野牦牛线粒体基因组序列测定及其系统进化

野牦牛属高寒地区的特有物种,是我国最珍贵的野生动物遗传资源之一,已被列为国家一级重点保护动物。对野牦牛mtDNA进行全序列测定和结构分析,并基于线粒体基因组序列对其系统发生进行了探讨。结果表明:(1)野牦牛线粒体基因组全序列的大小为16 322 bp,整个基因组由37个编码基因和D-loop区组成;22个tRNA基因序列长度为1 524 bp、2个RNA基因序列长度为2 528 bp、13个编码蛋白基因序列长度为11420 bp、D-loop区长度为892 bp。基因组中无间隔序列,基因间排列紧密,基因内无内含子。(2)野牦牛具有较丰富的遗传多样性。(3)分子系统发生关系显示牦牛为牛亚科中的一个独立属,即牦牛属(Poephagus),牦牛属包括家牦牛(Poephagus grunniens)和野牦牛(Poephagus mutus)2个种。野牦牛线粒体基因组全序列的获得和结构解析对研究牦牛的起源、演化和分类,以及野牦牛遗传资源的保护、开发和利用均具有重要的理论和实际意义。     

斐豹蛱蝶线粒体基因组全序列的测定和分析(英文)

该研究对斐豹蛱蝶(Argyreus hyperbius)(鳞翅目:蛱蝶科)线粒体基因组全序列进行了测定和初步分析。结果表明:斐豹蛱蝶线粒体基因全序列全长为15156bp,包含13个蛋白质编码基因、22个tRNA和2个rRNA基因以及1个非编码的A+T富集区,基因排列顺序与其它鳞翅目种类一致;线粒体全序列核苷酸组成和密码子使用显示出明显的A+T偏好(80.8%)和轻微的AT偏移(AT skew,?0.019)。基因组中共存在11个2～52bp不等的基因间隔区,总长96bp;以及14个1～8bp不等的基因重叠区,总长34bp。除COI以CGA作为起始密码子外,13个蛋白质编码基因中的其余12个基因是以ATN作为起始密码子。除COI和COII基因是以单独的一个T为终止密码子,其余11个蛋白质编码基因都是以TAA结尾的。除了缺少DHU臂的tRNASer(AGN),其余的tRNA基因都显示典型的三叶草结构。tRNA(AGN)和ND1之间的基因间隔区包含一个ATACTAA结构域,这个结构域在鳞翅目中是保守的。A+T富集区没有较大的多拷贝重复序列,但是包含一些微小重复结构:ATAGA结构域下游的20bp poly-T结构,ATTTA结构域后的(AT)9重复,以及位于tRNAMet上游的5bp poly-A结构等。这项研究所揭示的斐豹蛱蝶的线粒体基因组特征,不仅为认识蛱蝶科的遗传多样性贡献数据,而且对于该物种的保护生物学、群体遗传学、谱系地理及演化研究等具有重要意义。     

藏鸡和两种低地鸡线粒体全基因组序列 的测定和比对

藏鸡生活在青藏高原, 它对高原低氧具有一定的遗传适应能力. 丝羽乌骨鸡和寿光鸡都是低地鸡种, 缺乏对低氧环境的遗传适应能力. 鸡的线粒体基因组总长16785 bp, 共编码37个基因, 这些基因的编码产物均与线粒体的呼吸作用和氧化磷酸化相关, 它们的突变均有可能影响线粒体的功能, 因此均可被选为藏鸡低氧遗传适应的候选基因. 测定比较藏鸡和低地鸡的线粒体基因组, 可以为进一步研究藏鸡的低氧遗传适应机理提供线索. 本研究测定和比较了藏鸡、丝羽乌骨鸡和寿光鸡的线粒体全基因组序列, 发现藏鸡线粒体基因组总长为16784~16786 bp, 丝羽乌骨鸡为16785 bp, 寿光鸡为16784 bp; 比对后共发现突变(包括单核苷酸多态性(SNP)、单碱基缺失和插入)120个, 其中tRNA基因突变4个, rRNA基因突变10个, D-LOOP区突变39个, 基因间隔区突变1个, 编码蛋白质亚基的碱基突变66个(包括同义突变48个, 错义突变18个).     

藏鸡和两种低地鸡线粒体全基因组序列 的测定和比对

藏鸡生活在青藏高原, 它对高原低氧具有一定的遗传适应能力. 丝羽乌骨鸡和寿光鸡都是低地鸡种, 缺乏对低氧环境的遗传适应能力. 鸡的线粒体基因组总长16785 bp, 共编码37个基因, 这些基因的编码产物均与线粒体的呼吸作用和氧化磷酸化相关, 它们的突变均有可能影响线粒体的功能, 因此均可被选为藏鸡低氧遗传适应的候选基因. 测定比较藏鸡和低地鸡的线粒体基因组, 可以为进一步研究藏鸡的低氧遗传适应机理提供线索. 本研究测定和比较了藏鸡、丝羽乌骨鸡和寿光鸡的线粒体全基因组序列, 发现藏鸡线粒体基因组总长为16784~16786 bp, 丝羽乌骨鸡为16785 bp, 寿光鸡为16784 bp; 比对后共发现突变(包括单核苷酸多态性(SNP)、单碱基缺失和插入)120个, 其中tRNA基因突变4个, rRNA基因突变10个, D-LOOP区突变39个, 基因间隔区突变1个, 编码蛋白质亚基的碱基突变66个(包括同义突变48个, 错义突变18个).     

日本条螽线粒体基因组序列的测定与分析

日本条螽完整的线粒体基因组序列长16 281 bp,包括13个蛋白质编码基因、22个tRNA基因、2个r RNA基因和1个D-loop区,其基因次序和方向与祖先序列相同。该线粒体基因组排列紧凑,但在ND2和tRNA~(Trp)之间有一段长为650 bp的基因间隔区。为研究螽斯科的系统发育关系,本研究选取日本条螽及其它17个螽斯科物种线粒体基因组的蛋白质编码基因和r RNA基因序列构建贝叶斯系统发生树。     

柳紫闪蛱蝶线粒体基因组全序列及与相关蛱蝶类的比较分析(英文)

该文对柳紫闪蛱蝶Apaturailia(鳞翅目:蛱蝶科)的线粒体基因组全序列进行了测定,同时结合其它已知蛱蝶类的相应序列进行了比较分析。结果显示:柳紫闪蛱蝶的线粒体基因组(GenBankaccessionno.:JF437925)是一个15242bp的环状DNA分子,包含13个蛋白质编码基因、2个rRNA基因和22个tRNA基因。13个蛋白编码基因中,除了COI基因的起始密码子是CGA外,其余12个蛋白编码基因都具有标准的ATN起始密码子;柳紫闪蛱蝶与其它已测的10种蛱蝶在基因定位和排列顺序方面几乎相同,只是在非编码序列上存在细微的差异,其核苷酸的构成及密码子使用频率都处于鳞翅目昆虫的范围之内。22个的tRNA基因中,除了tRNASer(AGN)缺少DHU臂,其余的tRNA基因都显示为典型的三叶草结构。基因组共存在9处基因间重叠区(总长度为33bp)以及12个基因间隔区(总长为155bp,最长间隔是49bp,最短的是1bp)。在ND6和Cytb间的间隔区中还发现有(TA)23似微卫星结构。与其他蛱蝶类相似,403bp的AT富集区包含有ATAGA,ATTTA二个保守模块(一个21bp的poly-T,一个10bp的poly-A),以及二个似微卫星的重复结构((TA)10和(TA)7)。     

红足壮异蝽Urochela quadrinotata Reuter线粒体基因组的测定与分析

研究测定并分析了红足壮异蝽Urochela quadrinotata Reuter的线粒体基因组全序列。该线粒体基因组全长16585bp(GenBank登录号为JQ743678),A+T含量为75.4%,共编码35个基因,包括13个蛋白质基因、20个tRNA基因(两个tRNA基因,即tRNAIle和tRNAGln未被检测到)、2个rRNA基因及一段较长的非编码区(控制区,亦称A+T富含区)。基因排序与大部分昆虫的线粒体基因排列方式相同,没有发生基因重排。除tRNASer(AGN)的DHU臂无法形成典型的茎环结构,其余tRNA基因均能稳定形成典型的三叶草二级结构。预测了红足壮异蝽16S和12S rRNA的二级结果,分别包括6个结构域43个茎环和3个结构域27茎环。控制区含一个长1652bp的串联重复区域,由16个串联重复单元组成。     

高粱A1型细胞质雄性不育系与保持系线粒体基因组分析比较

线粒体基因组易位是导致作物细胞质雄性不育(Cytoplasmic male sterility,CMS)性状产生的重要遗传机制。比较高粱A1型细胞质雄性不育系与保持系线粒体基因组,寻找易位区为克隆高粱A1型细胞质雄性不育相关基因奠定基础。以高粱A1型细胞质雄性不育系Tx623A和其保持系Tx623B为试验材料,采用二代Illumina Hiseq结合三代PacBio测序技术,对2个样品的线粒体基因组进行组装,比较和分析不育系和保持系基因组结构和基因差异。高粱Tx623A和Tx623B线粒体基因组大小分别为449 727 bp和452 772 bp,预测编码开放阅读框(Open reading frame,ORFs)分别为147和145个,且两基因组特有基因分别为8个和6个。两线粒体基因组共线性比较分析,发现存在一个57 kb的基因组片段易位的结构变异(Structural variation,SV)区域,该易位区可能与A1型细胞质雄性不育有关。Tx623A和Tx623B线粒体基因组中易位区为高粱A1型细胞质雄性不育基因克隆提供了基因组信息。     

缅甸蟒线粒体基因组全序列测定与分析

采用LA-PCR(long and accurate PCR)、巢式PCR及TA克隆测序技术,首次获得缅甸蟒Python bivittatus线粒体基因组全序列(GenBank登录号NC_021479)。分析结果表明:缅甸蟒线粒体全长17 617 bp,与其它多数蛇类线粒体基因组结构相似,由13个蛋白编码区、2个rRNA、22个tRNA和双控区组成,基因间排列紧凑;与蟒属其它物种相比,缅甸蟒线粒体在氨基酸数目上存在增减现象;tRNA中tRNA-Cys长度最短,只有57 bp,二氢尿嘧啶环无配对的茎区;缅甸蟒在两个控制区各存在3个相同的串联重复,可能是造成个体间相差87~89 bp的原因。     

新西兰白兔线粒体基因组全序列的测定与分析

本研究旨在获得新西兰白兔(New Zealand white rabbit)线粒体DNA基因组全序列(mtDNA).根据GenBank已经公布的近缘物种穴兔mtDNA全基因组序列(GenBank登录号:AJ001588.1),设计12对可覆盖新西兰白兔mtDNA全序列的引物,通过PCR扩增、测序、拼接,获得新西兰白兔线粒体全序列,并分析其特点.新西兰白兔线粒体基因组全序列为17 418 bp,A+T含量高,为59.72％,蛋白编码基因数量为13个,rRNA基因数量为2个,tRNA基因数量为22个和1个非编码控制区(D-loop区),与其他兔属动物线粒体全基因组排列顺序一致.分析4种特殊的tRNA二级结构,发现tRNA-Ser(AGY)为二叶草型,缺失DHU臂,其余三种tRNA均为三叶草型.与其他哺乳动物线粒体基因组的D-loop区相比,新西兰白兔与格拉达野兔(Le-pus granatensis)核苷酸组成、编码偏好性和氨基酸组成较为相近.相比穴兔的线粒体基因组,新西兰白兔具有一定的保守性和异质性,该结果为其遗传种质资源保护和利用提供基础资料.     

Endotherms,ectotherms, and mitochondrial genome-size variation

The patterns of mitochondrial genomesize variation were investigated in endothermic and ectothermic species to examine the role that thermal habit might play in the evolution of animal mitochondrial DNA (mtDNA). Data on mtDNA size (the modal, largest, and smallest mtDNA reported within a species), the percent variation in mtDNA size (the difference in size between the largest and smallest mtDNAs divided by the model genome size for a given species), and the frequency of heteroplasmic individuals (those carrying more than one mtDNA length variant) were tabulated from the literature. Endotherms showed significantly less variation in mtDNA size and tended to have smaller mtDNAs than ectotherms. Further comparisons between endothermic and ectothermic vertebrates revealed that the largest genome and the percent variation in genome size were significantly smaller in the former than the latter. There was no difference between endothermic and ectotherms in the frequency of heteroplasmy. These data are discussed in light of two hypotheses: (1) more intense directional and purifying selection for small genome size in the cytoplasms of species with higher metabolic rates and (2) reduced mutation pressures generating mtDNA size variants in endotherms relative to those in ectotherms. The general trends are consistent with the selection hypothesis but in certain species mtDNA size variation appears to be governed by mutational pressures. To test these competing hypotheses further, comparative studies are proposed where mitochondrial genome size is quantified in sister taxa and tissue types with very different metabolic rates.     

中华卵索线虫线粒体基因组多态性分析

为完善昆虫病原索科线虫线粒体基因组全序列数据库, 更系统地研究其基因组特征和系统演化规律, 进而为发挥该线虫生防潜力打下基础, 我们开展了中华卵索线虫Ovomermis sinensis线粒体全基因组的研究。该研究通过线粒体基因组滚环复制及酶切图谱, 揭示了中华卵索线虫线粒体基因组具有种内遗传多态性, 即群体中单体线虫具有独特的酶切条带, 且条带累加之和变化范围较大, 为16.5～24.5 kb。为进一步了解线粒体基因组多态性特征及产生的分子机制, 采用两步长PCR方法对2条代表性成虫线粒体基因组进行了测序及拼接, 得其全长分别为18 864和16 777 bp。对这2条序列的比对表明, 线粒体基因组中位于ND2和ND4之间的可变区域, 不仅基因排列顺序不同, 且存在ND3基因重复现象, 这是导致中华卵索线虫线粒体基因组呈现多态性的主要原因。通过对以上研究结果的分析及与GenBank中已有的6种索科线虫线粒体基因组序列进行比对, 概括出其线粒体基因组基本特点: ①线粒体基因排列顺序各不相同;②部分线虫线粒体基因存在重复现象, 且重复次数不同;③线粒体基因组大小存在很大差异。     

Complete sequence of the mitochondrial genome of the tapeworm Hymenolepis diminuta: gene arrangements indicate that Platyhelminths are Eutrochozoans

Using "long-PCR," we amplified in overlapping fragments the complete mitochondrial genome of the tapeworm Hymenolepis diminuta (Platyhelminthes: Cestoda) and determined its 13,900-nt sequence. The gene content is the same as that typically found for animal mitochondrial DNA (mtDNA) except that atp8 appears to be lacking, a condition found previously for several other animals. Despite the small size of this mtDNA, there are two large noncoding regions, one of which contains 13 repeats of a 31-nt sequence and a potential stem-loop structure of 25 bp with an 11-member loop. Large potential secondary structures were identified also for the noncoding regions of two other cestode mtDNAS: Comparison of the mitochondrial gene arrangement of H. diminuta with those previously published supports a phylogenetic position of flatworms as members of the Eutrochozoa, rather than placing them basal to either a clade of protostomes or a clade of coelomates.     

The association between mitochondrial genetic variation and reduced colony fitness in an invasive wasp

Despite the mitochondrion's long‐recognized role in energy production, mitochondrial DNA (mtDNA) variation commonly found in natural populations was assumed to be effectively neutral. However, variation in mtDNA has now been increasingly linked to phenotypic variation in life history traits and fitness. We examined whether the relative fitness in native and invasive common wasp (Vespula vulgaris) populations in Belgium and New Zealand (NZ), respectively, can be linked to mtDNA variation. Social wasp colonies in NZ were smaller with comparatively fewer queen cells, indicating a reduced relative fitness in the invaded range. Interestingly, queen cells in this population were significantly larger leading to larger queen offspring. By sequencing 1,872 bp of the mitochondrial genome, we determined mitochondrial haplotypes and detected reduced genetic diversity in NZ. Three common haplotypes in NZ frequently produced many queens, whereas the four rare haplotypes produced significantly fewer or no queens. The entire mitochondrial genome for each of these haplotypes was sequenced to identify polymorphisms associated with fitness reduction. We found 16 variable sites; however, no nonsynonymous mutation that was clearly causing impaired mitochondrial function was detected. We discuss how detected variants may alter secondary structures, gene expression or mito‐nuclear interactions, or could be associated with nuclear‐encoded variation. Whatever the ultimate mechanism, we show reduced fitness and mtDNA variation in an invasive wasp population as well as specific mtDNA variants associated with fitness variation within this population. Ours is one of only a few studies that confirm fitness impacts of mtDNA variation in wild nonmodel populations.     

Variation of the mitochondrial genome in the evolution of Drosophila

The evidence on mitochondrial genome variation and its role in evolution of the genus Drosophila are reviewed. The mitochondrial genome is represented by a circular double-stranded DNA molecule 16 to 19 kb in length. The genome contains no introns involved in recombination. The entire mitochondrial genome can be arbitrarily divided into three parts: (1) protein-coding genes; (2) genes encoding rRNA and tRNA; and (3) the noncoding regulatory region (A + T region). The selective importance of mutations within different mtDNA regions is therefore unequal. In Drosophila, the content of the A + T pairs in mtDNA is extremely low and a pattern of nucleotide substitution is characterized by a low transition/transversion ratio (and a low threshold of mutation saturation). The deletions and duplications are of common occurrence in the mitochondrial genome. However, this genome lacks such characteristic for the nuclear genome aberrations as the inversions and transpositions. The phenomena of introgression and heteroplasmy provide an opportunity to study the adaptive role of the mitochondrial genome and its role in speciation. Analysis of evidence concerning mtDNA variation in different species of the genus Drosophila made it possible to ascertain data on phylogenetic relationships among species obtained by studying nuclear genome variation. In some species, mtDNA variation may serve as a reliable marker for population differentiation within a species, although evidence on the population dynamics of the mtDNA variation is very scarce.     

The Mitochondrial Genome of Xiphinema americanum sensu stricto (Nematoda: Enoplea): Considerable Economization in the Length and Structural Features of Encoded Genes

The complete sequence of the mitochondrial genome of the plant parasitic nematode Xiphinema americanum sensu stricto has been determined. At 12626bp it is the smallest metazoan mitochondrial genome reported to date. Genes are transcribed from both strands. Genes coding for 12 proteins, 2 rRNAs and 17 putative tRNAs (with the tRNA-C, I, N, S1, S2 missing) are predicted from the sequence. The arrangement of genes within the X. americanum mitochondrial genome is unique and includes gene overlaps. Comparisons with the mtDNA of other nematodes show that the small size of the X. americanum mtDNA is due to a combination of factors. The two mitochondrial rRNA genes are considerably smaller than those of other nematodes, with most of the protein encoding and tRNA genes also slightly smaller. In addition, five tRNAs genes are absent, lengthy noncoding regions are not present in the mtDNA, and several gene overlaps are present. [Reviewing Editor: Dr. Yues van de Peer] F. Lamberti: Deceased, 2004     

Small, repetitive DNAs contribute significantly to the expanded mitochondrial genome of cucumber

Closely related cucurbit species possess eightfold differences in the sizes of their mitochondrial genomes. We cloned mitochondrial DNA (mtDNA) fragments showing strong hybridization signals to cucumber mtDNA and little or no signal to watermelon mtDNA. The cucumber mtDNA clones carried short (30-53 bp), repetitive DNA motifs that were often degenerate, overlapping, and showed no homology to any sequences currently in the databases. On the basis of dot-blot hybridizations, seven repetitive DNA motifs accounted for >13% (194 kb) of the cucumber mitochondrial genome, equaling >50% of the size of the Arabidopsis mitochondrial genome. Sequence analysis of 136 kb of cucumber mtDNA revealed only 11.2% with significant homology to previously characterized mitochondrial sequences, 2.4% to chloroplast DNA, and 15% to the seven repetitive DNA motifs. The remaining 71.4% of the sequence was unique to the cucumber mitochondrial genome. There was <4% sequence colinearity surrounding the watermelon and cucumber atp9 coding regions, and the much smaller watermelon mitochondrial genome possessed no significant amounts of cucumber repetitive DNAs. Our results demonstrate that the expanded cucumber mitochondrial genome is in part due to extensive duplication of short repetitive sequences, possibly by recombination and/or replication slippage.     

Variation of the Mitochondral Genome in the Evolution of Drosophila

The evidence on mitochondrial genome variation and its role in evolution of the genus Drosophila are reviewed. The mitochondrial genome is represented by a circular double-stranded DNA molecule 16 to 19 kb in length. Mitochondrial genes lack introns and recombination. The entire mitochondrial genome can be arbitrarily divided into three parts: (1) protein-coding genes; (2) genes encoding rRNA and tRNA; and (3) the noncoding regulatory region (A + T region). The selective importance of mutations within different mtDNA regions is therefore unequal. In Drosophila, the content of the A + T pairs in mtDNA is extremely high and a pattern of nucleotide substitution is characterized by a low transition/transversion ratio (and a low threshold of mutation saturation). The deletions and duplications are of common occurrence in the mitochondrial genome. However, this genome lacks such characteristic for the nuclear genome aberrations as inversions and transpositions. The phenomena of introgression and heteroplasmy provide an opportunity to study the adaptive role of the mitochondrial genome and its role in speciation. Analysis of evidence concerning mtDNA variation in different species of the genus Drosophilamade it possible to ascertain data on phylogenetic relationships among species obtained by studying nuclear genome variation. In some species, mtDNA variation may serve as a reliable marker for population differentiation within a species, although evidence on the population dynamics of the mtDNA variation is very scarce.     

家马核基因组中线粒体核内插入序列分析

在各种真核生物核基因组中,存在一些由线粒体基因组转移进入核基因组中的DNA片段,这些被认为是分子化石的片段叫做线粒体核内插入序列(Numt)。由于Numt与真实的线粒体序列高度相似,因此它的存在必然会成为PCR扩增线粒体DNA的不利因素。利用已经公布的家马(Equus caballus)基因组序列(2007年9月公布,GenBank登录号为NC_009144-NC_009175)对家马Numt进行了深入分析,共发现200个可能的Numt,长度范围为29到3727bp,其中有10个的长度大于800bp。分析结果显示由于不存在线粒体控制区域的疑似Numt,因此对基于此区域的群体遗传学研究不会产生影响。本研究还发现在家马进化过程中,第1号和27号染色体更倾向于接受线粒体序列的转移。以上结果将为今后马科动物的研究提供重要的参考信息,有助于避免在线粒体DNA研究中由于Numt污染的存在而得出错误的实验结果。     

Animal mitochondrial DNA as a genetic marker in population and evolutionary biology

Animal mitochondrial DNA (mtDNA) is playing an increasingly important role as a genetic marker in population and evolutionary biology. The popularity of this molecule derives, in part, from the relative ease with which clearly homologous sequences can be isolated and compared. Simple sequence organization, maternal inheritance and absence of recombination make mtDNA an ideal marker for tracing maternal genealogies. Rapid rate of sequence divergence (at least in vertebrates) allows discrimination of recently diverged lineages. Studies of mtDNAs from a diversity of animal groups have revealed significant variation among taxa in mtDNA sequence dynamics, gene order and genome size. They have also provided important insights into population structure, geographic variation, zoogeography and phylogeny.