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首页> 外文期刊>Conservation genetics >A molecular screening method for unisexual Ambystoma using mitochondrial DNA
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A molecular screening method for unisexual Ambystoma using mitochondrial DNA

机译:一种利用线粒体DNA筛查单性畸胎的分子筛查方法

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摘要

The geographical range of unisexual Ambystoma overlaps with four bisexual species that also breed in spring ponds. Several of these species are of conservation concern, and both adults and larvae can be difficult to distinguish morphologically from unisexuals. Here we present a rapid molecular method for screening unisexuals, whose mtDNA is most similar to Ambystoma barbouri. A 258 bp segment of the cytochrome b gene was amplified in six Ambystoma species and exemplar unisexuals by PCR using taxon-specific primers. An internal 113 bp segment was amplified only in unisexuals and A. barbouri using Universal forward and Hybrid reverse primers. Multisequence alignment comparing the nucleotide sequence where Hybrid reverse primer anneals revealed nucleotide diversity in this region among Ambystoma species. This simple method for discriminating between unisexuals and bisexuals, excluding A. barbouri, can be applied prior to further research on these declining species.
机译:单性Ambystoma的地理范围与也在春天池塘中繁殖的四个双性恋物种重叠。这些物种中有几种是需要保护的,成虫和幼虫在形态上与单性者之间可能很难区分。在这里,我们提出了一种筛查单性恋者的快速分子方法,该方法的mtDNA最类似于Barbyi Ambystoma。通过使用分类群特异性引物的PCR,在六个Ambystoma物种和示例单性者中扩增了细胞色素b基因的258 bp片段。使用通用正向和杂交反向引物,仅在单性恋者和巴氏曲霉中扩增内部113 bp片段。多序列比对比较了杂交反向引物退火揭示了Ambystoma种之间该区域核苷酸多样性的核苷酸序列。这种区分单性恋者和双性恋者(巴氏曲霉除外)的简单方法,可以在对这些正在下降的物种进行进一步研究之前应用。

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