Mlinarec Jelena, Chester Mike, Siljak-Yakovlev Sonja, Papes Drazena, Leitch Andrew R, Besendorfer Visnja
Department of Molecular Biology, Biology Division, Faculty of Science, University of Zagreb, Horvatovac 102a, HR-10000, Zagreb, Croatia.
Chromosome Res. 2009;17(3):331-46. doi: 10.1007/s10577-009-9025-2. Epub 2009 Feb 18.
The structure, abundance and location of repetitive DNA sequences on chromosomes can characterize the nature of higher plant genomes. Here we report on three new repeat DNA families isolated from Anemone hortensis L.; (i) AhTR1, a family of satellite DNA (stDNA) composed of a 554-561 bp long EcoRV monomer; (ii) AhTR2, a stDNA family composed of a 743 bp long HindIII monomer and; (iii) AhDR, a repeat family composed of a 945 bp long HindIII fragment that exhibits some sequence similarity to Ty3/gypsy-like retroelements. Fluorescence in-situ hybridization (FISH) to metaphase chromosomes of A. hortensis (2n = 16) revealed that both AhTR1 and AhTR2 sequences co-localized with DAPI-positive AT-rich heterochromatic regions. AhTR1 sequences occur at intercalary DAPI bands while AhTR2 sequences occur at 8-10 terminally located heterochromatic blocks. In contrast AhDR sequences are dispersed over all chromosomes as expected of a Ty3/gypsy-like element. AhTR2 and AhTR1 repeat families include polyA- and polyT-tracks, AT/TA-motifs and a pentanucleotide sequence (CAAAA) that may have consequences for chromatin packing and sequence homogeneity. AhTR2 repeats also contain TTTAGGG motifs and degenerate variants. We suggest that they arose by interspersion of telomeric repeats with subtelomeric repeats, before hybrid unit(s) amplified through the heterochromatic domain. The three repetitive DNA families together occupy approximately 10% of the A. hortensis genome. Comparative analyses of eight Anemone species revealed that the divergence of the A. hortensis genome was accompanied by considerable modification and/or amplification of repeats.
染色体上重复DNA序列的结构、丰度和位置能够表征高等植物基因组的性质。在此,我们报告从园叶银莲花中分离出的三个新的重复DNA家族;(i)AhTR1,一个由554 - 561 bp碱基碱基长的EcoRV单体组成的卫星DNA(stDNA)家族;(ii)AhTR2,一个由743 bp长的HindIII单体组成的stDNA家族;以及(iii)AhDR,一个由945 bp长的HindIII片段组成的重复家族,该片段与Ty3/gypsy样逆转座子表现出一些序列相似性。对园叶银莲花(2n = 16)中期染色体进行荧光原位杂交(FISH)显示,AhTR1和AhTR2序列均与DAPI阳性的富含AT的异染色质区域共定位。AhTR1序列出现在居间的DAPI带处,而AhTR2序列出现在8 - 10个位于末端的异染色质块处。相比之下,AhDR序列如预期的Ty3/gypsy样元件一样分散在所有染色体上。AhTR2和AhTR1重复家族包括多聚A - 和多聚T - 序列、AT/TA基序以及一个五核苷酸序列(CAAAA),这些可能对染色质包装和序列同质性有影响。AhTR2重复序列还包含TTTAGGG基序和简并变体。我们认为它们是通过端粒重复序列与亚端粒重复序列的散布产生的,然后杂交单元通过异染色质区域进行扩增。这三个重复DNA家族总共占据了园叶银莲花基因组的约10%。对八个银莲花物种的比较分析表明,园叶银莲花基因组的分化伴随着重复序列的大量修饰和/或扩增。