉pima90-53的BAC文库为素材,采用两维筛库法从该文库中筛选BAC克隆,然后用BAC-FISH技术进行染色体定位检测.用10号染色体长臂末端的SSR引物BNL3563筛选到一个BAC克隆150D24,该克隆在四倍体陆地棉和海岛棉部分染色体着丝粒区有杂交信号,但信号强度不高.以二倍体D基因组为靶DNA进行FISH时,染色体着丝粒区有明显信号,但是以二倍体A,C,E等基因组棉种染色体为靶DNA进行FISH时,染色体着丝粒区未发现杂交信号.BAC克隆150D24可能含有D组特有卫星重复序列,可用作棉属D基因组棉种(包括四倍体棉种D亚组)的着丝粒FISH探针.
In order to screen the centromere probe, the centromeric region of the chromosome was labeled in cotton chromosome fluorescence in situ hybridization in order to construct the cotton boll line cytogenetics map. The cotton genetic linkage map was selected to be as close as possible to the centromeric region The BAC clone of pima90-53 from G. barbadense was used to screen the BAC clones from the library by two-dimensional screen library and then to detect the chromosomal location by BAC-FISH technique. The end of the SSR primer BNL3563 screened a BAC clone 150D24, the clone in tetraploid upland cotton and island cotton part of the chromosome centromere region hybridization signal, but the signal intensity is not high.Double diploid D genome as the target DNA FISH , There was a significant signal in the centromere region of chromosome, but no hybridization signal was found in the centromere region of chromosomes when FISH was carried out using genomic cotton chromosomes such as diploid A, C, E, etc. BAC clone 150D24 may contain D group Unique satellite repeats that can be used as centromeric FISH probes for cotton D genomic cotton (including tetraploid cotton D subgroups).