啮总目昆虫的线粒体基因组多样性及系统发育研究进展

啮总目包括啮虫目（皮虱和书虱）和虱目（羽虱和吸虱）,是农业和医学等领域具有重要经济意义和研究价值的类群,目前已鉴定和描述的物种超过10 000个。啮总目昆虫线粒体基因组的变异性在昆虫各类群中最为剧烈,这些变异包括基因组的结构、基因排序、基因含量和链上分布等诸多方面。本文全面分析和总结了啮总目昆虫裂化线粒体基因组的进化属性,并结合两侧对称动物线粒体基因组的裂化特征重构了线粒体基因组环裂化的过程。引入“线粒体基因组核型”的概念来描述动物线粒体基因组丰富的变异程度。动物线粒体的染色体有减小的趋势,而线粒体基因组的裂化正是体现这种趋势的一种重要策略。同时,总结和探讨了目前具有争议的啮总目主要类群间的系统发育关系。本综述为啮总目昆虫线粒体基因组学、啮总目系统发生关系以及两侧对称动物线粒体基因组进化模式的研究提供一个新的视角。     

基于太平洋甲胁虱裂化线粒体基因组推测甲胁虱属祖先线粒体核型

[目的]动物典型的单一染色体线粒体基因组在甲胁虱属Hoplopleura已裂化成多个线粒体微环染色体.本研究旨在通过测定太平洋甲胁虱Hoplopleura pacifica的线粒体基因组来推测甲胁虱属祖先线粒体核型.[方法]利用Illumina HiSeq X Ten高通量测序技术对太平洋甲胁虱裂化线粒体基因组进行测定...     

吸虱亚目trL1(tag)和trnL2(taa)基因间独立进化和协同进化的研究

[目的]吸虱亚目是真兽类动物的永久专性体表寄生虫.吸虱亚目的线粒体基因组发生了剧烈的裂化,形成了不同于以往典型单一大环的多个裂化微环.本文对17种吸虱以及外群尖叫虱Bothriometopus macrocnemis的trnL1(tag)和trnL2(taa)基因序列进行比较分析,探讨trnL1(tag)和trnL2(taa)基因间的独立进化和协同进化.[方法]对云南采集到的3科3属4种吸虱(弯多板虱Polyplax reclinata、锯多板虱Polyplax serrata、太平洋甲胁虱Hoplopleura pacifica和麝鼩钩板虱Ancistroplax crocidurae),用Illumina MiSeq PE250平台高通量测序后与GenBank中查找的其它13种吸虱及尖叫虱的trnL1(tag)和trnL2(taa)基因序列进行比较,采用最大简约法(Maximum parsimony,MP)分析17种吸虱trnL1(tag)和trnL2(taa)基因的进化关系.[结果]17种吸虱的trnL1(tag)和trnL2(taa)基因均形成典型的三叶草结构,trnL1(tag)和trnL2(taa)基因在虱属、阴虱属和猴虱属中有较长的等同序列;在甲胁虱属和钩板虱属中有很短的等同序列;在多板虱属、微胸虱属和血虱属中的等同序列介于二者之间.常见典型单一环状线粒体基因组物种的trnL1(tag)和trnL2(taa)基因等同序列的长度在6-10 bp之间.对17种吸虱的trnL1(tag)和trnL2(taa)基因序列进行同源性比较分析,发现不同属吸虱的trnL1(tag)和trnL2(taa)基因序列相似度差异较大,同属内吸虱的trnL1(tag)和trnL2(taa)基因序列相似度差异较小.基于17种吸虱及尖叫虱的trnL1(tag)和trnL taa)基因构建系统进化树,结果表明trnL1(tag)、trnL2(taa)及trnL1(tag)和trnL2(taa)基因间既有协同进化又有独立进化.[结论]trnL 1(tag)和trnL 2(taa)基因等同序列较长的吸虱易发生协同进化,等同序列较短的吸虱易发生独立进化.吸虱亚目的trnL tag)和trnL2(taa)基因协同进化是长期的或发生在远缘物种间,而独立进化是短期(两次重组事件间)的或发生在近缘物种间.吸虱亚目线粒体基因组的裂化模式可能影响trnL1(tag)和trnL2(taa)基因等同序列的长短.     

吸虱亚目非典型线粒体基因组研究进展

吸虱是真兽类哺乳动物体表的专性寄生虫,在全世界广泛分布,物种数量高达540种。近年来随着分子生物学的飞速发展,在NCBI中已收录15种吸虱线粒体基因组序列,其独特的非典型线粒体基因组发生剧烈的裂化现象,形成数目不等的微环染色体。本文综述了15种吸虱线粒体基因组的结构和组成、RNA基因、非编码区以及对吸虱祖先线粒体核型推测的方法和结果。探讨了不同种属间以及与其它昆虫的差异,提出今后对吸虱亚目线粒体基因组研究的展望。     

吸虱亚目trnL_1(tag)和trnL_2(taa)基因间独立进化和协同进化的研究

【目的】吸虱亚目是真兽类动物的永久专性体表寄生虫。吸虱亚目的线粒体基因组发生了剧烈的裂化,形成了不同于以往典型单一大环的多个裂化微环。本文对17种吸虱以及外群尖叫虱Bothriometopus macrocnemis的trnL_1(tag)和trnL_2(taa)基因序列进行比较分析,探讨trnL_1(tag)和trnL_2(taa)基因间的独立进化和协同进化。【方法】对云南采集到的3科3属4种吸虱(弯多板虱Polyplax reclinata、锯多板虱Polyplax serrata、太平洋甲胁虱Hoplopleura pacifica和麝鼩钩板虱Ancistroplax crocidurae),用IlluminaMiSeq PE250平台高通量测序后与GenBank中查找的其它13种吸虱及尖叫虱的trnL_1(tag)和trnL_2(taa)基因序列进行比较,采用最大简约法(Maximumparsimony,MP)分析17种吸虱trnL_1(tag)和trnL_2(taa)基因的进化关系。【结果】17种吸虱的trnL_1(tag)和trnL_2(taa)基因均形成典型的三叶草结构,trnL_1(tag)和trnL_2(taa)基因在虱属、阴虱属和猴虱属中有较长的等同序列;在甲胁虱属和钩板虱属中有很短的等同序列;在多板虱属、微胸虱属和血虱属中的等同序列介于二者之间。常见典型单一环状线粒体基因组物种的trnL_1(tag)和trn L2(taa)基因等同序列的长度在6-10 bp之间。对17种吸虱的trnL_1(tag)和trnL_2(taa)基因序列进行同源性比较分析,发现不同属吸虱的trnL_1(tag)和trnL_2(taa)基因序列相似度差异较大,同属内吸虱的trnL_1(tag)和trnL_2(taa)基因序列相似度差异较小。基于17种吸虱及尖叫虱的trnL_1(tag)和trnL_2(taa)基因构建系统进化树,结果表明trn L1(tag)、trn L2(taa)及trn L1(tag)和trn L2(taa)基因间既有协同进化又有独立进化。【结论】trnL_1(tag)和trnL_2(taa)基因等同序列较长的吸虱易发生协同进化,等同序列较短的吸虱易发生独立进化。吸虱亚目的 trnL_1(tag)和trnL_2(taa)基因协同进化是长期的或发生在远缘物种间,而独立进化是短期(两次重组事件间)的或发生在近缘物种间。吸虱亚目线粒体基因组的裂化模式可能影响trnL_1(tag)和trnL_2(taa)基因等同序列的长短。     

基于线粒体基因组序列的鞘翅目肉食亚目水生类群系统发育分析

【目的】明确肉食亚目(Adephaga)水生类群线粒体基因组的基本特征,并基于线粒体基因组序列分析肉食亚目水生类群的系统发育关系。【方法】基于Illumina HiSeq X Ten测序技术测定了圆鞘隐盾豉甲Dineutus mellyi和齿缘龙虱Eretes sticticus的线粒体全基因组序列,对其进行了基因注释,并对其tRNA基因二级结构进行了预测分析。加上已公布的鞘翅目(Coleoptera)肉食亚目水生类群17个种的线粒体基因组序列,对该类群共19个种线粒体的蛋白质编码基因(protein-coding genes, PCGs)开展了比较基因组学分析,包括AT含量、密码子偏好性、选择压力等。基于13个PCGs的氨基酸序列和核苷酸序列,利用最大似然法(ML)和贝叶斯法(BI)分别构建鞘翅目肉食亚目水生类群的系统发育关系,并通过FcLM分析进一步评估伪龙虱科(Noteridae)和瀑甲科(Meruidae)的系统发育位置。【结果】圆鞘隐盾豉甲和齿缘龙虱的线粒体基因组全长分别为16 123 bp(GenBank登录号: MN781126)和16 196 bp(GenBank登录号: MN781132),都包含13个PCGs、22个tRNA基因、2个rRNA基因和1个D-loop区(控制区)。19个肉食亚目水生类群线粒体基因组PCGs的碱基组成都呈现A+T偏好性,在密码子使用上也都偏向于使用富含A+T的密码子;在进化过程中13个PCGs的进化模式相同,都受到纯化选择。基于线粒体基因组13个PCGs的氨基酸序列的肉食亚目水生类群的系统发育关系为(豉甲科Gyrinidae+(沼梭甲科Haliplidae+((壁甲科Aspidytidae+(两栖甲科Amphizoidae+龙虱科Dytiscidae))+(水甲科Hygrobiidae+(瀑甲科Meruidae+伪龙虱科Noteridae)))))。【结论】研究结果表明,豉甲科是肉食亚目水生类群的基部类群,接下来是沼梭甲科和龙虱总科;伪龙虱科和瀑甲科互为姐妹群,并一起作为龙虱总科内部的一个分支;两栖甲科与龙虱科具有更近的亲缘关系。     

蚯蚓基因组学的研究进展: 基于全基因组及线粒体基因组

蚯蚓被喻为土壤中的“生态系统工程师”, 具有高度的多样性且在全世界都有分布, 被用作土壤健康的指示生物。蚯蚓具有极强的环境适应能力, 在不断适应的过程中促进了自身基因组的进化。本文对近年来蚯蚓全基因组以及线粒体基因组的研究进展进行了综述。蚯蚓全基因组的测序、拼装和分析为研究蚯蚓生态学、污染物对蚯蚓致毒的分子机制、免疫防御的分子机制、蚯蚓再生的分子机制等奠定基础。而线粒体基因组多应用于蚯蚓分子系统发育方面的研究, 目前已有多种蚯蚓通过线粒体基因组测序完成了物种的鉴定。本文建议今后重点开展以下几方面的研究: (1)针对现有的4种蚯蚓全基因组测序结果, 进一步进行比较基因组学、进化基因组学和功能基因组学的研究。(2)完善不同种蚯蚓的基因文库和表达序列标签。(3)建立线粒体基因组、全基因组与蚯蚓物种多样性的关联分析。     

乳酸菌微进化的研究进展

摘要:乳酸菌是食品工业中重要的微生物,乳酸菌微进化研究有助于深入解析其生物学功能与机制。随着分子生物学的发展,多位点序列分型(Multi-locus Sequence Typing,MLST)及基因组重测序(Whole-genome resequencing)等技术手段应运而生,使得从分子水平上阐述乳酸菌的系统发育和种群进化关系成为可能。MLST已被广泛用于乳酸菌遗传多样性和种群结构等微进化研究中,近期,测序成本的锐减使全基因组测序技术在乳酸菌微进化研究中的优势日益突显。本文对乳酸菌微进化的理论基础、研究方法和意义进行了阐述,并介绍了全基因组测序技术在乳酸菌微进化方面的应用,旨在为乳酸菌微进化分析研究提供新思路。     

瓦氏黄颡鱼线粒体全基因组序列分析及系统进化

鲿科鱼类种类繁多, 外形相似, 形态学分类较为困难。为了给鲿科鱼类乃至鲇形目鱼类的系统进化研究积累基础资料, 文章采用参照近缘物种线粒体基因组设计覆盖全基因组引物的方法, 利用16对引物对瓦氏黄颡鱼(Pelteobagrus vachelli)线粒体全基因组进行扩增, PCR产物转化到质粒后测序, 最终获得线粒体基因组全序列, 其全长为16 527 bp, 包括2个rRNA基因、22个tRNA基因、13个编码蛋白质基因和一个非编码控制区。瓦氏黄颡鱼(P. vachelli)线粒体基因组结构和基因排列顺序与现已公布的鲇形目鱼类完全一致, 序列分析表明, 与鲇形目其他种属间具有较高的同源性, 与拟鲿属的同源性最高(91%)。利用鲇形目共4科6属9种及3个外群的线粒体全基因组序列, 从线粒体基因组水平探讨了鲿科鱼类及其在鲇形目的系统进化地位, 结果表明: 鲿科鱼类的瓦氏黄颡鱼(P. vachelli)、黄颡鱼(Pelteobagrus fulvidraco)、光泽黄颡鱼(Pelteobagrus nitidus)及越南拟鲿(Pseudobagrus tokiensis)构成一单系群; 拟鲿属与黄颡鱼属的关系较近; 黄颡鱼属中瓦氏黄颡鱼(P. vachelli)与光泽黄颡鱼(P.nitidus)的关系近于黄颡鱼(P. fulvidraco)。     

动物线粒体基因组GC含量分析

碱基组成是指基因组中腺嘌呤、鸟嘌呤、胞嘧啶和胸腺嘧啶的相对含量,一般用鸟嘌呤和胞嘧啶的含量(GC含量)表示。前人的研究表明,不同类群的核基因组GC含量存在差异,然而,对于动物线粒体基因组碱基组成的研究却很少。为此,本文统计和分析了分属于20个门的609个动物线粒体基因组的碱基组成。结果表明,各门动物间线粒体基因组GC含量没有显著差异,不能反映各动物门间的进化关系。脊椎动物鱼纲、两栖纲、爬行纲、鸟纲和哺乳纲及其代表性目的线粒体基因组GC含量虽有一定差异,但并未发现明显的变化规律。     

Heteroplasmy in the Mitochondrial Genomes of Human Lice and Ticks Revealed by High Throughput Sequencing

The typical mitochondrial (mt) genomes of bilateral animals consist of 37 genes on a single circular chromosome. The mt genomes of the human body louse, Pediculus humanus, and the human head louse, Pediculus capitis, however, are extensively fragmented and contain 20 minichromosomes, with one to three genes on each minichromosome. Heteroplasmy, i.e. nucleotide polymorphisms in the mt genome within individuals, has been shown to be significantly higher in the mt cox1 gene of human lice than in humans and other animals that have the typical mt genomes. To understand whether the extent of heteroplasmy in human lice is associated with mt genome fragmentation, we sequenced the entire coding regions of all of the mt minichromosomes of six human body lice and six human head lice from Ethiopia, China and France with an Illumina HiSeq platform. For comparison, we also sequenced the entire coding regions of the mt genomes of seven species of ticks, which have the typical mitochondrial genome organization of bilateral animals. We found that the level of heteroplasmy varies significantly both among the human lice and among the ticks. The human lice from Ethiopia have significantly higher level of heteroplasmy than those from China and France (P_t<0.05). The tick, Amblyomma cajennense, has significantly higher level of heteroplasmy than other ticks (P_t<0.05). Our results indicate that heteroplasmy level can be substantially variable within a species and among closely related species, and does not appear to be determined by single factors such as genome fragmentation.     

The mitochondrial genome of the chimpanzee louse,Pediculus schaeffi: insights into the process of mitochondrial genome fragmentation in the blood-sucking lice of great apes

Background

Blood-sucking lice in the genera Pediculus and Pthirus are obligate ectoparasites of great apes. Unlike most bilateral animals, which have 37 mitochondrial (mt) genes on a single circular chromosome, the sucking lice of humans have extensively fragmented mt genomes. The head louse, Pediculus capitis, and the body louse, Pe. humanus, have their 37 mt genes on 20 minichromosomes. The pubic louse, Pthirus pubis, has its 34 mt genes known on 14 minichromosomes. To understand the process of mt genome fragmentation in the sucking lice of great apes, we sequenced the mt genome of the chimpanzee louse, Pe. schaeffi, and compared it with the three human lice.

Results

We identified all of the 37 mt genes typical of bilateral animals in the chimpanzee louse; these genes are on 18 types of minichromosomes. Seventeen of the 18 minichromosomes of the chimpanzee louse have the same gene content and gene arrangement as their counterparts in the human head louse and the human body louse. However, five genes, cob, trnS₁, trnN, trnE and trnM, which are on three minichromosomes in the human head louse and the human body louse, are together on one minichromosome in the chimpanzee louse.

Conclusions

Using the human pubic louse, Pt. pubis, as an outgroup for comparison, we infer that a single minichromosome has fragmented into three in the lineage leading to the human head louse and the human body louse since this lineage diverged from the chimpanzee louse ~6 million years ago. Our results provide insights into the process of mt genome fragmentation in the sucking lice in a relatively fine evolutionary scale.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1843-3) contains supplementary material, which is available to authorized users.     

Fragmented mitochondrial genomes are present in both major clades of the blood-sucking lice (suborder Anoplura): evidence from two Hoplopleura rodent lice (family Hoplopleuridae)

Background

The suborder Anoplura contains 540 species of blood-sucking lice that parasitize over 840 species of eutherian mammals. Fragmented mitochondrial (mt) genomes have been found in the lice of humans, pigs, horses and rats from four families: Pediculidae, Pthiridae, Haematopinidae and Polyplacidae. These lice, eight species in total, are from the same major clade of the Anoplura. The mt genomes of these lice consist of 9–20 minichromosomes; each minichromosome is 1.5–4 kb in size and has 1–8 genes. To understand mt genome fragmentation in the other major clade of the Anoplura, we sequenced the mt genomes of two species of rodent lice in the genus Hoplopleura (family Hoplopleuridae).

Results

We identified 28 mt genes on 10 minichromosomes in the mouse louse, Ho. akanezumi; each minichromosome is 1.7–2.7 kb long and has 1–6 genes. We identified 34 mt genes on 11 minichromosomes in the rat louse, Ho. kitti; each minichromosome is 1.8–2.8 kb long and has 1–5 genes. Ho. akanezumi also has a chimeric minichromosome with parts of two rRNA genes and a full-length tRNA gene for tyrosine. These two rodent lice share the same pattern for the distribution of all of the protein-coding and rRNA genes but differ in tRNA gene content and gene arrangement in four minichromosomes. Like the four genera of blood-sucking lice that have been investigated in previous studies, the Hoplopleura species have four minichromosomes that are only found in this genus.

Conclusions

Our results indicate that fragmented mt genomes were present in the most recent common ancestor of the two major clades of the blood-sucking lice, which lived ~75 million years ago. Intra-genus variation in the pattern of mt genome fragmentation is common in the blood-sucking lice (suborder Anoplura) and genus-specific minichromosomes are potential synapomorphies. Future studies should expand into more species, genera and families of blood-sucking lice to explore further the phylogenetic utility of the novel features associated with fragmented mt genomes.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-751) contains supplementary material, which is available to authorized users.     

The multipartite mitochondrial genome of Liposcelis bostrychophila: insights into the evolution of mitochondrial genomes in bilateral animals

Booklice (order Psocoptera) in the genus Liposcelis are major pests to stored grains worldwide and are closely related to parasitic lice (order Phthiraptera). We sequenced the mitochondrial (mt) genome of Liposcelis bostrychophila and found that the typical single mt chromosome of bilateral animals has fragmented into and been replaced by two medium-sized chromosomes in this booklouse; each of these chromosomes has about half of the genes of the typical mt chromosome of bilateral animals. These mt chromosomes are 8,530 bp (mt chromosome I) and 7,933 bp (mt chromosome II) in size. Intriguingly, mt chromosome I is twice as abundant as chromosome II. It appears that the selection pressure for compact mt genomes in bilateral animals favors small mt chromosomes when small mt chromosomes co-exist with the typical large mt chromosomes. Thus, small mt chromosomes may have selective advantages over large mt chromosomes in bilateral animals. Phylogenetic analyses of mt genome sequences of Psocodea (i.e. Psocoptera plus Phthiraptera) indicate that: 1) the order Psocoptera (booklice and barklice) is paraphyletic; and 2) the order Phthiraptera (the parasitic lice) is monophyletic. Within parasitic lice, however, the suborder Ischnocera is paraphyletic; this differs from the traditional view that each suborder of parasitic lice is monophyletic.     

Chimeric mitochondrial minichromosomes of the human body louse, Pediculus humanus: evidence for homologous and non-homologous recombination

The mitochondrial (mt) genome of the human body louse, Pediculus humanus, consists of 18 minichromosomes. Each minichromosome is 3 to 4 kb long and has 1 to 3 genes. There is unequivocal evidence for recombination between different mt minichromosomes in P. humanus. It is not known, however, how these minichromosomes recombine. Here, we report the discovery of eight chimeric mt minichromosomes in P. humanus. We classify these chimeric mt minichromosomes into two groups: Group I and Group II. Group I chimeric minichromosomes contain parts of two different protein-coding genes that are from different minichromosomes. The two parts of protein-coding genes in each Group I chimeric minichromosome are joined at a microhomologous nucleotide sequence; microhomologous nucleotide sequences are hallmarks of non-homologous recombination. Group II chimeric minichromosomes contain all of the genes and the non-coding regions of two different minichromosomes. The conserved sequence blocks in the non-coding regions of Group II chimeric minichromosomes resemble the "recombination repeats" in the non-coding regions of the mt genomes of higher plants. These repeats are essential to homologous recombination in higher plants. Our analyses of the nucleotide sequences of chimeric mt minichromosomes indicate both homologous and non-homologous recombination between minichromosomes in the mitochondria of the human body louse.     

Geographic distributions and origins of human head lice (Pediculus humanus capitis) based on mitochondrial data

Human head lice (Pediculus humanus capitis) are subdivided into 3 deeply divergent mitochondrial clades (Clades A, B, and C), each having unique geographical distributions. Determining the evolutionary history and geographic distribution of these mitochondrial clades can elucidate the evolutionary history of the lice as well as their human hosts. Previous data suggest that lice belonging to mitochondrial Clade B may have originated in North America or Asia; however, geographic sampling and sample sizes have been limited. With newly collected lice, we calculate the relative frequency, geographic distribution, and genetic diversity of louse mitochondrial clades to determine the geographic origin of lice belonging to Clade B. In agreement with previous studies, genetic diversity data support a North American origin of Clade B lice. It is likely that lice belonging to this mitochondrial clade recently migrated to other geographic localities, e.g., Europe and Australia, and, if not already present, may disperse further to occupy all geographic regions.