关键词: 黄麻, 单染色体, 分离, 未克隆文库

Abstract:

The technology of isolation, amplification and cloning of single chromosome is of significant value in plant genetic research, especially for those plants with poor genetic research basis. However, the technology has not been widely applied to plants because most plants have small chromosomes that are difficult to be distinguished. To investigate the feasibility and possibility of applying the technology to plants with small chromosomes, we experimented on the construction of uncloned single chromosomal DNA libraries using jute as a model, which has typical small chromosomes. We successfully isolated 80 single chromosomes from cultivar Minyin-1 of jute (Corchours olitorius L.) by micromanipulation with glass needles and amplified individual chromosomes by two-round LAM-PCR or DOP-PCR, from which DNA fragments ranging 250–2 000 bp or 100–600 bp were obtained. Southern hybridization with a-³²P-dCTP labeled genomic DNA confirmed that the PCR products were really from the jute genome. Hence, we acquired uncloned DNA libraries of single chromosomes of jute. A total of 50 uncloned single-chromosome DNA libraries were established. Our research suggests that isolation individual small chromosomes with glass needles and amplification of DNA fragments from them with LAM-PCR or DOP-PCR for constructing uncloned single chromosome DNA libraries in plants is feasible. Issues concerning the application of uncloned single-chromosome DNA libraries are discussed.

Key words: Jute, Single chromosome, Isolation, Uncloned library