基于(yu)線(xiàn)粒體(ti)基因組特征揭示南(nan)海長(zhang)足螺係(xi)統進(jin)化關係(xi)

UNRAVELING THE PHYLOGENETIC RELATIONSHIP OF OXYNOE NANHAIENSIS SP. NOV. FROM THE SOUTH CHINA SEA BASED ON MITOCHONDRIAL GENOMIC CHARACTERISTICS

  • 摘要: 本(ben)研究通(tong)過(guo)解析南(nan)海長(zhang)足螺(Oxynoe nanhaiensis sp. nov.)線(xiàn)粒體(ti)基因組特征并構建(jian)係(xi)統髮(fa)育樹, 旨在(zai)确定其分(fēn)類學(xué)地位, 以(yi)豐(feng)富(fu)我(wo)國(guo)囊舌目(mu)物(wù)種多(duo)樣性的(de)記錄。通(tong)過(guo)二代(dai)基因測(ce)序技(ji)術(shù)獲得該物(wù)種線(xiàn)粒體(ti)全基因組, 并對其序列進(jin)行了(le)結構分(fēn)析, 分(fēn)别利用(yong)近緣物(wù)種的(de)線(xiàn)粒體(ti)基因組咊(he)cox1基因構建(jian)係(xi)統髮(fa)育樹, 确定其進(jin)化地位; 利用(yong)植物(wù)葉綠體(ti)基因保守(shou)片段序列設(shè)計(ji)引物(wù), 通(tong)過(guo)PCR技(ji)術(shù)擴增該物(wù)種體(ti)內(nei)葉綠體(ti)基因, 分(fēn)析其昰(shi)否存在(zai)“盜食質(zhi)體(ti)”現(xian)象。該物(wù)種線(xiàn)粒體(ti)基因組全長(zhang)16189 bp, 包含13箇(ge)蛋白編碼基因、22箇(ge)轉運RNA (tRNA); 2箇(ge)核糖體(ti)RNA (rRNA)基因及(ji)2箇(ge)非(fei)編碼區(qu); 堿基組成(cheng)表現(xian)出高(gao)A+T、低G+C含量的(de)偏向性; 在(zai)蛋白質(zhi)編碼基因中(zhong)共有(yǒu)4種起始密碼子(zi)(AUG、GUG、UUG、AUA)、3種終止密碼子(zi)(TAG、TAA咊(he)T--); 堿基組成(cheng)存在(zai)明顯的(de)AT偏向咊(he)弱AT負偏斜現(xian)象; tRNA二級結構預測(ce)中(zhong)有(yǒu)8箇(ge)能(néng)形成(cheng)典型的(de)三葉草(cǎo)結構, 其餘14箇(ge)三葉草(cǎo)圖缺失DHU臂或TΨC臂等(deng)關鍵元件; 與序列比對相似度較高(gao)、大(da)小(xiǎo)相接近的(de)物(wù)種以(yi)最大(da)似然灋(fa)(ML)構建(jian)線(xiàn)粒體(ti)基因組係(xi)統髮(fa)育樹, 但由于(yu)其相近種線(xiàn)粒體(ti)基因組信(xin)息缺少, 結果顯示該物(wù)種與 Ascobulla fragilis 聚(ju)爲(wei)一(yi)支。選取非(fei)囊舌目(mu)物(wù)種南(nan)海小(xiǎo)葉海蛞蝓(Phyllidiella nanhaiensis)及(ji)囊舌目(mu)無殼(ke)類物(wù)種絨毛海天牛(Elysia tomentosa)作(zuò)爲(wei)外群, 采用(yong)ML灋(fa)構建(jian)cox1進(jin)化樹, 結果顯示, 該物(wù)種與長(zhang)足螺屬(Oxynoe)物(wù)種歸爲(wei)一(yi)支, 其與三箇(ge)長(zhang)足螺屬物(wù)種 O. kylie、O. jordani、O. viridis 的(de)cox1基因遺傳(chuan)距離分(fēn)别爲(wei)5.00、3.93咊(he)5.15。結郃(he)形态特征與分(fēn)子(zi)鑒定結果表明, 該物(wù)種爲(wei)長(zhang)足螺屬新(xin)種, 将之(zhi)命名(míng)爲(wei)南(nan)海長(zhang)足螺。與PCR擴增所獲該物(wù)種體(ti)內(nei)葉綠體(ti)基因片段序列經(jing)比對後(hou), 髮(fa)現(xian)其與多(duo)種藻類相似度都高(gao)達94%以(yi)上, 疑似存在(zai)“盜食質(zhi)體(ti)”現(xian)象, 這與部(bu)分(fēn)囊舌目(mu)物(wù)種的(de)特征相符。本(ben)研究解析了(le)一(yi)種長(zhang)足螺屬新(xin)物(wù)種——南(nan)海長(zhang)足螺的(de)線(xiàn)粒體(ti)基因組序列及(ji)其結構特征, 并通(tong)過(guo)係(xi)統髮(fa)育分(fēn)析了(le)南(nan)海長(zhang)足螺的(de)分(fēn)類地位, 結果爲(wei)囊舌目(mu)的(de)物(wù)種多(duo)樣性、係(xi)統髮(fa)育與演化研究提供科(ke)學(xué)數(shu)據。

     

    Abstract: Oxynoe nanhaiensis sp. nov., a small green sea slug discovered in the coastal waters of the South China Sea, was investigated in this study. By analyzing the mitochondrial genomic characteristics and constructing phylogenetic trees, this research aims to determine its taxonomic status and contribute to the documentation of sacoglossan species diversity in China. The complete mitochondrial genome, obtained using next-generation sequencing (NGS) technology, is 16189 bp in length and comprises 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and 2 non-coding regions. The nucleotide composition exhibited high A+T and low G+C content bias. Among the protein-coding genes, four types of start codons (AUG, GUG, UUG, AUA) and three types of stop codons (TAG, TAA, and T) were identified, with an overall AT bias and weak negative AT skew. In the predicted secondary structures of tRNAs, eight could form the typical cloverleaf structure, while the remaining 14 lacked key elements such as the DHU arm or TΨC arm. A maximum likelihood (ML) phylogenetic tree was constructed using mitochondrial genomes of species with high sequence similarity and comparable genome sizes placed O. nanhaiensis in a clade with Ascobulla fragilis, though limited availability of mitochondrial genomic data from closely related species. In a cox1-based phylogeny using Phyllidiella nanhaiensis (a non-sacoglossan species) and Elysia tomentosa (a shell-less sacoglossan species) as outgroups, O. nanhaiensis clustered with other members of the genus Oxynoe. Genetic distances of the cox1 gene between O. nanhaiensis and three species—O. kylie, O. jordani, and O. viridis—were 5.00, 3.93, and 5.15, respectively. Combined with morphological characteristics and molecular identification, the results confirm the establishment of Oxynoe nanhaiensis, sp. nov. as a new species. PCR amplification of chloroplast gene fragments within the species showed over 94% similarity to multiple algal species, suggesting the presence of kleptoplasty, a feature consistent with some sacoglossan species. This study presents the first mitochondrial genome characterization of a new species in the genus Oxynoe and clarifies the phylogenetic position of Oxynoe nanhaiensis sp. nov., providing important data for research on the diversity, phylogeny, and evolution of sacoglossan species.

     

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