Pagnotta Mario Augusto, Mondini Linda, Porceddu Enrico
Department of Agrobiology and Agrochemistry, University of Tuscia, Viterbo, Italy.
Mol Genet Genomics. 2009 Sep;282(3):245-55. doi: 10.1007/s00438-009-0462-6. Epub 2009 Jun 19.
A real-time PCR approach was adopted and optimized to estimate and compare, through a relative quantification, the copy number of WIS2-1A and BARE-1 retrotransposons. The aim of this approach was to identify and quantify the presence of these retrotransposons in Triticum and Aegilops species, and to understand better the genome organization of these retroelements. The species were selected to assess and compare the evolution of the different types of genomes between the more recent species such as the diploid Triticum monococcum, tetraploid T. dicoccon and hexaploid T. spelta, and the corresponding genome donors of the ancient diploids Aegilops (Ae. speltoides, Ae. tauschii, Ae. sharonensis and Ae. bicornis) and T. urartu. The results of this study indicated the presence of great variation in copy number both within and among species, and the existence of a non-linear relationship between retrotransposon copy number and ploidy level. For WIS2-1A, as expected, T. monococcum showed the lowest copy number which instead was similar in T. dicoccon and T. spelta; also T. urartu (AA), Ae. speltoides (BB) and Ae. tauschii (DD) showed a higher WIS2-1A copy number. Similar results were observed for BARE-1 retroelements except for Ae. tauschii which as in T. monococcum showed lower retroelements content; a similar content for T. dicoccon and T. urartu, whereas a higher number was found in T. spelta and Ae. speltoides. The results presented here are in accord with previous studies and contribute to unravelling the structure and evolution of polyploidy and repetitive genomes.
采用并优化了实时聚合酶链反应(PCR)方法,通过相对定量来估计和比较WIS2-1A和BARE-1逆转座子的拷贝数。该方法的目的是鉴定和定量这些逆转座子在小麦属和山羊草属物种中的存在情况,并更好地了解这些逆转元件的基因组组织。选择这些物种是为了评估和比较不同类型基因组在较新物种(如二倍体一粒小麦、四倍体二粒小麦和六倍体斯卑尔脱小麦)以及古代二倍体山羊草(拟斯卑尔脱山羊草、节节麦、沙伦山羊草和双角山羊草)和乌拉尔图小麦相应基因组供体之间的进化情况。本研究结果表明,物种内部和物种之间的拷贝数存在很大差异,并且逆转座子拷贝数与倍性水平之间存在非线性关系。对于WIS2-1A,正如预期的那样,一粒小麦的拷贝数最低,而二粒小麦和斯卑尔脱小麦的拷贝数相似;乌拉尔图小麦(AA)、拟斯卑尔脱山羊草(BB)和节节麦(DD)的WIS2-1A拷贝数也较高。除了节节麦,其逆转元件含量与一粒小麦一样较低,而二粒小麦和乌拉尔图小麦的含量相似,斯卑尔脱小麦和拟斯卑尔脱山羊草的含量较高外,BARE-1逆转元件也观察到了类似的结果。这里呈现的结果与先前的研究一致,有助于阐明多倍体和重复基因组的结构与进化。