葎草单染色体的显微分离及DOP-PCR技术体系建立

Establishment of DOP-PCR Amplification System for Microdissecting Single Chromosome in Humulus japonicas

葎草是具有XX/XY1Y2性染色体系统的雌雄异株植物,是研究植物性染色体演化的模式材料之一.利用染色体显微分离技术从葎草根尖有丝分裂中期分裂相中将单条染色体进行了显微分离及DOP-PCR(Degenerate oligonucleotide primer-PCR)扩增,并构建了单染色体DOP-PCR扩增产物的荧光探针,对葎草根尖有丝分裂中期分裂相染色体进行了荧光原位杂交,其结果表明荧光信号分布在所有的染色体上,表明所建立的技术体系能够成功分离葎草单染色体并进行DNA扩增.本研究结果为进一步进行葎草X,Y染色体的细胞及分子生物学研究提供了技术支持.

Humulus japonicas is dioecious plants with XX/XY1Y2 sex chromosome system,which is one of model materials for studying sex chromosome evolution.In this study,the single chromosome of Humulus japonicaswas microdissected and amplified from roots tip on mitosis metaphases.Then,the DOP-PCR products amplified from the single chromosome were labeled with Alexa Flour-488 by nick translation method and hybridized to the chromosomes of female Humulus japonicasplants.The fluorescence signals were distributed on all chromosomes.The results showed that the single chromosome of Humulus japonicaswas successfully microdissected and amplified,which found a basis for microdissect the X or Y chromosome of H.japonicas.