Vasiliev V A, Korsunenko A V, Pereshkolnik S L, Mazanaeva L F, Bannikova A A, Bondarenko D A, Peregontsev E A, Semyenova S K
Genetika. 2014 Oct;50(10):1200-15.
Based on polymorphism of the 12S rRNA gene and RAPD markers, differentiation of 122 tortoise individuals belonging to the three species of genus Testudo (T. kleinmanni, T. marginata, and T. graeca), six subspecies of T. graeca (T. g. nikolskii, T. g. pallasi, T. g. armeniaca, T. g. zarudnyi, T. g. terrestris, T. g. ibera), and two subspecies of the Central Asian tortoise Agrionenemys horsfieldii (A. h. horsfieldii, A. h. kazakhstanica) was performed. For comparison, 32 known sequences of 12S rRNA gene (392 bp) from tortoises of the two genera inhabiting the territories of Europe, Asia, and Africa were used. In the populations of A. horsfieldii, a total of six haplotypes; including three newly described variants, were identified. In the examined tortoises of the genus Testudo, eleven 12S rRNA haplotypes were identified. One new haplotype was detected in T. kleinmanni. Among the eight subspecies of T. graeca, eight haplotypes were identified, with four newly described ones. The reported RAPD markers generally supported the reconstructions obtained with the use of the mitochondrial marker. Similarly to the 12S rRNA-based reconstructions, two independent clusters included representatives of the two genera, Agrionemys and Testudio. Among the latter, representatives of T. marginata and T. kleinmanni, as well as T. graeca, with high statistical support values, formed two reciprocally monophyletic groups. Compared to the mitochondrial markers, RAPDs more statisticallysignificantly discriminated the sample of T. g. terrestris and the four subspecies, T. g. ibera, T. g. armeniaca, T. g. pallasi, and T. g. nikolskii. In almost all cases except the representatives of T. g. ibera, the representatives of each of four subspecies formed individual subclusters. The geographical haplotype distribution patterns and possible evolutionary scenario of the origin and dispersal of tortoises of the two genera are discussed.
基于12S rRNA基因多态性和随机扩增多态性DNA（RAPD）标记，对122只陆龟个体进行了区分，这些个体分属于陆龟属的三个物种（埃及陆龟、缘翘陆龟和希腊陆龟）、希腊陆龟的六个亚种（希腊陆龟尼古拉耶夫亚种、希腊陆龟帕拉斯亚种、希腊陆龟亚美尼亚亚种、希腊陆龟扎鲁德尼亚种、希腊陆龟草原亚种、希腊陆龟伊比利亚亚种）以及中亚陆龟（四爪陆龟）的两个亚种（四爪陆龟指名亚种、四爪陆龟哈萨克斯坦亚种）。为作比较，使用了来自欧洲、亚洲和非洲地区的两个陆龟属陆龟的32个已知的12S rRNA基因序列（392 bp）。在四爪陆龟种群中，共鉴定出六个单倍型，包括三个新描述的变异型。在被检测的陆龟属陆龟中，鉴定出11个1,2S rRNA单倍型。在埃及陆龟中检测到一个新的单倍型。在希腊陆龟的八个亚种中，鉴定出八个单倍型，其中四个是新描述的。所报道的RAPD标记总体上支持了使用线粒体标记所获得的系统发育重建。与基于12S rRNA的系统发育重建类似，两个独立的类群包含了四爪陆龟属和陆龟属的代表。在后者中，缘翘陆龟和埃及陆龟以及希腊陆龟的代表在高统计支持值下形成了两个相互单系的类群。与线粒体标记相比，RAPD在统计学上更能显著区分希腊陆龟草原亚种以及希腊陆龟伊比利亚亚种、希腊陆龟亚美尼亚亚种、希腊陆龟帕拉斯亚种和希腊陆龟尼古拉耶夫亚种这四个亚种的样本。除希腊陆龟伊比利亚亚种的代表外，在几乎所有情况下，四个亚种中的每一个亚种的代表都形成了单独的亚类群。讨论了两个陆龟属陆龟的地理单倍型分布模式以及可能的起源和扩散进化情景。
