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野生甜菜Beta procumbens基因组中卫星DNA的差异及分散重复序列的散布

Divergence of satellite DNA and interspersion of dispersed repeats in the genome of the wild beet Beta procumbens.

作者信息

Dechyeva Daryna, Gindullis Frank, Schmidt Thomas

机构信息

Plant Molecular Cytogenetics Group, Institute of Crop Science and Plant Breeding, Christian-Albrechts-University of Kiel, Olshausenstr. 40, D-24118 Kiel, Germany.

出版信息

Chromosome Res. 2003;11(1):3-21. doi: 10.1023/a:1022005514470.

Abstract

Several repetitive sequences of the genome of Beta procumbens Chr. Sm., a wild beet species of the section Procumbentes of the genus Beta have been isolated. According to their genomic organization, the repeats were assigned to satellite DNA and families of dispersed DNA sequences. The tandem repeats are 229-246 bp long and belong to an AluI restriction satellite designated pAp11. Monomers of this satellite DNA form subfamilies which can be distinguished by the divergence or methylation of an internal restriction site. The satellite is amplified in the section Procumbentes, but is also found in species of the section Beta including cultivated beet (Beta vulgaris). The existence of the pAp11 satellite in distantly related species suggests that the AluI sequence family is an ancient component of Beta genomes and the ancestor of the diverged satellite subfamily pEV4 in B. vulgaris. Comparative fluorescent in-situ hybridization revealed remarkable differences in the chromosomal position between B. procumbens and B. vulgaris, indicating that the pAp11 and pEV4 satellites were most likely involved in the expansion or rearrangement of the intercalary B. vulgaris heterochromatin. Furthermore, we describe the molecular structure, and genomic and chromosomal organization of two repetitive DNA families which were designated pAp4 and pAp22 and are 1354 and 582 bp long, respectively. The families consist of sequence elements which are widely dispersed along B. procumbens chromosomes with local clustering and exclusion from distal euchromatic regions. FISH on meiotic chromosomes showed that both dispersed repeats are colocalized in some chromosomal regions. The interspersion of repeats of the pAp4 and pAp22 family was studied by PCR and enabled the determination of repeat flanking sequences. Sequence analysis revealed that pAp22 is either derived from or part of a long terminal repeat (LTR) of an Athila-like retrotransposon. Southern analysis and FISH with pAp4 and pAp22 showed that both dispersed repeats are species-specific and can be used as DNA probes to discriminate parental genomes in interspecific hybrids. This was tested in the sugar beet hybrid PRO1 which contains a small B. procumbens chromosome fragment.

摘要

已分离出野生甜菜物种平卧甜菜(Beta procumbens Chr. Sm.)基因组中的几个重复序列,平卧甜菜属于甜菜属平卧组。根据其基因组结构,这些重复序列被归为卫星DNA和分散DNA序列家族。串联重复序列长度为229 - 246 bp,属于一种名为pAp11的AluI限制性卫星。这种卫星DNA的单体形成亚家族,可通过内部限制性位点的差异或甲基化来区分。该卫星在平卧组中被扩增,但在甜菜组的物种中也有发现,包括栽培甜菜(Beta vulgaris)。远缘物种中存在pAp11卫星表明,AluI序列家族是甜菜基因组的一个古老组成部分,也是栽培甜菜中分化的卫星亚家族pEV4的祖先。比较荧光原位杂交显示,平卧甜菜和栽培甜菜在染色体位置上存在显著差异,这表明pAp11和pEV4卫星很可能参与了栽培甜菜居间异染色质的扩增或重排。此外,我们描述了两个重复DNA家族的分子结构、基因组和染色体组织,它们分别被命名为pAp4和pAp22,长度分别为1354 bp和582 bp。这些家族由沿着平卧甜菜染色体广泛分散的序列元件组成,具有局部聚类且排除在远端常染色质区域之外。减数分裂染色体上的荧光原位杂交显示,这两个分散重复序列在一些染色体区域共定位。通过PCR研究了pAp4和pAp22家族重复序列的散布情况,并确定了重复序列侧翼序列。序列分析表明,pAp22要么来源于类似Athila的逆转录转座子的长末端重复序列(LTR),要么是其一部分。用pAp4和pAp22进行的Southern分析和荧光原位杂交表明,这两个分散重复序列都是物种特异性的,可作为DNA探针来区分种间杂种中的亲本基因组。这在含有一小段平卧甜菜染色体片段的甜菜杂交种PRO1中得到了验证。

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