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长臂缨鲆核糖体RNA基因序列多态性特征分析
引用本文:杨敏,孔晓瑜,时伟,龚理. 长臂缨鲆核糖体RNA基因序列多态性特征分析[J]. 热带海洋学报, 2019, 38(1): 55-66. DOI: 10.11978/2018038
作者姓名:杨敏  孔晓瑜  时伟  龚理
作者单位:1. 中国科学院热带海洋生物资源与生态重点实验室, 中国科学院南海海洋研究所, 广东 广州 5103012. 中国科学院大学, 北京 1000493. 浙江海洋大学, 海洋科学与技术学院, 国家海洋设施养殖工程技术研究中心, 浙江 舟山 316022
基金项目:国家自然科学基金项目(31272273)
摘    要:为了解鲽形目Pleuronectiformes鲆科Bothidae长臂缨鲆Crossorhombus kobensis (Jordan & Starks, 1906) 核糖体RNA基因的序列多态性特征, 本研究共获得该鱼类包括18S、5.8S、ITS1和ITS2全长及28S部分序列的128条克隆序列。经序列比对、聚类分析以及重组检测, 结果显示5.8S (158bp) 无长度变异, 而其他4个基因片段则表现出较高的长度多态性, 并可分为不同序列类型: 18S (1856~1893 bp) 有4种序列类型A、B、C和R; 28S (967~974bp) 和ITS1 (407~ 505bp) 均有3种类型A、B和R; ITS2 (423~447 bp)存在2种类型A和B。此外5个基因片段在碱基组成中均表现出GC偏倚, 并且ITS2 (71.14%)>ITS1 (65.37%)>28S (62.22%)>5.8S (57.67%)>18S (54.95%)。对具有不同序列类型的18S、28S和ITS进行真、假基因推断时, 通常的判别特征不足以提供有力依据, 因此增加了与4种鲆科近缘鱼类长冠羊舌鲆Arnoglossus macrolophus、青缨鲆Crossorhombus azureus、大鳞短额鲆Engyprosopon grandisquama以及冠毛鲆Lophonectes gallus相应基因片段的比对。各基因片段的插入/缺失以及特异性碱基差异位点比对结果显示: 18S和28S的短序列类型A与4种鲆科鱼类序列一致, 而其他序列类型则不同; ITS1序列类型A与4种鲆科鱼类在类型B的缺失位点均无缺失, 因此推测18S、28S和ITS1的A类型为真基因, 而其他类型为假基因。ITS2的A和B类型在差异位点上与4个鲆科鱼类不存在一致性, 没有足够的依据对两个类型做出真、假基因的推断。长臂缨鲆核糖体RNA基因中, 5.8S序列最为保守遵循协同进化的方式, 而其他4个基因片段为非协同进化的方式。

关 键 词:核糖体RNA基因  长臂缨鲆  多态性  假基因  非协同进化  重组  
收稿时间:2018-04-09
修稿时间:2018-06-07

Analysis of polymorphism characteristics of ribosomal RNA genes in Crossorhombus kobensis (Pleuronectiformes: Bothidae)
Min YANG,Xiaoyu KONG,Wei SHI,Li GONG. Analysis of polymorphism characteristics of ribosomal RNA genes in Crossorhombus kobensis (Pleuronectiformes: Bothidae)[J]. Journal of Tropical Oceanography, 2019, 38(1): 55-66. DOI: 10.11978/2018038
Authors:Min YANG  Xiaoyu KONG  Wei SHI  Li GONG
Affiliation:1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, China2. University of Chinese Academy of Sciences, Beijing 100049, China3. National Engineering Research Center for Facilitated Marine Aquaculture, Marine Science and Technology College, Zhejiang Ocean University, Zhoushan, 316022, China
Abstract:To better understand the polymorphism characteristics of ribosomal RNA genes of Crossorhombus kobensis (Jordan & Starks, 1906) from Bothidae, Pleuronectiformes, a total of 128 clone sequences were obtained, including full-length sequences of 18S, ITS1, 5.8S, and ITS2 and partial fragments of 28S. After sequence alignments, clustering analyses and recombination detection, the results showed that only 5.8S (158 bp) had no length variation, while the other four gene fragments showed high length polymorphism and resulted in several distinct types: 18S (1856-1893 bp) with four types of Types A, B, C, and R; 28S (967-974) and ITS1 (407-505 bp) both had three types of Types A, B and R; ITS2 (423-447 bp) had two types of Types A and B. All five gene fragments showed GC-bias, and ITS2 (71.14%) > ITS1 (65.37%) > 28S (62.22%) > 5.8S (57.67%) > 18S (54.95%). The current characteristics criteria were not sufficient to provide strong evidence for the inference of functional gene or pseudogene of 18S, 28S and ITS sequences. Therefore, comparison with each of corresponding gene fragment of four affinis species from family Bothidae was conducted, Arnoglossus macrolophus, Crossorhombus azureus, Engyprosopon grandisquama, and Lophonectes gallus. The alignment showed that the indels and differential sites of Type A sequences of both 18S and 28S were the similar as those of the four species; and Type A of ITS1, as well as the four species, had no fragment deletion at the missing loci of Type B. Therefore, Type A sequences of 18S, 28S and ITS1 were speculated as functional genes, while the other types were putative pseudogenes. As for ITS2, the divergence loci of Type A and Type B compared to each of the four species had no consistency, and there was no evidence to infer the status of ITS2. In this study, 5.8S rDNA is the most conserved gene, suggesting a concerted evolution, while non-concerted evolution was confirmed in other four genes because of high intra-individual polymorphism.
Keywords:ribosomal RNA gene  Crossorhombus kobensis  polymorphism  pseudogene  non-concerted evolution  recombination  
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