Pak J Biol Sci. 2022 Jan;25(8):732-740. doi: 10.3923/pjbs.2022.732.740.
<b>Background and Objective:</b> Cytochrome c oxidase gene subunit-I (COI) has conserved and variable regions and along with the 658 nucleotide base pairs at the '5 of these have been used as animal barcode, species identification and evolutionary studies of several vertebrates, especially Anura. This research was conducted to characterize the nucleotides of the COI sequence gene of (<i>Limnonectes cf. grunniens</i>) that live sympatrically with <i>L. modestus</i> on several headwater streams in Kendari, Southeast Sulawesi. <b>Materials and Methods:</b> This research is explorative, samples of frogs were obtained from the Lahundape and Moramo headwater streams. A total of 16 frogs were sampled and the genomic DNA of frog samples was extracted and, then amplified using the PCR method. The next steps are sequencing and analysis using MEGA 7. <b>Results:</b> The result showed nucleotides along 688 to 705 base pairs. There were two haplotypes of <i>L. cf. grunniens</i> and three <i>L. modestus</i>. <i>L. cf. grunniens </i>consist of 32.6% Thymine (Uracil), 32.3% Cytisine, 17.9% Adenine and 17.9% Guanine. While <i>L. modestus</i> is 37.6% Thymine, 26.0% Cytosine, 20.7% Adenine and 15.8% Guanine. Based on Kimura-2 parameter, the genetic distance between genera ranges from 0.25-0.26 while the genetic distance between species is 0.00-0.01. <b>Conclusion:</b> Phylogeny trees based on partial sequences of COI frog genes showed that <i>L. cf. grunniens</i> and <i>L. modestus</i> are monophyletic with bootstrapped values ranging from 86-100% and differentiated between species. There is a genetic variability of COI sequences of <i>Limnonectes cf. grunniens</i> and <i>L. modestus</i> from Kendari, Southeast Sulawesi.
细胞色素 c 氧化酶亚基 I(COI)具有保守和可变区域,与这些区域的 5'端的 658 个核苷酸碱基一起,已被用作动物条形码、几种脊椎动物的物种鉴定和进化研究。本研究旨在对与 <i>L. modestus</i> 共生的 (<i>Limnonectes cf. grunniens</i>) 的 COI 序列基因的核苷酸进行特征描述,这些 (<i>Limnonectes cf. grunniens</i>) 生活在东南苏拉威西肯达里的几条源头溪流中。材料与方法:本研究为探索性研究,从 Lahundape 和 Moramo 源头溪流中获取了青蛙样本。共采集了 16 只青蛙,提取青蛙样本的基因组 DNA,然后使用 PCR 方法进行扩增。下一步是使用 MEGA 7 进行测序和分析。结果:结果显示了 688 到 705 个碱基对的核苷酸。有两种 <i>L. cf. grunniens</i> 单倍型和三种 <i>L. modestus</i> 单倍型。<i>L. cf. grunniens </i> 由 32.6%胸腺嘧啶(尿嘧啶)、32.3%胞嘧啶、17.9%腺嘌呤和 17.9%鸟嘌呤组成。而 <i>L. modestus</i> 则是 37.6%胸腺嘧啶、26.0%胞嘧啶、20.7%腺嘌呤和 15.8%鸟嘌呤。基于 Kimura-2 参数,属间的遗传距离在 0.25-0.26 之间,种间的遗传距离在 0.00-0.01 之间。结论:基于 COI 青蛙基因部分序列构建的系统发育树表明,<i>L. cf. grunniens</i> 和 <i>L. modestus</i> 是单系的,bootstrap 值范围为 86-100%,并在种间分化。来自东南苏拉威西肯达里的 <i>Limnonectes cf. grunniens</i> 和 <i>L. modestus</i> 的 COI 序列存在遗传变异性。