title: DOP-PCR based painting of rye chromosomes in a wheat background.
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.
authors.affiliation: a Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, Chaoyang District, Beijing 100101, China.
authors.forename: Chuanliang
authors.initials: C
authors.lastname: Deng
authors.forename: Lili
authors.initials: L
authors.lastname: Bai
authors.forename: Shufen
authors.initials: S
authors.lastname: Li
authors.forename: Yingxin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xiang
authors.initials: X
authors.lastname: Li
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Chen
authors.forename: Richard R-C
authors.initials: RR
authors.lastname: Wang
authors.forename: Fangpu
authors.initials: F
authors.lastname: Han
authors.forename: Zanmin
authors.initials: Z
authors.lastname: Hu
mesh_terms: D020223:Chromosome Painting / Q000379:methods*; D032461:Chromosomes, Plant; D015342:DNA Probes; D017404:In Situ Hybridization, Fluorescence; D042282:Microdissection; D016133:Polymerase Chain Reaction; D012434:Secale / Q000235:genetics*; D014908:Triticum / Q000235:genetics*
chemical_list: D015342:DNA Probes
keywords: 1R; 1RS; Secale cereale; chromosome painting; marquage des chromosomes; microdissection