基于2种核蛋白编码基因序列研究16种对虾系统发育关系

为了研究对虾科(Penaeidae)对虾的系统发育关系,基于2种细胞核蛋白编码基因钾钠腺苷三磷酸α亚基(NaK)基因和磷酸烯醇丙酮酸激酶(PEPCK)基因,使用邻接(neighbor-jointing,NJ)法构建了9属16种对虾的系统发育树.研究结果有力地支持了1983年Burkenroad提出的三群体分类系统,除新对虾属处于进化树最先起源的位置外,系统发育树形成3个进化枝,其一为对虾属、明对虾属、滨对虾属、囊对虾属和沟对虾属,其二为仿对虾属和鹰爪虾属,其三为赤虾属,分别对应为Penaeini、Trachypenaeini和Parapenaeini 3个群体.基于核基因的系统发育树很好地解决了线粒体基因构建的系统发育树中对虾属和明对虾属进化关系不明确的问题,赤虾属的进化关系也很好地符合了形态学分类结果,因此可以作为对虾系统进化研究的重要补充手段与其他的研究方法联合应用.     

常用基因序列在摇蚊科昆虫系统发育研究中的应用进展

摇蚊科隶属于双翅目长角亚目,现行分为11亚科.本文整理了常用核基因和线粒体基因在摇蚊科系统发育研究中的应用情况:摇蚊科中的9亚科(不含智利摇蚊亚科和乌桑巴摇蚊亚科),除前寡角摇蚊亚科外,其余均为单系;直突摇蚊亚科与摇蚊亚科的亲缘关系较近,寡脉摇蚊亚科与长足摇蚊亚科为姊妹群.在摇蚊亚科中,摇蚊族(Chironomini)、长跗摇蚊族(Tanytarsini)都是单系群.常用核基因和线粒体基因在摇蚊科昆虫系统发育研究中,18SrDNA和28SrDNA适合研究从界到科所有高级阶元的系统发育关系,对于部分类群可以进行属间分析;CAD基因能较好地解决从亚科到属之间各分类阶元的系统发育关系;ITS和EF-1α可以解决种或属间的系统发育关系;COⅠ和COⅡ适合研究属、种及种以下阶元的系统发育关系,但不能很好地解决较高的阶元如族间、亚科以及科的系统发育关系;16SrDNA适用于种、属阶元水平的系统学研究,但不适用于种内;Cytb适合研究种内到种间甚至科间的系统发育关系.     

寄蝇亚科部分物种线粒体COl基因DNA条形码分类

DNA条形码分类(DNA Barcoding)是以生物线粒体DNA为工具来鉴定物种的新方法,是外来物种快速鉴定、物种分类与系统发育和进化分析的基础.研究以我国辽宁地区常见的寄蝇亚科(双翅目:寄蝇科)2族4属6种为对象,扩增和测定COI基因部分序列,对得到数据用MEGA4软件分析计算,构建该类群分子系统发育树.分析表明:...     

细胞色素b基因序列变异与东亚鲿科鱼类系统发育

采用PCR技术获得了中国（鱼尝）科鱼类线粒体DNA细胞色素b基因1138bp全序列.所得序列与GenBank中分布于日本、韩国和俄罗斯的（鱼尝）科2属9种鱼类同一基因序列排序,并选用鲇形目鲇科的大口鲇、钝头（鱼危）科的鳗尾（鱼央）和伦氏（鱼央）以及脂鲤目的断线脂鲤作外类群.分析了细胞色素b基因序列,计算了Kimura双因子遗传距离和（鱼尝）科鱼类线粒体DNA的进化速率,用最大简约法和邻接法构建了（鱼尝）科鱼类分子系统树,得出如下结论:(1)线粒体DNA序列分析显示所测定的鲇形目鱼类细胞色素b基因序列中,存在3bp的缺失;(2)系统发育分析显示（鱼尝）科构成一单系类群;（鱼艹）属为较为特化的属,拟（鱼尝）属为一明显的类群,而黄颡鱼属与（鱼危）属关系较为混乱;(3)东亚（鱼尝）科鱼类线粒体DNA的进化速率约为每百万年0.18%～0.30%.本文是中国鲇形目（鱼尝）科鱼类线粒体DNA细胞色素b基因序列的首次报道.     

细胞色素b基因序列变异与东亚 科鱼类系统发育

采用PCR技术获得了中国 科鱼类线粒体DNA细胞色素b基因1138bp全序列，所得序列与GenBank中分布于日本，韩国和俄罗斯的 科2属9种鱼类同一基因序列排序，并选用鲇形目鲇科的大口鲇，钝头Wei科的鳗尾 以及脂鲤目的断线脂鲤作外类群，分析了细胞色素b基因序列，计算了Kimura双因子遗传距离和 科鱼类线粒体DNA的进化速率，用最是约法和邻接法构建了 科鱼类分子系统树，得出如下结论：（1）线粒体DNA序列分析显示所测定的鲇形目鱼类细胞色素b基因序列中，存在3bp的缺失。（2）系统发育分析示 科构成一单系类群； 属为较为特化的属，拟 属为一明显的类群，而黄颡鱼属与Wei属关系较为混乱；（3）东亚 科鱼类线粒体DNA的进化速率约为每百万年0.18%-0.30%,本文是中国鲇形目 科鱼类线粒体DNA细胞色素b基因序列的首次报道。     

小五台山蟹蛛DNA条形码分子鉴定

为了探讨DNA条形码技术在蟹蛛科蜘蛛物种鉴定中的可行性,本研究基于线粒体COI基因序列,使用邻接法和贝叶斯法构建系统发育树,ABGD(automatic barcode gap discovery)软件对样本进行划分,对小五台山25种蟹蛛110个样本进行DNA条形码分子鉴定.结果表明:邻接法和贝叶斯法构建的系统发育树聚类结果与ABGD软件划分结果以及形态分类鉴定结果相一致.据此笔者认为DNA条形码作为一种有效的分子鉴定工具可以应用到蟹蛛科蜘蛛物种鉴定中.     

兰州鲇线粒体基因组序列结构及系统发育分析

利用GenBank中欧洲鲇的线粒体基因组序列, 设计出6对引物, 通过PCR扩增产物直接测序和引物行走(primer walking)法测定了兰州鲇的线粒体基因组序列, 并对其进行结构分析。兰州鲇线粒体基因组序列全长16524 bp, 包括37个基因(2个rRNA基因、22个tRNA基因和13个蛋白质编码基因)和1个非编码区。用最大似然法对鲇形目11种鲇鱼线粒体基因组的13个蛋白质编码基因的核苷酸序列进行系统发育分析。结果显示, 兰州鲇首先跟其他鲇科鱼类聚为一支, 形成一个单系群, 位于系统发育关系树的中部。     

14种帘蛤科贝类线粒体基因组特征与系统进化分析

利用从genbank数据库摘录了帘蛤科Veneridae的14种贝类的线粒体基因组全序列,分析其特征,并研究帘蛤科贝类分子系统进化关系。结果显示,A+T碱基含量高于G+C碱基含量,线粒体基因组表现出明显的碱基偏倚现象。对蛋白质编码基因分析,ATG并不是唯一的起始密码子,终止密码子有TAA和TAG两种;密码子组成表现出亲缘关系近的密码子组成差异较小,而亲缘关系远的差异较大。研究还发现14个物种线粒体基因排列顺序存在差异,有基因重排现象和ATP8基因缺失现象,可能是和物种生活环境有关。选取巴非蛤属的和蔼巴非蛤,真曲巴非蛤,织锦巴非蛤,波纹巴非蛤来研究帘蛤科贝类线粒体基因组选择压力,结果表明其12种蛋白质编码基因的Ka/Ks均介于0和1之间,表明基因受到纯化选择作用。基于线粒体基因组序列计算的遗传距离和构建的NJ系统发育树所得结论与传统形态分类基本一致,说明线粒体基因适合作为帘蛤科系统发育的研究手段。     

基于NCBI单基因ITS序列的散囊菌目系统发育学探讨

为了从分子水平探讨散囊菌目的系统发育关系,利用NCBI的Taxonomy,SIF等数据库中的散囊菌目物种信息及GenBank中已有的rDNA ITS序列,采用邻接法(Neighbor-Joining,NJ)构建散囊菌目属一级水平的系统发育树.结果表明:散囊菌目的4个科中的27个属聚为2大枝1小枝,单基因ITS序列的分析结果与散囊菌目的4个科的分类结果不完全一致.     

位点特异性频率模型在昆虫分子系统发育分析中的应用

根据位点特异性频率模型(site-specific rate model),利用PAUP软件,对线粒体COI基因序列不同密码子位点的数据模块分别进行了豉甲科和水生肉食亚目在亚科或科水平上的系统发育学分析,结果表明第二密码子数据模块获得的系统分析结果与形态学分析结果一致。表明位点特异性频率模型在线粒体COI基因作为标记来进行昆虫分子系统发育分析上获得了更好的效果。     

Phylogenetic positions of some genera and species of the family Buccinidae (Gastropoda: Mollusca) from China based on ribosomal RNA and COI sequences

A phylogenetic analysis of members of the family Buccinidae was conducted using 18S rRNA gene, 28S rRNA gene and the mitochondrial cytochrome oxidase Ⅰ gene. We studied 18 species of Buccinidae that belong to eight different genera and inhabit the China coastal seas. We analyzed the patterns of divergence between an outgroup and basal ingroup taxa, the monophyly of the genus Neptunea, and the position of one unnamed species within the Buccinidae. A phylogenetic tree (neighbor-joining (NJ) method) was reconstructed based on the sequences of 18S rRNA, 28S rRNA and COI, with Rapana venosa as outgroup. The NJ tree indicated that the 18 species could be divided into five groups. The genus Buccinum was monophyletic, whereas Neptunea was shown to be paraphyletic since it included Siphonalia subdilatata and Neptunea sp., a new species. This novel species otherwise clustered consistently with Neptunea cumingi in three other phylogenetic trees, showing a low genetic distance and divergence percentage of sequences belonging to the genus Neptunea. A smaller genetic distance and a smaller difference of 18S rRNA, 28S rRNA and COI sequences between Neptunea cumingii and Neptunea arthritica cumingii confirmed them to be the same species.     

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.     

Testing relative evolutionary rates and estimating divergence times among six genera of Rhizophoraceae using cpDNA and nrDNA sequences

The molecular phylogenetic trees of 10 species representing 6 genera of the family Rhizophoraceae have been constructed using the sequences of chloroplast genesmatK andrbcL as well as the ITS regions of nuclear ribosomal DNA. Relative-rate tests between lineages in these phylogenetic trees have been performed. On the basis of the results of the relative-rate tests and related molecular evolutionary rate data, the divergence times between the lineages are estimated as follows: (i) the first divergence time in these genera is 132.25 million years ago (mya); (ii) the average divergence time between two tribes, i.e. inland Legnotideae (exceptCarallia brachiata) and mangrove Rhizophoreae, is 64.13 mya; and (iii) the average divergence time between two inland species,C. garciniaefolia andC. pectinifolia, is 19.92 mya.     

基于16S rRNA序列的鳃金龟亚科5族种类的系统发育关系

测定了鳃金龟亚科部分种类的线粒体16SrRNA部分序列,运用MEGA 4.0、PAUP*4.0b10和MrBayes等软件,对5个族29个代表种的序列变异和系统关系进行了研究.序列变异分析结果显示:绢金龟族Sericini、哦鳃金龟族Hopliini、鳃金龟族Melolonthini的族内遗传距离分别为7.7%,11.3%,10.6%,族间遗传距离在12.6%～19.2%.最大似然树(ML)和贝叶斯树(BI)结果表明,所有种类分别聚集在所属的族下,哦鳃金龟族、绢金龟族分别为单系.齿爪鳃金龟属Holotrichia、云鳃金龟属Polyphylla、胸突鳃金龟属Hoplosternus、Hilyotrogus、双绺鳃金龟属Amphimallon、Hoplochelus、婆鳃金龟属Brahmina和皱鳃金龟属Trematodes聚在鳃金龟族分支下,没有形成明显的根鳃金龟族Rhizotrogini分支.齿爪鳃金龟属的非单系性与前人研究结果一致.皱鳃金龟属Trematodes和婆鳃金龟属Brahmina的代表种为姐妹种.绢金龟族的绢金龟属Serica和玛绢金龟属Maladera多数种类聚在本属分支下.证明16SrRNA序列可探讨鳃金龟亚科高级阶元的系统发育关系.     

井冈山大学绶草群落植物区系与物种多样性研究

采用样地调查方法,对分布于井冈山大学的国家Ⅱ级保护植物绶草群落的物种组成、区系成分和物种多样性进行了研究。结果表明：井冈山大学绶草群落物种组成简单,共有维管植物16科20属20种,其中蕨类植物和裸子植物均有1科1属1种,被子植物14科18属18种。井冈山大学绶草群落植物科的区系地理成分较简单,以世界分布为主,所占比例高达87.5%;属的区系地理成分较复杂,有7个分布区类型3个变型,区系成分以热带性成分占优势,温带性成分和世界分布所占比例仍较大。井冈山大学绶草群落垂直结构简单,除Pielou指数较高外,其它3种物种多样性指数均较低,群落物种丰富度、Simpson指数、Shannon-wiener指数和Pielou指数的平均值分别为6.20、0.78、2.36和0.91;10个样地间的各种多样性指数测值存在一定的差异,4种多样性指数在绶草群落中的变化趋势不一致。加强绶草自然生长环境和群落物种多样性的保护、合理开发利用是绶草的保护利用策略。     

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.     

广西5种常见星虫动物的线粒体基因组特征和系统进化分析

为探明广西北部湾星虫动物的线粒体基因组遗传变异和基因序列特征,采用高通量测序测定广西北部湾5种常见星虫动物的线粒体基因组,并对其基因序列特征、遗传变异、系统进化进行分析。结果显示,星虫动物线粒体基因组具有典型的无脊椎动物线粒体基因组的特征,基因排列高度保守,特别是其13个蛋白质编码基因（PCGs）。此外,星虫动物线粒体基因组的基因均编码在H链上,并存在3个高度保守的基因排列区块,与环节动物和螠虫动物线粒体基因组特征较为相似。cox1、cox2和cob等3个基因进化速率最慢、遗传变异水平最低,适合作为星虫动物种属系统进化研究以及不同种间生物条形码构建的分子标记。nad6、nad4、nad5和nad2等4个基因的遗传变异水平较高（大于60%）,变异位点数量较多,适合作为星虫动物种群遗传多样性研究的分子标记。星虫动物线粒体13个蛋白质编码基因的Ka/Ks比值均低于1（0.058 2-0.726 6）,其中cox1、cox3和cob等3个基因的Ka/Ks比值最低（小于0.1）,表明在星虫动物线粒体遗传进化过程中,这3个蛋白质编码基因承受强烈的自然选择压力和功能束缚。基于线粒体基因组蛋白质编码基因系统进化树的研究结果表明,星虫动物可分为方格星虫纲和革囊星虫纲两个进化分支,星虫动物与环节动物的进化地位、亲缘关系较近,而与软体动物的亲缘关系较远。本研究结果不仅为广西北部湾特色星虫动物渔业资源多样性调查和保护提供分子遗传数据,也为星虫动物系统进化研究提供了科学参考。     

Molecular identification and phylosenetic position of Cuora yunnanensis

The Yunnan box turtle （Cuora yunnanensis, Boulenger, 1906）, which has drawn much attention in conservation biology, was regarded as extinct since it was previously known only from 12 specimens collected in Yunnan, China, before 1908. Recently, live specimens have been discovered which are suggested to be C. yunnanensis. To determine whether the newly discovered specimens are really C. yunnanensis, we have established a molecular phylogeny, with a 1725-bp fragment of mitochondrial DNA, using samples from three live individuals of C. yunnanensis, together with sequence data from a museum specimen of C. yunnanensis （MNHN 1907.10） and other members of the genus Cuora. We found that the three newly discovered individuals and the old museum specimen of C. yunnanensis are very similar both in morphology and in mitochondrial DNA sequence, suggesting that the three new individuals are the very C. yunnanensis, and thus the species is not extinct. Our phylogenetic analysis also demonstrates that C. yunnanensis is not of recent hybrid origin, but rather represents a distinct evolutionary lineage.     

Phylogeny of fireflies (Coleoptera: Lampyridae) inferred from mitochondrial 16S ribosomal DNA，with references to morphological and ethological traits

We sequenced partial mitochondrial 16S ribosomal DNA (16S rDNA) of 18 firefly species from Southwest of China. Combined with homologous sequences previously reported, phylogenetic trees including Japanese, Korean and Chinese species were reconstructed by neighbor-joining, maximum parsimony and Bayesian methods. All reconstructions agree on most nodes of the trees. Monophyly of Lampyridae is not supported because Rhagophthalmus ohbai in Rhagophthalmidae is included within it. Lamprigera, a genus placed unreliably in Lampyrinae, shows a close relationship to Amydetinae. Monophyly of Luciolinae is not supported because Pristolycus sagulatus (Lampyrinae) is included within it. In the Luciolinae, monophyly of Curtos and Hotaria is well established, respectively, but both morphological and molecular data continue to indicate that Luciola is not monophyletic and its subdivision is indeed necessary. Within the Lampyrinae, both Pyrocoelia and Diaphanes are not monophyletic, but monophyly of Pyrocoelia+Diaphanes is well supported. Phylogeny of Diaphanes is discussed for the first time. Generic placement of a newly discovered species (Diaphanes pectinealis Li et Liang) sharing some characters of Pyrocoelia and Diaphanes challenges the delimitation of these two similar genera. With references to the firefly mating systems, we suggest that more emphases should be placed on those sexually selected characters such as antennal structure in taxonomy of Lampyridae.     

滇西二高温温泉中Thermus高温菌的16S rRNA

 通过对滇西二温泉纯培养和免培养法获得的Thermus菌株序列进行16SrDNA系统发育及16SrRNA二级结构比较,发现二温泉中至少存在Thermus属的2个潜在新种,它们在系统发育树上形成1个独立的地理簇群分支,它们的16SrRNAhelix10二级结构也与其它地区的Thermus属菌株不同,形成明显的地域进化特征.