暗纹东方鲀（Takifugu obscurus）性别分化的组织学及芳香化酶基因表达研究

采用组织切片技术和实时定量PCR（qRT-PCR）方法研究了暗纹东方鲀性别分化过程中性腺发育的组织学及芳香化酶基因CYP19A的表达变化.结果显示：孵化后11d的仔鱼切片中可看到原始生殖细胞（PGC）;孵化后17d可看到隆起的性腺原基（GA）;孵化后26d发育成为原始性腺（PG）并从体腔膜上游离出来.卵巢分化时间早于精...     

暗纹东方鲀线粒体若干tRNA基因的克隆及序列分析

利用分离纯化的暗纹东方鲀肝脏mtDNA为模板，按照红鳍东方纯（Takifugu rubripes）mtDNA序列设计合成特异引物进行PCR扩增，首次克隆并测定了暗纹东方纯mtDNA的Cytb、COⅠ、COⅡ、COⅢ、D-loop等5个重要基因及其侧翼的8个tRNA基因。上述基因均已在GenBank登录。8个tRNA基因含有69-72个碱基。推定了各个tRNA的二级结构并进行了初步的序列分析。结果表明：8个tRNA基因具有较为典型的三叶草型结构，各臂的碱基配对率大多数较高，稳定性较好.     

暗纹东方鲀线粒体12S rRNA基因的克隆及序列分析

利用提取纯化的暗纹东方纯肝脏的mtDNA为模板,按照红鳍东方纯mtDNA序列设计合成特异引物进行PCR扩增,首次克隆并测定了暗纹东方纯mtDNA的12S rRNA基因.同时运用DNA分析软件,对暗纹东方纯与GenBank中选取的几种同目鱼类的相应序列进行比较分析,显示暗纹东方纯与这些鱼类的12S rRNA基因具有较高的同源性.利用分子生物学相关软件,对暗纹东方纯及与其同目的5种鱼的12S rRNA基因序列建立NJ和MP分子进化树,并与传统的分类地位进行比较,结果表明与传统的分类地位基本吻合.     

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．     

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.     

Molecular phylogeny and biogeography of the highly specialized grade schizothoracine fishes （Teleostei： Cyprinidae） inferred from cytochrome b sequences

We recovered the phylogenetic relationships among 23 species and subspecies of the highly special-ized grade schizothoracine fishes distributing at 36 geographical sites in the Tibetan Plateau and its surrounding regions by analyzing sequences of cytochrome b genes. Furthermore, we estimated the possible divergent times among lineages based on a historical geological isolation event in the Tibetan Plateau. The molecular data revealed that the highly specialized grade schizothoracine fishes were not a monophyletic group, but were the same as genera Gymnocypris and Schizogypsis. Our results indi-cated that the molecular phylogenetic relationships apparently reflected their geographical and historical associations with drainages, namely species from the same and adjacent drainages clustered together and had close relationships. The divergence times of different lineages were well consistent with the rapid uplift phases of the Tibetan Plateau in the late Cenozoic, suggesting that the origin and evolution of schizothoracine fishes were strongly influenced by environment changes resulting from the upheaval of the Tibetan Plateau.     

Reconsideration of the phylogenetic positions of three stichotrichous genera Holosticha, Anteholosticha and Pseudokeronopsis （Spirotrichea： Ciliophora） inferred from complete SSU rRNA gene sequences

The small subunit rRNA (SSU rRNA) gene for six marine stichotrichs, Pseudokeronopsis carnea, P. flava, Holosticha heterofoissneri, H. diademata, H. bradburyae and Anteholosticha manca, was sequenced and characterized. Using this molecular information, the phylogenetic positions of three related genera, Pseudokeronopsis, Holosticha and Anteholosticha were determined. The results indicate that the family Urostylidae is clearly separated from the family Pseudokeronopsidae. The present phylogenetic analyses unambiguously placed Pseudokeronopsis close to Holosticha, and support the conclusion that these genera should be regarded as the members of the order Urostylidae within the stichotrich clade. Furthermore, the results of this study also support the monophyly of the genus Pseudokeronopsis,the redefinition of Holosticha s. str. and the placement of Anteholosticha in a clade separate from Holosticha.     

Phylogenetic identification and microbial diversity in snow of the summit （8201 m） of Cho Oyu Mountain,Tibet

The bacterial diversity and abundance in snow of the summit （8201 m） of Cho Oyu mountain, Tibet, were analyzed by 16S rRNA gene sequencing followed by scanning electronic microscopy analysis. Most of bacteria were found to be of spherical or oval shape （〉95%）. Bacterial 16S rDNA sequences were classified into 5 genera （Caulobacter, Ralstonia, Cupriavidus, Pelomonas and Pseudomonas）. Gammaproteobacteria were the most abundant （91.25%） among the library that consists of 594 clones. The sequences found in this study are highly similar to those previously retrieved from other cold environments, such as ice core, sea ice, permafrost and snow. The results showed that the cold and barren environments strongly influence the survival of bacteria. The high similarity among sequences retrieved from snow sample and other places, such as ocean, soil and water, suggested that the bacteria in snow, soil and water environments have the same origin.     

Phylogenetic investigation on five genera of tintinnid ciliates （Ciliophora, Choreotrichia）, based on the small subunit ribosomal RNA gene sequences

The phylogenetic relationship among tintinnid ciliates are relatively poorly studied based on molecular data. In the present work, seven species belonging to five genera of the order Tintinnida (Amphorellopsis acuta, Codonellopsis nipponica, Favella taraikaensis, Stenosemella nivalis, Tintinnopsis beroidea, Tintinnopsis cylindrica and Tintinnopsis lohmanni) were analyzed using the information on their small subunit ribosomal RNA gene sequences. Phylogenetic trees were constructed using Bayesian inference (BI), maximum parsimony (MP), neighbor-joining (NJ), and least-squares (LS) methods. Generally, similar topologies were revealed with high or moderate supports, in which the main results show that (1) all tintinnids analyzed belong to a single assemblage; (2) congeners in Tintinnopsis do not cluster together, which indicates that the lorica-based definition for this genus is not consistent with the SSU rRNA phylogeny; (3) A. acuta groups with Tintinnidium mucicola but not with Eutintinnus, indicating that the traditional family Tintinnidae might be a paraphyletic group; (4) Stenosemella and Codonellopsis are clearly most related and possibly even merged into one genus regarding their similar morphology and molecular analyses, and possession of a hyaline collar is the only characteristic of the genus.     

Investigations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodontidae)

The phylogenetic relationships within the genus Takifugu Abe, 1949 (Tetraodontiformes, Tetraodonti- dae) remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this ge- nus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that (1) the genus Takifugu is a monophyletic assemblage; (2) the genus is divided into 6 subgroups based on the mo- lecular data; and (3) there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our re- sults also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

中国南方沿海13种石斑鱼类的分子系统进化关系分析

石斑鱼类(Groupers)是世界性海洋经济鱼类,也是中国南方沿海省份重要的海水增养殖对象,其种类繁多,由于缺乏明显的种间外部形态区别特征,石斑鱼的分类一直是鱼类系统分类学的一个难题,存在着较多的争议和混淆。为了从分子水平揭示石斑鱼类的系统进化关系,应用线粒体细胞色素b基因(Cyt b)及两个核糖体RNA基因(16S和12S)序列联合构建系统进化树对中国近海石斑鱼亚科3属13种进行种类鉴定及亲缘关系分析,探讨了石斑鱼类的分子系统进化。结果表明:①线粒体基因序列在石斑鱼类种间存在较大的变异,种内变异极小,是进行石斑鱼类种类鉴定较为适宜的分子标记;②所研究的3属13种石斑鱼类中,石斑鱼属与九棘鲈属的亲缘关系较近,与鳃棘鲈属的较远;③赤点石斑鱼和青石斑鱼、蜂巢石斑鱼与鲑点石斑鱼、斜带石斑鱼与棕斑石斑鱼两两间具有较近的亲缘关系。     

淡水豚类mtDNA 16S rRNA基因的系统发生

ｍｔＤＮＡ １６Ｓ ｒＲＮＡ基因我分析支持将现生淡水豚４个属归入不同的科,即白暨豚科（Ｌｉｐｏｔｉｉｄａｅ）、恒河豚科（Ｐｌａｔａｎｉｓｔｉｄａｅ）、亚河豚科（Ｉｎｉｉｄａｅ）和弗西豚科（Ｐｏｎｔｏｐｏｒｉｄａｅ）。基于邻接法的系统发生分析显示淡水豚类由白暨豚＋恒河豚和弗西豚＋亚河豚两个单系组成,但两个单系之间并无姊妹关系。淡水豚类是并系的。１６Ｓ ｒＲＮＡ基因的系统发生树与ｍｔＤＮＡ细胞色素ｂ基     

四川黑熊线粒体12S和16S rRNA基因的克隆及序列分析

利用哺乳动物线粒体基因的保守序列设计引物,采用PCR方法,首次从亚洲黑熊四川亚种（Ursus thibetanus mupinensis）的肌肉组织总DNA中扩增出了线粒体12S rRNA和16S rRNA基因并进行了序列测定及分析.结果表明,四川黑熊12S rRNA基因长965 bp;16S rRNA基因长1 580 bp.通过进一步的序列分析表明,四川黑熊的12S rRNA和16S rRNA基因有较高的进化速率,与美洲黑熊、棕熊、北极熊、眼镜熊及大熊猫的相应基因相比较有较大差异,其中与美洲黑熊的同源性相对较高.     

基于16S rRNA和POMC基因序列的中国北方林蛙属动物系统发生关系

基于16S rRNA和POMC基因部分序列研究了中国北方林蛙属动物的系统发生关系.利用ClustalX软件对序列进行对位排列后,长度为1148bp,内群变异位点259,简约位点110.利用MP法、ML法和BI法构建系统进化树,结果显示林蛙属物种构成单系群,其中东北种群聚为一支,中国林蛙聚为一支,支持东北林蛙为有效物种.桓仁林蛙和核型为2n=26的林蛙属物种聚为一支,因此不支持核型为2n=24的林蛙形成单系群.桓仁林蛙、昆嵛林蛙、黑龙江林蛙单独聚为一支,表明三者之间的亲缘关系较近.     

8种唇口目苔藓动物的分子系统发生--基于28S rRNA基因的证据

对我国沿海较为常见的8种唇口目苔藓动物的28S rRNA基因的部分序列进行了PCR扩增和序列测定。用排序软件进行序列比对后，以腕足动物为外组群，运用MEGA遗传分析软件以邻接法（NJ）和最大似然法（MP）构建了系统发生树。结果表明，有囊亚目的4种苔藓动物（刺轴拟缘孔苔虫、假缘孔苔虫、拟迷误裂孔苔虫、仿分胞苔虫）构成一个单系群；无囊亚目则是一个复系数，其中2种草苔虫（匍茎草苔虫、多室草苔虫）为单系发生，而大室膜孔苔虫和艳丽琥珀苔虫的系统发生位置尚需进一步探讨。     

中国6种白色羽鹭科鸟类的系统归属研究

对黄嘴白鹭(Egretta eulophotes)、岩鹭(E.sacra)、大白鹭(E.alba)、中白鹭(E.intermedia)、白鹭(E.garzetta)和牛背鹭(Bubulcus ibis)6种白色羽鹭科鸟类和其它8种鹭科鸟类的线粒体12S rRNA基因分析比对及建立系统树.结果显示:黄嘴白鹭和岩鹭的亲缘关系最近,它们与白鹭聚为一支,三者间遗传距离小于1.05%,应归于同一属,即白鹭属;大白鹭、中白鹭与白鹭属种类遗传距离大于9.52%,而与鹭属种类的遗传距离在4.89%～5.46%之间,可见它们与鹭属的亲缘关系更近,应从白鹭属分离出来;大白鹭、中白鹭和牛背鹭三者与鹭属种类的遗传距离都小于5.46%,亲缘关系比较接近,综合(开鸟)亚科外群的遗传距离分类标准(6.74%～7.70%),支持将大白鹭、中白鹭和牛背鹭归入鹭属的结论.