Rohilla Manoj S, Tiwari Pramod K
School of Studies in Zoology, Jiwaji University, Gwalior, India.
J Appl Genet. 2008;49(2):167-82. doi: 10.1007/BF03195610.
DNA-based identification of species for phylogenetic analysis as well as forensic identification is widely being carried out with the help of polymerase chain reaction (PCR). In this study, a successful effort has been made to identify 5 species of Indian freshwater turtles, including 3 hard-shell turtles (Geoemydidae), i.e. Kachuga dhongoka, K. kachuga and Geoclemys hamiltoni, and 2 species of soft-shell turtles (Trionychidae), i.e. Aspideretes gangeticus and Lissemys punctata punctata, by using a well-optimized PCR-RFLP method. The analysis of nucleotide sequence variations in the PCR-amplified mitochondrial cyt-b genes (encoding cytochrome b) from the 5 species revealed its usefulness in the taxonomic differentiation of these species. On the basis of cyt-b sequence data and the PCR-RFLP pattern, a phylogeny was developed to resolve the genetic relationships between these species, living in the same habitat type. In comparison, the PCR-RFLP of mitochondrial 16S rDNA genes appeared less decisive in analysing phylogenetic relationships or even in species differentiation. Further, the molecular method (PCR-RFLP) developed here is simple, rapid, reliable and reproducible; hence it can be routinely applied for species identification, essential for conservation and management of endangered chelonian species.
借助聚合酶链反应（PCR），基于DNA的物种鉴定已广泛应用于系统发育分析以及法医鉴定。在本研究中，通过使用优化良好的PCR-RFLP方法，成功鉴定了5种印度淡水龟，其中包括3种硬壳龟（地龟科），即印度孔雀龟、印度棱背龟和哈米顿氏龟，以及2种软壳龟（鳖科），即恒河鳖和斑点池龟。对这5个物种PCR扩增的线粒体细胞色素b基因（编码细胞色素b）的核苷酸序列变异分析表明，其在这些物种的分类鉴别中具有实用性。基于细胞色素b序列数据和PCR-RFLP模式，构建了系统发育树以解析生活在相同栖息地类型的这些物种之间的遗传关系。相比之下，线粒体16S rDNA基因的PCR-RFLP在分析系统发育关系甚至物种鉴别方面显得不太具有决定性。此外，此处开发的分子方法（PCR-RFLP）简单、快速、可靠且可重复；因此，它可常规应用于物种鉴定，这对于濒危龟类物种的保护和管理至关重要。
