凤尾蕨科植物rbcL基因的适应性进化分析

为深入理解蕨类植物辐射式物种分化的分子适应机制,在时间框架下,采用位点模型和分支-位点模型对凤尾蕨科植物rbcL基因的进化式样进行了分析.通过比较模型M1a/M2a和M7/M8,在氨基酸水平上共鉴定出6个正选择位点:1491、251M、255V、282F、359S和375F,其中位点282F对维持Rubisco功能有重要作用.分别检验凤尾蕨科的附生分支和水蕨类分支发现,前者不具适应性进化位点,而后者有两个位点(230A和247C)经历正选择.相对于荫蔽的光条件,水生生境可能对RbcL亚基的选择作用更强.另外,基于UCLD分子钟模型估算出的风尾蕨科各分支分化时间表明,该科物种丰富度的辐射式增长发生在新生代渐新世,推测古、始新世最热事件可能对物种分化的形成也产生一定作用.这对认识薄囊蕨类如何应对被子植物兴起导致的陆地生态系统改变具重要意义.     

大黄属(蓼科) 植物ｎｄｈＦ 基因的适应性进化

大黄属(Rheum L.)是蓼科(Polygonaceae)中一个高度分化的大属,广泛分布在亚洲和欧洲的高山和沙漠地区,全世界约60种,其中在青藏高原及其邻近地区发现了约40种。该属种的高度分化曾被推测是第三纪末青藏高原的快速隆升以及第四纪气候的反复变化所引发的适应性辐射导致。为进一步了解大黄属植物辐射式物种分化的分子适应机制,该研究选取34个形态上多样化的大黄属物种,利用系统发育分析软件,在时间框架下采用位点模型和分支模型对大黄属的叶绿体ndhF基因进行了适应性进化分析。结果表明：大黄属植物的分子进化系统树呈现短而平行的辐射式分支式样,显示出典型的物种快速辐射多样化特征;用位点模型检验ndhF基因是否存在经受正向选择(ω>1)时,在氨基酸水平上共鉴定出3个NDHF亚基的正选择位点(188H,465H,551L),对NDHF亚基的二级结构进行分析后发现编码的188H氨基酸位于α螺旋上。大黄属植物可能通过这些结构域的适应性进化,适应青藏高原的快速隆升以及第四纪气候的反复变化而引发的陆地生态系统改变。该研究结果可为今后对该属植物的实验分析提供首选位点。     

念珠藻属植物nifH基因的适应性进化分析

念珠藻(Nostoc)固氮过程关键在于固氮酶的催化,而固氮酶复合物中的铁蛋白(NifH)是由高度保守的nifH基因编码的,该基因是进化史上现存最古老的功能基因之一。该研究选取念珠藻属及近缘类群的nifH基因序列共40条,采用最大似然法构建系统发育树;运行PAML4.9软件,对nifH基因编码蛋白进行生物信息学分析,并使用分支模型、位点模型和分支-位点模型检测该基因的选择位点,探讨nifH基因的适应性进化特征。结果表明:(1)最大似然树显示内类群中该研究物种共分为6个分支(A、B、C、D、E和F),其中D和E是2个大的分支,每个大分支中又各包含2个特殊的小分支A、F和B、C,其中F分支包含新疆古尔班通古特沙漠采集到的9株念珠藻,A分支包含F分支及该研究测定序列的4株葛仙米,B分支包含本研究测定序列的4株地皮菜和3株未定种的念珠藻,C分支包含NCBI数据库中下载的5株念珠藻、鱼腥藻序列和本研究测定序列的1株念珠藻。(2)在所分析的3种进化模型中,仅通过分支-位点模型检测出14个统计学上显著的正选择位点,即1F、2S、3S、4T、5A、6F、7F、8I、9S、10C、17I、27Y、29D和31R位点,表明念珠藻属植物的nifH基因发生了适应性变化,分支-位点模型是研究藻类基因适应性进化较好的模型。     

丙型肝炎病毒5个基因的正选择位点研究

按照时间和亚型筛选丙型肝炎病毒5个主要基因(E1、E2、NS3、NS5A和NS5B)的正选择位点,并分析其变化规律。研究结果表明这些基因在总体上均受到纯化选择压力(平均ω<1),且大小依次是E2相似文献   

蕨类植物叶绿体rps4基因的适应性进化分析

在原核生物和植物叶绿体中,RPS4(ribosomal protein small subunit4)在核糖体30S小亚基形成起始过程中发挥重要作用;该蛋白在植物中由叶绿体rps4基因编码。为验证蕨类植物在白垩纪适应被子植物兴起而发生分化的观点,本文以23种蕨类植物为研究对象,利用分支模型、位点模型和分支位点模型对其叶绿体rps4基因进化适应性进行分析。分支模型检测到4个可能存在正选择的分支;位点模型和分支位点模型虽然没有检测出正选择位点,但是位点模型检测出了85个负选择位点。通过研究我们仅仅得出a、b两个代表水龙骨类的分支处于正选择压力下,这与水龙骨类在白垩纪发生辐射式演化的理论相一致。同时rps4基因处于强烈的负选择压力这一事实表明该基因的功能与结构已经趋于稳定。     

CVNH结构域的进化分析和选择压力检测

蓝藻抗病毒蛋白-N(Cyanovirin-N,CV-N)具有广谱抗病毒活性,其同源物构成CVNH(Cyanovirin-N homology)蛋白家族,并且家族成员的抗人类免疫缺陷病毒结构域在进化上非常保守。文章通过重建基因树对CVNH结构域的"零散分布"特点作了更为细致的了解,发现在黑曲霉、费氏曲菌、产黄青霉、粗糙脉孢霉、蓝杆藻和水蕨等物种中存在多份该结构域拷贝。在此基础上,分别采用机理式模型(Mechanistic model)和MEC模型(Mechanistic-empirical combination model)对CVNH结构域序列位点进行适应性进化分析,结果显示:1)两类模型均未检测到统计上显著的正选择位点;2)净化选择对CVNH起主导作用;3)MEC模型更适合所研究的数据。进一步使用"支-特异"模型和"支-位点"模型对蓝杆菌菌株7822和7424的祖先分支进行检测,发现该分支经历过适应性进化,并且鉴定出6个正选择位点(34L、63L、13H、76C、78K和80I)。     

蕨类植物psbD基因的适应性进化和共进化分析

D2蛋白是植物光系统Ⅱ复合体（PSⅡ）核心蛋白之一,由叶绿体psbD基因编码。为了深入理解核心薄囊蕨类植物在阴生环境下的“辐射”式演化,我们对12种蕨类植物的psbD基因进行了克隆和测序,然后联合已公布的其他8种蕨类植物的psbD序列,基于ω值（非同义替换率ds和同义替换率ds的比值）探讨了该基因经受的选择压力。发现D2蛋白在大多数分支和位点受到强烈的负选择,但是树蕨类分支的psbD进化速率低且ω值较高。借助多种模型进行的共进化分析显示,树蕨类D2蛋白的168R、245H和272M两两组成具有共进化关系的氨基酸位点对。     

裸子植物中光敏色素PHY-PAS1 结构域的适应性进化

光敏色素是一类红光/远红光受体, 在植物种子萌发到成熟的整个生长发育过程中均起重要的调节作用。光敏色素PHYPAS1 结构域存在于光敏色素基因家族的所有成员中, 对调节发色团的光谱特性和光信号转导非常关键。光敏色素基因家族通过基因重复产生, 而基因重复可能与物种形成有关。PHYP基因是裸子植物光敏色素基因家族发生第1次重复后产生的, 并且以单拷贝形式存在。为了研究不同裸子植物PHYP基因编码蛋白的PHY-PAS1结构域在进化过程中是否受到相同的选择压力以及是否发生了适应性进化, 该研究利用分支模型、位点模型以及分支-位点模型对裸子植物31条PHYP基因序列编码蛋白的PHY-PAS1结构域所受到的选择压力进行了分析。结果表明, 在由PHY-PAS1结构域序列构建的系统树中, 多数分支处于强烈的负选择压力下 (w<1); 有14个分支处于正选择压力下 (w>1), 其中13个分支发生在属内种间; 与之相比, 在较为古老的谱系中相对缺少这种正选择压力。     

CVNH 结构域进化分析及选择压力检测

蓝藻抗病毒蛋白-N(Cyanovirin-N,CV-N)具有广谱抗病毒活性;其同源物构成 CVNH 蛋白家族,并且家族成员的抗人类免疫缺陷病毒结构域在进化上非常保守。本研究通过重建基因树对 CVNH 结构域的“零散分布”特点作了更为细致的了解,发现在黑曲霉、费氏曲菌、产黄青霉、粗糙脉孢霉、蓝杆藻和水蕨等物种中该结构域存在多份拷贝。在此基础上,分别采用机理式模型和 MEC 模型对 CVNH 结构域序列位点进行适应性进化分析,结果显示：（1）两类模型均未检测到统计上显著的正选择位点;（2）净化选择对 CVNH 起主导作用;（3）MEC 模型更适合所研究数据。进一步使用“支-特异”模型和“支-位点”模型对蓝杆菌菌株7822和7424的祖先分支进行检测,发现该分支经历过适应性进化,并且鉴定出6 个正选择位点(34L、63L、13H、76C、78K 和 80I)。这些结果对后续的 CVNH 功能验证和借助基因工程手段改良蛋白的抗病毒活性具重要意义。     

裸子植物中光敏色素PHY-PAS1结构域的适应性进化

光敏色素是一类红光／远红光受体,在植物种子萌发到成熟的整个生长发育过程中均起重要的调节作用。光敏色素PHY-PAS1结构域存在于光敏色素基因家族的所有成员中,对调节发色团的光谱特性和光信号转导非常关键。光敏色素基因家族通过基因重复产生,而基因重复可能与物种形成有关。PHYP基因是裸子植物光敏色素基因家族发生第1次重复后产生的,并且以单拷贝形式存在。为了研究不同裸子植物PHYP基因编码蛋白的PHY-PAS1结构域在进化过程中是否受到相同的选择压力以及是否发生了适应性进化,该研究利用分支模型、位点模型以及分支．位点模型对裸子植物31条PHYP基因序列编码蛋白的PHY-PAS1结构域所受到的选择压力进行了分析。结果表明,在由PHY-PAS1结构域序列构建的系统树中,多数分支处于强烈的负选择压力下（ω〈1）：有14个分支处于正选择压力下（ω〉1）,其中13个分支发生在属内种间;与之相比,在较为古老的谱系中相对缺少这种正选择压力。     

Assessing allometric models to predict vegetative growth of mango (Mangifera indica; Anacardiaceae) at the current-year branch scale

? Premise of the study: Accurate and reliable predictive models are necessary to estimate nondestructively key variables for plant growth studies such as leaf area and leaf, stem, and total biomass. Predictive models are lacking at the current-year branch scale despite the importance of this scale in plant science. ? Methods: We calibrated allometric models to estimate leaf area and stem and branch (leaves + stem) mass of current-year branches, i.e., branches several months old studied at the end of the vegetative growth season, of four mango cultivars on the basis of their basal cross-sectional area. The effects of year, site, and cultivar were tested. Models were validated with independent data and prediction accuracy was evaluated with the appropriate statistics. ? Key results: Models revealed a positive allometry between dependent and independent variables, whose y-intercept but not the slope, was affected by the cultivar. The effects of year and site were negligible. For each branch characteristic, cultivar-specific models were more accurate than common models built with pooled data from the four cultivars. Prediction quality was satisfactory but with data dispersion around the models, particularly for large values. ? Conclusions: Leaf area and stem and branch mass of mango current-year branches could be satisfactorily estimated on the basis of branch basal cross-sectional area with cultivar-specific allometric models. The results suggested that, in addition to the heteroscedastic behavior of the variables studied, model accuracy was probably related to the functional plasticity of branches in relation to the light environment and/or to the number of growth units composing the branches.     

Evaluation of an improved branch-site likelihood method for detecting positive selection at the molecular level

Detecting positive Darwinian selection at the DNA sequence level has been a subject of considerable interest. However, positive selection is difficult to detect because it often operates episodically on a few amino acid sites, and the signal may be masked by negative selection. Several methods have been developed to test positive selection that acts on given branches (branch methods) or on a subset of sites (site methods). Recently, Yang, Z., and R. Nielsen (2002. Codon-substitution models for detecting molecular adaptation at individual sites along specific lineages. Mol. Biol. Evol. 19:908-917) developed likelihood ratio tests (LRTs) based on branch-site models to detect positive selection that affects a small number of sites along prespecified lineages. However, computer simulations suggested that the tests were sensitive to the model assumptions and were unable to distinguish between relaxation of selective constraint and positive selection (Zhang, J. 2004. Frequent false detection of positive selection by the likelihood method with branch-site models. Mol. Biol. Evol. 21:1332-1339). Here, we describe a modified branch-site model and use it to construct two LRTs, called branch-site tests 1 and 2. We applied the new tests to reanalyze several real data sets and used computer simulation to examine the performance of the two tests by examining their false-positive rate, power, and robustness. We found that test 1 was unable to distinguish relaxed constraint from positive selection affecting the lineages of interest, while test 2 had acceptable false-positive rates and appeared robust against violations of model assumptions. As test 2 is a direct test of positive selection on the lineages of interest, it is referred to as the branch-site test of positive selection and is recommended for use in real data analysis. The test appeared conservative overall, but exhibited better power in detecting positive selection than the branch-based test. Bayes empirical Bayes identification of amino acid sites under positive selection along the foreground branches was found to be reliable, but lacked power.     

Kakusan4 and Aminosan: two programs for comparing nonpartitioned, proportional and separate models for combined molecular phylogenetic analyses of multilocus sequence data

Proportional and separate models able to apply different combination of substitution rate matrix (SRM) and among-site rate variation model (ASRVM) to each locus are frequently used in phylogenetic studies of multilocus data. A proportional model assumes that branch lengths are proportional among partitions and a separate model assumes that each partition has an independent set of branch lengths. However, the selection from among nonpartitioned (i.e., a common combination of models is applied to all-loci concatenated sequences), proportional and separate models is usually based on the researcher's preference rather than on any information criteria. This study describes two programs, 'Kakusan4' (for DNA sequences) and 'Aminosan' (for amino-acid sequences), which allow the selection of evolutionary models based on several types of information criteria. The programs can handle both multilocus and single-locus data, in addition to providing an easy-to-use wizard interface and a noninteractive command line interface. In the case of multilocus data, SRMs and ASRVMs are compared at each locus and at all-loci concatenated sequences, after which nonpartitioned, proportional and separate models are compared based on information criteria. The programs also provide model configuration files for mrbayes, paup*, phyml, raxml and Treefinder to support further phylogenetic analysis using a selected model. When likelihoods are optimized by Treefinder, the best-fit models were found to differ depending on the data set. Furthermore, differences in the information criteria among nonpartitioned, proportional and separate models were much larger than those among the nonpartitioned models. These findings suggest that selecting from nonpartitioned, proportional and separate models results in a better phylogenetic tree. Kakusan4 and Aminosan are available at http://www.fifthdimension.jp/. They are licensed under gnugpl Ver.2, and are able to run on Windows, MacOS X and Linux.     

Long branch effects distort maximum likelihood phylogenies in simulations despite selection of the correct model

The aim of our study was to test the robustness and efficiency of maximum likelihood with respect to different long branch effects on multiple-taxon trees. We simulated data of different alignment lengths under two different 11-taxon trees and a broad range of different branch length conditions. The data were analyzed with the true model parameters as well as with estimated and incorrect assumptions about among-site rate variation. If length differences between connected branches strongly increase, tree inference with the correct likelihood model assumptions can fail. We found that incorporating invariant sites together with Γ distributed site rates in the tree reconstruction (Γ+I) increases the robustness of maximum likelihood in comparison with models using only Γ. The results show that for some topologies and branch lengths the reconstruction success of maximum likelihood under the correct model is still low for alignments with a length of 100,000 base positions. Altogether, the high confidence that is put in maximum likelihood trees is not always justified under certain tree shapes even if alignment lengths reach 100,000 base positions.     

Multiple hypothesis testing to detect lineages under positive selection that affects only a few sites

Detection of positive Darwinian selection has become ever more important with the rapid growth of genomic data sets. Recent branch-site models of codon substitution account for variation of selective pressure over branches on the tree and across sites in the sequence and provide a means to detect short episodes of molecular adaptation affecting just a few sites. In likelihood ratio tests based on such models, the branches to be tested for positive selection have to be specified a priori. In the absence of a biological hypothesis to designate so-called foreground branches, one may test many branches, but a correction for multiple testing becomes necessary. In this paper, we employ computer simulation to evaluate the performance of 6 multiple test correction procedures when the branch-site models are used to test every branch on the phylogeny for positive selection. Four of the methods control the familywise error rates (FWERs), whereas the other 2 control the false discovery rate (FDR). We found that all correction procedures achieved acceptable FWER except for extremely divergent sequences and serious model violations, when the test may become unreliable. The power of the test to detect positive selection is influenced by the strength of selection and the sequence divergence, with the highest power observed at intermediate divergences. The 4 correction procedures that control the FWER had similar power. We recommend Rom's procedure for its slightly higher power, but the simple Bonferroni correction is useable as well. The 2 correction procedures that control the FDR had slightly more power and also higher FWER. We demonstrate the multiple test procedures by analyzing gene sequences from the extracellular domain of the cluster of differentiation 2 (CD2) gene from 10 mammalian species. Both our simulation and real data analysis suggest that the multiple test procedures are useful when multiple branches have to be tested on the same data set.     

The Arp2/3 complex nucleates actin filament branches from the sides of pre-existing filaments

Regulated assembly of actin-filament networks provides the mechanical force that pushes forward the leading edge of motile eukaryotic cells and intracellular pathogenic bacteria and viruses. When activated by binding to actin filaments and to the WA domain of Wiskott-Aldrich-syndrome protein (WASP)/Scar proteins, the Arp2/3 complex nucleates new filaments that grow from their barbed ends. The Arp2/3 complex binds to the sides and pointed ends of actin filaments, localizes to distinctive 70 degrees actin-filament branches present in lamellae, and forms similar branches in vitro. These observations have given rise to the dendritic nucleation model for actin-network assembly, in which the Arp2/3 complex initiates branches on the sides of older filaments. Recently, however, an alternative mechanism for branch formation has been proposed. In the 'barbed-end nucleation' model, the Arp2/3 complex binds to the free barbed end of a filament and two filaments subsequently grow from the branch. Here we report the use of kinetic and microscopic experiments to distinguish between these models. Our results indicate that the activated Arp2/3 complex preferentially nucleates filament branches directly on the sides of pre-existing filaments.     

Tree Branching: Leonardo da Vinci's Rule versus Biomechanical Models

This study examined Leonardo da Vinci''s rule (i.e., the sum of the cross-sectional area of all tree branches above a branching point at any height is equal to the cross-sectional area of the trunk or the branch immediately below the branching point) using simulations based on two biomechanical models: the uniform stress and elastic similarity models. Model calculations of the daughter/mother ratio (i.e., the ratio of the total cross-sectional area of the daughter branches to the cross-sectional area of the mother branch at the branching point) showed that both biomechanical models agreed with da Vinci''s rule when the branching angles of daughter branches and the weights of lateral daughter branches were small; however, the models deviated from da Vinci''s rule as the weights and/or the branching angles of lateral daughter branches increased. The calculated values of the two models were largely similar but differed in some ways. Field measurements of Fagus crenata and Abies homolepis also fit this trend, wherein models deviated from da Vinci''s rule with increasing relative weights of lateral daughter branches. However, this deviation was small for a branching pattern in nature, where empirical measurements were taken under realistic measurement conditions; thus, da Vinci''s rule did not critically contradict the biomechanical models in the case of real branching patterns, though the model calculations described the contradiction between da Vinci''s rule and the biomechanical models. The field data for Fagus crenata fit the uniform stress model best, indicating that stress uniformity is the key constraint of branch morphology in Fagus crenata rather than elastic similarity or da Vinci''s rule. On the other hand, mechanical constraints are not necessarily significant in the morphology of Abies homolepis branches, depending on the number of daughter branches. Rather, these branches were often in agreement with da Vinci''s rule.     

Additive Uncorrelated Relaxed Clock Models for the Dating of Genomic Epidemiology Phylogenies

Phylogenetic dating is one of the most powerful and commonly used methods of drawing epidemiological interpretations from pathogen genomic data. Building such trees requires considering a molecular clock model which represents the rate at which substitutions accumulate on genomes. When the molecular clock rate is constant throughout the tree then the clock is said to be strict, but this is often not an acceptable assumption. Alternatively, relaxed clock models consider variations in the clock rate, often based on a distribution of rates for each branch. However, we show here that the distributions of rates across branches in commonly used relaxed clock models are incompatible with the biological expectation that the sum of the numbers of substitutions on two neighboring branches should be distributed as the substitution number on a single branch of equivalent length. We call this expectation the additivity property. We further show how assumptions of commonly used relaxed clock models can lead to estimates of evolutionary rates and dates with low precision and biased confidence intervals. We therefore propose a new additive relaxed clock model where the additivity property is satisfied. We illustrate the use of our new additive relaxed clock model on a range of simulated and real data sets, and we show that using this new model leads to more accurate estimates of mean evolutionary rates and ancestral dates.     

Nutritional resources as positional information for morphogenesis in the stony coral Stylophora pistillata

We are interested in deciphering the mechanisms for morphogenesis in the Red Sea scleractinian coral Stylophora pistillata with the help of mathematical models. Previous mathematical models for coral morphogenesis assume that skeletal growth is proportional to the amount of locally available energetic resources like diffusible nutrients and photosynthetic products. We introduce a new model which includes factors like dissolved nutrients and photosynthates, but these resources do not serve as building blocks for growth but rather provide some kind of positional information for coral morphogenesis. Depending on this positional information side branches are generated, splittings of branches take place and branch growth direction is determined. The model results are supported by quantitative comparisons with experimental data obtained from young coral colonies.