中国吸虱研究X.甲胁虱属分种检索表及一新种记述（吸虱亚目：甲胁虱科）

本文编撰我国甲胁虱属２２种吸虱的检索表,并描述采自云南的解氏甲胁虱,新种ＨｏｐｌｏｐｌｅｕｒａｘｉｅｉＣｈｉｎ,ｓｐ．ｎｏｖ。     

太平洋甲胁虱（虱目：甲胁虱科）在其主要宿主黄胸鼠体表的空间分布格局

太平洋甲胁虱是寄生于黄胸鼠体表的一种主要吸虱昆虫 ,在云南广泛分布。应用Iwao直线回归方法及其随机偏离度检验对太平洋甲胁虱在黄胸鼠不同个体间的空间分布格局进行了研究。根据Iwao直线回归方法 ,建立了M =12 .10 +4.76M (r=0 .75 ,P <0 .0 1)的回归方程 ,所得到的α与 β值 (α =12 .10 ,β =4 .76 )均明显高于判定界线值 0和 1。对α与β值进行随机偏离度检验 ,F =6 .0 7(P <0 .0 5 ) ,由此判定太平洋甲胁虱在黄胸鼠不同个体间的空间分布格局为聚集型分布 ,这说明太平洋甲胁虱对黄胸鼠的寄生是不均匀的 ,存在聚集并有形成大小不一的吸虱个体群的趋势。     

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。     

中国的甲胁虱属吸虱(虱目:甲胁虱科)

甲胁虱属(Hoplopleura)是吸虱中种类最多的一属,主要寄生啮齿动物。Ferris(1951)在其吸虱专著中记录了49种。近二十多年来,新种陆续发现,至今已记录的共90余种。本文根据我们以在收集的标本,结合文献资料加以整理,报道我国甲胁虱属共11种,其中包括一新种和3新纪录。     

虱目裂化线粒体基因组研究进展

虱目是哺乳类和鸟类体表的专性寄生虫。在虱科、阴虱科、长角鸟虱科和兽羽虱科的某些寄生虱种中发现了线粒体基因组裂化现象, 其线粒体基因组裂化成了多个环状的线粒体染色体, 如体虱(Pediculus humanus)、头虱(pediculus capitis)和阴虱(Pthirus pubis)的线粒体基因组分别裂化形成20个、20个和14个微环染色体。微环染色体可能是基因删除和同源重组的结果, 关于线粒体基因组裂化的具体原因和机制, 目前并不清楚, 推测可能是进化选择或随机遗传漂变的结果或与线粒体单链DNA结合蛋白的缺失有关。鉴于线粒体基因组裂化研究对于深入理解线粒体的起源和进化方面具有重要意义, 文章以虱目裂化线粒体基因组为主线, 列举了动物裂化线粒体基因组和裂化特征, 阐述了虱目裂化线粒体基因组的研究现状, 分析了虱目线粒体基因组裂化的类型、原因和机制, 并对该领域未来的研究方向进行了展望。     

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.     

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。     

入侵害虫甘薯凹胫跳甲的鉴定及线粒体基因组分析

【目的】基于形态学鉴定和分子生物学技术确认甘薯凹胫跳甲Chaetocnema confinis是否入侵中国大陆,测定甘薯凹胫跳甲线粒体基因组序列,分析基因组结构及其系统发育关系。【方法】应用显微镜观察从广东不同地点采集的甘薯凹胫跳甲成虫的形态特征,并扩增cox1基因DNA序列进行分子鉴定;利用Illumina MiSeq测序平台对甘薯凹胫跳甲线粒体基因组进行测序、拼装、注释和特征分析;基于亲缘关系相近种属的线粒体基因组序列进行共线性分析和构建系统发育树,分析基因重排和系统发育关系。【结果】形态和分子鉴定结果表明大陆甘薯上发现的跳甲为甘薯凹胫跳甲。甘薯凹胫跳甲线粒体基因组序列大小为15 685 bp,包括有13个蛋白质编码基因、2个rRNA基因、22个tRNA基因和1个非编码控制区;这37个基因之间排列紧凑,间隔总长度101 bp,排列顺序与模式昆虫Drosophila yakuba线粒体基因排列顺序相同。甘薯凹胫跳甲线粒体基因组A+T含量为77.3%,具有明显的AT偏向性。13个蛋白质编码基因的起始密码子均为ATN。在22个tRNA基因中除trnS1的DHU臂缺失,trnD, trnG, trnN和trnT的二级结构中缺少TψC环外,其余17个都能形成典型的三叶草式二级结构,另trnK的反密码子突变为UUU,trnS1的反密码子突变为UCU。甘薯凹胫跳甲的控制区片段长度仅有60 bp,是目前已报道的昆虫线粒体基因组中最短的控制区。基于线粒体基因组的系统发育分析表明,甘薯凹胫跳甲与跳甲亚科(Alticinae)黄曲条跳甲Phyllotreta striolata亲缘关系最近。【结论】甘薯凹胫跳甲已经入侵到中国大陆。本研究获得了甘薯凹胫跳甲的线粒体基因组序列,为防控甘薯凹胫跳甲和分析叶甲科(Chrysomelidae)各种属间的系统发育关系奠定了基础。     

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。     

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 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.     

绿眼赛茧蜂线粒体基因组全序列测定和分析

【目的】测定绿眼赛茧蜂Zele chlorophthalmus线粒体基因组全序列,分析其基因组结构及茧蜂科(Braconidae)部分类群的系统发育关系。【方法】利用Illumina MiSeq二代测序技术对绿眼赛茧蜂的线粒体基因组进行测序,对基因组序列进行拼装、注释,分析其结构特点和碱基组成;基于22种茧蜂科昆虫的COX1蛋白编码基因序列,应用最大似然法(ML)和邻接法(NJ)构建系统发育树,分析绿眼赛茧蜂与茧蜂科其他昆虫的系统发育关系。【结果】绿眼赛茧蜂线粒体基因组全长16 661 bp(GenBank登录号: MG822749),包含13个蛋白质编码基因、22个tRNA基因和2个rRNA基因,共37个基因,以及1个控制区。线粒体基因组有明显的核苷酸组成的偏倚,AT偏正,GC偏负,其A+T含量为82.83%。基因排列顺序与推测的昆虫祖先的序列不完全一致,tRNA基因7处发生重排。13个蛋白质编码基因均以ATN为起始密码子,以TAA为终止密码子。在22个tRNA基因二级结构中,除tRNA^His(H)缺失TΨC环和tRNA^Cys(C)仅剩二氢尿嘧啶(DHU)臂和反密码子臂外,其余tRNA基因均能形成典型的三叶草结构。基于COX1蛋白编码序列的系统发育分析结果显示,与绿眼赛茧蜂亲缘关系最近的是同属于赛茧蜂属的雪跗赛茧蜂Z. niveitarsis。【结论】本研究首次获得绿眼赛茧蜂线粒体基因组全序列。结果表明绿眼赛茧蜂隶属于优茧蜂亚科(Euphorinae)赛茧蜂属,并支持赛茧蜂属的单系性。     

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.     

土蜂科线粒体基因组序列测定和分析

【目的】本研究旨在通过针尾部(Aculeata)昆虫线粒体基因组系统发育分析认知土蜂科(Scoliidae)的单系性及系统发育位置。【方法】利用Illumina Hiseq2500二代测序技术测序土蜂科3属5种的线粒体基因组,并进行注释和分析;基于针尾部昆虫36个线粒体基因组13个蛋白质编码基因(protein-coding genes, PCGs)和2个rRNA基因序列采用最大似然法(maximum likelihood, ML)和贝叶斯法(Bayesian inference, BI)法构建系统发育树。【结果】新测序的土蜂科5个线粒体基因组为五带波壁土蜂Colpa quinquecincta线粒体基因组(GenBank登录号：OM103696),齿石波壁土蜂Colpa tartara线粒体基因组(GenBank登录号：OM103697),厚大长腹土蜂Megacampsomeris grossa线粒体基因组(GenBank登录号：OM103796),台湾大长腹土蜂Megacampsomeris formosensis线粒体基因组(GenBank登录号：OM142776)和斯式土蜂Sc...     

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.