Advances in the study of helminth mitochondrial genomes and their associated applications

Helminths, including flatworms and roundworms, are abundant organisms that have a variety of life histories. Of these, the genera Schistosoma, Echinococcus, Trichinella are notable parasites of veterinary and medical importance, and cause substantial socio- economic losses throughout China and the rest of the world. Genetic markers in the mitochondrial (mt) genome have proven use- ful for systematic, ecological, evolutionary and population studies, and the growth of mt genomic research has increased in the last two decades. Technological improvements, such as the long-polymerase chain reaction method and high-throughput se- quencing have allowed minute amounts of DNA from single worms, biopsy samples or microscopic organisms to be used for whole mt genome characterization. To facilitate the retrieval, annotation and analyses of mitochondrial features, multiple data- bases and specific software have also been designed and established. This review focuses on current progress, applications and perspectives regarding helminth mt genomics. To date, the complete mt genomes for 93 species of helminths have been sequenced and analyzed. Analyses of the mt genes, including gene content, arrangement, composition and variation have revealed unique features among the helminths when compared with other metazoans. This provides important data concerning their functional and comparative mitochondrial genomics, molecular taxonomy and characterization, population genetics and systematics, and evolu- tionary history. Moreover, mt genome data for parasitic helminths are important for diagnosis, epidemiology and ecology of in- fections. Mitochondrial genome data offer a rich source of markers for the systematics and population genetics of socioeconomi- cally important parasitic helminths of humans and other animals.     

Synonymy of Rhynchophorus ferrugineus (Olivier), 1790 and R. vulneratus (Panzer), 1798 (Coleoptera,Curculionidae, Rhynchophorinae)

Morphological, molecular-genetic and breeding data were collected to investigate the species status of the Asian palm weevils, Rhynchophorus ferrugineus (Olivier) and R. vulneratus (Panzer) (Coleoptera: Curculionidae). These weevils are distinguished by characteristic colouring of the pronota and elytra, but naturally occurring colour intermorphs were observed. Contrary to the literature, quantitative measurements of the concavity of subgenal sutures and of pronotal shape indicated no differences between the two species. Larvae did not differ significantly in labral characteristics. Random amplified polymorphic DNA (RAPD) banding patterns were identical for nine of 14 primers, indicating that these weevils are very closely related. Sequences of the cytochrome oxidase gene for 201 base pairs read were identical for R. ferrugineus and R. vulneratus, but the congener R. bilineatus differed from them by 10%, suggesting divergence of these lineages about 5?million years ago. Hybrid F1s were obtained from all heterospecific crosses, and one surviving hybrid F1 female produced viable eggs. Previous studies have revealed no pheromonal differences. On the basis of this evidence, R. ferrugineus and R. vulneratus should be considered colour morphs of the same species and be synonymized under the name Rhynchophorus ferrugineus (Olivier), with the common name Asian palm weevil.     

扬子鳃蛭线粒体基因组全序列测定及其系统学地位——基于Pacbio和Illumina技术

利用Illumina和Pacbio测序技术对扬子鳃蛭（Ozobranchus jantseanus）线粒体基因组全序列进行测序及注释，并与NCBI数据库中蛭纲（Hirudinea）其他物种的线粒体全序列的排列方式和系统进化关系进行了比较，以确定其系统学地位.结果显示：获得的线粒体基因组序列长度为14 868 bp，蛋白编码基因存在2种起始密码子，分别为ATG和ATT；终止密码子共有4种，分别为TAA，TAG，TA和T；蛭纲10个物种的线粒体基因组全序列基因排序可分为2种类型，其差异表现在tRNAs的互换和长非编码区的长度的不同；扬子鳃蛭所属的吻蛭目（Rhynchobdellida）均为b型，绝大部分无吻蛭目（Arhynchobdellida）的排列方式为a型.基于蛭纲物种的13个蛋白编码基因(PCGs)的Neighbor-Joining（NJ）树进化分析显示，扬子鳃蛭与其他吻蛭目物种聚类.     

云南大理白族和汉族弱精子症患者精子mtDNA 4977 bp缺失相关性研究

探讨云南大理白族和汉族弱精子症患者精子线粒体DNA(mtDNA) 4977 bp缺失的相关性。收集云南大理白族弱精子症患者137例,汉族弱精子症患者121例,正常对照组大理白族和白族精子活力正常人各120例,提取精液基因组DNA,采用聚合酶链式反应(PCR)技术进行精子mtDNA4977bp缺失的研究。结果显示,120例正常对照的大理白族精子活力正常人中有10例(8.33%)存在mtDNA4977 bp缺失;而137例大理白族弱精子症患者中有89例(64.96%)存在mtDNA 4977 bp缺失。两组4977 bp缺失频率比较有极显著性差异(P 0.01)。120例正常对照的汉族精子活力正常人中有7例(5.83%)存在mtDNA 4977 bp缺失;121例大理白族弱精子症患者中有85例(70.25%)存在mtDNA 4977 bp缺失。两组4977 bp缺失频率比较差异有极显著性(P 0.01)。同时发现大理白族弱精子症患者与汉族弱精子症患者mtDNA 4977 bp缺失频率无显著性差异(P 0.05)。大理白族和汉族人群mt DNA 4977 bp缺失与弱精子症的发病有明显的关联,提示mtDNA 4977 bp缺失在弱精子症的发生中可能起重要作用。     

芪参补气药茶清除线粒体活性氧研究

以黄芪和党参为原料,根据中医学原理组方制成芪参补气药茶(ACT),探索ACT清除线粒体活性氧作用及促进健康的机制。分别用AlCl₃比色法和硫酸苯酚法测定ACT的功能因子总黄酮、总糖及总多糖含量。以还原型辅酶Ⅰ/吩嗪硫酸甲酯(NADH/PMS)为超氧阴离子(O₂·^－)生成系统,过氧化氢(H₂O₂)/Fe²⁺体系为羟自由基(·OH)生成系统,分别用氮蓝四唑(NBT)还原法和Fenton反应显色法测定ACT清除O₂·^－及·OH的能力;用Na₂S₂O₃滴定法测定ACT清除H₂O₂的能力。ACT总糖质量分数为(26±1.3) mg·mL^-1,总多糖质量分数为(12.5±0.8) mg·mL^-1,总黄酮质量分数为(121±8.5) μg·mL^-1。ACT可一定程度上清除O₂·^－、·OH和H₂O₂。ACT具有温和的抗氧化作用,从而维持机体氧化与抗氧化平衡,这是ACT促进健康的潜在机制。     

The complete mitochondrial genome and phylogenetic analysis of forest musk deer (Moschus berezovskii)

The complete mitochondrial genome sequence of the forest musk deer (Moschus berezovskii) was determined. Sequence analysis revealed that the organization of the mitochondrial genome in the forest musk deer was consistent with that of other reported mammals. Molecular phylogenetic analysis of the forest musk deer and 22 other species of the order Cetartiodactyla was conducted using maximum parsimony and Bayesian inference methods based on the 12 heavy‐strand protein‐coding genes, RNA genes and their combination. Both the consensus phylogenetic trees and the topologies test supported the monophyly of Cervidae, Bovidae and Cetancodonta, and placed Moschidae as a sister group to Cervidae/Bovidae.     

News from a small island – first record of a freshwater shrimp (Decapoda,Atyidae, Caridina) from Peleng,Banggai Islands,Indonesia

The Indonesian island Sulawesi is well known for its fascinating fauna. Among the atyid and palaemonid freshwater shrimps described from Sulawesi are also a number of endemic species. In contrast, freshwater shrimps have never been reported from the nearby and smaller Peleng Island. Here we describe Caridina thomasi sp. nov. as a first record of an atyid freshwater shrimp from Peleng and compare it with atyids from Sulawesi and other localities. The new species is probably endemic to the island and shows some characters never described in the genus Caridina before. However, a molecular phylogeny of atyid shrimps from the Indo‐West Pacific based on mitochondrial DNA (16S) places C. thomasi within a clade of typical species of Caridina. We tentatively assign the new species to the genus Caridina and discuss the possibility of future changes in the classification of species in this genus.     

黑线姬鼠线粒体DNA控制区的序列测定及分析

以黑线姬鼠肝脏组织为材料提取线粒体DNA，并以此为模版定向扩增D-loop区．克隆后测序得到的全序列大小为852bp．选取其相对保守序列与欧洲大陆同种黑线姬鼠进行序列比较，发现2个同种不同域的黑线姬鼠亲缘关系很近．通过比较突变位点出现的位置可以推断黑线姬鼠线粒体DNA控制区可分ETAS，central，CSB3个区域．由中心区最为保守这一原则，推测出central区大体位置，central区两侧则为ETAS和CSB区．     

Present status and development on biological nitrogen fixation research in China

This presentation introduces the advances in biological nitrogen fixation research abroad, in particular,describes the great progress and achievements on its research in China as follows: collection of rhizobial resources and establishment of the largest database of Rhizobium in China, correction and development of Rhizobium taxonomy in international; discovery of a couple of nif genes, identification and unification of linkage among the nif gene operons of Klebsiella pneumoniae, finding of regulative mechanism of positive regulation nif gene and its sensitivity to oxygen,temperature; finding of the activity of nodulation gene nodD3 product in Sinorhizobium meliloti which is not controlled by flavonoid produced from its host alfalfa; finding of the association between expression of genes coding the products for carbon utilization and nitrogen metabolism and their regulations; chemical synthesis of nodulation factor of Sinorhizobium meliloti; constructions of engineered nitrogen fixers and utilization in practice based on the research of gene expression and regulation; chemical simulation of the structure and function of nitrogenase and bringing forward the model of nitrogenase active center for the first time in international and synthesis of model compounds which were paid attention by colleagues abroad. Finally, the development of nitrogen fixation research in China in future has been put forward, suggesting that the nif gene regulation and its role in providing crops with nitrogen element, signal transduction and molecular interactions between Rhizobium and legume, coupling between carbon and nitrogen metabolisms, nitrogen fixation and photosynthesis, and functional genomics of nitrogen-fixing nodule symbiosis, etc., would be actively worked on.     

动物线粒体DNA的RFLP在家养动物遗传多样性研究中的应用(综述)

对线粒体ＤＮＡ限制性片段长度多态性（ＲＦＬＰ）在家养动物遗传多样性研究中的应用现状进行了综述,并对其应用前景进行了展望。众多的研究表明,线粒体ＤＮＡ的ＲＦＬＰ可以作为一个可靠的母性遗传标记用于家养动物起源分化及其亲缘关系的研究方面。我国有丰富的能够适应各种不同生态条件的优良畜禽品种遗传资源,因此利用线粒体ＤＮＡ的ＲＦＬＰ研究其遗传多样性对于探讨其起源分化以及今后合理的利用和保存将有十分重要的理论和实践意义。