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ARS Home » Pacific West Area » Logan, Utah » Forage and Range Research » Research » Publications at this Location » Publication #303501

Research Project: Develop Improved Plant Genetic Resources to Enhance Pasture and Rangeland Productivity in the Semiarid Regions of the Western U.S.

Location: Forage and Range Research

Title: DOP-PCR-based chromosome painting of rye (Secale cereale) and wheat-rye hybrid 1R and 1RS chromosomes

Author
item DENG, C - Chinese Academy Of Sciences
item BAI, L - Chinese Academy Of Sciences
item LI, S - Chinese Academy Of Sciences
item ZHANG, Y - Chinese Academy Of Sciences
item LI, X - Chinese Academy Of Sciences
item CHEN, Y - Chinese Academy Of Sciences
item Wang, Richard
item HAN, F - Chinese Academy Of Sciences
item HU, Z - Chinese Academy Of Sciences

Submitted to: Genome
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 10/29/2014
Publication Date: 11/3/2014
Citation: Deng, C.L., Bai, L.L., Li, S.F., Zhang, Y.X., Li, X., Chen, Y.H., Wang, R., Han, F.P., Hu, Z.M. 2014. DOP-PCR-based chromosome painting of rye (Secale cereale) and wheat-rye hybrid 1R and 1RS chromosomes. Genome. 57:473-479.

Interpretive Summary: There is an abundance of repetitive DNA sequences in plant genomes in contrast to those of human and animal species. Plant genomes diverged by various mechanisms to maintain their identifies for ensuring chromosomal behaviors and seed production. One of the mechanisms is homogenization of genome-specific repetitive sequences over all chromosomes of a genome. We have previously demonstrated the presence of genome-specific CAPS sequences on all seven chromosomes of two wheatgrass species and one perennial rye species. Now, the present study shows the technique of DOP-PCR with a micro-dissected chromosome is an efficient method to enrich genome-specific repetitive sequences. The products of this new technique can be used to (1) characterize the source genome, 2) paint all chromosomes of that genome in cytogenetic stocks, (3) isolate various families of repetitive sequences for further studies, and (4) assess genome relationships in plant systematic research, etc. Development of this technique adds a new molecular tool to the arsenal of plant scientists.

Technical Abstract: The chromosome painting is an efficient tool for chromosome research. In oeder to determine whether the chromosome painting techniques can be used to identify rye genome in wheat genetic background, 1R and 1RS chromosomes were microdissected from rye (Secale cereale L. var. King ll) and wheat-rye addition line 1RS, respectively. Subsequently, the 1R and 1RS DNAs were amplified by DOP-PCR (Degenerate oligonucleotide primed - polymerase chain reaction) from the dissected chromosomes. The DOP-PCR products of 1R and 1RS DNAs were used as the probe to hybridize the metaphase chromosomes of rye, wheat-rye addition lines 1R and 1RS, translocation metaphase chromosomes of rye, wheat-rye addition lines 1R and 1RS translocation line 1RS, 1BL and the allohexaploid triticale. The results showed that 1) the hybridization signal distribution patterns on rye chromosomes using 1R derived DOP-PCR products as the probe were similar to those using 1RS derived DOP-PCR products as the probe; 2) 1R and/or 1RS could not be distinguished from other rye chromosomes solely by the hybridization patterns using 1R and/or 1RS derived DOP-PCR products as the probe; 3) the rye chromosome and/or rye chromosome fragments could be clearly identified in wheat-rye hybrids using either 1R or 1RS derived DOP-PCR products as the probe and could be more accurate than using GISH (Genomic in situ hybridization). Our results suggested that 1) 1R and/or 1RS derived DOP-PCR products should contain many repetitive DNA sequences, 2) the distribution pattern of amplified repetitive DNA sequences should be similar on different rye chromosomes and 3) the amplified repetitive DNA sequences should be R-genome specific and can be used to identify rye chromosomes and chromosome fragments in wheat-rye hybrids. Our research widened the application range of chromosome painting in plants.