高原鳅属Triplophysa鱼类的分子系统发育和生物地理学研究

高原鳅属Triplophysa鱼类广泛分布于亚洲中部的高原地区,是青藏高原鱼类区系重要的组成部分.它们对高原环境表现出极强的适应性,是已知世界上分布海拔最高的鱼类.采用线粒体DNA细胞色素b基因序列分析了青藏高原及其邻近地区13个水系30个地点22种高原鳅属鱼类的分子系统发育关系.结果表明高原鳅属鱼类不是一个单系群,赫氏鳅亚属Hedinichthys的种类嵌入到其他条鳅亚科鱼类分支中,而依据第二性征划分的高原鳅亚属Triplophysa是一个单系群.在高原鳅亚属中,最先发生分歧的是分布于黄河上游的似鲶高原鳅T.(T.)siluroides,其次是分布于南盘江上游的抚仙高原鳅T.(T.)fuxianensis.将骨质鳔囊和鳔后室的形态特征图解到分子系统发育树中时,并没有呈现出共近离征的改变.高原鳅亚属内种间系统发育关系在一定程度上反映了地理关系的远近,即相同和相邻水系的物种之间通常具有更近的亲缘关系;进而在生物地理过程中反映了青藏高原东部和西北部水系与青藏高原主体及东南部水系在9.5-7.2 Ma出现分离,这一事件可能与晚新生代青藏高原在8 Ma左右发生的地质构造事件及气候重大转型相关.     

高原鳅属Triplophysa鱼类的分子系统发育和生物地理学研究

高原鳅属Triplophysa鱼类广泛分布于亚洲中部的高原地区，是青藏高原鱼类区系重要的组成部分.它们对高原环境表现出极强的适应性,是已知世界上分布海拔最高的鱼类. 采用线粒体DNA细胞色素b基因序列分析了青藏高原及其邻近地区13个水系30个地点22种高原鳅属鱼类的分子系统发育关系.结果表明高原鳅属鱼类不是一个单系群，赫氏鳅亚属Hedinichthys的种类嵌入到其他条鳅亚科鱼类分支中，而依据第二性征划分的高原鳅亚属Triplophysa是一个单系群.在高原鳅亚属中，最先发生分歧的是分布于黄河上游的似鲶高原鳅T.(T.) siluroides, 其次是分布于南盘江上游的抚仙高原鳅T.(T.) fuxianensis. 将骨质鳔囊和鳔后室的形态特征图解到分子系统发育树中时，并没有呈现出共近离征的改变.高原鳅亚属内种间系统发育关系在一定程度上反映了地理关系的远近，即相同和相邻水系的物种之间通常具有更近的亲缘关系；进而在生物地理过程中反映了青藏高原东部和西北部水系与青藏高原主体及东南部水系在9.5—7.2Ma出现分离，这一事件可能与晚新生代青藏高原在8Ma左右发生的地质构造事件及气候重大转型相关.     

基于线粒体Cytb基因的黄海、东海小黄鱼（Larimichthys polyactis）群体遗传结构

采用线粒体DNA细胞色素b基因全序列分析技术研究了黄海、东海小黄鱼（Larimich—thyspolyactis）的群体遗传结构．在所分析的9个取样点177个个体中,共检测到137个单倍型．9个群体呈现出高的单倍型多样性（h=0．956—1．00）和低的核苷酸多样性（π=0．0037-0．0058）．单倍型邻接关系树的拓扑结构比较简单,没有明显的地理谱系结构．分子方差分析和FST显示小黄鱼的遗传变异均来自群体内个体间,而群体问无显著遗传分化．Exact检验表明单倍型在两两群体间分布频率的差异是不显著的．中性检验和核苷酸不配对分析均表明黄海、东海的小黄鱼经历了群体扩张,扩张时间约为78—138ka前．研究结果表明,黄海、东海小黄鱼群体间具有高度的基因交流,是一个随机交配的群体．较强的扩散能力,黄海、东海的海洋环流以及近期的群体扩张可能是造成黄海、东海小黄鱼群体间遗传同质性较高的原因．     

高原鳅属鱼类线粒体全基因组序列结构特征及其系统发育信息

为了深入开展高原鳅属(Triplophysa)鱼类的分类鉴定、系统进化等研究,利用生物信息学的方法分析了37种高原鳅属鱼类的线粒体全基因组序列及系统发育信息.通过Clustal X对线粒体DNA(mtDNA)全基因组序列进行对比,然后用MEGA7分析mtDNA的序列差异,并用邻接法生成系统进化树.结果发现:(1)高原鳅...     

从Cytb基因序列探讨蝽类部分昆虫的系统发生

通过对蝽总科18种昆虫线粒体DNA细胞色素6基因部分序列进行PCR扩增和序列测定,比较其同源性,统计密码子使用频率并应用生物学软件构建系统发育树．在获得的432bp序列中,碱基T,C,A和G的平均含量分别为37．4％,18．8％,31．8％和12．0％,表现出强烈的AT偏向性．就每个氨基酸密码子采看,第3位点的A＋T含量较高,达到83．6％．谊序列片段中共有215个核苷酸位点发生变异（约占49．8％）,种间变异较大．在氨基酸组成上,共编码144个氨基酸。其中有47个发生变异。变异率为32．7％．碱基替换主要发生在密码子第3位点,转换略多于颠换．以筛豆龟蝽（Megacopta cribraria）为外群构建的系统发育树支持荔蝽、盾蝽作为独立的科与蝽科并列,滴蝽从蝽亚科划出归属于舌盾蝽亚科,但蝽科内各亚科间及蝽亚科各族间的系统进化关系与传统的形态分类结果不完全一致,还有待于进一步研究．     

蚁科弓背蚁属部分种的分子系统学研究

对弓背蚁属12种昆虫的线粒体细胞色素b(Cyt b)基因部分序列进行测定.获得了长度为487 bp的序列片段.基于Cytb基因序列数据,采用邻接法(NJ)、最大简约法(MP)和最大相似法(ML)分析其系统发育关系,研究结果表明,弓背蚁属12种蚂蚁分成4支:第一个支系包括杂色弓背蚁C.variegates、拟光腹弓背蚁C.pseudoirritans、黄斑弓背蚁C.albosparsus和弓背蚁C.sp1;第二个支系由尼科巴弓背蚁C.nicobarensis、弓背蚁C.sp2和日本弓背蚁C.japonicus组成;第三个支系包含哀弓背蚁C.dolendus、少毛弓背蚁C.spanis与平和弓背蚁C.mitis;第四个支系由弓背蚁C.sp和浅毛弓背蚁C.albivillosus组成,属内种间的分类关系明确.遗传距离的分子数据支持将未定种弓背蚁C.sp定为弓背蚁属的新种.     

绒螯蟹线粒体细胞色素b基因的RFLP分析

对中国大陆4个水系的8个地理种群绒螯蟹样本的线粒体细胞色素b基因片段进行了PCR扩增，并应用6种限制性内切酶对该PCR产物进行RFLP分析．在应用的6种内切酶中，HinfⅠ，RsaⅠ和EcoRⅤ酶切该PCR片段后，在某些地区绒螯蟹之间表现出限制性片段长度多态性，为多态性的内切酶，但在本研究中尚未发现种群特异的RFLP标记．应用3种多态性的限制性内切酶对8个地理种群的绒鳌蟹个体样本的线粒体细胞色素b基因片段进行RFLP分析，共检测到5种复合限制性酶切类型，即AAA型、BBB型、ABA型、BAB型、ACA型．依据群体间的净遗传距离绘制的UPGMA分子系统树显示，合浦绒螯蟹形成了独立的一支．     

海南产花鳗鲡细胞色素b基因的克隆及序列分析

线粒体细胞色素b基因是目前研究鱼类分子系统进化的重要分子标记.笔者以日本鳗鲡线粒体基因组序列为模板设计并合成引物进行PCR扩增,并将扩增产物经琼脂糖凝胶电泳和纯化后,克隆到pMD18-T载体、测序,成功得到全长为1 140 bp的海南产花鳗鲡细胞色素b(Cyt b)基因序列.将该基因全长序列递交到Genebank数据库,获得序列登录号为EF690363.用DNA分析软件MEGA2比较了海南产花鳗鲡与递交到GenBank中的8种分布在中国东南沿海、日本海域及南太平洋海域的鳗鲡属(Anguillia)细胞色素b基因的地理差异,并构建了系统进化树.结果显示:日本产和夏威夷产花鳗的地理差异小于海南产花鳗与它们之间的地理差异.     

基于线粒体Cytb基因探讨我国日本囊对虾4个地理群体的遗传结构及种群分化

利用线粒体细胞色素b基因序列分析了4个不同地理群体(北海群体、陵水群体、惠来群体和厦门群体)的野生日本囊对虾(Marsupenaeus japonicus)的群体遗传结构.以特异引物进行PCR扩增,获得了日本囊对虾530 bp的基因片段序列.序列分析结果表明,在120个日本囊对虾个体中,共检测到75个多态位点,获得62个单倍型,其中有44个简约信息位点,占整段序列的8.3%.各单倍型变异无碱基的插入或缺失,全部为转换或颠换,碱基频率含量(fA+fT)为59.4%,大于(fG+fC)(40.6%).核苷酸多态性以惠来群体最高,其它3个群体较低.群体内遗传距离为0.45%～2.60% ,群体间遗传距离在0.50%～5.30%之间.采用分子方差分析(AMOVA)方法,结果显示,群体间遗传变异占总变异的69.9%,群体内的遗传变异占总变异的31.1%,群体间变异是总变异的主要来源.构建的4个群体分子系统树表明,北海群体、陵水群体和部分惠来群体由于亲缘关系较近而聚在一起,而厦门群体则和部分惠来群体聚成另一支,两支的平均遗传距离为5.17%,分化接近亚种水平,据此认为我国东南近海的日本囊对虾可以分为2个种群,2个种群的分布区域在广东惠来海域附近存在重叠.     

稀有鮈鲫β-肌动蛋白基因片段的克隆及其同源性分析

采用RT-PCR方法,从稀有鮈鲫肌肉组织中分离和克隆出β-肌动蛋白基因cDNA部分序列,长度为1029bp,编码343个氨基酸残基．序列分析表明,该cDNA序列与其他物种昏肌动蛋白基因同源性非常高．RT-PCR能够检出该基因在稀有鮈鲫肌肉、眼、脑、心脏、肝、肠、鳃、睥、卵巢等组织中广普表达,在胚胎不同发育时期持续恒量表达．并基于已知的鱼类β-肌动蛋白基因序列构建了进化树．     

六种鱼巴亚科鱼类线粒体细胞色素b基因序列分析

 测定了鱼巴亚科云南光唇鱼(Acrossocheilus yunnanensis)、宽头四须鱼巴(Barbodes laticeps)、抚仙金线鱼巴(Sinocyclocheilus tingi)和滇池金线鱼巴(Sinocyclocheilus grahami)4种鱼类线粒体细胞色素b基因DNA序列402bp,结合已知的云南倒刺鱼巴(Spinibarb denticulatus yunnanensis)和细尾长臀鱼巴(Mystacoleucus lepturus)的同源序列,组成1个6种代表鱼巴亚科5属鱼类的数据集.选用已知的云南鲴(Xenocypris yunnanensis)作为外群,采用邻接法、最大简约法和最大似然法构建了分子系统树.结果显示:倒刺鱼巴属(Spinibarbus)和光唇鱼属(Acrossocheilus)有较近的亲缘关系,四须鱼巴属(Barbodes)和长臀鱼巴属(Mystacoleucus)有较近的亲缘关系,而金线鱼巴属(Sinocyclocheilus)和它们的关系不能确定.上述结果与它们的地理分布基本吻合.     

六种鱼巴亚科鱼类线粒体细胞色素b基因序列分析

测定了鱼巴亚科云南光唇鱼(Acrossocheilus yunnanensis)、宽头四须鱼巴(Barbodes laticeps)、抚仙金线鱼巴(Sinocyclocheilus tingi)和滇池金线鱼巴(Sinocyclocheilus grahami)4种鱼类线粒体细胞色素b基因DNA序列402bp,结合已知的云南倒刺鱼巴(Spinibarb denticulatus yunnanensis)和细尾长臀鱼巴(Mystacoleucus lepturus)的同源序列,组成1个6种代表鱼巴亚科5属鱼类的数据集.选用已知的云南鲴(Xenocypris yunnanensis)作为外群,采用邻接法、最大简约法和最大似然法构建了分子系统树.结果显示:倒刺鱼巴属(Spinibarbus)和光唇鱼属(Acrossocheilus)有较近的亲缘关系,四须鱼巴属(Barbodes)和长臀鱼巴属(Mystacoleucus)有较近的亲缘关系,而金线鱼巴属(Sinocyclocheilus)和它们的关系不能确定.上述结果与它们的地理分布基本吻合.       

Sequences of cytochrome b gene for primitive cyprinid fishes in East Asia and their phylogenetic concerning

1140 bp of cytochrome b gene were amplified and sequenced from 14 species of primitive cyprinid fishes in East Asia. Aligned with other ten cytochrome b gene sequences of cyprinid fish from Europe and North America retrieved from Gene bank, we obtained a matrix of 24 DNA sequences. A cladogram was generated by the method of Maximum likelihood for the primitive cyprinid fishes. The result indicated that subfamily Leuciscinae and Danioninae do not form a monophyletic group. In the subfamily Danioninae, Opsariichthys biden and Zacco platypus are very primitive and form a natural group and located at the root. But the genera in subfamily Danioninae are included in different groups and have not direct relationship. Among them, Aphyocypris chinensis and Yaoshanicus arcus form a monophyletic group. Tanichthys albonubes and Gobiocypris rarus have a close relation to Gobioninae. The genus Danio is far from other genera in Danioninae. In our cladogram, the genera in Leuciscinae were divided into two groups that have no direct relationship. The genera in Leuciscinae distributed in Europe, Sibera and North America, including Leuciscus, Rutilus, Phoxinus, N. crysole, Opsopoeodus emilae, form a monophyletic group. And the Leuciscinae in southern China including Ctenopharyngodon idettus, Mylopharyngodon piceus, Squalibarbus and Ochetobius elongatus have a common origination.     

从细胞色素b基因序列分析羚牛分子系统进化

目的分析羚牛分子系统进化,解决多年来关于羚牛分类地位及其与麝牛关系的争论。方法应用聚合酶链式反应(PCR)分别扩增羚牛、绵羊、山羊细胞色素b基因,并对其全序列(1140bp)进行测定。结合GenBank检索序列,对9种偶蹄类动物(麝牛、绵羊、羚牛等)、1种奇蹄类动物(斑马)的细胞色素b基因序列差异进行分析,并基于序列差异构建分子系统树。结果羚牛与羊亚科的动物亲缘关系最近(序列差异分别为9.085%和11.652%),与其他动物亲缘关系较远(序列差异为12.344%~23.333%),与麝牛的差异达到了13.658%。羚牛与绵羊分歧的时间约在360万年前,而与麝牛的分歧时间约在550万年前。结论将羚牛归入羊亚科较为合理,羚牛和麝牛形态和行为上很强的相似性可能是趋同进化的结果。     

Molecular phylogenetic status of Siberian roe deer （Capreolus pygargus） based on mitochondrial cytochrome b from Jeju Island in Korea

In order to elucidate the genetic diversity and taxonomic status of Siberian roe deer （Capreolus pygar- gus） from the Korean peninsula, partial sequences of the mitochondrial cytochrome b gene were analyzed. We col- lected 57 roe deer samples from Mainland and Jeju Island between July 2004 and June 2009. A total of 50 sequences （1,070 bp） were obtained, and seven haplotypes were defined. Five haplotypes were found from Mainland, and two were discovered from Jeju Island. The genetic diver- sity was lower in the Jeju Island population than in the Mainland population. In addition, both the consensus neighbor-joining tree and maximum parsimony tree showed an identical topology between the Mainland and Jeju Island clades. Based on the results, two subspecies of the Korean roe deer were confirmed. Siberian roe deer in Jeju Island showed a distinct difference in the cytochrome b gene compared with other Siberian roe deer, being aunique native species inhabiting only Jeju Island in Korea. Because Siberian roe deer in Jeju Island can be classified at subspecies level, the new subspecies in Jeju was named Capreolus pygargus jejuensis in this study. For this new subspecies, reasonable conservation and management effort is needed in the future.     

羚牛细胞色素b基因序列分析和系统进化研究

应用聚合酶链式反应（PCR）分别扩增了羚牛、绵羊、山羊、黄牛细胞色素b基因,并对其全序列（1140bp）进行了测定。其中羚牛细胞色素b基因序列属首次报道。通过对8种偶蹄类动物细胞色素b基因序列差异分析和基于序列差异所构建的分子系统树,发现羚牛与羊亚科的动物亲缘关系最近,与其他动物亲缘关系较远。表明将羚牛放入羊亚科较为合理,序列差异分析还表明羚羊约在距今500万年前（上新世）从牛类动物中分化出来,牛类分化时间约在600万年前的中新世（Miocene）。     

Mitochondrial cytochrome b sequences variation of Protura and molecular systematics of Apterygota

Mitochondrial cytochrome b genes of about 450 bp fragments from 3 proturan species, 5 collembolan species and 2 dipluran species have been sequenced. The number of nucleotide substitutions and Kimura 2-parameter distances have also been calculated, and a series of molecular phylogenetic trees reconstructed by using parsimony and distance methods. The proturan, collembolan and dipluran species have evolved monophyletic groups. The results suggest that Protura and Collembola are sister groups, while Diplura is more or less demonstrating a closer phylogenetic relationship to the pterygotan insects. The phylogeny and their systematic position of protura and other groups are also discussed.     

Mitochondrial cytochrome b sequence variations and phylogeny of the East Asian bagrid catfishes

The mitochondrial DNA cytochrome b gene was sequenced from 8 bagrid catfishes in China. Aligned with cytochrome b sequences from 9 bagrid catfishes in Japan, Korea and Russia retrieved from GenBank, and selected Silurus meridionalis, Liobagrus anguillicauda, Liobagrus reini and Phenacogrammus interruptus as outgroups, we constructed a matrix of 21 DNA sequences. The Kimura's two-parameter distances were calculated and molecular phylogenetic trees were constructed by using the maximum parsimony (MP) and neighbor-joining (NJ) methods. The results show that (i) there exist 3-bp deletions of mitochondrial cytochrome b gene compared with cypriniforms and characiforms; (ii) the molecular phylogenetic tree suggests that bagrid catfishes form a monophyletic group, and the genus Mystus is the earliest divergent in the East Asian bagrid catfishes, as well as the genus Pseudobagrus is a monophyletic group but the genus Pelteobagrus and Leiocassis are complicated; and (iii) the evolution rate of the East Asian bagrids mitochondrial cytochrome b gene is about 0.18%～0.30% sequence divergence per million years.     

Application of the phylogenetic analysis in mitochondrial disease study

Mitochondrial disease currently received an increasing concern. However, the case-control design commonly adopted in this field is vulnerable to genetic background, population stratification and poor data quality. Although the phylogenetic analysis could help solve part of these problems, it has not received adequate attention. This paper is a review of this method as well as its application in mitochondrial disease study.     

Phylogenetic relationships of host insects of Cordyceps sinensis inferred from mitochondrial Cytochrome b sequences

This study used the sequence of the mitochondrial Cytochrome b(Cytb)to estimate phylogenetic relationships among host Hepialidae insects of Cordyceps sinensis.Genome DNA of host insect was extracted from the dead larva head part of 18 cordyceps populations and 2 species of Hepialus,and the Cytb fragment of host insect was amplified with PCR technique.The nucleotide sequence alignments and their homologous sequences of 24 species host Hepialidae insects of Cordyceps sinensis were obtained from GenBank and were used to construct phylogenetic trees based on neighbor-joining method.The results showed that genus Bipectilus diverged earlier than genus Hepialus and Hepialiscus.Hepialus host insects of Cordyceps sinensis have multitudinous species with different morphological characteristics and geographical distributions.The interspecific genetic differentiations are obvious in Hepialus.Thus,the genus Hepialus might be considered as polyphyletic origin.Cytb sequences have abundant variations among the host insects of Cordyceps sinensis on spe- cific and generic level.The divergence rate of Cytb sequences among the species in Hepialus ranged from 0.23% to 9.24%,except that Hepialus pratensis and Hepialus jinshaensis have the same sequence.Cytb sequence can be used for species identification of host insects of Cordyceps sinensis,but further confirmation in more host insect species is needed.To obtain the Cytb sequence of host insect by ampli- fying DNA extracted from the head part of dead larva in cordyceps turns out to be an effective and accurate approach,which will be useful for studies on phylogeny and genetic structure of host insects of cordyceps populations,especially for analyzing relationships between C. sinensis and its host insects.