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甜菜属中一个差异扩增重复DNA序列家族揭示了甜菜亚种和栽培品种的遗传变异。

A family of differentially amplified repetitive DNA sequences in the genus Beta reveals genetic variation in Beta vulgaris subspecies and cultivars.

作者信息

Kubis S, Heslop-Harrison J S, Schmidt T

机构信息

Norman Borlaug Institute For Plant Science Research, DeMontfort University, Scraptoft, Leicester, United Kingdom.

出版信息

J Mol Evol. 1997 Mar;44(3):310-20. doi: 10.1007/pl00006148.

Abstract

Members of a highly abundant restriction satellite family have been isolated from the wild beet species Beta nana. The satellite DNA sequence is characterized by a conserved RsaI restriction site and is present in three of four sections of the genus Beta, namely Nanae, Corollinae, and Beta. It was not detected in species of the evolutionary old section Procumbentes, suggesting its amplification after separation of this section. Sequences of eight monomers were aligned revealing a size variation from 209 to 233 bp and an AT content ranging from 56.5% to 60.5%. The similarity between monomers in B. nana varied from 77.7% to 92.2%. Diverged subfamilies were identified by sequence analysis and Southern hybridization. A comparative study of this repetitive DNA element by fluorescent in situ hybridization and Southern analyses in three representative species was performed showing a variable genomic organization and heterogeneous localizations along metaphase chromosomes both within and between species. In B. nana the copy number of this satellite, with some 30,000 per haploid genome, is more than tenfold higher than in Beta lomatogona and up to 200 times higher than in Beta vulgaris, indicating different levels of sequence amplification during evolution in the genus Beta. In sugar beet (B. vulgaris), the large-scale organization of this tandem repeat was examined by pulsed-field gel electrophoresis. Southern hybridization to genomic DNA digested with DraI demonstrated that satellite arrays are located in AT-rich regions and the tandem repeat is a useful probe for the detection of genetic variation in closely related B. vulgaris cultivars, accessions, and subspecies.

摘要

已从野生甜菜物种矮甜菜(Beta nana)中分离出一个高度丰富的限制性卫星家族的成员。该卫星DNA序列的特征是具有一个保守的RsaI限制性位点,并且存在于甜菜属四个组中的三个组中,即矮生组(Nanae)、花冠组(Corollinae)和甜菜组(Beta)。在进化上较古老的匍匐组(Procumbentes)的物种中未检测到,这表明该卫星家族在该组分离后发生了扩增。对八个单体的序列进行比对,发现其大小在209至233 bp之间变化,AT含量在56.5%至60.5%之间。矮甜菜中单体之间的相似性在77.7%至92.2%之间变化。通过序列分析和Southern杂交鉴定出了分化的亚家族。通过荧光原位杂交和Southern分析对三个代表性物种中的这种重复DNA元件进行了比较研究,结果表明其基因组组织可变,并且在物种内部和物种之间沿中期染色体的定位存在异质性。在矮甜菜中,这种卫星的拷贝数约为每个单倍体基因组30,000个,比长柄甜菜(Beta lomatogona)高十多倍,比普通甜菜(Beta vulgaris)高多达200倍,这表明甜菜属在进化过程中序列扩增的水平不同。在甜菜(Beta vulgaris)中,通过脉冲场凝胶电泳研究了这种串联重复序列的大规模组织。用DraI消化基因组DNA后的Southern杂交表明,卫星阵列位于富含AT的区域,并且该串联重复序列是检测密切相关的普通甜菜品种、种质和亚种中遗传变异的有用探针。

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