Two Complete Mitochondrial Genomes From Leuctridae (Plecoptera: Nemouroidea): Implications for the Phylogenetic Relationships Among Stoneflies

The family-level relationships within Plecoptera have been a focused area of research for a long time. Its higher classification remains unstable, and the phylogenetic relationships within Plecoptera should be re-examined. Here, we sequenced and analyzed two complete mitochondrial genomes (mitogenomes) of Paraleuctra cercia and Perlomyia isobeae of the family Leuctridae. We reconstructed the phylogeny of Plecoptera based on 13 protein-coding genes (PCGs) from published stoneflies. Our results showed that the Bayesian inference and maximum-likelihood tree had similar topological structures except for the positions of two families, Peltoperlidae and Scopuridae. The Plecoptera is divided into two clades, the suborder Antarctoperlaria and the suborder Arctoperlaria. The two suborders subsequently formed two groups, Eusthenioidea and Gripopterygoidea, and Euholognatha and Systellognatha, which is consistent with the results of morphological studies. In addition, the Leuctridae is the earliest branch within the superfamily Nemouroidea. But the monophyly of Perloidea and Pteronarcyoidea are still not well supported.     

The complete mitochondrial genome of Rondotia menciana (Lepidoptera: Bombycidae)

The mulberry white caterpillar, Rondotia menciana Moore (Lepidoptera: Bombycidae) is a species with closest relationship with Bombyx mori and Bombyx mandarina, and the genetic information of R. menciana is important for understanding the diversity of the Bombycidae. In this study, the mitochondrial genome (mitogenome) of R. menciana was amplified by polymerase chain reaction and sequenced. The mitogenome of R. menciana was determined to be 15,301 bp, including 13 protein-coding genes (PCGs), 2 ribosomal RNA genes, 22 transfer RNA genes, and an AT-rich region. The A+T content (78.87%) was lower than that observed for other Bombycidae insects. All PCGs were initiated by ATN codons and terminated with the canonical stop codons, except for coxII, which was terminated by a single T. All the tRNA genes displayed a typical clover-leaf structure of mitochondrial tRNA. The length of AT-rich region (360 bp) of R. menciana mitogenome is shorter than that of other Bombycidae species. Phylogenetic analysis showed that the R. menciana was clustered on one branch with B. mori and B. mandarina from Bombycidae.     

白边大叶蝉线粒体基因组测序与分析

采用引物步移法测得白边大叶蝉Kolla paulula(Walker)线粒体基因组90%左右的序列,并分析了该叶蝉的线粒体基因组特征。基于23个物种(半翅目)的蛋白编码基因序列,以最大似然法和贝叶斯法构建了头喙亚目系统发育树。已测得序列长度为13 579 bp(AT:73.33%),其中包含了13个蛋白编码基因、21个t RNA基因和1个r RNA基因。除了ND5基因使用GTT作为起始密码子外,其他所有蛋白编码基因均使用ATN作为起始密码子;除了CO II使用不完整的T作为终止密码子,其他所有蛋白编码基因均使用TAA或TAG作为终止密码子。21个t RNA基因中,除了t RNASer(AGN)缺失1个稳定的茎环结构外,其他所有t RNA基因均能形成典型的三叶草二级结构。系统发育分析结果表明,进化树明显可以被分为沫蝉总科+(角蝉总科+蜡蝉总科);白边大叶蝉属于角蝉总科的叶蝉科,白边大叶蝉线粒体基因组特征与其他叶蝉科昆虫相同。     

Characterization and Phylogenetic Analysis of the Complete Mitochondrial Genome of Laelia suffusa (Lepidoptera: Erebidae,Lymantriinae)

In this study, the complete mitochondrial genome of a white tussock moth, Laelia suffusa (Walker, 1855) (Lepidoptera: Erebidae, Lymantriinae), was sequenced and annotated. The genome sequence was 15,502 bp in length and comprised 13 PCGs, 2 rRNAs, 22 tRNAs, and a single noncoding control region (CR). The nucleotide composition of the genome was highly A + T biased, accounting for 79.04% of the whole genome and with a slightly positive AT skewness (0.015). Comparing the gene order with the basal species of Lepidoptera, a typical trnM rearrangement was detected in the mitogenome of L. suffusa. Besides, the trnM rearrangement was found at the head of trnI and trnQ, rather than at the back. The 13 PCGs used ATN as their start codons, except for the cox1 which used CGA. Out of the 22 tRNAs, only 1 tRNA (trnS1) failed to fold in a typical cloverleaf secondary structure. The conserved motif ‘ATAGA + poly-T’ was detected at the start of the control region which was similar to other Lepidoptera species. In total, 10 overlapping regions and 19 intergenic spacers were identified, ranging from 1 to 41 and 2 to 73 bp, respectively. Phylogenetic analysis showed that Lymantriinae was a monophyletic group with a high support value and L. suffusa was closely related to tribe Orgyiini (Erebidae, Lymantriinae). Moreover, the phylogenetic relationship of Noctuoidea (Lepidoptera) species was reconstructed using two datasets (13 PCGs and 37 genes) and these supported the topology of (Notodontidae + (Erebidae + (Nolidae + (Euteliidae + Noctuidae)))).     

The Complete Mitochondrial Genome of Brachmia macroscopa (Lepidoptera: Gelechiidae) and Its Related Phylogenetic Analysis

The sweet potato leaf folder, Brachmia macroscopa, is an important pest in China. The complete mitogenome, which consists of 13 protein-coding genes (PCGs), 22 transfer RNA genes, two ribosomal RNA genes, and an A + T-rich region, was sequenced and found to be 15,394 bp in length (GeneBank no. KT354968). The gene order and orientation of the B. macroscopa mitogenome were similar to those of other sequenced lepidopteran species. All of the PCGs started with ATN as the canonical start codon except for cox1, which started with CGA. In regard to stop codons, most PCGs stopped at TAA except for cox2, which stopped at TA, and nad4, which stopped at a single T. Thirteen PCGs of the available species (33 taxa) were used to demonstrate phylogenetic relationships. The ditrysian cluster was supported as a monophyletic clade at high levels by using maximum likelihood and Bayesian methods. The apoditrysian group, covering the Gelechioidea, formed a monophyletic clade with a bootstrap value of 88% and a posterior probability of 1.00. The superfamily Gelechioidea was supported as a monophyletic lineage by a posterior probability of 1.00.     

野牦牛及青海地方牦牛品种全线粒体基因组母系遗传多样性、分化及系统发育分析

旨在从分子水平上探究野牦牛及青海地方牦牛品种的母系遗传多样性、群体遗传结构、亲缘关系和遗传背景。本研究在测定青海省4个地方牦牛品种(即青海高原、环湖、雪多和玉树牦牛)22条全线粒体基因组(Mitogenome)序列的基础上,从GenBank下载了已公布的野牦牛及上述4个地方牦牛品种的142条相应序列,使用BioEdit 7.2.5、Arlequin 3.11和Network 10.1等软件对共计164条线粒体基因组序列进行综合分析。结果显示：1)根据序列间核苷酸变异共确定了115种单倍型,其中野牦牛和青海地方牦牛品种分别拥有22种和93种单倍型;在野牦牛和青海高原、环湖、雪多、玉树牦牛中分别检测到22、26、18、23、19种特有的单倍型。遗传多样性分析显示,野牦牛单倍型多样度最高(0.992 8±0.014 4),且高于4个青海地方牦牛品种的单倍型多样度(0.973 1±0.007 7);4个青海地方牦牛品种单倍型多样度大小依次为：雪多牦牛(0.988 5±0.012 6)、玉树牦牛(0.975 8±0.018 7)、青海高原牦牛(0.973 0±0.016 6)和环湖牦牛(0.939 3±0.027 8)。2)野牦牛与环湖牦牛之间的固定分化指数值(F_ST值)最大(0.041 2),分化程度最高,而与玉树牦牛间的F_ST值最小(-0.008 8),分化程度最低。青海4个地方牦牛品种中,雪多牦牛与青海高原牦牛之间F_ST值最大(0.035 8),分化程度最高,而雪多牦牛与环湖牦牛间F_ST值最小(0.011 2),分化程度最低。3)聚类分析显示,4个青海地方牦牛品种各自为1类,存在明显的母系遗传差异。相比而言,环湖牦牛和雪多牦牛聚类较近,青海高原牦牛和玉树牦牛聚类较近,而野牦牛与玉树牦牛聚类关系更近,各品种(群体)间的聚类结果与其分化程度、地理分布一致。4)系统发育分析表明,115种单倍型分布在3个大的母系遗传分支(即Mt-Ⅰ、Mt-Ⅱ和Mt-Ⅲ),其中Mt-Ⅰ支系所占比例为72.17%,由A、B、E和F 4种单倍型组构成;Mt-Ⅱ支系包括C、D和H 3种单倍型组,占26.09%;而Mt-Ⅲ支系只包含G单倍型组,由雪多牦牛和野牦牛所拥有,所占比例为1.74%,提示牦牛有3个母系起源。综上所述,野牦牛和青海4个地方牦牛品种均具有丰富的母系遗传多样性,其多样性水平由高到低依次为野牦牛、雪多牦牛、玉树牦牛、青海高原牦牛和环湖牦牛。青海4个地方牦牛品种间及与野牦牛间的遗传分化程度均较弱,但各自拥有特有的母系遗传信息,存在明显的母系遗传差异。野牦牛和青海家牦牛品种由3个母系支系组成,推测牦牛有3个母系起源。     

Mitochondrial DNA diversity and origin of indigenous pigs in South China and their contribution to western modern pig breeds

Indigenous pigs in South China are valuable genetic resources with many specific and unique characters, which have played an important role in the establishment of some western modern pig breeds. However, the origin and genetic diversity of indigenous pigs in South China have not been fully understood. In the present study, we sequenced 534 novel mitochondrial DNA(mt DNA) D-loop and assembled 54 complete mitogenome sequences for all 17 indigenous pig breeds from Fujian, Guangdong, Guangxi and Hainan in South China. These data were analyzed together with previously published homologous sequences relevant to this study. We found that all 13 coding genes of the mitogenomes were under purifying selection, but ND1 had the most variable sites and CYTB contained the most non-synonymous SNPs. Phylogenetic analysis showed that all indigenous pigs in South China were clustered into the D haplogroup with D1 a1, D1 b, D1 c and D1 e sub-haplogroups found to be dominant. Haplotype and nucleotide diversities of D-loop sequences ranged from 0.427 to 0.899 and from 0.00342 to 0.00695, respectively, among which all pigs in Guangdong had the lowest diversity. The estimates of pairwise FST, gene flow(Nm) and genetic distance(Da) indicated that most of these indigenous pig breeds differentiated from each other significantly(P0.05). Among the western modern breeds, Berkshire and Yorkshire had significant Asian matrilineal footprints from indigenous pigs in South China, especially the Spotted pigs distributed in Guangdong and Guangxi. The neutrality test(Fu's F_S) indicated that indigenous pigs from Fujian and Guangxi had gone through recent population expansion events(P0.05). It is concluded that indigenous pigs in South China were most likely derived from the Mekong region and the middle and downstream regions of Yangtze River through Guangxi and Fujian. Our findings provide a complete and in-depth insight on the origin and distribution pattern of maternal genetic diversity of indigenous pigs in South China.     

对虾科物种线粒体基因组特征和系统发育分析

本研究综合分析了对虾科（Penaeidae）21个物种线粒体基因组的全序列,发现其线粒体基因组的长度为15893~16071 bp,A+T含量为64.59%~70.61%。Ka/Ks分析表明,对虾科物种线粒体13个蛋白质编码基因（protein-coding genes, PCGs）中,atp8基因的Ka/Ks最高,表明在对虾科中atp8基因受到了较弱的选择压力;在差异位点的分析中,发现nd5和rrnL基因的差异位点比例较高,是理想的分子标记,可用于分析对虾不同群体之间的遗传多样性;在密码子的使用中,被编码的氨基酸均体现相似的偏好性。同时,本研究采用ML（maximum likelihood）和BI（bayesian inference）方法构建系统发育树,结果显示,这两种方法构建的系统发育树拓扑结构完全一致,且同属物种也都归为一类或者单独归为一支。本研究为快速鉴定对虾科生物提供了可靠的分子标记,为分析对虾科物种遗传多样性提供理论依据。     

十足目异尾次目线粒体基因组比较分析及重排研究进展

近年来，线粒体基因组被广泛应用于物种鉴定、群体遗传、系统演化及适应性进化等领域。十足目异尾次目是一类体型介于虾类和蟹类之间高度特化的甲壳动物，在海洋生物系统演化研究中具有十分重要的意义。然而，与短尾次目相比，人们对异尾次目线粒体基因组关注度明显不足；迄今为止对该类群线粒体基因组及基因重排现象仍缺乏系统全面的了解。本研究综述了异尾次目线粒体基因组的研究现状，并对GenBank数据库已公布的26种异尾次目线粒体基因组全序列进行了比较分析，总结了该类群线粒体基因组的基本特征。在此基础上，首次分析了该类群线粒体基因组常见的重排类型及可能的重排机制。比较发现异尾次目线粒体基因组均发生了重排，表现为15种不同重排类型；这些重排事件都可以用复制-随机丢失模型和线粒体内重组模型进行合理的解释。综述还对重排现象在异尾次目系统发育中的应用进行了探讨，以期为异尾次目线粒体基因组进化及系统发育研究提供科学依据。     

笛鲷属线粒体基因组编码序列的系统进化分析能力评估

运用通用引物长距PCR(Long-PCR)和常规PCR相结合的方法测定了鲈形目笛鲷科的4种笛鲷属鱼类(孟加拉笛鲷、四带笛鲷、千年笛鲷和马拉巴笛鲷)和军曹鱼科的军曹鱼线粒体DNA基因组全序列(GenBank序列号分别为FJ171339,FJ416614,FJ824741,FJ824742 和NC_011219),得出所用全序列测定体系的方法通用性较强,操作简单。线粒体基因组的比对分析表明,测定的mtDNA基因组的绝大部分区段与GenBank中现有的脊椎动物的序列有较高的同源性。以军曹鱼外群结合GenBank中近缘笛鲷鱼类(勒氏笛鲷、蓝点笛鲷和黑带鳞鳍梅鲷)进行的聚类分析中,勒氏笛鲷与黑带鳞鳍梅鲷的聚类关系近于同属物种,与形态学分类存在矛盾。通过单个基因和基因拼接序列比较,综合考虑邻位连接法构建系统进化树的置信度和序列的信息量,对13种蛋白质编码基因在属内种间的系统进化分析能力进行了评估,将基因分成不同的4等:很好的为ATPase6和cox2;好的序列为ND2和cox1;差的为ND6、ND3和ATPase8;包括Cyt b在内的其余6种基因为中等。分析还揭示出序列长度的增加可以提高系统进化树的置信度,且属内物种间比较时序列长度的影响小于高级阶元。