Complete cox 1 gene and ITS sequence of Hirudinaria manillensis and molecular evolution analysis.

IF 3.9 2区综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES

Scientific Reports Pub Date : 2025-10-10 DOI:10.1038/s41598-025-19661-9

Wenyi Wang, Chaojie Yang, Guangzong He, Weitao Zhao, Jinshen Yang, Yaojun Yang

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Abstract

Hirudinaria manillensis is an important blood-sucking medicinal leech widely distributed in Southeast Asia and southern China. It has been shown to treat cardiovascular and cerebrovascular diseases. Molecular identification technology has developed rapidly, and the results are accurate and stable, which can help us to obtain rich biological information of samples for genetic evolution analysis. This study designed 3 pairs of primers for complete cox 1 gene. After sequencing by PCR products and splicing by SeqMan software, the full-length sequence of cox 1 gene was obtained by annotation of open reading frame (ORFs) on MITOS2 platform. The cox 1 gene with a length of 1536nt was obtained, and the species was identified as Hirudinaria manillensis. The Hd value and FST value of cox 1 gene were 0.858 and 0.10938, which were higher than the Hd value of ITS (0.314) and the FST value close to 0. The nucleotide diversity and GST values of the two genes were very low, and the Tajima's D values of the neutral test were negative, without statistical significance. The genetic distance calculated by the Kimura-2 parameter method was very low. According to the biological information provided by the samples in this study, the population size of Hirudinaria manillensis in southern China was on the rise, but stable genetic diversity was not formed, and there was no obvious population differentiation and geographical isolation. This may be due to the mature artificial breeding technology in recent years, which enabled Hirudinaria manillensis to migrate from the traditional gathering places and breed in a large number in the new environment and migration site. At present, there are many molecular barcoding data of cox 1 gene in the NCBI database, but there is no complete cox 1 gene sequence as a reference, and the related ITS sequence information is more limited, indicating that there is still a lot of space for gene research of Hirudinaria manillensis.

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水蛭cox 1全基因、ITS序列及分子进化分析。

马尼拉水蛭是一种重要的吸血药用水蛭，广泛分布于东南亚和中国南方。它已被证明可以治疗心脑血管疾病。分子鉴定技术发展迅速，结果准确、稳定，可以帮助我们获得丰富的样品生物信息，用于遗传进化分析。本研究设计了3对cox 1全基因引物。PCR产物测序，SeqMan软件剪接后，在MITOS2平台上通过开放阅读框（orf）注释获得cox 1基因的全长序列。获得cox 1基因，基因长度为1536nt，鉴定为水蛭属。cox 1基因的Hd值和FST值分别为0.858和0.10938，均高于ITS的Hd值（0.314）和FST值接近0。两个基因的核苷酸多样性和GST值都很低，中性检验的田岛D值为阴性，无统计学意义。Kimura-2参数法计算的遗传距离很低。根据本研究样本提供的生物学信息，中国南方水蛭种群规模呈上升趋势，但没有形成稳定的遗传多样性，没有明显的种群分化和地理隔离。这可能是由于近年来成熟的人工育种技术，使得水蛭能够从传统的聚集地迁徙到新的环境和迁徙地点大量繁殖。目前NCBI数据库中cox 1基因的分子条形码数据较多，但没有完整的cox 1基因序列作为参考，相关ITS序列信息较为有限，说明水蛭的基因研究仍有很大的空间。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Scientific Reports Natural Science Disciplines-

CiteScore

7.50

自引率

4.30%

发文量

19567

审稿时长

3.9 months

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