Biology Centre of the Czech Academy of Sciences, Institute of Plant Molecular Biology, České Budějovice, 37005, Czech Republic.
University of South Bohemia, Faculty of Science, České Budějovice, 37005, Czech Republic.
Sci Rep. 2018 Apr 11;8(1):5838. doi: 10.1038/s41598-018-24196-3.
Satellite DNA, a class of repetitive sequences forming long arrays of tandemly repeated units, represents substantial portions of many plant genomes yet remains poorly characterized due to various methodological obstacles. Here we show that the genome of the field bean (Vicia faba, 2n = 12), a long-established model for cytogenetic studies in plants, contains a diverse set of satellite repeats, most of which remained concealed until their present investigation. Using next-generation sequencing combined with novel bioinformatics tools, we reconstructed consensus sequences of 23 novel satellite repeats representing 0.008-2.700% of the genome and mapped their distribution on chromosomes. We found that in addition to typical satellites with monomers hundreds of nucleotides long, V. faba contains a large number of satellite repeats with unusually long monomers (687-2033 bp), which are predominantly localized in pericentromeric regions. Using chromatin immunoprecipitation with CenH3 antibody, we revealed an extraordinary diversity of centromeric satellites, consisting of seven repeats with chromosome-specific distribution. We also found that in spite of their different nucleotide sequences, all centromeric repeats are replicated during mid-S phase, while most other satellites are replicated in the first part of late S phase, followed by a single family of FokI repeats representing the latest replicating chromatin.
卫星 DNA 是一类形成串联重复单位长阵列的重复序列,代表了许多植物基因组的很大一部分,但由于各种方法学障碍,其特征仍描述不足。在这里,我们展示了野豌豆(Vicia faba,2n = 12)的基因组,它是植物细胞遗传学研究中一种历史悠久的模式生物,包含了一组多样化的卫星重复序列,其中大多数直到现在的研究才被揭示出来。我们使用下一代测序技术结合新的生物信息学工具,重建了 23 个新的卫星重复序列的共识序列,这些序列代表了基因组的 0.008-2.700%,并在染色体上绘制了它们的分布。我们发现,除了具有数百个核苷酸长的单体的典型卫星外,V. faba 还包含大量具有异常长单体(687-2033 bp)的卫星重复序列,这些重复序列主要位于着丝粒周围区域。使用 CenH3 抗体的染色质免疫沉淀,我们揭示了具有染色体特异性分布的七个重复序列的丰富的着丝粒卫星多样性。我们还发现,尽管它们的核苷酸序列不同,但所有着丝粒重复序列都在中期 S 期复制,而大多数其他卫星则在晚期 S 期的前半部分复制,随后是一个代表最新复制染色质的 FokI 重复家族。