Phylogenetic Diversity of Sequences of Cyanophage Photosynthetic Gene psbA in Marine and Freshwaters

Many cyanophage isolates which infect the marine cyanobacteria Synechococcus spp. and Prochlorococcus spp. contain a gene homologous to psbA, which codes for the D1 protein involved in photosynthesis. In the present study, cyanophage psbA gene fragments were readily amplified from freshwater and marine samples, confirming their widespread occurrence in aquatic communities. Phylogenetic analyses demonstrated that sequences from freshwaters have an evolutionary history that is distinct from that of their marine counterparts. Similarly, sequences from cyanophages infecting Prochlorococcus and Synechococcus spp. were readily discriminated, as were sequences from podoviruses and myoviruses. Viral psbA sequences from the same geographic origins clustered within different clades. For example, cyanophage psbA sequences from the Arctic Ocean fell within the Synechococcus as well as Prochlorococcus phage groups. Moreover, as psbA sequences are not confined to a single family of phages, they provide an additional genetic marker that can be used to explore the diversity and evolutionary history of cyanophages in aquatic environments.     

系统发育分析指示细菌向Apicoplast的水平基因转移

顶复合器门的原生动物(Apicomplexan protozoa)含有一个高度退化的质体样(plastid-like)细胞器,定名为apicoplast。来自apicoplast的c／pC基因和它在其他质体和细菌中的同源物用来重建apicoplast的系统发育史。使用邻接法(Neighbor-Joining)、最小进化法(Minimum Evolution)、最大简约法(Maximum Pamimony)和最大似然法(Maximum Likelihood)建立进化树。此外为了避免由于序列之间相似的碱基组成而引起的虚假聚类,建立了基于LogDet距离的核苷酸NJ树;以及为了避免由于在核苷酸和氨基酸水平上的突变饱和而引起的长分枝吸引(Long Branch Attraction,LBA),建立了基于非饱和位点的核苷酸和氨基酸序列的系统发育树。重建结果强有力地支持apicoplast和细菌B．buigorferi之间的单系(monophyly)起源关系,也强化了apicoplasst属于混合基因组的假设,并且提供了对这个混合基因组起源的新的认识。     

DNA聚合酶x家族的系统发育分析

随着DNA聚合酶x家族成员数量的增加,家族内部的系统发育需要重新检查,来自病毒和细胞的DNA聚合酶x家族成员顺序第一次被汇编在一起,进行系统发育分析。分析显示：真核生物DNA聚合酶beta(polβ)可能起源于病毒基因的水平转移;DNA聚合酶mu(polμ)基因仅存在于哺乳动物基因中,是脱氧核苷酸末端转移酶（TdT）的重复基因;DNA聚合酶lambda(polλ)可能是polμ和TdT的祖先基因,但在某些物种的进化过程中发生了基因丢失。     

Phylogenetic Diversity of Marine Cyanophage Isolates and Natural Virus Communities as Revealed by Sequences of Viral Capsid Assembly Protein Gene g20

In order to characterize the genetic diversity and phylogenetic affiliations of marine cyanophage isolates and natural cyanophage assemblages, oligonucleotide primers CPS1 and CPS8 were designed to specifically amplify ca. 592-bp fragments of the gene for viral capsid assembly protein g20. Phylogenetic analysis of isolated cyanophages revealed that the marine cyanophages were highly diverse yet more closely related to each other than to enteric coliphage T4. Genetically related marine cyanophage isolates were widely distributed without significant geographic segregation (i.e., no correlation between genetic variation and geographic distance). Cloning and sequencing analysis of six natural virus concentrates from estuarine and oligotrophic offshore environments revealed nine phylogenetic groups in a total of 114 different g20 homologs, with up to six clusters and 29 genotypes encountered in a single sample. The composition and structure of natural cyanophage communities in the estuary and open-ocean samples were different from each other, with unique phylogenetic clusters found for each environment. Changes in clonal diversity were also observed from the surface waters to the deep chlorophyll maximum layer in the open ocean. Only three clusters contained known cyanophage isolates, while the identities of the other six clusters remain unknown. Whether or not these unidentified groups are composed of bacteriophages that infect different Synechococcus groups or other closely related cyanobacteria remains to be determined. The high genetic diversity of marine cyanophage assemblages revealed by the g20 sequences suggests that marine viruses can potentially play important roles in regulating microbial genetic diversity.     

绵羊Cytb基因序列多态性及系统进化研究

以8个中国地方绵羊品种和1个外来品种的20个个体为研究对象,通过对Cytb基因的全序列测定,结果表明：绵羊的单倍型多样性为97．1％。所有序列的平均碱基组成为27．1％T,28．5％C,31．4％A及13．0％G,G＋C含量为41．5％,核苷酸多样性为0．602％。在所有序列中共检测到43个变异位点,其中包括40处转换和3处颠换。Fu’S中性检验表明差异不显著（0．10〉P〉0．05）,说明绵羊群体未发生群体扩张事件。NJ、ME及UPGMA聚类结果均表明,我国绵羊可分为3个单倍型组,这提示我国绵羊有3个母系起源。     

基于线粒体Cytb基因分析高原鳅种群遗传多样性与系统发育关系

以高原鳅(Triplophysa)属鱼类8个种群共75个个体的线粒体Cytb基因为标记,通过生物信息学分析高原鳅属种群遗传多样性和系统发育关系.结果表明,全长1 140bp的Cytb基因序列中共检测到527个多态位点,界定了61个单倍型,A+T含量(55.9％)高于G+C含量(44.2％).8个种群单倍型多样度(Hd)...     

基于孕激素受体基因的类人猿亚目的分子系统学研究

通过对类人猿亚目中部分种类的孕激素受体基因进行分析,重建类人猿亚目的 系统发育关系.扩增并测定了来源于14个属的类人猿亚目物种的孕激素受体编码区序列,并基于这一序列数据,分别采用邻接法、最大简约法和最大似然法重建了系统发育关系.除了阔鼻下目,3种方法构建的系统发生树的拓扑结构类似且各节点支持率高.重建的人猿超科和猴超科内部亲缘关系支持多数人所认可的分类系统.本研究为黑猩猩和人的姐妹群关系提供了证据,提示黑猩猩比大猩猩或其他猿猴更接近人类.阔鼻下目中蜘蛛猴科、卷尾猴科和僧面猴科三者之间的系统发育关系在本研究中未得到很好辨析.     

DNA聚合酶X家族系统发育树的重建

随着DNA聚合酶X家族成员数量的增加,特别是两个昆虫痘病毒(entomopoxvirus,EPV)成员的加入,家族内部的系统发育关系需要重新检查。总共37个来自不同物种的DNA聚合酶x家族成员的核心结构域序列被分析。结果显示：系统发育树呈现的家族内部系统发育关系基本上与家族成员的物种分布状态相吻合,其中,病毒成员与真核生物DNA聚合酶beta(polβ)亚群构成姐妹群,提示病毒成员可能起源于真核细胞的相应基因。亚群的歧异时间、基因结构比较和同线基因保守性分析显示,哺乳动物DNA聚合酶基因(Pol M)可能起源于脱氧核苷酸末端转移酶基因(TdT)最近发生的基因重复。     

金粉蕨属的分子系统学研究——基于5种叶绿体DNA序列片段

金粉蕨属（Onychium Kaulfuss）隶属广义凤尾蕨科中的凤尾蕨亚科。迄今为止,该属属下分组及种间界定等仍有诸多问题亟待解决。本研究选取5个叶绿体DNA 序列片段 （rbcL/atpA/matK/trnL-trnF/trnG-trnR）,采用最大似然法（ML）和贝叶斯法（BI）构建金粉蕨属的系统发育树。结果表明：（1）金粉蕨属的9个成员被分置于两大支上。其中野雉尾金粉蕨（Onychium japonicum(Thunberg) Kunze）、西藏金粉蕨（O.tibeticum Ching & S.K.Wu）、木坪金粉蕨（O.moupinense Ching）、湖北金粉蕨（O.moupinense var. ipii(Ching) K.H.Shing）、栗柄金粉蕨（O.japonicum var. lucidum(D.Don) Christ）、黑足金粉蕨（O.cryptogrammoides Christ）、繁羽金粉蕨（O.plumosum Ching）聚为一支;而金粉蕨（O.siliculosum(Desvaux) C.Christensen）和蚀盖金粉蕨（O.tenuifrons Ching）则聚为另一支,可为该属的属下分组提供分子系统学证据;（2）野雉尾金粉蕨与栗柄金粉蕨在系统树中并没有聚在一起,而是被其它类群分割开来,不支持将后者作为野雉尾金粉蕨的变种,建议将栗柄金粉蕨提升为种的等级;（3）系统树上木坪金粉蕨与湖北金粉蕨的样本聚在一个细支上,支持《中国植物志》将湖北金粉蕨作为木坪金粉蕨变种的分类处理;（4）西藏金粉蕨与野雉尾金粉蕨聚在一起,并得到较高的支持,说明两者的关系近缘。     

Analysis of the Genetic Diversity and the Phylogenetic Evolution of Chinese Sheep Based on Cyt b Gene Sequences

The complete sequences of Cyt b gene from 20 individuals belonging to eight Chinese indigenous sheep breeds and one foreign breed were studied. The results showed that the hapolotype diversity of Chinese sheep breeds was 97.1%. The mean nucleotide composition of all the sequences was 27.1% T, 28.5% C, 31.4% A, and 13.0% G. The nucleotide diversity was 0.602%. A total of 43 mutation sites were detected, including 40 transitions and 3 transversions. Fu's test of selective neutrality showed that the sheep populations had no population demographic expansion (0.10 > P > 0.05). The different clustering methods, namely neighbor-joining, minimum evolution, and unweighted pair group method with arithmetic means, all showed a similar result, which indicated that Chinese local sheep had three maternal resources.     

Community-Level Analysis of psbA Gene Sequences and Irgarol Tolerance in Marine Periphyton

This study analyzes psbA gene sequences, predicted D1 protein sequences, species relative abundance, and pollution-induced community tolerance in marine periphyton communities exposed to the antifouling compound Irgarol 1051. The mechanism of action of Irgarol is the inhibition of photosynthetic electron transport at photosystem II by binding to the D1 protein. The metagenome of the communities was used to produce clone libraries containing fragments of the psbA gene encoding the D1 protein. Community tolerance was quantified with a short-term test for the inhibition of photosynthesis. The communities were established in a continuous flow of natural seawater through microcosms with or without added Irgarol. The selection pressure from Irgarol resulted in an altered species composition and an inducted community tolerance to Irgarol. Moreover, there was a very high diversity in the psbA gene sequences in the periphyton, and the composition of psbA and D1 fragments within the communities was dramatically altered by increased Irgarol exposure. Even though tolerance to this type of compound in land plants often depends on a single amino acid substitution (Ser₂₆₄→Gly) in the D1 protein, this was not the case for marine periphyton species. Instead, the tolerance mechanism likely involves increased degradation of D1. When we compared sequences from low and high Irgarol exposure, differences in nonconserved amino acids were found only in the so-called PEST region of D1, which is involved in regulating its degradation. Our results suggest that environmental contamination with Irgarol has led to selection for high-turnover D1 proteins in marine periphyton communities at the west coast of Sweden.     

两种缘毛类纤毛虫胞质Hsp70基因序列的克隆及其系统发育分析

克隆得到2种缘毛类纤毛虫——钟形钟虫(Vorticella campanula)和螅状独缩虫(Carchesium polypinum)的胞质Hsp70基因部分序列,长度均为438bp,编码146个氨基酸。以细菌为外类群,利用最大似然法和邻接法构建包括其他5种纤毛虫在内的共26个物种的Hsp70基因氨基酸序列系统发育树,其拓扑结构显示：V．campanula和C．polypinum聚在一起,并与另2种寡膜纲的嗜热四膜虫(Tetrahymena thermophila)及草履虫(Paramecium tetraurelia)聚为姊妹枝,提示了缘毛类纤毛虫为单系,且隶属于寡膜纲的系统发育地位。     

DNA 序列在蕨类分子系统学研究中的应用

在分子系统学研究中, 目的基因或者基因片段的选择是最关键的一步, 由于进化速率的差异, 不同的DNA序列适用于不同分类阶元的系统发育研究。本文综述了目前蕨类分子系统发育研究中常用的DNA序列分析, 它们分别来自叶绿体基因组、核基因组和线粒体基因组, 着重阐明叶绿体基因在蕨类分子系统学研究中的应用。本文还简要介绍了分子系统学研究中常见的问题及解决方法(如内类群和外类群的选择, 适宜DNA片段的选择策略), 总结了目前蕨类植物分子系统学研究所取得的进展和研究现状, 展望了当今国际蕨类分子系统学的研究趋势。     

DNA序列在植物系统学研究中的应用

植物DNA序列由于进化速率上的差异而适用于不同分类阶元的系统发育研究,因此,针对某一特定的系统学问题选择相应合适的分子片段是分子系统学研究中最为关键的一步。在前人研究的基础上,主要讨论了目前分子系统发育和进化研究中一些常用的DNA序列的适用范围,包括nrDNA的18S基因及ITS等非编码区,cpDNA的编码基因（rbcI、matK、ndhF、atpB）及非编码区序列（rpL16、rpoC1、rps16、trnL-F和trnT-L）和应用较少的mtDNA。研究表明,18S、rbcI等编码基因及mtDNA一般适用于较高分类阶元甚至整个种子植物谱系间的系统发育的探讨,而ITS及cpDNA的非编码区序列等因其较快的进化速率多用于较低分类阶元的系统关系研究。     

碎米蕨属的系统学研究——基于4种叶绿体DNA序列片段

碎米蕨属(Cheilosoria Trevis.)隶属碎米蕨类, 由于形态上的趋同进化, 使得该类群的系统分类一直存在争议。该研究对该碎米蕨属植物的4种叶绿体DNA序列片段(rbcL/matK/rps4/rps4-trnS)进行PCR扩增和序列分析, 再结合其它相关类群, 用贝叶斯法和最大似然法构建系统树并探讨其系统发育关系。结果表明, 碎米蕨属不是一个单系类群, 旧世界分布的碎米蕨属植物(薄叶碎米蕨除外)均聚在亚洲Cheilanthes群内, 与粉背蕨属(Aleuritopteris Fée)等类群形成不同亚支。该属孢子形态具有明显异质性。薄叶碎米蕨(Cheilosoria tenuifolia (Burm. f.) Trevis.)与亚洲其它碎米蕨属植物的系统位置相距甚远, 且与隐囊蕨(Notholaena hirsuta (Poir.) Desv.)聚为完全支持的分支, 两者可能均为大洋洲起源, 并属于另一类群。美洲和旧世界分布的碎米蕨属植物关系较远, 二者可能代表了不同的演化路线。     

DNA序列在蕨类分子系统学研究中的应用

在分子系统学研究中,目的基因或者基因片段的选择是最关键的一步,由于进化速率的差异,不同的DNA序列适用于不同分类阶元的系统发育研究.本文综述了目前蕨类分子系统发育研究中常用的DNA序列分析,它们分别来自叶绿体基因组、核基因组和线粒体基因组,着重阐明叶绿体基因在蕨类分子系统学研究中的应用.本文还简要介绍了分子系统学研究中常见的问题及解决方法(如内类群和外类群的选择.适宜DNA片段的选择策略),总结了目前蕨类植物分子系统学研究所取得的进展和研究现状,展望了当今国际蕨类分子系统学的研究趋势.     

利用CoI基因序列对雀科鸟类的分子系统发育关系初探

基于线粒体DNA(mtDNA)中Col基因的部分序列(1300bp)对雀形目雀科(Fringillidae)36种鸟类进行系统发育分析.对数据集构建NJ树、Baycs树和ML树.对建树结果进行分析,发现铁爪鸦(Calcarius lapponicus)与鸦属(Emberiza)鸟类的亲缘关系比其他雀科的鸟类更近;支持蓝鹀(Latoucheornis siemsseni)隶属于鹀属的观点:证实了黄颈拟蜡嘴雀(Mycerobas affinis)与黑尾蜡嘴雀(Eophona migratorius)之间紧密的亲缘关系;发现长尾雀(Uragus sibiricus)和朱鸦(Urocynchramus pylzowi)之间亲缘关系很远,而与朱雀属(Carpodacus)有较近的亲缘关系:结果支持雀类与鸦类的亚科级分类水平.     

Evolutionary Relationship of DNA Sequences in Finite Populations

With the aim of analyzing and interpreting data on DNA polymorphism obtained by DNA sequencing or restriction enzyme technique, a mathematical theory on the expected evolutionary relationship among DNA sequences (nucleons) sampled is developed under the assumption that the evolutionary change of nucleons is determined solely by mutation and random genetic drift. The statistical property of the number of nucleotide differences between randomly chosen nucleons and that of heterozygosity or nucleon diversity is investigated using this theory. These studies indicate that the estimates of the average number of nucleotide differences and nucleon diversity have a large variance, and a large part of this variance is due to stochastic factors. Therefore, increasing sample size does not help reduce the variance significantly. The distribution of sample allele (nucleomorph) frequencies is also studied, and it is shown that a small number of samples are sufficient in order to know the distribution pattern.     

十字花科植物CYP86MF基因同源序列的克隆与进化分析

为了从分子水平阐明十字花科植物间的亲缘进化关系,给植物种质资源的创建提供理论依据,试验根据课题组已报道的CYP86MF基因编码的氨基酸保守区域设计特异引物,运用PCR技术分别从十字花科6个属11个物种中分离克隆到了CYP86MF基因的同源序列,经比较分析,结果表明：这些同源序列的相似性达80%以上,所推导的氨基酸序列相似性达70%以上,且两者种间差异分别为1.0% ~ 5.7%和2.6% ~ 7.3%,属间差异分别是5.6% ~ 22.5%和7.3% ~ 31.2%;由氨基酸序列构建的分子系统树可知,在亲缘进化关系上芸薹属与萝卜属较近,其他依次为蔊菜属、拟南芥属、荠菜属,而与诸葛菜属最远。因此,CYP86MF基因的核苷酸及其可能编码的氨基酸序列差异属间较种间大,它可用于属间的分类等级研究,而难用于属以下的分类等级研究。