Frello S, Heslop-Harrison J S
Section of Botany, The Royal Agricultural and Veterinary University, Copenhagen, Denmark.
Genome. 2000 Oct;43(5):902-9. doi: 10.1139/g00-044.
Eight clones of repetitive DNA were isolated from Crocus vernus Hill. The genomic organization of the clones was analyzed by in situ hybridization to C. vernus and Southern hybridization to a range of Crocus and other species. Seven clones were used for in situ hybridization. Sequence analysis showed that all eight clones were nonhomologous, and thus represented eight different sequence-families. In situ hybridization showed that six were dispersed in high copy numbers on all chromosomes of the C. vernus genome, whereas one was localized proximal to the secondary constriction, at the NOR (nucleolar organizer region) and was not further analyzed, as it was considered part of the 18S-25S rDNA repeat. Except for short palindromes, none of the sequences showed notable internal structures. Clone pCvKB4 showed homology to the reverse transcriptase gene of Ty1-copia-like retrotransposons; the others showed no homology to known sequences. When used as probes for Southern hybridization, four showed a ladder of 3-4 bands superimposed by irregular patterns, indicating organization in short tandem arrays. Each clone had a unique distribution among Crocus species (12-16 species analyzed with each clone) and six species of Iridaceae, Liliaceae, and Amaryllidaceae; all seven investigated sequences were Iridaceae specific and four were Crocus specific. The species distribution of these seven clones showed notable discrepancies with the taxonomic subdivision of the genus at the subgenus, section, and series levels. The results suggest that the phylogeny and taxonomic structure of the genus Crocus might need reconsideration. The analysis of repetitive DNA as a major and rapidly evolving part of the genome could contribute to the study of species relationships and evolution.
从番红花(Crocus vernus Hill)中分离出八个重复DNA克隆。通过对番红花进行原位杂交以及对一系列番红花属和其他物种进行Southern杂交,分析了这些克隆的基因组组织。七个克隆用于原位杂交。序列分析表明,所有八个克隆均非同源,因此代表八个不同的序列家族。原位杂交显示,六个克隆以高拷贝数分散在番红花基因组的所有染色体上,而其中一个位于次缢痕近端的核仁组织区(NOR),由于其被认为是18S - 25S rDNA重复序列的一部分,因此未作进一步分析。除了短回文序列外,这些序列均未显示出明显的内部结构。克隆pCvKB4与Ty1 - copia类逆转录转座子的逆转录酶基因具有同源性;其他克隆与已知序列无同源性。当用作Southern杂交探针时,四个克隆显示出由不规则模式叠加的3 - 4条带的梯状条带,表明其以短串联阵列形式组织。每个克隆在番红花属物种(每个克隆分析12 - 16个物种)以及鸢尾科、百合科和石蒜科的六个物种中具有独特的分布;所有七个研究序列均为鸢尾科特异性,四个为番红花属特异性。这七个克隆的物种分布在亚属、组和系水平上与该属的分类细分存在显著差异。结果表明,番红花属的系统发育和分类结构可能需要重新考虑。将重复DNA作为基因组中主要且快速进化的部分进行分析,可能有助于研究物种关系和进化。