额尔齐斯河流域不同来源哲罗鲑形态及COⅠ基因比较研究

为对哲罗鲑(Hucho)的引种放流和土著物种的保护提供科学依据, 研究采集了数尾来源于天然水域的哲罗鲑(Hucho BHB和Hucho BEJ), 与引进的哲罗鲑(Hucho HLJ)一起进行了形态特点观察描述, 同时对COⅠ基因扩增测序并从GenBank下载哲罗鲑属COⅠ基因构建系统发育树、计算遗传距离进行分子生物学比较研究。结果显示, Hucho BEJ和Hucho HLJ形态特点相一致, Hucho BHB与其他两种来源的哲罗鲑在体色、斑点大小等均有差异。基于COⅠ基因的系统发育树显示几种不同来源的哲罗鲑均与太门哲罗鲑(H. taimen)聚为一个大的分枝, 但在这个分枝下, Hucho BEJ和Hucho HLJ与俄罗斯阿穆尔河(中国称黑龙江)的哲罗鲑聚为一枝, 而Hucho BHB单独形成一支。基于COⅠ基因的遗传矩阵显示, Hucho BEJ和Hucho HLJ与俄罗斯阿穆尔河的哲罗鲑遗传距离较近(0—0.0044), 而Hucho BHB与Hucho BEJ、Hucho HLJ及俄罗斯阿穆尔河的哲罗鲑遗传距离较远(0.0057—0.0082)。这说明Hucho BHB在形态和基因序列上与Hucho HLJ及俄罗斯阿穆尔河的哲罗鲑具有较大差异, 可能是不同的生态地理类型, Hucho BEJ和Hucho HLJ在形态和基因序列上基本一致, 我们推测Hucho BEJ是放流或养殖场逃逸的哲罗鲑。     

濒危物种川陕哲罗鲑在汉江上游太白河再发现

正川陕哲罗鲑(Hucho bleekeri)隶属于鲑形目(Salmoniformes)鲑科(Salmonidae)哲罗鲑属,是我国特有物种,主要分布在29°~33°N秦岭以南长江流域北侧一级支流岷江水系和汉江水系的上游山区激流环境中,是欧亚大陆鲑鳟鱼类中分布纬度最低的种类。20世纪60年代以前,川陕哲罗鲑资源还较丰富,在岷江上游的干流、大渡河、青衣江及其附属支流均有分布。20世纪60年代以后,受环境恶化、过度捕捞等影响,资源量锐减,分布区域不断缩     

贵州荔波发现弱须洞鳅

洞鳅属（Troglonectes Zhang, Zhao & Tang 2016）鱼类为中国特有的小型洞穴淡水鱼类,主要分布在贵州和广西喀斯特地区的地下河流中。2018至2019年在贵州省荔波县境内进行洞穴生物多样性调查时采集到4号洞鳅属标本,经过形态比较,与弱须洞鳅（Troglonectes barbatus）相似;基于线粒体16S rRNA和Cyt b重建的系统发育树及遗传距离分析显示,4号洞鳅属标本与来自模式产地的弱须洞鳅高度支持聚为一支;利用Kimura双参数模型计算基于16S rRNA和Cyt b的遗传距离,这些标本与弱须洞鳅之间的遗传距离分别为0.4%和0.2%,小于洞鳅属物种之间的遗传距离（16S rRNA遗传距离为2.4%,Cyt b遗传距离为6.1%）。本次研究使用16S rRNA和Cyt b分子标记研究洞鳅属部分物种的系统发育关系和遗传距离。综合形态和分子系统发育分析结果,确定采集自贵州省荔波县小七孔景区的4号标本为隶属于鲤形目条鳅科洞鳅属的弱须洞鳅,系贵州省洞穴鱼类新记录种。     

基于线粒体COI、COII和Cyt b基因的蚁属13种的分子系统学研究（膜翅目：蚁科：蚁亚科）

以线粒体mtDNA中的COI、COII及Cyt b基因序列为分子标记,对蚁属13种进行分析研究。结果表明,蚁属线粒体基因组成具有昆虫基因的普遍特征,如有明显的A+T偏向性,A、T密码子使用频率高等特点。基于蚁属线粒体基因进行遗传距离分析,并结合构建系统发育树,结果表明蚁属内各物种间遗传距离相差较小,遗传距离值基本在0.1以内,遗传距离较远的种发生在北京凹头蚁和掘穴蚁之间,蚁属内各种间分化与形态学研究结果基本一致。     

基于线粒体COⅡ基因的中国蝽科分子系统学研究(半翅目,异翅亚目)

扩增并测定了我国蝽科4亚科8属11种昆虫线粒体COⅡ基因585 bp的序列,对序列的碱基组成、转换颠换、遗传距离等进行分析,探讨了COⅡ基因在该科的分子进化机制.并基于COⅡ基因序列数据,分别采用邻接法(NI)、最大简约法(MP)和贝叶斯推论法(BI)建立蝽科分子系统发育关系.研究结果表明,蝽科昆虫COⅡ基因A T含量平均为71.7%,存在较强的A T含量偏向性,氨基酸的变异率为27.2%;亚科间的遗传距离介于0.168～0.242之间,大于亚科内属种间的遗传距离,蝽科与盾蝽科2外群之间遗传距离最大,两科之间存在明显的间断.分子系统发育树表明,短喙蝽亚科为蝽科中较为原始的类群,分化较早,益蝽亚科与舌盾蝽亚科关系较近,形成一对姐妹群,蝽科中捕食性种类--益蝽亚科是较为特化的类群,它是由植食性种类分化而来.蝽科4亚科间的分子系统发育关系为Phyllocephalinae (Pentatominae (Asopinae Podopinae).     

基于COⅠ基因的中国广头叶蝉亚科系统发育初探

广头叶蝉亚科Macropsinae广泛分布于世界各地,其内部分类系统较混乱,部分族属级分类地位一直存在争议。本文选取23种叶蝉样本(内群22种+外群1种),分析其线粒体COⅠ基因部分片段的序列组成和遗传距离,基于最大似然法和贝叶斯法构建广头叶蝉亚科系统发育树。结果发现,COⅠ基因片段序列A+T含量(68. 3%)明显较G+C含量(31. 7%)高;遗传距离在种间、属间和亚科间的差异有统计学意义;系统发育树较好地支持了横皱叶蝉属Oncopsis和尖尾叶蝉属Pedionis的单系性,但广头叶蝉属Macropsis和暗纹叶蝉属Pediopsoides较为混乱。     

舌鳎亚科鱼类单系起源和同种异名的线粒体DNA证据

舌鳎亚科鱼类因形态特征的特殊性,其分类及系统发育关系一直存在争议.本研究测定了中国沿海14种舌鳎亚科鱼类线粒体DNA的16S rRNA和Cyt b基因的部分片段.两个基因构建的舌鳎亚科系统发育树结果显示,中国沿海舌鳎亚科鱼类为明显的单系群,但舌鳎亚科鱼类内部的系统发育关系与形态分类划分的亚属并不完全一致,日本须鳎(Paraplagusis japonica)与其他舌鳎属(Cynoglossus)种类并未形成不同的分支.虽然舌鳎属内的舌鳎亚属(Cynoglossus)、拟舌鳎亚属(Cynoglossoides)和三线舌鳎亚属(Areliscus)均可以聚为独立分支,但长吻红舌鳎(C.lighti)与短吻红舌鳎(C.joyneri)、短吻三线舌鳎(C.abbreviatus)与紫斑舌鳎(C.purpureomaculatus)、半滑舌鳎(C.semilaevis)与窄体舌鳎(C.gracilis)及褐斑三线舌鳎(C.trigrammus)这三组物种可能存在同种异名现象.这一结果提示,基于形态学对舌鳎亚科的种属分类鉴定尚存在不足,线粒体DNA的系统发育关系可为其分类的修订提供有意义的参考和佐证.     

基于线粒体COⅠ基因的沙鳅亚科鱼类DNA条形码及其分子系统发育研究

选择线粒体COⅠ基因作为分子标记,进行沙鳅亚科鱼类(Botiinae)DNA条形码及其分子系统发育研究。研究获得了沙鳅亚科7属19种共131个个体的COⅠ基因序列,利用MEGA5.0软件分析了沙鳅亚科鱼类COⅠ基因的序列特征,计算了种内及种间遗传距离。沙鳅亚科鱼类的分子系统发育关系的重建分别采用NJ法和Bayesian法。研究发现,沙鳅亚科COⅠ基因的碱基组成为:A 24.4%、T 29.5%、G 18.0%、C 28.1%。沙鳅亚科鱼类种内平均遗传距离为0.002±0.000,种间平均遗传距离为0.148±0.008。DNA条形码研究结果显示,所分析的19种沙鳅鱼类各自分别聚成单系分支,表明COⅠ基因在本研究中具有100%的物种鉴别率。同时,系统发育分析支持各属的单系性,并且结果显示沙鳅亚科鱼类聚为两个分支,其中一支由薄鳅属和副沙鳅属构成,另一分支则包括:(沙鳅属、色鳅属)和[中华沙鳅属、(缨须鳅属、安彦鳅属)]。因此,COⅠ基因可以作为有效的分子标记对沙鳅亚科进行DNA条形码研究以及分子系统发育研究。     

中国缘蝽科部分种类细胞色素b基因序列及系统进化研究

以线粒体细胞色素b(Cyt b)基因作为分子标记,首次对缘蝽科4亚科14种昆虫进行序列测定,获得Cyt b基因412 bp的序列片段,该片段中碱基T、C、A、G的平均含量分别为34.2%、11.4%、35.7 %和18.7 %,A T平均含量为69.9 %,明显高于G C含量(30.1 %).密码子第3位点的A T含量高达82.8%,亚科间序列变异大,有193个核苷酸位点发生变异,碱基替换多发生于第3位点.以筛豆龟蝽为外群构建系统发育树,表明在亚科级关系上,姬缘蝽亚科最原始,蛛缘蝽亚科次之,巨缘蝽亚科和缘蝽亚科亲缘关系较近,为较进化种类.     

不同流域细鳞鱼染色体遗传多态性分析

细鳞鱼Brachymystax lenok属鲑形目(Salmoniformes)、鲑科(Salmonidae)、细鳞鱼属(Brachymystax);地方名为细鳞鱼(东北地区),该鱼在我国东北地区的乌苏里江、鸭绿江、牡丹江、松花江等流域广泛分布[1-3].     

Evidence supporting the paraphyly of Hucho (Salmonidae) based on ribosomal DNA restriction maps

Relationships among the less derived genera in the subfamily Salmoninae were investigated by phylogenetic analysis of data from restriction maps of the ribosomal DNA using Osmerus mordax (rainbow smelt), Coregonus hoyi (bloater), and Thymattus arcticus (Arctic grayling) as outgroups. Analysis of the results suggests that Hucho h. taimen is more closely related to Brachymystax lenok (lenok) than to Hucho perryi (Sakhalin taimen) and that B. lenok and Hucho h. taimen (Eurasian taimen) form the most ancestral clade of the Salmoninae.     

基于线粒体控制区和微卫星标记探讨秦岭细鳞鲑物种有效性

文章基于线粒体控制区基因序列和微卫星标记比较秦岭细鳞鲑(Brachymystax tsinlingensis Li)、黑龙江流域的尖吻细鳞鲑(Brachymystax lenok Pallas)和钝吻细鳞鲑(Brachymystax tumensis Mori)的分子遗传差异,为澄清其分类地位争议提供分子证据。结果表明:(1)扩增217个样本的mtDNA D-loop区序列,共获得45个单倍型,类群间无共享单倍型;基于单倍型构建的系统进化树显示三个细鳞鲑类群呈独立的支系;(2)基于14个呈多态性位点的遗传分化结果表明,秦岭细鳞鲑与尖吻或钝吻细鳞鲑之间的遗传距离均大于尖吻细鳞鲑和钝吻细鳞鲑之间的遗传距离;(3)基于线粒体D-loop和多态性微卫星位点计算出的遗传分化系数(F_ST)都远高于0.25表明三个类群间的遗传分化程度极高。这些结果表明,秦岭细鳞鲑与黑龙江流域细鳞鲑之间遗传分化程度高,结合前期发现秦岭细鳞鲑与黑龙江细鳞鲑类群有明显形态分化的研究结果及它们之间地理隔离已久的现状,研究初步判定秦岭细鳞鲑为独立物种,并建议以Brachymystax tsinlinge...     

秦岭细鳞鲑群体遗传结构

秦岭细鳞鲑(Brachymystax lenok tsinlingensis)是秦岭地区特有鱼类,近年来由于环境恶化及人类活动的加剧,已对其造成严重影响,种群处于濒危状态,因此研究秦岭细鳞鲑的群体遗传结构、演化历史、分布动态等对其进行有效保护具有重要意义。本研究用线粒体D-loop区序列对秦岭地区6个群体(n=112)进行了遗传结构和群体演化分析。D-loop区扩增出的891bp序列在112个个体中,检测到42个变异位点,共26个单倍型;碱基序列总的单倍型多样度较高为0.883,核苷酸多样度为0.00799。AMOVA分析显示,60.05%的分子差异位于群体内,39.95%的分子差异位于群体间,Fst值统计检验表明,除那布大河群体与漳河群体和千河群体之间差异不显著之外,其余两两群体之间Fst值统计检验均为显著。系统树和单倍型网络图分析表明,6个地理群体的单倍型按照渭河上游和渭河中游两个河段形成两个大的类群,且5个群体共享一个单倍型H2,表明这些群体具有相同的演化历史,为同一个祖先群体演化而来。中性检验和歧点分布显示,秦岭细鳞鲑种群大小保持相对稳定,未经历明显的种群扩张。同时建议将渭河上游秦岭细鳞鲑群体作为一个整体进行重点保护。     

Molecular systematics of Salmonidae: combined nuclear data yields a robust phylogeny

The phylogeny of salmonid fishes has been the focus of intensive study for many years, but some of the most important relationships within this group remain unclear. We used 269 Genbank sequences of mitochondrial DNA (from 16 genes) and nuclear DNA (from nine genes) to infer phylogenies for 30 species of salmonids. We used maximum parsimony and maximum likelihood to analyze each gene separately, the mtDNA data combined, the nuclear data combined, and all of the data together. The phylogeny with the best overall resolution and support from bootstrapping and Bayesian analyses was inferred from the combined nuclear DNA data set, for which the different genes reinforced and complemented one another to a considerable degree. Addition of the mitochondrial DNA degraded the phylogenetic signal, apparently as a result of saturation, hybridization, selection, or some combination of these processes. By the nuclear-DNA phylogeny: (1) (Hucho hucho, Brachymystax lenok) form the sister group to (Salmo, Salvelinus, Oncorhynchus, H. perryi); (2) Salmo is the sister-group to (Oncorhynchus, Salvelinus); (3) Salvelinus is the sister-group to Oncorhynchus; and (4) Oncorhynchus masou forms a monophyletic group with O. mykiss and O. clarki, with these three taxa constituting the sister-group to the five other Oncorhynchus species. Species-level relationships within Oncorhynchus and Salvelinus were well supported by bootstrap levels and Bayesian analyses. These findings have important implications for understanding the evolution of behavior, ecology and life-history in Salmonidae.     

Evolutionary and taxonomic relationships among Far-Eastern salmonid fishes inferred from mitochondrial DNA divergence

Mitochondrial DNA (mtDNA) restriction analysis was used to examine the evolutionary and taxonomic relationships among 11 taxa of the subfamily Salmoninae. The genera Brachymystax and Hucho were closely related, diverging by sequence divergence estimates of 3.1%. Because the mtDNA sequence divergence between blunt- and sharp-snouted forms of Brachymystax (2.24%) was similar to divergence level of Brachymystax and Hucho , then taking into account the distinct morphological, ecological and allozyme differences between them, it is possible to recognize these forms as two separate species. The subgenus Parahucho formed a very distinct group differing by 6.35–7.08% (sequence divergence estimate) from both Brachymystax and Hucho and must be considered as a valid genus. The UPGMA and neighbour-joined phenograms showed that the five genera studied are divided into two main groupings: (1) Hucho, Brachymystax and Salvelinus ; and (2) Oncorhynchus and Parahucho species. The mtDNA sequence divergence estimates between these groupings were about 8.1%. However, the subsequent bootstrap analysis of mtDNA RFLP data did not support the monophyly of the latter grouping. The concordance of morphological and mtDNA phylogenetic patterns is discussed.     

黑龙江流域两种细鳞鲑的形态学比较及其分类地位初探

对比研究了分布于我国黑龙江流域具钝吻和尖吻两种形态特征的细鳞鲑,其主要形态学性状的显著差异达到种间分化水平.细鳞鲑属Brachymystax在中国应有两个种,分别为图们江细鳞鲑B.tumensis(具钝吻特征)和细鳞鲑B.lenok(具尖吻特征).     

Phylogenetic relationships of Sakhalin taimen Parahucho perryi inferred from PCR-RFLP analysis of mitochondrial DNA

RFLP analysis of three amplified mtDNA fragments (Cytb/D-loop, ND1/ND2, and ND3/ND4L/ND4) was performed in the following taxa: Parahucho perryi, Hucho taimen, Brachymystax lenok, B. tumensis, Salmo salar, Salvelinus leucomaenis, and S. levanidovi. For mtDNA of P. perryi, a substantial decrease in the haplotype and nucleotide diversity was observed as a result of random genetic drift, caused by a reduction in the effective population size. Nucleotide divergence estimates between the mtDNA haplotypes were determined. Sakhalin taimen P. perryi was found to be approximately equally diverged from S. salar and from the charrs of the genus Salvelinus, by 11.0 and 10.0%, respectively. The divergence between P. perryi and H. taimen constituted 14.6%, between P. perryi and lenoks of the genus Brachymystax, 14.2%, and between H. taimen and Brachymystax, 7.7%. The analysis of possible phylogenetic relationships of the mtDNA from P. perryi among the group of taxa examined confirmed validity of the genus Parahucho. Phylogenetic reconstructions performed showed that robustness of the trees constructed for the complex of phylogenetically informative characters over three mtDNA fragments was considerably higher than that of the trees constructed for individual genes.     

不同脂肪源对细鳞鲑生长、脂质代谢及抗氧化性能的影响

配制4种含有鱼油(FO)、菜籽油(CO)、棕榈油(PaO)和大豆油(SO)的饲料投喂初始体重为(22.19±1.10) g的细鳞鲑幼鱼52d, 探讨不同脂肪源对细鳞鲑(Brachymystax lenok)幼鱼生长、脂质代谢及抗氧化性能的影响。结果显示, 增重率、特定生长率和饲料效率均以鱼油和大豆油组最高, 棕榈油组次之, 菜籽油组最低, 其中大豆油组与鱼油组无显著差异(P>0.05)。不同脂肪源对细鳞鲑幼鱼肌肉粗成分无显著影响(P>0.05)。摄食大豆油的细鳞鲑可以明显提升肌肉中的22:6n-3含量, 表明细鳞鲑具有将18:3n-3转化为22:6n-3的能力, n-3 系列脂肪酸是细鳞鲑起主要作用的必需脂肪酸。与鱼油相比, 植物油脂导致了细鳞鲑血清中甘油三酯含量的显著升高以及胆固醇含量的显著降低, 肝脏中ACC1和FAS的mRNA表达量明显下调, 以及Δ6 Fad的mRNA表达量的明显上调(P<0.05)。投喂不同脂肪源饲料的细鳞鲑肝脏ROS、MDA、GPx含量以及TBARS值发生明显变化(P<0.05)。研究结果表明鱼油和大豆油可作为细鳞鲑幼鱼饲料的脂肪源, 而棕榈油和菜籽油不适宜作为细鳞鲑幼鱼的单一脂肪源。     

基于形态学差异探讨秦岭细鳞鲑亚种有效性问题

秦岭细鳞鲑(Brachymystax lenok tsinlingensis Li)的亚种分类地位自命名以来一直饱受争议。研究运用单因素方差分析、主成分分析、判别分析、聚类分析及差异系数检验法, 对秦岭细鳞鲑和黑龙江流域的尖吻细鳞鲑(B. lenok lenok)和钝吻细鳞鲑(B. tumensis)的5项可数性状和34项标准化后的比例性状进行统计分析。单因素方差分析结果表明, 尖吻细鳞鲑、钝吻细鳞鲑和秦岭细鳞鲑在32项比例性状和5项可数性状上存在极显著差异(P<0.01); 主成分分析结果显示, 贡献率较大的前3个主成分的累计贡献率为92.779%; 以逐步判别分析方法选用14个判别效果较好的比例性状构建了3个细鳞鲑类群的判别函数, 综合判别率为99.4%。基于欧式距离矩阵法构建的形态学聚类图显示, 秦岭细鳞鲑和尖吻细鳞鲑距离较近, 而与钝吻细鳞鲑距离较远。据Mayr 75%亚种识别和划分规则, 34项比例性状和5项可数性状中, 尖吻细鳞鲑和秦岭细鳞鲑第一鳃弓外鳃耙数目的差异系数大于1.28, 而钝吻细鳞鲑和秦岭细鳞鲑有31项比例性状和2项可数性状的差异系数高于1.28。据分析结果, 结合秦岭细鳞鲑地理隔离的事实及其与尖吻细鳞鲑和钝吻细鳞鲑的形态差异, 推断秦岭细鳞鲑与黑龙江流域内的尖吻细鳞鲑的形态差异程度至少已达亚种水平。     

Mitochondrial cytochrome b sequence variation in Korean salmonids

The complete 1141 bp mitochondrial cytochrome b sequences were determined for lenok Brachymystax lenok , cherry salmon Oncorhynchus masou masou , Ishikawa's cherry salmon O. m. ishikawai , chum salmon O. keta , rainbow trout O. mykiss , and an albino mutant of rainbow trout. Common substitutions detected in these species were transitional mutations. There were no significant differences in the intraspecific variation of the cytochrome b genes. Interspecific divergences were greater than intraspecific variation. The level of variation ranged from 8·026–15·686%. The cherry salmon was closer to chum salmon than to rainbow trout, and the lenok was the most distantly related species.