DOP-PCR based painting of rye chromosomes in a wheat background.
DOP-PCR based painting of rye chromosomes in a wheat background.
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DOI：
10.1139/gen-2014-0110
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发表时间：
2014-11
期刊：
影响因子：
3.1
通讯作者：
C. Deng;Lili Bai;Shu-Fen Li;Yingxin Zhang;Xiang Li;Yuhong Chen;R. C. Wang;F. Han;Zanmin Hu-Zanm
中科院分区：
文献类型：
--
作者：
C. Deng;Lili Bai;Shu-Fen Li;Yingxin Zhang;Xiang Li;Yuhong Chen;R. C. Wang;F. Han;Zanmin Hu-Zanm
为了确定染色体涂染技术在黑麦基因组元件鉴定中的适用性，我们对黑麦的1 R和1 RS染色体进行了显微解剖。变种King II）和小麦-黑麦异附加系1 RS。利用简并寡核苷酸引物-聚合酶链反应（DOP-PCR）技术，以1 R和1 RS为探针，与黑麦、小麦-黑麦异附加系1 R和1 RS、易位系1 RS、1BL和异源六倍体小黑麦的中期染色体进行杂交。结果表明：（1）1 R和（或）1 RS的DOP-PCR产物在黑麦染色体上的杂交信号分布模式与1 RS和（或）1 RS的杂交信号分布模式相似，不能仅通过1 R和（或）1 RS的DOP-PCR产物的杂交信号来区分1 R和（或）1 RS与其它黑麦染色体;（iii）用1 R或1 RS的DOP-PCR产物作探针，可以清楚地鉴定小麦-黑麦杂种中的黑麦染色体和（或）黑麦染色体片段，而且在非端粒区的鉴定比基因组原位杂交（GISH）更准确。结果表明，1 R和（或）1 RS来源的DOP-PCR产物含有大量重复序列，在不同黑麦染色体上具有相似性，具有R基因组特异性，可用于小麦-黑麦杂种中黑麦染色体和染色体片段的鉴定。本研究拓宽了植物染色体彩绘的应用范围。
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.