Phylogenetic analyses of some genera in Oedipodidae （Orthoptera： Acridoidea） based on 16S mitochondrial partial gene sequences

Basedonthe 16S mitochondrial partial gene sequences of 29 genera, containing 26 from Oedipodidae and one each from Tanaoceridae, Pyrgomorphidae and Tetrigidae （as outgroups）, the homologus sequences were compared and phylogenetic analyses were performed. A phylogenetic tree was inferred by neighbor-joining （N J）. The results of sequences compared show that： （i） in a total of 574 bp of Oedipodidae, the number of substituted nucleotides was 265 bp and the average percentages ofT, C, A and G were 38.3%, 11.4%, 31.8% and 18.5%, respectively, and the content of A＋T （70.1%） was distinctly richer than that of C＋G （29.9%）; and （ii） the average nucleotide divergence of 16S rDNA sequences among genera of Oedipodidae were 9.0%, among families of Acridoidea were 17.0%, and between superfamilies （Tetrigoidea and Acridoidea） were 23.9%, respectively. The phylogenetic tree indicated： （i） the Oedipodidae was a monophyletic group, which suggested that the taxonomic status of this family was confirmed; （ii） the genus Heteropternis separated from the other Oedipodids first and had another unique sound-producing structure in morphology, which is the type-genus of subfamily Heteropterninae; and （iii） the relative intergeneric relationship within the same continent was closer than that of different continents, and between the Eurasian genera and the African genera, was closer than that between Eurasians and Americans.     

Demographic parameters of cotton aphid, Aphis gossypii Glover （Homoptera： Aphididae） on five cotton cultivars

Life table parameters of the cotton aphid, Aphis gossypii Glover, were estimated on five cotton cultivars （‘ Sealand＇ ,‘Siokra＇ ,＇ Vararnin＇ ,‘ Bakhtegan＇ and ‘ Sahel＇ ）. Demographic parameters of the cotton aphid were assessed at 27.5 ± 1 ℃, 65% ＋ 10% RH and a photoperiod of 14：10 （L： D）h. The shortest developmental time for the nymphal stages was 5.5 days on ‘Siokra＇ and the longest was 6.1 days on‘ Sealand＇. The highest offspring per female was 29.6 on ‘Sahel＇ and the lowest was 15.3 on ‘Sealand＇. The rm values varied from 0.272 on ‘Sealand＇ to 0.382 （day^-1） on ‘Varamin＇. Jackknife estimates of the A. gossypii parameters on these cultivars indicated the greatest developmental rate and fecundity on ‘ Varamin＇ and the poorest on ‘ Sealand＇ cultivar.     

Differentiation of non-pylori Helicobacter species based on PCR–restriction fragment length polymorphism of the 23S rRNA gene

Phenotypic identification of non-pylori Helicobacter species has always been problematic and time-consuming in comparison with many other bacteria. We developed a rapid two-step identification assay based on PCR–restriction fragment length polymorphism (PCR–RFLP) analysis of the 23S rRNA gene for differentiating between non-pylori Helicobacter species. A new genus-specific primer pair based on all available complete and partial 23S rRNA sequences of Helicobacter species was designed. In silico restriction analysis of variable regions of the 23S rRNA gene suggested SmaI and HindIII endonucleases would provide a good level of differentiation. Analysis of the obtained 23S rRNA RFLP patterns divided all Helicobacter study strains into three species groups (groups A–C) and 12 unique restriction patterns. Wolinella succinogenes also gave a unique pattern. Our proposed PCR–RFLP method was found to be as a valuable tool for routine identification of non-pylori Helicobacter species from human or animal samples.     

Resolution of phylogenetic relationships of the major subfamilies of the Delphacidae （Homoptera： Fulgoroidea） using the mitochondrial ribosomal DNA

Delphacid relationships from the genus level to the subfamily have been completely resolved （among those taxa examined） using sequence data from the 3＇ end of the 12S gene. Monophyly of the non-asiracine subfamilies was strongly supported and the asiracine Ugyops was placed in the most basal position of the tree. Support levels for monophyly of the Delphacini increased after weighting transversions more heavily than transitions and after removing the cixiid outgroup from the dataset. Among the Delphacini, Conomelus and Megamelus were more closely related to each other than either was to Chloriona. These results are in agreement with the tree based on morphological characters. However, in contrast to morphological data our results strongly supported a sister group relationship between the Stenocraninae and the Kelisiinae. Although the 12S gene fragment gave some information about the species relationships within Chloriona, neither this fragment nor the 5＇ end of the 16S gene appear to be very useful for this level. Molecular evolutionary patterns provided evidence that there has been a shift in base composition from T to A during the early evolution of the non-Asiracinae. The non-Asiracinae also had comparatively fast substitution rates and these two observations are possibly correlated. In the ‘ modem＇ delphacid Chloriona, the AT content was comparatively low in regions free of constraints but this was not the case for ‘ non-modem＇ delphacids. The tRNA for valine has been translocated elsewhere, probably before the Delphacidae and Cixiidae diverged from each other.     

Comparison of bacterial communities in the Solimões and Negro River tributaries of the Amazon River based on small subunit rRNA gene sequences

The microbiota of the Amazon River basin has been little studied. We compared the structure of bacterial communities of the Solim?es and Negro Rivers, the main Amazon River tributaries, based on analysis of 16S rRNA gene sequences. Water was sampled with a 3-L Van Dorn collection bottle; samples were collected at nine different points/depths totaling 27 L of water from each river. Total DNA was extracted from biomass retained by a 0.22-μm filter after sequential filtration of the water through 0.8- and 0.22-μm filters. The 16S rRNA gene was amplified by PCR, cloned and sequenced, and the sequences were analyzed with the PHYLIP and DOTUR programs to obtain the operational taxonomic units (OTUs) and to calculate the diversity and richness indices using the SPADE program. Taxonomic affiliation was determined using the naive Bayesian rRNA Classifier of the RDP II (Ribosomal Database Project). We recovered 158 sequences from the Solim?es River grouped into 103 OTUs, and 197 sequences from the Negro River library grouped into 90 OTUs by the DOTUR program. The Solim?es River was found to have a greater diversity of bacterial genera, and greater estimated richness of 446 OTUs, compared with 242 OTUs from the Negro River, as calculated by ACE estimator. The Negro River has less bacterial diversity, but more 16S rRNA gene sequences belonging to the bacterial genus Polynucleobacter were detected; 56 sequences from this genus were found (about 30% of the total sequences). We suggest that a more in-depth investigation be made to elucidate the role played by these bacteria in the river environment. These differences in bacterial diversity between Solim?es and Negro Rivers could be explained by differences in organic matter content and pH of the rivers.     

Polyphasic taxonomic studies of lactic acid bacteria associated with Tunisian fermented meat based on the heterogeneity of the 16S–23S rRNA gene intergenic spacer region

The objective of this work was to investigate the structure and diversity of lactic acid bacteria (LAB) communities in traditionally fermented meat collected from different areas of Tunisia. A polyphasic study, which involves phenotypic tests and ribosomal DNA-based techniques, was used to identify Gram-positive and catalase-negative isolates. PCR amplification of the 16S–23S rDNA ISR of 102 isolates and other reference LAB strains gave (1) one type of rrn operon (M-ISR) for lactococci, (2) two types of rrn operon (S-ISR and M-ISR) for enterococci, (3) two types of rrn operon (S-ISR and L-ISR) for Lactobacilli, and (4) three PCR amplicons (S-ISR, M-ISR, and L-ISR) obtained for Pediococcus spp. and Weissella genus. The clustering and comparison of ISR–RFLP profiles given by the isolates with those given by reference LAB strains, allowed their identification as Lactococcus lactis, Enterococcus faecium, Enterococcus faecalis, Enterococcus sanguinicola, Enterococcus hawaiiensis, Lactobacillus sakei, Lactobacillus curvatus, Lactobacillus plantarum, Lactobacillus alimentarius, Pediococcus pentosaceus, and Weissella confusa. Combined 16S–23S rDNA ISR and RFLP patterns can be considered as a good potential target for a rapid and reliable differentiation between isolates of LAB and provided further information on the organization of their rrn operons.     

Water Striders： The Biomechanics of Water Locomotion and Functional Morphology of the Hydrophobic Surface （Insecta： Hemiptera-Heteroptera）

Water striders are insects living on the water surface, over which they can move very quickly and rarely get wetted. We measured the force of free walking in water striders, using a hair attached to their backs and a 3D strain gauge. The error was calculated by comparing force and data derived from geometry and was estimated as 13%. Females on average were stronger （1.32 mN） than males （0.87 mN）, however, the ratio of force to weight was not significantly different. Compared with other lighter species, Aquarius paludum seems stronger, but the ratio of force to weight is actually lower. A. paludum applies about 0.3 mN.cm^-1 to 0.4 mN.cm 1 with its mid-legs, thus avoiding penetrating the surface tension layer while propelling itself rapidly over the water surface. We also investigated the external morphology with SEM. The body is covered by effectively two layers of macro-and micro-hairs, which renders them hydrophobic. The setae are long （40 μm-60μm） and stiff, being responsible for waterproofing, and the microtrichia are much smaller （〈10μm）, slender, and flexible, holding a bubble over the body when submerged.     

Detection of food source by PCR analysis of the gut contents of Aldrichina grahami （Aldrich） （Diptera： Calliphoridae） during post-feeding period

The last meal of sarcophagous maggots may be useful in identifying the species on whose flesh they have fed （the＂host＂species）. The DNA profile of the host species may indeed be detectable in the＂last meal＂. In this paper, mitochondrial DNA analysis of gut contents was used to identify the prior host of post-feeding larvae of Aldrichina grahami （Aldrich） （Diptera： Calliphoridae）. A modified logistic equation was fitted to estimate the probability of identifying the host under five different constant temperatures （16, 20, 24, 28 and 32 ℃）. Our results shows that the detected time ranged from a maximum of 24 h at 32℃ to 42 h at 16℃ and a minimum of 12 h at 32~C to 30 h at 16℃. Furthermore, the host detection time was also calculated to give the maximal time after larval hatching from the egg. These results indicate that, in criminal cases where the maggots stray from the corpse, the last meal of the larvae should not be overlooked as potentially critical evidence.     

Assessing the sensitivity of Melanoplus frigidus （Orthoptera： Acrididae） to different weather conditions＂ A modeling approach focussing on development times

The temperature and soil moisture conditions as well as vegetation patterns were studied to describe the habitat and to model the life cycle of Melanoplusfrigidus, a true alpine grasshopper of the Scandes. In the low alpine belt of the Norwegian Scandes the species colonizes only the warmest microhabitats with maximum soil surface temperatures of 31℃. Vegetation of these habitats consists of shrub-rich heath dominated by Vaccinium myrtillus and Calluna vulgaris. Using continuously measured temperature data, the development times for four different seasons were modeled and related to field observations. The maximum delay of adult molt was estimated to amount to 3 weeks, the delay being determined by the variation in spring temperature conditions between different years. The possibilities of using M. frigidus as an indicator organism of climate change effects on alpine zoo-coenoses of the Scandes are discussed.     

Development of setal patterns in embryonic and adult aphids of the Hormaphidinae （Hemiptera： Aphididae）

The elementary traits and diversity of embryonic setal patterns of 31 species of 22 genera, in three tribes of Hormaphidinae are studied. The embryonic setal patterns to every species are provided. Based on the parsimony principle and the developmental principle, these patterns are analyzed in each tribe following the terminology of Richards （1965）. It is suggested that the setal patterns of abdominal tergites are the most important characters in embryonic and adult setal patterns. In conclusion, neoteny is a phylogenetic trend in the Hormaphidinae. Nipponaphidini and Hormaphidini have more typical neoteny in each genus and more diversity in different types of adult viviparous females than Cerataphidini, by showing more frequent and various protopatterns or mesopatterns.     

基于线粒体Cytb基因序列探讨中国斑翅蝗科部分种类的系统发育关系

基于斑翅蝗科14种的线粒体Cytb基因462 bp序列,使用MEGA2和PAUP4.0b软件包进行分析,显示Cytb基因序列具有明显的高A、T偏向性和距离依赖的TS/TV值.采用邻接法(NJ)、最大似然法(MP)和极似然法(ML)分别构建斑翅蝗科4亚科8属的系统发育树,不同算法构建的系统发育树均支持分为四大分支(亚科):A(绿纹蝗属)、B(束颈蝗属)、C((飞蝗属+车蝗属)+小车蝗属)、D((痂蝗属+异痂蝗属)+皱膝蝗属);红胫小车蝗和黄胫小车蝗作为两个种比较合适;痂蝗亚科和异痂蝗亚科应该合并为一个亚科;飞蝗亚科与斑翅蝗亚科的分类关系尚未解决,需要进一步的研究进行分析验证.     

利用线粒体16S rRNA基因全序列分析直翅目主要类群的系统发生关系

为了构建稳健的直翅目主要类群间的系统发生关系并探讨16S rRNA基因序列在构建直翅目昆虫不同分类阶元系统发生关系时的可行性、功效以及性能,文章测定了直翅目4总科9科18种昆虫的16S rRNA基因全序列,联合已知该基因全序列的其他40种昆虫,构建了直翅目主要类群之间的系统发生关系,并分析了16SrRNA基因全序列的系统发生性能和功效。结果表明,直翅目昆虫的16S rRNA基因全长平均为1 310 bp;除生活方式特化的蚤蝼总科和蝼蛄总科的地位无法确定外,直翅目其他主要类群系统发生关系比较稳定;蝗总科下除了斑翅蝗科和槌角蝗科外,剑角蝗科、斑腿蝗科、网翅蝗科都不是单系群,且用不同的方法构建的系统发生树中聚类情况完全一致,各科间遗传距离差异不大,建议将其合为一科;锥头蝗科、瘤锥蝗科和癞蝗科间的遗传距离差异也不大;在构建系统发生树时,16S rRNA基因环区的信息量要比茎区的大;16S rRNA基因可以构建可靠的直翅目属与种水平和目与亚目高级阶元的系统发生关系,但对科和总科阶元缺乏足够的分辨力。     

基于12S,16S和28S rRNA基因序列的中国小毛瓢虫属系统发育分析

【目的】小毛瓢虫属Scymnus Kugelann昆虫主要捕食蚜虫、蚧虫等害虫,是一类经济上重要的天敌昆虫。目前针对小毛瓢虫属的系统发育研究尚属空白,亚属之间的系统演化关系尚不明确,为了建立合理的分类系统,亟需对小毛瓢虫属的亲缘关系进行研究和探讨。【方法】以华南农业大学馆藏的小毛瓢虫属5亚属共44种为研究对象,采用PCR技术对12S, 16S和28S rRNA基因的部分序列进行扩增;运用MEGA 7.0分析了小毛瓢虫属内12S, 16S和28S rRNA基因的碱基组成,基于K2P模型计算了小毛瓢虫属44种的种间遗传距离;采用最大似然法(maximum-likelihood, ML)和贝叶斯推断法(Bayesian-inference, BI)构建该属的系统发育树。【结果】扩增获得小毛瓢虫属44种的12S rRNA基因序列平均长度为356 bp, 16S rRNA基因序列平均长度为351 bp, 28S rRNA基因序列平均长度为315 bp;序列分析表明,12S rRNA基因的A, T, G和C平均含量分别为38.8%, 43.5%, 11.9%和5.8%, 16S rRNA基因的A, T, G和C平均含量分别为37.6%, 40.3%, 14.4%和7.7%, 28S rRNA基因的A, T, G和C平均含量分别为26.7%, 18.3%, 31.4%和23.5%;基于联合序列分析的种间遗传距离为0.004～0.276,平均遗传距离为0.115。系统发育分析结果表明,小毛瓢虫属为单系起源,而小毛瓢虫亚属Scymnus(Scymnus) Kugelann、毛瓢虫亚属Scymnus(Neopullus) Sasaji、小瓢虫亚属Scymnus(Pullus) Mulsant和拟小瓢虫亚属Scymnus(Parapullus) Yang均为并系起源。【结论】基于12S, 16S和28S rRNA基因序列的小毛瓢虫属系统发育分析显示传统的形态学分类体系与基于分子数据分析的结果部分不一致,这表明应该对该属内各亚属的鉴别特征进行全面检视,筛选并确立各亚属的形态指标,同时也表明该属内的亚属分类单元需重新厘定。     

九种蟋蟀mtDNA-16S rRNA基因序列及其系统进化(直翅目:蟋蟀科)

蟋蟀科5属9个种的线粒体16S rRNA基因部分序列被测定或从Gen Bank获得,比较其同源性,计算核苷酸使用频率,并构建NJ和MP分子系统树。在获得的449bp的序列中A、T、C和G碱基含量分别为31．8％、36．9％、9．9％和21．4％,A T平均含量为68．7％。研究结果表明：所研究的5属9种蟋蟀聚成3个聚类簇,斗蟋属先与灶蟋属汇合,再与棺头蟋属构成聚类簇Ⅰ;油葫芦属黑脸油葫芦和北京油葫芦与蟋蟀属的家蟋相聚构成聚类簇Ⅱ;蟋蟀属的田蟋单独构成聚类簇Ⅲ。     

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。     

拟诺卡氏菌16S rRNA,gyrB,sod和rpoB基因的系统发育分析

为了更好地了解拟诺卡氏菌属(Nocardiopsis)各物种间的系统发育关系,该属现有有效描述种的gyrB,sod和rpoB基因的部分序列被测定,结合16S rRNA基因,对拟诺卡氏菌属进行了系统发育重建。研究发现拟诺卡氏菌属gyrB,sod和rpoB基因的平均相似性分别为87.7%、87.3%和94.1%,而16S rRNA基因的平均相似性则达到96.65%,3个看家基因均比16S rRNA具有更高的分歧度。比较基于不同基因的系统树发现,由gyrB基因得到的系统树拓扑结构与16S rRNA得到的结构在亚群上基本一致。因此,gyrB基因在拟诺卡氏菌属的系统分类上比16S rRNA基因更具优越性。     

Phylogenetic relationships of the Japanese minnows,Pseudorasbora (Cyprinidae), as inferred from mitochondrial 16S rRNA gene sequences

The phylogenetic relationships among threePseudorasbora fishes (Cyprinidae, Sarcocheiichthyinae) occurring in Japan (P. parva, P. pumila pumila andP. pumila subsp. sensu Nakamura [1963]) were inferred from nucleotide sequences of the mitochondrial 16S rRNA gene. The sequences. of 1240 bp, were determined and compared for 22 specimens from 2–8 populations for each taxon, with a singlePungtungia herzi specimen as an outgroup. A total of 171 sites (13.8%) were variable among the specimens, but only 0–2 sites within each population. The phylogenetic relationships estimated by neighbor-joining, maximum-parsimony and maximum-likelihood methods confirmed a sister relationship between the twoP. pumila subspecies, with a high level of confidence. However, their genetic distinction from each other (4.1±0.4SD % sequence difference on average) was at a level similar to that between them andP. parva (5.9±0.5%). The geographic distribution of the twoP. pumila subspecies, which are separated by the Fossa Magna region, suggests that the genetic divergence of the two subspecies originated from a vicariant process separating the freshwater ichthyofaunas of eastern and western Honshu.Pseudorasbora parva populations were divided into two genetic groups (1.8±0.2% sequence difference), one group comprising continental and part of the Japanese populations, and the other the remaining Japanese populations. This suggests that at least two genetically divergent lineages had been originally distributed in Japan, but a strong possibility remains that the present situation has resulted from artificial transplantation.     

Phylogenetic analysis of the genus Aquaspirillum based on 16S rRNA gene sequences

Phylogenetic analysis of 15 species of the genus Aquaspirillum based on 16S rRNA gene (rDNA) sequences indicated that the genus Aquaspirillum is phylogenetically heterogeneous and the species could be divided into four groups as follows: Aquaspirillum serpens, the type species of this genus, A. dispar and A. putridiconchylium are situated in the family Neisseriaceae; members of the second group, A. gracile, A. delicatum, A. anulus, A. giesbergeri, A. sinuosum, A. metamorphum and A. psychrophilum, are included in the family Comamonadaceae; the two members of the third group, A. arcticum and A. autotrophicum, are included in the family Oxalobacteriaceae; and members of the fourth group, A. polymorphum, A. peregrinum, and A. itersonii, are included in the alpha-subdivision of Proteobacteria. Thus, phylogenetic studies indicated that all the species excepting A. serpens, the type species, should be transferred to distinct genera.