Souto R P, Fernandes O, Macedo A M, Campbell D A, Zingales B
Departamento de Bioquímica, Universidade de São Paulo, Brazil.
Mol Biochem Parasitol. 1996 Dec 20;83(2):141-52. doi: 10.1016/s0166-6851(96)02755-7.
Parasitic protozoa within the taxon Trypanosoma cruzi are considered to be derived from multiple clonal lineages, and show broad genetic diversity as a result of propagation with little or no genetic exchange. We have analyzed a wide sample of T. cruzi isolates from vertebrate and invertebrate hosts by PCR amplification of a ribosomal RNA gene sequence, a mini-exon gene sequence and random amplified polymorphic DNA (RAPD). Amplification of the distinct rDNA and mini-exon gene sequences indicated a dimorphism within both of the tandemly-repeated genes: 125 or 110 bp products for rDNA and 300 or 350 bp products for the mini-exon. Within individual isolates, one of three associations was observed: the 125 bp rDNA product with the 300 bp mini-exon product (defined as group 1), the 110 bp rDNA product with the 350 bp mini-exon product (defined as group 2) and the presence of both rDNA amplification products with the mini-exon group 1 product (group 1/2). The RAPD analysis showed variability between individual isolates, however, tree analysis clearly indicated the presence of two major branches. Interestingly, the rDNA/mini-exon group 2 isolates correlated precisely with one branch of the RAPD-derived tree; group 1 and group 1/2 isolates correlated with the other branch. Our studies show a clear division of T. cruzi into two major lineages presenting a high phylogenetic divergence. Hypotheses are discussed to explain the origin of the two lineages as well as isolates that are hybrid for group 1 and 2 rDNA markers.
锥虫属内的寄生原生动物克氏锥虫被认为源自多个克隆谱系,并且由于繁殖过程中很少或没有基因交换而表现出广泛的遗传多样性。我们通过对核糖体RNA基因序列、小外显子基因序列和随机扩增多态性DNA(RAPD)进行PCR扩增,分析了来自脊椎动物和无脊椎动物宿主的大量克氏锥虫分离株样本。不同的核糖体DNA(rDNA)和小外显子基因序列的扩增表明,这两个串联重复基因均存在二态性:rDNA的产物为125或110 bp,小外显子的产物为300或350 bp。在单个分离株中,观察到三种关联之一:125 bp的rDNA产物与300 bp的小外显子产物(定义为第1组),110 bp的rDNA产物与350 bp的小外显子产物(定义为第2组),以及两种rDNA扩增产物与小外显子第1组产物同时存在(第1/2组)。RAPD分析显示单个分离株之间存在变异性,然而,树形分析清楚地表明存在两个主要分支。有趣的是,rDNA/小外显子第2组分离株与RAPD衍生树的一个分支精确相关;第1组和第1/2组分离株与另一个分支相关。我们的研究表明,克氏锥虫明显分为两个主要谱系,呈现出高度系统发育分歧。讨论了解释这两个谱系以及第1组和第2组rDNA标记杂交分离株起源的假说。