Molecular Data and Karyotype Revealed Two Distinct Species of Domesticated Water Buffaloes in the Philippines

Therese Patricka Cailipan, Alexander Paraguas, Aivhie Jhoy Cuanang, Nelvie Fatima Jane Soliven, John Gregor Roño, Francis Fontanilla, Emerson Servo, Ernelea Cao, Ian Kendrich Fontanilla, Lilian Villamor
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Abstract

The six species of the genus Bubalus include the lowland and mountain anoa, Asian wild buffalo, tamaraw, swamp buffalo, and riverine buffalo. Few studies were conducted on the matrilineal phylogeny and genetic diversity of Bubalus species. In addition, there is controversy regarding the identity of the two domesticated water buffaloes in the Philippines – the introduced riverine and the native swamp buffalo. Thus, this study aimed to determine the genetic distance and phylogenetic relationship of Bubalus species using the mitochondrial cytochrome b (cytb) variation and to re-evaluate the taxonomic species designation of Bubalus bubalis. A total of499 Philippine cytb sequences from this study (n = 179), as well as other Asian countries (n = 319)and Syncerus caffer (n = 1), from NCBI were included in the analysis. The highest interspecies genetic distance (d) (0.040) was between the tamaraw and lowland anoa, whereas the lowest genetic distance (0.020) was between mountain and lowland anoa. Moreover, the phylogenetic tree indicated 52 haplotypes that delineated three groups, including lowland anoa, mountain anoa, and tamaraw with two domesticated buffaloes. The latter group further separated into three species – tamaraw, riverine, and swamp buffaloes. The analyses of genetic distance,phylogeny, and median-joining network structure based on cytb confirmed that the swamp buffaloes from Occidental Mindoro and South Cotabato, Philippines belong to maternal lineage B. Karyotype analysis consistently confirms that riverine buffaloes possess 50 chromosomes, whereas swamp buffaloes have 48 chromosomes, aligning with previous findings. Moreover, the swamp and riverine buffaloes appear to be distinct species, with the swamp buffalo having a closer affinity to the tamaraw than to the riverine buffalo. We, therefore, propose to revive the species assignment of the Bubalus kerabau Fitzinger, 1860 as a distinct species to the swamp buffalo. Furthermore, a common maternal ancestry was confirmed between the tamaraw and two domesticated buffaloes – the swamp and riverine. Research findings provided valuable information on understanding and monitoring the matrilineal genetic diversity of the genus Bubalus toward conservation and management.
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菲律宾两种不同驯化水牛的分子数据和核型分析
水牛属的六种包括低地和山地水牛、亚洲野生水牛、塔玛罗水牛、沼泽水牛和河流水牛。目前,国内外对黄菖蒲属植物母系系统发育和遗传多样性的研究较少。此外,关于菲律宾两种驯化水牛的身份也存在争议——引进的河水牛和本地沼泽水牛。因此,本研究旨在利用线粒体细胞色素b (cytb)的变异来确定黄颡鱼种间的遗传距离和系统发育关系,并重新评价黄颡鱼的分类学种名。来自本研究的499个菲律宾cytb序列(n = 179),以及来自NCBI的其他亚洲国家(n = 319)和Syncerus caffer (n = 1)被纳入分析。柽柳与低地柽柳的种间遗传距离最高(0.040),山地与低地柽柳的种间遗传距离最低(0.020)。系统发育树共鉴定出52个单倍型,分别划分为低地野水牛、山地野水牛和与2头驯化水牛的塔玛罗水牛3个类群。后者进一步分为三个物种——塔玛罗水牛、河流水牛和沼泽水牛。遗传距离、系统发育和基于cytb的中间连接网络结构分析证实,来自菲律宾西民都罗岛和南哥打巴托岛的沼泽水牛属于母系b。核型分析一致证实,河流水牛有50条染色体,而沼泽水牛有48条染色体,与先前的研究结果一致。此外,沼泽水牛和河流水牛似乎是不同的物种,沼泽水牛与塔玛罗的关系比与河流水牛的关系更密切。因此,我们建议恢复Bubalus kerabau Fitzinger, 1860的物种分配,作为沼泽水牛的一个独特物种。此外,证实了塔玛罗和两只驯养水牛——沼泽水牛和河流水牛之间有共同的母系祖先。研究结果为了解和监测黄颡鱼属植物母系遗传多样性提供了有价值的信息,有助于黄颡鱼属植物的保护和管理。
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