基因树冲突与系统发育基因组学研究

随着越来越多的基因序列被运用于系统发育重建中,随之产生的基因树冲突已成为分子系统发育研究中日益突出的问题.因此,在分子系统发育研究中,应正确理解基因树和物种树之间的差异,充分注意和分析基因树冲突的原因,正确解释分子系统发育的结果.本文通过一些典型实例分析了在多基因系统发育研究中引发基因树冲突的三类主要原因:随机误差、系统误差和生物学因素.在此基础上,对近年来兴起的系统发育基因组学进行了介绍,并以稻属Oryza研究为例,阐述了系统发育基因组学方法在解决基因树冲突以及系统发育研究中的优势和应用价值,并进一步探讨了解决基因树冲突的策略和方法,以期为分子系统发育研究提供一些肩示和帮助.     

常用系统发育树构建算法和软件鸟瞰

系统发育树又称进化树、生命树等,在达尔文的"进化论"一书中首次出现,之后系统发育树的重构被广大生物学家所接受。该文阐述了构建系统发育树的基本流程,对目前用于构建系统发育树的四类算法(距离法、最大简约法、最大似然法和贝叶斯法)进行了详细地分析和比较,并介绍了一些常用系统发育树构建和分析软件(PHYLIP、MEGA、MrBayes)的特点。     

全长rDNA簇在蝽类昆虫生命树构建中的应用前景(昆虫纲,半翅目)

在昆虫纲中,生命树计划正在以目级阶元中的分类单元为单位逐步推进.针对这一大的背景,以及高级阶元和种级阶元分子系统学研究间脱节现状,提出以rDNA簇为一组分子标记、并且在未来高级阶元系统发育研究中将目前选取少量代表类群的做法改为将尽量多的物种包含到一棵系统发育树中的建议.其中首先简要介绍了rDNA簇的结构及其中各分子标记在分子系统学研究中的应用价值,随后以蝽类昆虫为例,有针对性地总结了rDNA簇中不同基因序列已有数据的丰富程度及其在分子系统学研究中的应用情况.首次给出了蝽类昆虫28S rRNA近全长序列的二级结构模型.基于对18S rRNA和28S rRNA二级结构研究所积累的认识,强调了18S rDNA和28S rDNA内部不同区段之间变异模式和应用价值的差异,论证了未来在生命树构建中深化对rDNA簇中各基因进行联合应用的合理性和可行性.     

共同祖先原则和系统发育树的解读

系统发育树是基于共同祖先原则,用于描述生物传代谱系的常用表达方式.可以直观地表达分类群(物种或基因)之间的亲缘关系和进化方向,但这两方面在生物学研究和教学中却常常被误读.通过具体实例,阐释了如何应用共同祖先原则正确解读系统发育树的方法.     

等位基因杂合子现象及其在系统发育学分析中的研究概述

系统发育研究已是澄清所有进化历史问题的必由之路。选择合适的分子标记以及最大限度地挖掘和利用其所包含的系统发育信息是构建可靠的系统发育树的关键。等位基因杂合子(Intra-individual allele heterozygotes, IIAHs)是核基因内含子作为系统发育研究中的分子标记时常常出现的现象。如何挖掘并利用其中所包含的系统发育信息成为近年来系统发育学的研究热点。文章从此现象的产生、杂合子的分离以及现有的研究方法3个方面详尽概述, 阐述了IIAHs及其在系统发育分析中的最新研究进展。     

基于ITS与 trn L-F序列探讨槭树科的系统发育

报道了槭树科41种(其中槭属39种)植物的 trn L-F和ITS序列(其中部分种的ITS序列为重新测定),以期通过分子手段对槭树科内部尤其是复杂的槭属的系统发育关系进行重建.以无患子科和七叶树科为外类群,基于对57个种单独的ITS序列(包括从GenBank下载的16种的序列)、41种 trn L-F序列及41种两者序列的联合数据,分别采用最大简约法(Maximum Parsimony Method)和邻接法(Neighbor-Joining Method)对槭树科的系统发育进行了分析.结果显示,整个槭树科为一单系类群;金钱槭位于槭树科的基部;但由于云南金钱槭( Dipteronia dyerana )聚在了槭属内部,认为金钱槭属和槭属均可能是非单系类群;槭属内组间关系的支持率普遍较低,但多数组的组内关系得到了较好的支持.将两个片段结合比单独的ITS或 trn L-F分析能更好地解决槭属内部的系统关系,其中sect.Palmata 和sect.Microcarpa ,sect.Platanoidea 、sect.Lithocarpa 和sect.Macrophylla ,sect.Integrifolia 、sect.Trifoliata 和sect.Pentaphylla ,以及sect.Acer 、sect.Goniocarpa 和sect.Saccharina (sensu Ogata)的组间亲缘关系得到了一定的支持,但对其中部分组的划分可能应做进一步调整.重新评价了徐廷志系统中对sect.Rubra 和sect.Saccharodendron 的处理.     

中国麻风树种质资源SSR遗传多样性的研究初报

麻风树（Jatropha curcas）是世界上最有潜力的能源植物之一。本研究针对来自全国6省区的219份麻风树种质资源,应用SSR分子标记对其进行了初步的遗传多样性分析。根据公共数据库中的所有序列设计出了20对SSR及4对eSSR引物,这些引物均能很好地扩增麻风树DNA片段。在上述24个SSR标记中,有8个（33.33%）探测到麻风树群体的DNA多态性。这24个SSR标记共扩增出36个位点,平均每个标记扩增1.5个位点,其中12个（33.33%）位点显现出多态性。这表明中国麻风树是一个具有中等SSR多态性水平的物种,其多态百分率为33.3%。     

蜘蛛分子系统学研究进展

蜘蛛是地球上最古老和物种最丰富的生物类群之一,有化石记录最早可追溯至泥盆纪(距今约三亿八百万年).对蜘蛛的系统学研究长期以传统的形态分类为主,随着分子生物学技术的发展,蜘蛛分子系统学研究也取得了长足进展:部分类群依据分子数据或分子结合形态的系统发育关系得以解决,并建立了相应的系统发育树; 所选择的分子标记除来自于线粒体DNA外(如:COI、12S rDNA和16S rDNA等)还有核DNA(如:18S rDNA、28S rDNA和Hitone 3等); 最新的分析方法和分析软件也得到运用.这些工作为蜘蛛的生命之树研究奠定了重要基础.本文对蜘蛛分子系统学研究所涉及到的问题进行了分析讨论,以期为蜘蛛分子系统学的深入开展提供基础资料.     

树鼩PSEN1全长编码序列的克隆及分子特征分析

目的 获取树鼩早老素蛋白-1(PSEN1)的全长编码序列并进行分子特征分析。方法 以树鼩脑组织总RNA为材料,通过RT-PCR、RACE-PCR扩增和序列拼接获得PSEN1基因全长编码序列,进而通过DNAMAN、MEGA等生物信息学软件对其序列和分子特征进行分析。qRT-PCR和Western Blot进一步分析PSEN1在树鼩各个组织的表达模式。结果 克隆鉴定了树鼩PSEN1基因,其cDNA的开放阅读框全长1128 bp,编码375个氨基酸。通过系统发育谱系树、氨基酸序列对比分析,发现树鼩PSEN1与小鼠、大鼠等相比更接近人类和非人灵长类动物。qRT-PCR和Western Blot的结果表明,树鼩PSEN1在脑组织的表达量明显高于其他脏器组织。结论 通过克隆树鼩PSEN1基因序列并进行分析,为今后进一步深入研究该基因功能和建立相关疾病动物模型提供理论基础。     

植物生命之树重建的现状、问题和对策建议

生命之树的概念源自1859年达尔文的《物种起源》, 但利用分子数据重建植物生命之树的研究则在20世纪90年代才开始兴起。近年来, 随着测序技术、分析方法和计算能力的快速发展, 植物生命之树重建研究取得了显著成果。本文首先概述了当前以及未来很长一段时间内植物生命之树重建工作的重点, 包括植物属级和种级水平的系统发育研究、植物系统发育基因组学研究、分子和形态数据联合分析、包括灭绝与现存植物类群的生命之树重建, 以及超大植物生命之树重建等5个方面; 然后简要概括国内植物生命之树重建研究的现状, 指出了我国在植物生命之树重建领域发展中所存在的问题, 并从“类群研究体系、学科评价体系、国家顶层设计, 以及拓展国际合作”等方面对学科未来的发展提出了一些对策建议。     

Genetic Algorithm-Based Maximum-Likelihood Analysis for Molecular Phylogeny

A heuristic approach to search for the maximum-likelihood (ML) phylogenetic tree based on a genetic algorithm (GA) has been developed. It outputs the best tree as well as multiple alternative trees that are not significantly worse than the best one on the basis of the likelihood criterion. These near-optimum trees are subjected to further statistical tests. This approach enables ones to infer phylogenetic trees of over 20 taxa taking account of the rate heterogeneity among sites on practical time scales on a PC cluster. Computer simulations were conducted to compare the efficiency of the present approach with that of several likelihood-based methods and distance-based methods, using amino acid sequence data of relatively large (5–24) taxa. The superiority of the ML method over distance-based methods increases as the condition of simulations becomes more realistic (an incorrect model is assumed or many taxa are involved). This approach was applied to the inference of the universal tree based on the concatenated amino acid sequences of vertically descendent genes that are shared among all genomes whose complete sequences have been reported. The inferred tree strongly supports that Archaea is paraphyletic and Eukarya is specifically related to Crenarchaeota. Apart from the paraphyly of Archaea and some minor disagreements, the universal tree based on these genes is largely consistent with the universal tree based on SSU rRNA. Received: 4 January 2001 / Accepted: 16 May 2001     

Ancient phylogenetic relationships

Traditional views on deep evolutionary events have been seriously challenged over the last few years, following the identification of major pitfalls affecting molecular phylogeny reconstruction. Here we describe the principally encountered artifacts, notably long branch attraction, and their causes (i.e., difference in evolutionary rates, mutational saturation, compositional biases). Additional difficulties due to phenomena of biological nature (i.e., lateral gene transfer, recombination, hidden paralogy) are also discussed. Moreover, contrary to common beliefs, we show that the use of rare genomic events can also be misleading and should be treated with the same caution as standard molecular phylogeny. The universal tree of life, as described in most textbooks, is partly affected by tree reconstruction artifacts, e.g. (i) the bacterial rooting of the universal tree of life; (ii) the early emergence of amitochondriate lineages in eukaryotic phylogenies; and (iii) the position of hyperthermophilic taxa in bacterial phylogenies. We present an alternative view of this tree, based on recent evidence obtained from reanalyses of ancient data sets and from novel analyses of large combination of genes.     

The nature of the last universal ancestor and the root of the tree of life, still open questions.

The nature of the last universal ancestor to all extent cellular organisms and the rooting of the universal tree of life are fundamental questions which can now be addressed by molecular evolutionists. Several scenarios have been proposed during the last years, based on the phylogenies of ribosomal RNA and of duplicated proteins, which suggest that the last universal ancestor was either an RNA progenote or an hyperthermophilic prokaryote. We discuss these hypotheses in the light of new data on the evolution of DNA metabolizing enzymes and of contradictions between different protein phylogenies. We conclude that the last universal ancestor was a member of the DNA world already containing several DNA polymerases and DNA topoisomerases. Furthermore, we criticize current data which suggest that the rooting of the universal tree of life is located in the eubacterial branch and we conclude that both rooting the universal tree and the nature of the last universal ancestor are still open questions.     

Mosaic nature in the skeleton of East Asian crocodylians fills the morphological gap between “Tomistominae” and Gavialinae

Crocodylian systematics has long been confounded by conflicting hypotheses of higher level relationships—although molecular data sets strongly supported the sister-taxon relationship of Tomistoma and Gavialis, morphological data sets placed Gavialis as sister to all other living taxa. One of the perceived difficulties in interpreting morphological character evolution on the molecular tree is the extensive character reversal occurring in Gavialinae, the mechanism of which has yet to be explained. Here, we provide evidence of gavialine-specific atavistic characters from East Asian “tomistomines” Penghusuchus pani and Toyotamaphimeia machikanensis. These taxa exhibit a mosaic assembly of “tomistomine” and gavialine features, which fill the gap between the two longirostrine groups. Although the parsimony analysis of morphological data (69 taxa, 254 characters) still supports the previous morphological hypothesis, the alternative tree that was forced to fit the molecular hypothesis was insignificantly (5/954 steps; 0.52%) longer than the unconstrained tree, suggesting that morphological evolution can also be interpreted on the molecular tree. Although the problem of stratigraphic gaps remains, future studies may be directed to resolving the interrelationships within Gavialoidea, a large longirostrine group of crocodylians, in the molecular tree context.     

Identifying species of moths (Lepidoptera) from Baihua Mountain,Beijing, China,using DNA barcodes

DNA barcoding has become a promising means for the identification of organisms of all life‐history stages. Currently, distance‐based and tree‐based methods are most widely used to define species boundaries and uncover cryptic species. However, there is no universal threshold of genetic distance values that can be used to distinguish taxonomic groups. Alternatively, DNA barcoding can deploy a “character‐based” method, whereby species are identified through the discrete nucleotide substitutions. Our research focuses on the delimitation of moth species using DNA‐barcoding methods. We analyzed 393 Lepidopteran specimens belonging to 80 morphologically recognized species with a standard cytochrome c oxidase subunit I (COI) sequencing approach, and deployed tree‐based, distance‐based, and diagnostic character‐based methods to identify the taxa. The tree‐based method divided the 393 specimens into 79 taxa (species), and the distance‐based method divided them into 84 taxa (species). Although the diagnostic character‐based method found only 39 so‐identifiable species in the 80 species, with a reduction in sample size the accuracy rate substantially improved. For example, in the Arctiidae subset, all 12 species had diagnostics characteristics. Compared with traditional morphological method, molecular taxonomy performed well. All three methods enable the rapid delimitation of species, although they have different characteristics and different strengths. The tree‐based and distance‐based methods can be used for accurate species identification and biodiversity studies in large data sets, while the character‐based method performs well in small data sets and can also be used as the foundation of species‐specific biochips.     

Taxonomic identification of algae (morphological and molecular): species concepts,methodologies, and their implications for ecological bioassessment

Algal taxonomy is a key discipline in phycology and is critical for algal genetics, physiology, ecology, applied phycology, and particularly bioassessment. Taxonomic identification is the most common analysis and hypothesis‐testing endeavor in science. Errors of identification are often related to the inherent problem of small organisms with morphologies that are difficult to distinguish without research‐grade microscopes and taxonomic expertise in phycology. Proposed molecular approaches for taxonomic identification from environmental samples promise rapid, potentially inexpensive, and more thorough culture‐independent identification of all algal species present in a sample of interest. Molecular identification has been used in biodiversity and conservation, but it also has great potential for applications in bioassessment. Comparisons of morphological and molecular identification of benthic algal communities are improved by the identification of more taxa; however, automated identification technology does not allow for the simultaneous analysis of thousands of samples. Currently, morphological identification is used to verify molecular taxonomic identities, but with the increased number of taxa verified in algal gene libraries, molecular identification will become a universal tool in biological studies. Thus, in this report, successful application of molecular techniques related to algal bioassessment is discussed.     

Maximum likelihood Jukes-Cantor triplets: analytic solutions

Maximum likelihood (ML) is a popular method for inferring a phylogenetic tree of the evolutionary relationship of a set of taxa, from observed homologous aligned genetic sequences of the taxa. Generally, the computation of the ML tree is based on numerical methods, which in a few cases, are known to converge to a local maximum on a tree, which is suboptimal. The extent of this problem is unknown, one approach is to attempt to derive algebraic equations for the likelihood equation and find the maximum points analytically. This approach has so far only been successful in the very simplest cases, of three or four taxa under the Neyman model of evolution of two-state characters. In this paper we extend this approach, for the first time, to four-state characters, the Jukes-Cantor model under a molecular clock, on a tree T on three taxa, a rooted triple. We employ spectral methods (Hadamard conjugation) to express the likelihood function parameterized by the path-length spectrum. Taking partial derivatives, we derive a set of polynomial equations whose simultaneous solution contains all critical points of the likelihood function. Using tools of algebraic geometry (the resultant of two polynomials) in the computer algebra packages (Maple), we are able to find all turning points analytically. We then employ this method on real sequence data and obtain realistic results on the primate-rodents divergence time.     

Shrinkage Effect in Ancestral Maximum Likelihood

Ancestral maximum likelihood (AML) is a method that simultaneously reconstructs a phylogenetic tree and ancestral sequences from extant data (sequences at the leaves). The tree and ancestral sequences maximize the probability of observing the given data under a Markov model of sequence evolution, in which branch lengths are also optimized but constrained to take the same value on any edge across all sequence sites. AML differs from the more usual form of maximum likelihood (ML) in phylogenetics because ML averages over all possible ancestral sequences. ML has long been know to be statistically consistent - that is, it converges on the correct tree with probability approaching 1 as the sequence length grows. However, the statistical consistency of AML has not been formally determined, despite informal remarks in a literature that dates back 20 years. In this short note we prove a general result that implies that AML is statistically inconsistent. In particular we show that AML can 'shrink' short edges in a tree, resulting in a tree that has no internal resolution as the sequence length grows. Our results apply to any number of taxa.     

Placing cryptic,recently extinct,or hypothesized taxa into an ultrametric phylogeny using continuous character data: A case study with the lizard Anolis roosevelti

In recent years, enormous effort and investment has been put into assembling the tree of life: a phylogenetic history for all species on Earth. Overwhelmingly, this progress toward building an ever increasingly complete phylogeny of living things has been accomplished through sophisticated analysis of molecular data. In the modern genomic age, molecular genetic data have become very easy and inexpensive to obtain for many species. However, some lineages are poorly represented in or absent from tissue collections, or are unavailable for molecular analysis for other reasons such as restrictive biological sample export laws. Other species went extinct recently and are only available in formalin museum preparations or perhaps even as subfossils. In this brief communication we present a new method for placing cryptic, recently extinct, or hypothesized taxa into an ultrametric phylogeny of extant taxa using continuous character data. This method is based on a relatively simple modification of an established maximum likelihood (ML) method for phylogeny inference from continuous traits. We show that the method works well on simulated trees and data. We then apply it to the case of placing the Culebra Island Giant Anole (Anolis roosevelti) into a phylogeny of Caribbean anoles. Anolis roosevelti is a “crown‐giant” ecomorph anole hypothesized to have once been found throughout the Spanish, United States, and British Virgin Islands, but that has not been encountered or collected since the 1930s. Although this species is widely thought to be closely related to the Puerto Rican giant anole, A. cuvieri, our ML method actually places A. roosevelti in a different part of the tree and closely related to a clade of morphologically similar species. We are unable, however, to reject a phylogenetic position for A. roosevelti that places it as sister taxon to A. cuvieri; although close relationship with the remainder of Puerto Rican anole species is strongly rejected by our method.     

Allopolyploid evolution in Geinae (Colurieae: Rosaceae) – building reticulate species trees from bifurcating gene trees

A previous phylogenetic study of paralogous nuclear low-copy granule-bound starch synthase (GBSSI) gene sequences from polyploid and diploid species in Geinae indicated that the clade has experienced two major allopolyploid events in its history. These were estimated to have occurred several million years ago. In this extended study we test if the reticulate phylogenetic hypothesis for Geinae can be maintained when additional sequences are added. The results are compatible with the hypothesis and strengthen it in minor aspects. We also attempt to identify extant members of one of the inferred ancestral lineages of the allopolyploids. On the basis of previous molecular phylogenies, one specific group has been proposed to be the descendants of this taxon. However, none of the additional paralogues belong to this ancestral lineage. A general method is proposed for converting a bifurcating gene tree, with multiple paralogous low-copy gene sequences from allopolyploid taxa, into a reticulate species tree.