Garcia-Longoria Luz, Hellgren Olof, Bensch Staffan
Departamento de Biología Animal, Universidad de Extremadura, E-06071 Badajoz, Spain.
Malar J. 2014 Jun 18;13:239. doi: 10.1186/1475-2875-13-239.
Malaria parasites need to synthesize chitinase in order to go through the peritrophic membrane, which is created around the mosquito midgut, to complete its life cycle. In mammalian malaria species, the chitinase gene comprises either a large or a short copy. In the avian malaria parasites Plasmodium gallinaceum both copies are present, suggesting that a gene duplication in the ancestor to these extant species preceded the loss of either the long or the short copy in Plasmodium parasites of mammals. Plasmodium gallinaceum is not the most widespread and harmful parasite of birds. This study is the first to search for and identify the chitinase gene in one of the most prevalent avian malaria parasites, Plasmodium relictum.
Both copies of P. gallinaceum chitinase were used as reference sequences for primer design. Different sequences of Plasmodium spp. were used to build the phylogenetic tree of chitinase gene.
The gene encoding for chitinase was identified in isolates of two mitochondrial lineages of P. relictum (SGS1 and GRW4). The chitinase found in these two lineages consists both of the long (PrCHT1) and the short (PrCHT2) copy. The genetic differences found in the long copy of the chitinase gene between SGS1 and GRW4 were higher than the difference observed for the cytochrome b gene.
The identification of both copies in P. relictum sheds light on the phylogenetic relationship of the chitinase gene in the genus Plasmodium. Due to its high variability, the chitinase gene could be used to study the genetic population structure in isolates from different host species and geographic regions.
疟原虫需要合成几丁质酶才能穿过在蚊子中肠周围形成的围食膜,以完成其生命周期。在哺乳动物疟原虫物种中,几丁质酶基因包括一个长拷贝或一个短拷贝。在禽疟原虫鸡疟原虫中,两个拷贝都存在,这表明在这些现存物种的祖先中发生的基因复制先于哺乳动物疟原虫中长拷贝或短拷贝的丢失。鸡疟原虫不是鸟类中分布最广、危害最大的寄生虫。本研究首次在最普遍的禽疟原虫之一残疟原虫中寻找并鉴定了几丁质酶基因。
鸡疟原虫几丁质酶的两个拷贝均用作引物设计的参考序列。使用疟原虫属的不同序列构建几丁质酶基因的系统发育树。
在残疟原虫的两个线粒体谱系(SGS1和GRW4)的分离株中鉴定出编码几丁质酶的基因。在这两个谱系中发现的几丁质酶由长拷贝(PrCHT1)和短拷贝(PrCHT2)组成。SGS1和GRW4之间几丁质酶基因长拷贝中发现的遗传差异高于细胞色素b基因观察到的差异。
在残疟原虫中鉴定出两个拷贝,为疟原虫属中几丁质酶基因的系统发育关系提供了线索。由于其高度变异性,几丁质酶基因可用于研究来自不同宿主物种和地理区域的分离株中的遗传种群结构。