Ciavaglia Sherryn, Donnellan Stephen, Henry Julianne, Linacre Adrian
School of Biological Sciences, Flinders University, Adelaide, SA, Australia,
Forensic Sci Med Pathol. 2014 Sep;10(3):295-305. doi: 10.1007/s12024-014-9573-4. Epub 2014 Jun 12.
In this paper we report on the identification of a section of mitochondrial DNA that can be used to identify the species of protected and illegally traded pythons of the genus Morelia. Successful enforcement of wildlife laws requires forensic tests that can identify the species nominated in the relevant legislation. The potentially degraded state of evidentiary samples requires that forensic investigation using molecular genetic species identification is optimized to interrogate small fragments of DNA. DNA was isolated from 35 samples of Morelia spilota from which the complete cytochrome b was sequenced. The ND6 gene was also sequenced in 32 of these samples. Additional DNA sequences were generated from 9 additional species of Morelia. The sequences were aligned by Geneious and imported into MEGA to create phylogenetic trees based on the entire complex of approximately 1,706 base pairs (bp). To mimic degraded DNA, which is usually found in forensic cases, short sub-sections of the full alignment were used to generate phylogenetic trees. The sub-sections that had the greatest DNA sequence information were in parts of the cytochrome b gene. Our results highlight that legislation is presently informed by inadequate taxonomy. We demonstrated that a 278 bp region of the cytochrome b gene recovered the topology of the phylogenetic tree found with the entire gene sequence and correctly identified species of Morelia with a high degree of confidence. The locus described in this report will assist in the successful prosecution of alleged illegal trade in python species.
在本文中，我们报告了一段线粒体DNA的鉴定情况，该线粒体DNA可用于鉴定受保护的以及非法交易的莫瑞蟒属蟒蛇的物种。野生动物法律的成功实施需要能够鉴定相关立法中指定物种的法医检测。证据样本可能处于降解状态，这就要求优化使用分子遗传物种鉴定的法医调查，以便检测小片段DNA。从35个斯氏莫瑞蟒样本中分离出DNA，并对其完整的细胞色素b进行测序。其中32个样本还对ND6基因进行了测序。另外从莫瑞蟒属的9个其他物种中生成了额外的DNA序列。这些序列通过Geneious软件进行比对，并导入到MEGA软件中，以基于大约1706个碱基对（bp）的整个复合体构建系统发育树。为了模拟法医案件中通常发现的降解DNA，使用完整比对的短片段来生成系统发育树。细胞色素b基因部分区域具有最大的DNA序列信息。我们的结果突出表明，目前的立法依据的分类学并不充分。我们证明，细胞色素b基因的一个278 bp区域恢复了用整个基因序列所发现的系统发育树的拓扑结构，并以高度的可信度正确鉴定了莫瑞蟒属的物种。本报告中描述的基因座将有助于成功起诉涉嫌蟒蛇物种的非法贸易。
