DOP–PCR based painting of rye chromosomes in a wheat background

Author:

Deng Chuanliang12,Bai Lili1,Li Shufen12,Zhang Yingxin13,Li Xiang12,Chen Yuhong1,Wang Richard R.-C.4,Han Fangpu1,Hu Zanmin1

Affiliation:

1. Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101, China.

2. Henan Normal University, Xinxiang 453007, China.

3. Graduate University of Chinese Academy of Sciences, Beijing 100049, China.

4. USDA-ARS, FRRL, Utah State University, Logan, UT 84322-6300, USA.

Abstract

To determine the appropriateness of chromosome painting for identifying genomic elements in rye, we microdissected the 1R and 1RS chromosomes from rye (Secale cereale L. var. King II) and wheat–rye addition line 1RS, respectively. Degenerate oligonucleotide primed – polymerase chain reaction (DOP–PCR) amplification of 1R and 1RS products from dissected chromosomes were used as probes to hybridize to metaphase chromosomes of rye, wheat–rye addition lines 1R and 1RS, translocation line 1RS.1BL, and allohexaploid triticale. The results showed that (i) 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; (ii) 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; (iii) rye chromosomes 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 in the nontelomeric region than using genomic in situ hybridization (GISH). Our results suggested that 1R- and (or) 1RS-derived DOP–PCR products contain many repetitive DNA sequences, are similar on different rye chromosomes, are R-genome specific, and can be used to identify rye chromosomes and chromosome fragments in wheat–rye hybrids. Our research widens the application range of chromosome painting in plants.

Publisher

Canadian Science Publishing

Subject

Genetics,Molecular Biology,General Medicine,Biotechnology

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