Sequence of mitochondrial DNA cytochrome oxidase II inCryptopygus nanjiensis and Phylogeny of Apterygota

The mitochondrial cytochrome oxidase II (Co II) from four different apterygotensCryptopygus nanjiensis (Collembola),Neanura latior (Collembola),Gracilentulus maijiawensis (Protura) andLepidocampa weberi (Diplura) were sequenced. Their A+T content, number of nucleotide substitutions, TV/TV ratio, and Tamura-Nei’s distance were calculated. A series of phylogenetic trees were constructed by parsimony and distance methods using a crustaceanArtemia franciscana as outgroup. Finally the evolutionary trend A+T content of CO II genetic divergence and phylogenetic relationship of apterygotan groups were discussed.     

五种真蝽(Pentatoma Olivier) mtDNA-COII基因序列分析及系统发育关系初探(半翅目:蝽科)

测定并比较分析了真蝽属(Pentatoma)5种昆虫：褐真蝽P．semiannulata、斜纹真蝽P．illuminata、红足真蝽P．rufipes、H本真蝽P．japonica和金绿真蝽P．metallifera以及益蝽亚科2种外群种类(并蝽Pinthaeus humeralis和蠋蝽Arma custos)mtDNA—COII基因部分序列,依据分子数据建立了该5种昆虫的系统发育关系。结果显示,5种真蝽COII基因A T平均含量(73％)高丁弹尾目、缨尾目和直翅目昆虫,低于双翅目、鳞翅目及膜翅目昆虫,而与缨翅目的蓟马和半翅目的突角长蝽Oncopeltus fasciatus非常接近。系统发育结果显示,红足真蝽与日本真蝽亲缘关系最近,形成一个姊妹群,斜纹真蝽与金绿真蝽亲缘关系较近,也形成一个姊妹群,褐真蝽为较早分化的种类。     

Detection and phylogenetic analysis of Wolbachia in Collembola

Wolbachia are obligatory, cytoplasmatically inherited alpha-Proteobacteria which are known for infecting the reproductive tissues of many arthropods. Their prevalence in the large group of Collembola, however, is not known, except for PCR detection in the parthenogenetically reproducing species Folsomia candida (Order: Entomobryomorpha; Family: Isotomidae). In this study, fluorescence in situ hybridization on microscopic sections of F. candida specimens indicated that Wolbachia-related bacteria were restricted to tissues of the ovary and brain. PCR with primers designed to detect 16S rRNA genes of Wolbachia were positive with specimens from all of five geographically independent F. candida breeding stocks and with three parthenogenetic species from another order (Poduromorpha; Family Tullbergiidae), i.e. Mesaphorura italica, M. macrochaeta and Paratullbergia callipygos. In contrast, negative results were obtained with the two sexually reproducing species, Isotoma viridis (Isotomidae) and Protaphorura fimata (Poduromorpha; Onychiuridae). The ftsZ gene of Wolbachia could be PCR-amplified from all Wolbachia-positive hosts with the exception of M. macrochaeta. The phylogenetic distances of the ftsZ and 16S rRNA gene sequences reflected the phylogenetic distances of the host organisms but the sequences of Wolbachia were relatively closely related, indicating that Wolbachia infections took place after the Collembola had diversified. Our study confirms a monophyletic branch (supergroup E) of Collembola colonizing Wolbachia and indicates that this group is a sister group of supergroup A, the latter harbouring a high diversity of host organisms within the group of insects.     

Molecular phylogeny of Collembola inferred from ribosomal RNA genes

The classification of taxa within Collembola (Springtails, Hexapoda) has been controversial. In this study, we combined complete 18S rRNA gene with partial 28S rRNA gene (D7-D10) sequences to investigate the phylogeny of Collembola. About 2500 aligned sites of thirty species representing 29 genera from14 families of Collembola were analyzed, including one species of Neelipleona from which no sequence has been reported previously.The phylogenetic trees were obtained by different methods (maximum parsimony, maximum likelihood, and Bayesian analysis). Our results supported the monophyly of two of the four taxonomic groups of Collembola summarized by Deharveng [Deharveng, L., 2004. Recent advances in Collembola systematics. Pedobiologia 48, 415–433.], namely of Poduromorpha and of Symphypleona. Within Poduromorpha, Neanuridae was monophyletic with high support, but Hypogastruridae was not. Entomobryomorpha was paraphyletic, as the Tomoceroidea (Tomoceridae and Oncopoduridae) was found to be apart from the other entomobryomorphs. In the latter Isotomoidea and Entomobryoidea joined into a group with moderate support. Within Symphypleona, the phylogenetic relationship [(Sminthuridae + Bourletiellidae) + Sminthurididae] was consistent with traditional morphological studies. Neelipleona grouped with Symphypleona in all trees, with moderate support in the ML and Bayesian analyses.     

Distribution and diversity of springtails (Hexapoda: Collembola) on the altitudinal gradient of a temperate forest in Mexico

Mountain ecosystems have shown slow mineralization activity due to weather conditions, and to some groups with arthropods with special roles. The Collembola is an important group for litter fragmentation, showing different distribution patterns. The objective of the present study was to determine the diversity of Collembola along a volcano altitudinal gradient. For this, four sampling expeditions evaluated four altitudinal levels (I = 2 753, II = 3 015, III = 3 250 and IV = 3 687 masl) in Iztaccihuatl Volcano, from November 2003, and March, June and August 2004. Shannon diversity (H'), Pielou evenness (J') and Simpson dominance (1/lambda) indices were calculated. The similarity between the associations of springtails between the sampling sites was evaluated by a cluster analysis using the Pearson correlation coefficient, as distance and the unpaired arithmetic averages (UPGMA) as amalgamation method. A total of 24 075 springtails, distributed in 12 families, 46 genera and 86 species was collected. The higher species abundance was found at the altitudinal area II. The lowest diversity value was recorded in IV, while the higher diversity values were found in III. Significant differences between Shannon indices were found between zones II-III (t(0.01, 187) = 4.11, p < 0.05) and between III-IV (t(0.01, 187) = 3.8, p < 0.05) according to modified t-test. When considering sampling dates, no significant differences were found. The dendrogram showed that in composition the level I is more homogeneous throughout the year. In conclusion, a statistically significant seasonal variation in springtail abundances was not found, but it was observed that, the lower altitude (I) resulted more homogeneous along the studied period, followed by level II and III. There is a particular assemblage of springtails community in each altitudinal area studied; in general, the sites with low slope resulted more diverse in Collembola communities. These results show that there are important factors such as altitude, vegetation type and microhabitat heterogeneity that may affect the distribution of springtails communities along an altitudinal gradient.     

The complete mitochondrial DNA sequence of the basal hexapod Tetrodontophora bielanensis: evidence for heteroplasmy and tRNA translocations.

We present the complete 15,455-nt mitochondrial DNA sequence of the springtail Tetrodontophora bielanensis (Arthropoda, Hexapoda, Collembola). The gene content is typical of most metazoans, with 13 protein-coding genes (PCGs), 2 genes encoding for ribosomal RNA subunits, and 22 tRNA genes. The nucleotide sequence shows the well-known A+T bias typical of insect mtDNA; its A+T content is lower (72.7%) than that observed in other insect species, but still higher than that in other arthropodan taxa. The bias appears to be uniform across the whole molecule, unlike other insect taxa, which show increased A+T content in the so-called A+T-rich region. However, the bias is slightly higher in the third codon positions of the PCGs (81.4%). Anomalous initiation codons have been observed in the nad2 and the cox1 genes. In the latter, the ATTTAA hexanucleotide is suggested to be involved in the initiation signaling. All tRNAs could be folded into the typical cloverleaf secondary structure, but the tRNA for cysteine appears to be missing the DHU arm. Long tandemly repeated regions (193 nt) were found in the A+T-rich region, which in turn was shown to have the possibility of forming a complex array of secondary structures. One of these structures encompassed the junction between the repeats. The A+T-rich region was also interesting in that it showed heteroplasmy in the number of repeats. Three haplotypes were found, possessing 2, 3, and 4 identical repeats, respectively. The order of protein coding and rRNA genes in the molecule was determined and was identical to that of all insects studied so far. However, two tRNA translocations were found which were unprecedented among Arthropoda. These involved the trnQ, which was found between the rrnS and the A+T-rich region, and the trnS(ucn), which was located between trnM and trnI. A preliminary phylogenetic analysis based on the amino acid sequence of the PCGs failed to find support for the monophyly of Hexapoda.     

Brain organization in Collembola (springtails)

Arthropoda is comprised of four major taxa: Hexapoda, Crustacea, Myriapoda and Chelicerata. Although this classification is widely accepted, there is still some debate about the internal relationships of these groups. In particular, the phylogenetic position of Collembola remains enigmatic. Some molecular studies place Collembola into a close relationship to Protura and Diplura within the monophyletic Hexapoda, but this placement is not universally accepted, as Collembola is also regarded as either the sister group to Branchiopoda (a crustacean taxon) or to Pancrustacea (crustaceans + hexapods). To contribute to the current debate on the phylogenetic position of Collembola, we examined the brains in three collembolan species: Folsomia candida, Protaphorura armata and Tetrodontophora bielanensis, using antennal backfills, series of semi-thin sections, and immunostaining technique with several antisera, in conjunction with confocal laser scanning microscopy and three-dimensional reconstructions. We identified several neuroanatomical structures in the collembolan brain, including a fan-shaped central body showing a columnar organization, a protocerebral bridge, one pair of antennal lobes with 20-30 spheroidal glomeruli each, and a structure, which we interpret as a simply organized mushroom body. The results of our neuroanatomical study are consistent with the phylogenetic position of Collembola within the Hexapoda and do not contradict the hypothesis of a close relationship of Collembola, Protura and Diplura.     

Ferrovibrio denitrificans gen. nov., sp. nov., a novel neutrophilic facultative anaerobic Fe(II)-oxidizing bacterium

A neutrophilic Fe(II)-oxidizing bacterium was isolated from the redox zone of a low-salinity spring in Krasnodar krai (Russia), at the FeS-Fe(OH)(3) interface deposited at the sediment surface. The cells of strain Sp-1 were short, thin motile vibrioids with one polar flagellum dividing by binary fission. The optimal values and ranges for pH and temperature were pH 6.2 (5.5-8) and 35?°C (5-45?°C), respectively. The organism was a facultative anaerobe. Strain Sp-1 was capable of organotrophic, lithoheterotrophic and mixotrophic growth with Fe(II) as an electron donor. The denitrification chain was 'disrupted'. Oxidation of Fe(II) was coupled to reduction of NO3 - to NO2 - or of N(2) O to N(2) , as well as under microaerobic conditions, with O(2) as an electron acceptor. The DNA G+C content was 64.2?mol%. According to the results of phylogenetic analysis, the strain was 10.6-12% remote from the closest relatives, members of the genera Sneathiella, Inquilinus, Oceanibaculum and Phaeospirillum within the Alphaproteobacteria. Based on its morphological, physiological and taxonomic characteristics, together with the results of phylogenetic analysis, strain Sp-1 is described as a member of a new genus Ferrovibrio gen. nov., with the type species Ferrovibrio denitrificans sp. nov. and the type strain Sp-1(T) (=?LMG 25817(T) =?VKM B-2673(T) ).     

Molecular phylogenetic study of the Iranians based on polymorphic markers

The application of polymorphic markers in construction of phylogenetic trees has been documented. Five polymorphic markers located in the PAH gene region including PAH-BglII, PAH-PvuII(A), PAH-EcoRI, PAH-MspI and PAH-STR were selected for analysis of phylogenetic relationships of the Iranians with 15 other populations of the world. The lowest genetic distance was observed between the Iranians and populations residing in Adygei (an ethnic group of the Russian Caucasus), Russia and Druze (a Middle Eastern group). However, East Asian populations including Han, Japanese and Cambodians, Khmer or the Oceanians (Melanesian, Nasioi) showed high genetic distance with the Iranians. The data suggested that the Iranians might have relatively close evolutionary history with the populations residing in Russia rather than East Asian populations. This study provided the first new molecular insight into the evolutionary history of the Iranian population.     

Neutral lipid fatty acid composition as trait and constraint in Collembola evolution

Functional traits determine the occurrence of species along environmental gradients and their coexistence with other species. Understanding how traits evolved among coexisting species helps to infer community assembly processes. We propose fatty acid composition in consumer tissue as a functional trait related to both food resources and physiological functions of species. We measured phylogenetic signal in fatty acid profiles of 13 field‐sampled Collembola (springtail) species and then combined the data with published fatty acid profiles of another 24 species. Collembola fatty acid profiles generally showed phylogenetic signal, with related species resembling each other. Long‐chain polyunsaturated fatty acids, related to physiological functions, demonstrated phylogenetic signal. In contrast, most food resource biomarker fatty acids and the ratios between bacterial, fungal, and plant biomarker fatty acids exhibited no phylogenetic signal. Presumably, fatty acids related to physiological functions have been constrained during Collembola evolutionary history: Species with close phylogenetic affinity experienced similar environments during divergence, while niche partitioning in food resources among closely related species favored species coexistence. Measuring phylogenetic signal in ecologically relevant traits of coexisting species provides an evolutionary perspective to contemporary assembly processes of ecological communities. Integrating phylogenetic comparative methods with community phylogenetic and trait‐based approaches may compensate for the limitations of each method when used alone and improve understanding of processes driving and maintaining assembly patterns.     

云南省三种典型气候带凋落物层弹尾类多样性分布格局

弹尾类是土壤动物中常见的优势类群,其作为土壤微食物网的重要组成部分,参与凋落物分解、土壤团聚体形成等重要生态过程。以往对弹尾类分布格局的研究通常关注其在不同海拔梯度或者不同生境类型下的分布情况,但在不同气候带下弹尾类多样性沿纬度分布格局仍不清楚。为讨论不同气候带下弹尾类沿纬度的分布格局及其潜在的环境影响机制,于2017年10月(雨季末期)在云南省同一经度(E 101°)分布的三种典型气候带设置海拔梯度样带:热带雨林(西双版纳,800 m、1000 m、1200 m、1400 m)、亚热带常绿阔叶林(哀牢山,2000 m、2200 m、2400 m、2600 m)、亚高山针叶林(丽江玉龙雪山,3200 m、3400 m、3600 m、3800 m),采集凋落物层弹尾类并调查分析土壤温度、土壤含水量、凋落物厚度、土壤pH、土壤容重及土壤孔隙度等环境因子。利用Berlese-Tullgren法收集土壤动物,共获得弹尾类19150只,隶属于10科29属,其中符■属(39.9%,等节■科)数量最多,其余优势属为棘■属(21.7%,棘■科)和球角■属(10.1%,球角■科),这3个属合计占总体的71.7%。凋落物层弹尾类的密度在亚高山针叶林明显高于热带雨林和亚热带常绿阔叶林,弹尾类的属数排序由多到少依次为热带雨林、亚高山针叶林、亚热带常绿阔叶林。通过最小二乘回归法对弹尾类多样性的海拔格局进行回归分析,得出弹尾类的丰富度指数(Margalef′s指数、Menhinick′s指数)、多样性指数(Shannon-Wiener多样性指数、Simpson多样性指数)和均匀度指数(Pielou均匀度指数)沿热带雨林、亚热带常绿阔叶林、亚高山针叶林呈单调递减格局。通过全模型子集回归筛选最佳环境模型表明,温度是影响弹尾类多样性沿不同气候带分布格局的主要环境因子。本研究为预测不同气候带下弹尾类多样性如何响应环境变化提供参考。     

<Emphasis Type="Italic">Paenibacillus ginsengisoli</Emphasis> sp. nov., a novel bacterium isolated from soil of a ginseng field in Pocheon province,South Korea

A Gram-positive, aerobic or facultative anaerobic, motile, spore-forming bacterial strain, designated Gsoil 1638T, was isolated from a soil sample of a ginseng field in Pocheon province (South Korea), and was characterized taxonomically by using a polyphasic approach. It grew well on nutrient agar medium, utilized a fairly narrow spectrum of carbon sources and tolerated 10% NaCl. The isolate was positive for catalase and oxidase tests but negative for the degradation of macromolecules such as casein, collagen, starch, chitin, CM-cellulose, xylan and DNA. The G + C content of the genomic DNA was 50.7 mol%. The predominant isoprenoid quinone was menaquinone 7 (MK-7). The major fatty acids were anteiso-C15:0 (44%) and C16:0 (25%). Comparative 16S rRNA gene sequence analysis showed that strain Gsoil 1638T fell within the radiation of the cluster comprising Paenibacillus species and joined Paenibacillus anaericanus DSM 15890T with a bootstrap value of 100%. These two strains shared 99.5% 16S rRNA gene sequence similarity with each other. The phylogenetic distance from any other validly described species within the genus Paenibacillus was less than 96.2%. DNA-DNA relatedness value between strain Gsoil 1638T and its closest phylogenetic neighbor, Paenibacillus anaericanus, was 62%. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain Gsoil 1638T (= KCTC 13931T = LMG 23406T = CCUG 52472T) was classified in the genus Paenibacillus as the type strain of a novel species, for which the name Paenibacillus ginsengisoli sp. nov. is proposed.     

Fatty acid compositions of Collembola: unusually high proportions of C20 polyunsaturated fatty acids in a terrestrial invertebrate

To examine the C(20) polyunsaturated fatty acid (PUFA) compositions of Collembola, we raised five species of Collembola on yeast diets, and then quantified body mass, neutral lipid fatty acid (NLFA) and phospholipid fatty acid (PLFA) compositions. PLFA content was always less than 5% of dry weight, but NLFA content varied from 5.9% to 29.6% of dry weight, depending upon species. Combined C(20) PUFA proportions of up to 9.2% and 48% were observed in the NLFA and PLFA fractions, respectively, resulting in total C(20) PUFA proportions of up to 19.4% of the total fatty acid compositions of Collembola. C(20) PUFAs were also detected in Collembola specimens from a deciduous woodland at proportions up to 29.7% of the total fatty acid composition. Terrestrial invertebrates generally contain <4% and <22% C(20) PUFAs in PLFAs and NLFAs, respectively; therefore, these results demonstrate that Collembola often possess the highest proportions of C(20) PUFAs yet observed in terrestrial invertebrates. The biochemical reasons for such high C(20) PUFA proportions, which were biosynthesised by the Collembola since these components were absent from the yeast diets, remain unclear. The distinctive fatty acid compositions of Collembola may be useful in soil food web studies utilising fatty acids as biomarkers of trophic behaviour.     

跳虫系统进化的研究进展

跳虫是弹尾纲（Collembola）的俗称,在所有六足动物中化石年代最早,因此跳虫是六足动物起源及进化研究中非常重要的类群。跳虫的起源、分类地位和系统关系等问题,对于阐明六足动物甚至节肢动物各大类群的系统关系非常关键,日益成为相关学者关注和争论的焦点。本文就跳虫形态学和分子系统学方面的研究工作进行了综述。     

基于线粒体ND2基因的中国斑翅蝗科部分种类分子系统学研究（直翅目：蝗总科）

本文的目的是通过对斑翅蝗科部分种类的线粒体ND2基因进行分析,重建斑翅蝗科昆虫的系统发育关系,并探讨分子系统发育关系和传统分类结果的异同。扩增并测定了我国斑翅蝗科10属16种蝗虫的线粒体ND2全基因1 023 bp的序列,对序列的碱基组成、转换颠换、系统发育信号等进行了分析。并基于ND2全基因序列数据,分别采用邻接法（NJ）、最简约法（MP）、最大似然法（ML）和贝叶斯法重建了10属16种蝗虫的系统发育关系。结果表明：斑翅蝗科蝗虫ND2全基因A+T含量平均为74.6%;痂蝗亚科和异痂蝗亚科没能得到区分,建议合并为一个亚科;而斑翅蝗亚科和飞蝗亚科的分类地位还存在争议。     

Amino acid sequence of the dimeric hemoglobin (Hb I) from the deep-sea cold-seep clam Calyptogena soyoae and the phylogenetic relationship with other molluscan globins

The deep-sea cold-seep clam Calyptogena soyoae has two homodimeric hemoglobins (Hbs I and II) in erythrocytes. The complete amino acid sequence of Hb I has been determined. It is composed of 144 amino acid residues, has a high content of hydrophobic residues, and a calculated molecular weight of 16,350 including a heme group. The sequence of Calyptogena Hb I showed high homology (42% identity) with that of Calyptogena Hb II (Suzuki, T., Takagi T. and Ohta, S. (1989) Biochem. J. 260, 177-182), although it has a long insertion of seven residues in the C-terminal region compared with Hb II. On the other hand, it showed low homology (12-20% identity) with other molluscan globins. As well as Hb II, Calyptogena Hb I lacked the N-terminal extension of 7-9 residues characteristic of molluscan intracellular hemoglobins, and the distal (E7) histidine was replaced by glutamine. A phylogenetic tree was constructed from 13 molluscan globins belonging to the five families Aplysiidae, Galeodidae, Potamididae, Arcidae and Vesicomyidae. The globin sequences of Calyptogena (Vesicomyidae) were found to be rather distant from other globin sequences, suggesting that they might conserve a primitive form of molluscan globins.     

New insight into the systematics of Tomoceridae (Hexapoda,Collembola) by integrating molecular and morphological evidence

Tomoceridae is common but among the most problematic groups of Collembola. Its position within Collembola and the relationships within the family remain obscure. This also extends to the generic division of the subfamily Tomocerinae that remains controversial. This study examines these issues by integrating both molecular and morphological evidence. Our molecular phylogeny based on rDNA sequences supports the monophyly of Tomoceridae and the sister relationship between Tomocerinae and Lepidophorellinae. Reconstructions and tree topology tests constraining monophyly did not resolve the relationships between Tomoceridae and other collembolan groups. We also examined the morphology of the first instar (primary) larvae, which has significant phylogenetic value among higher Collembola. Mapping primary chaetotaxy onto our molecular phylogeny provided further evidence for the unique position of Tomoceridae within Entomobryomorpha and Collembola. The monophyly and subfamilial classification within Tomoceridae were validated here, whereas its position among Collembola will need further studies in a broader consideration across the major collembolan orders. Within Tomocerinae, the monophyly of Pogonognathellus was demonstrated, but the status of Tomocerus and Tomocerina is still to be resolved.     

小蔗螟Cytb基因序列的分析研究(鳞翅目:螟蛾总科:草螟亚科)

扩增并测定了小蔗螟线粒体细胞色素b(Cyt b)基因,其序列全长1146 bp,利用分子生物学软件分析了包括小蔗螟在内的4目(鳞翅目、双翅目、鞘翅目、缨尾目)共19条Cyt b基因序列.分析结果显示:鳞翅目昆虫AT含量明显低于其它昆虫,其中小蔗螟AT含量为75.1%;AT含量与A或T碱基使用的偏倚没有明显的相关性;碱基替换主要发生在密码子第三位,并且颠换率远大于转换率;密码子使用频率与基因碱基组成AT偏倚有相关性;相比于核苷酸数据,氨基酸所显示的遗传距离更为准确;使用Cyt b基因构建的系统进化树支持昆虫的单源进化学说.     

A novel copper(II) coordination at His186 in full-length murine prion protein

To explore Cu(II) ion coordination by His¹⁸⁶ in the C-terminal domain of full-length prion protein (moPrP), we utilized the magnetic dipolar interaction between a paramagnetic metal, Cu(II) ion, and a spin probe introduced in the neighborhood of the postulated binding site by the spin labeling technique (SDSL technique). Six moPrP mutants, moPrP(D143C), moPrP(Y148C), moPrP(E151C), moPrP(Y156C), moPrP(T189C), and moPrP(Y156C,H186A), were reacted with a methane thiosulfonate spin probe and a nitroxide residue (R1) was created in the binding site of each one. Line broadening of the ESR spectra was induced in the presence of Cu(II) ions in moPrP(Y148R1), moPrP(Y151R1), moPrP(Y156R1), and moPrP(T189R1) but not moPrP(D143R1). This line broadening indicated the presence of electron-electron dipolar interaction between Cu(II) and the nitroxide spin probe, suggesting that each interspin distance was within 20 Å. The interspin distance ranges between Cu(II) and the spin probes of moPrP(Y148R1), moPrP(Y151R1), moPrP(Y156R1), and moPrP(T189R1) were estimated to be 12.1 Å, 18.1 Å, 10.7 Å, and 8.4 Å, respectively. In moPrP(Y156R1,H186A), line broadening between Cu(II) and the spin probe was not observed. These results suggest that a novel Cu(II) binding site is involved in His186 in the Helix2 region of the C-terminal domain of moPrP^C.