关键词: 黑麦, 1R染色体, 显微分离, PCR扩增, rDNA探针

Abstract: The chromosome 1R of rye was microdissected and isolated under reverse-microscope with a manipulator from the cells of root tip at mitotic metaphase. Single chromosome 1R was then transferred into 0.5 ml centrifuge tube. After the treatment with Protease K, the DNA of chromosome 1R was digested with restriction enzyme Sau3A, then the Sau3A linker-adaptor was ligated to the ends of the DNA fragments, which was subsequently amplified by means of linker-adapter polym erase chain reaction (LA-PCR) with one of chains of the linker-adapter as primer. By dot blotting with biotinlatyled rDNA of wheat and genome DNA of rye as probes, it was confirmed that the amplified products by LA-PCR are derived from the DNA of chromosome 1R in rye. This work would facilitated to construct the sub-genomic DNA library of chromosome 1R and to screen the specific probes of chromosome 1R in rye.

Key words: rye, chromosome 1R, microdissection, LA-PCR, rDNA