慈姑属4种同域分布植物的人工种间杂交

泽泻科（Alismataceae）是现存最原始的单子叶植物类群之一,慈姑属（Sagittaria）是泽泻科相对进化的分类群,地域分布广泛,其物种大多集中分布于北温带,且有种间同域分布现象。为了探究同域分布物种之间的杂交可能性,我们通过选取同域分布且在系统进化树上具有代表性的冠果草（S.guayanensis subsp. lappula）、小慈姑（S. potamogetonifolia）、利川慈姑（S. lichuanensis）和野慈姑（S. trifolia）为研究对象,调研中国数字植物标本馆（CVH）的标本信息和已有文献,分析这4种慈姑的地理分布格局,并分别以这4种慈姑为母本进行人工授粉实验以检验种间杂交的可能性。基于对1 172份4种慈姑属腊叶标本的分析,表明这4种慈姑在福建省武夷山地区存在同域分布情况。通过对4种慈姑之间的人工授粉实验结果分析表明,冠果草、小慈姑、利川慈姑和野慈姑异种授粉后均出现心皮膨大现象,所有异种授粉产生的种子均不可育,败育胚种子比例各不相同,且该比例与父本和母本在系统进化树上的位置有关。     

Comprehensive molecular phylogenetic analysis and evolution of the genus Phyllactinia (Ascomycota: Erysiphales) and its allied genera

Phyllactinia is a unique genus within the Erysiphales (Ascomycota) having a partly endo-parasitic nature of the mycelium within the host plant tissues. We constructed phylogenetic trees for the genus Phyllactinia and its allied genera based on a total of 120 nucleotide sequences of the 28S rDNA and ITS regions to discuss their phylogenetic relationships with special references to host plants, biogeography, evolutionary dating, and taxonomy. The analysis of the Erysiphales confirmed the monophyly of the endo-parasitic genera, i.e. Leveillula, Phyllactinia, and Pleochaeta. Phyllactinia specimens used in this study were divided into six distinctive groups and three subgroups. Interestingly, Leveillula, an obligately endo-parasitic genus of the Erysiphales, grouped together with Phyllactinia, although this was not significantly supported by the Kishino–Hasegawa and Shimodaira–Hasegawa tests. This suggests that the evolution within this group of fungi occurred from partial endo-parasitism to obligate endo-parasitism. The host range of Phyllactinia is mostly confined to woody plants, especially deciduous trees. Betulaceae, Fagaceae, Ulmaceae, Moraceae, and Rosaceae may have close connections to the divergence of the groups and subgroups of Phyllactinia concerned. Most of these plant families are known as major members of the boreotropical flora of the Tertiary, which suggests an early Tertiary origin of this genus. A comparison of the phylogenies of hosts and parasites revealed that host range expansion at higher taxonomic levels (higher than family level) is independent of the phylogeny of plants. Conversely, host range expansions in lower taxonomic levels (infrafamilial or infrageneric) tend to occur within a single family or genus. An estimation of the evolutionary timing using a molecular clock approach suggested that Phyllactinia split from Pleochaeta about 60 M years ago (Ma) in the early Tertiary and divergence of the six major clades of Phyllactinia occurred between 5 and 40 Ma during the Oligocene and Miocene. Divergence within the major clades and within Leveillula occurred maybe from more than 5 Ma onwards during the Pliocene and Quaternary. This is the first comprehensive phylogenetic study of Phyllactinia and other endo-parasitic genera of the Erysiphales.     

Phylogeny and divergence times of some racerunner lizards (Lacertidae: Eremias) inferred from mitochondrial 16S rRNA gene segments

Eremias, or racerunners, is a widespread lacertid genus occurring in China, Mongolia, Korea, Central Asia, Southwest Asia and Southeast Europe. It has been through a series of taxonomic revisions, but the phylogenetic relationships among the species and subgenera remain unclear. In this study, a frequently studied region of the mitochondrial 16S rRNA was used to (i) reassess the phylogenetic relationships of some Eremias species, (ii) test if the viviparous species form a monophyletic group, and (iii) estimate divergence time among lineages using a Bayesian relaxed molecular-clock approach. The resulting phylogeny supports monophyly of Eremias sensu Szczerbak and a clade comprising Eremias, Acanthodactylus and Latastia. An earlier finding demonstrating monophyly of the subgenus Pareremias is corroborated, with Eremias argus being the sister taxon to Eremias brenchleyi. We present the first evidence that viviparous species form a monophyletic group. In addition, Eremias przewalskii is nested within Eremias multiocellata, suggesting that the latter is likely a paraphyletic species or a species complex. Eremias acutirostris and Eremias persica form a clade that is closely related to the subgenus Pareremias. However, the subgenera Aspidorhinus, Scapteira, and Rhabderemias seem not to be monophyletic, respectively. The Bayesian divergence-time estimation suggests that Eremias originated at about 9.9 million years ago (with the 95% confidence interval ranging from 7.6 to 12 Ma), and diversified from Late Miocene to Pleistocene. Specifically, the divergence time of the subgenus Pareremias was dated to about 6.3 million years ago (with the 95% confidence interval ranging from 5.3 to 8.5 Ma), which suggests that the diversification of this subgenus might be correlated with the evolution of an East Asian monsoon climate triggered by the rapid uplift of the Tibetan Plateau approximately 8 Ma.     

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。     

Pleistocene speciation of freshwater crabs (Crustacea: Potamidae: Geothelphusa) from northern Taiwan and southern Ryukyus, as revealed by phylogenetic relationships

The phylogenetic relationship among freshwater crab species of Geothelphusa from northern Taiwan and the Yaeyama Group of islands (including Iriomote and Ishigaki) in the southern Ryukyus was studied using the mitochondrial genes 16S rRNA and COI. Our results support the hypothesis that speciation of Geothelphusa among these islands was the result of cyclic glaciations and interglaciations during the Pleistocene. Two main clades, one the Taiwan Group (containing several clades, including most Taiwanese Geothelphusa species except Geothelphusa miyazakii but including Geothelphusa minei from Yaeyama), was estimated to be separated from its sister group, the southern Ryukyus-northern Taiwan (SRN) clade (including G. miyazakii, Geothelphusa shokitai, Geothelphusa fulva and G. marginata from northern Taiwan, the Pinnacle Islands [=Diaoyutai Islands or Senkaku Islands] and Yaeyama) at about 5.3 million years ago (mya). G. shokitai was separated from others within the SRN clade at 2.4 mya, but was probably derived from G. miyazakii in northern Taiwan. The ancestor of G. miyazakii is hypothesised to have dispersed from ancestors in Yaeyama and then isolated at 2.0 mya during the Pleistocene interglaciations. This is similar to the speciation of G. minei in Yaeyama at 1.5 mya, except that its ancestors originated from north-eastern Taiwan. Four clades of freshwater crabs are present in the Fushan Botanical Garden, located in the mountainous area of north-eastern Taiwan, which might be due to the historical rearrangements of the drainage and proximity of the various river origins.     

Molecular phylogeny and evolution of the genus Neoerysiphe (Erysiphaceae, Ascomycota)

The genus Neoerysiphe belongs to the tribe Golovinomyceteae of the Erysiphaceae together with the genera Arthrocladiella and Golovinomyces. This is a relatively small genus, comprising only six species, and having ca 300 species from six plant families as hosts. To investigate the molecular phylogeny and evolution of the genus, we determined the nucleotide sequences of the rDNA ITS regions and the divergent domains D1 and D2 of the 28S rDNA. The 30 ITS sequences from Neoerysiphe are divided into three monophyletic groups that are represented by their host families. Groups 1 and 3 consist of N. galeopsidis from Lamiaceae and N. galii from Rubiaceae, respectively, and the genetic diversity within each group is extremely low. Group 2 is represented by N. cumminsiana from Asteraceae. This group also includes Oidium baccharidis, O. maquii, and Oidium spp. from Galinsoga (Asteraceae) and Aloysia (Verbenaceae), and is further divided into four subgroups. N. galeopsidis is distributed worldwide, but is especially common in western Eurasia from Central Asia to Europe. N. galii is also common in western Eurasia. In contrast, the specimens of group 2 were all collected in the New World, except for one specimen that was collected in Japan; this may indicate a close relationship of group 2 with the New World. Molecular clock calibration demonstrated that Neoerysiphe split from other genera of the Erysiphaceae ca 35–45 M years ago (Mya), and that the three groups of Neoerysiphe diverged between 10 and 15 Mya, in the Miocene. Aloysia citriodora is a new host for the Erysiphaceae and the fungus on this plant is described as O. aloysiae sp. nov.     

Molecular evidence for species level divergence in African Nile Crocodiles Crocodylus niloticus (Laurenti, 1786)

Relationships of the newly discovered dwarf crocodiles from Mauritania were inferred from mitochondrial 12S sequences. Specimens from 13 different Crocodylus niloticus populations (from East Africa, West Africa and Madagascar) were compared. Additional representatives of the genus Crocodylus (one from Africa and one from Australia), the African genus Osteolaemus and the South American alligatorid Paleosuchus palpebrosus (as outgroup) were included in the analysis. Maximum-likelihood and Bayesian analyses yielded relationships that were strikingly different from currently prevailing phylogenetic hypotheses. Both analyses consistently revealed two groups, one consisting of the monophyletic West- and Central African populations and the other of a paraphyletic group containing the East African and Madagascan populations. High genetic divergence between those groups indicates separation on the species level. Furthermore ‘C’ cataphractus is clearly shown not to be a member of the genus Crocodylus. The resulting nomenclatural changes are discussed. To cite this article: A. Schmitz et al., C. R. Palevol 2 (2003).     

两种象甲幼虫肠道微生物组成及对高单宁食物的适应

【目的】象甲是栎属植物橡子中主要的寄生昆虫,但其适应高单宁食物(橡子)的肠道微生物基础尚待揭示。本研究分析了蒙古栎和辽东栎橡子中两种柞栎象(Curculio arakawai和C.dentipes)幼虫的肠道菌群结构和多样性,试图阐明柞栎象幼虫适应高单宁食物的肠道微生物基础。【方法】分别提取蒙古栎和辽东栎橡子中象甲幼虫各50头的肠道DNA,利用Illumina MiSeq技术对肠道菌群的V3–V4区序列进行16S rRNA测序,统计样品操作分类单元(OTU)数量,分析物种组成丰度、α多样性和β多样性。【结果】结果表明,可用于物种分类的OTU分别有2054和2308个,C. arakawai和C. dentipes共有的OUT 481个。在两种柞栎象C. arakawai和C. dentipes肠道菌群中,共注释到的主要分类阶元有27个门、145个科和274个属。变形菌门(Proteobacteria)、拟杆菌门(Bacteroidetes)和厚壁菌门(Firmicutes)在两种象甲肠道菌群中占优势;假单胞菌属Pseudomonas(63.8%)、沙雷氏菌属Serratia(6%)和不动杆菌属Acinetobacter (5.2%)是C. arakawai肠道中的主要类群,而沙雷氏菌属Serratia (32%)、拉恩菌属Rahnella(24.2%)、气单胞菌属Aeromonas(6.8%)和立克次体属Rickettsia(6.6%)在C.dentipes肠道菌群中占主导优势。C. arakawai和C. dentipes肠道菌群α多样性无显著差异,β多样性则差异显著。具有单宁酶活性的肠道细菌,如粘质沙雷菌Serratia marcescens、乳球菌Lactococcus lactis、假单胞菌Pseudomonas spp.在C. arakawai和C. dentipes之间差异不显著。【结论】寄生在蒙古栎和辽东栎橡子中的C. arakawai和C.dentipes肠道菌群组成迥异,这可能与遗传因素和食物特点有关。具有单宁酶活性的粘质沙雷氏菌Serratia marcescens和乳球菌Lactococcus lactis等菌类可能是两种象甲幼虫适应高单宁食物的肠道微生物基础。     

Genetic relatedness among fish species of Genus Channa using mitochondrial DNA genes

The family Channidae is represented by 26 species, out of which 23 species are found in Asia. However, the taxonomy and phylogeny of the Channid fishes found in India are poorly understood. In the present study, eight species of Channa (Channa striata, Channa punctatus, Channa marulius, Channa gachua, Channa stewartii, Channa aurantimaculata, Channa barca and Channa bleheri) were investigated using partial sequences of 16S rRNA and Cytochrome c Oxidase subunit I (COI) of mitochondrial genes to differentiate among the eight species and study their relationships. The sequence analysis of the genes revealed two distinct groups, which are genetically distant from each other and exhibit identical phylogenetic resolution. The partial sequences of both the genes provided sufficient phylogenetic information to distinguish all the eight species of Channa.     

Complex estimates of evolutionary relationships in Tarentola mauritanica (Reptilia: Gekkonidae) derived from mitochondrial DNA sequences

Mitochondrial DNA (12S rRNA, 16S rRNA) sequences were analysed within Tarentola mauritanica and other selected species of Tarentola. Several highly genetically distinct lineages occur in North Africa, revealing phylogroups in southern and central Morocco, northern Morocco, Algeria, Tunisia, and Libya. A single haplotype characterizes populations across Spain, Portugal, Italy, Menorca, Crete, and Tunisia raising the possibility of an anthropogenic introduction followed by rapid population expansion throughout southern Europe. T. mauritanica is paraphyletic with respect to T. angustimentalis, a Canary islands endemic. The high genetic diversity observed across North Africa suggests T. mauritanica may represent a species complex.     

Genetic differences among three species of the genus Trichiurus (Perciformes: Trichiuridae) based on mitochondrial DNA analysis

The mitochondrial DNA segment encoding the 16S ribosomal RNA (16S rRNA) gene sequence (ca. 600 bp) was compared among Trichiurus sp. 2 (sensu Nakabo, 2002) (obtained from various areas of Japan), T. japonicus Temminck and Schlegel (collected from various localities within Japan), and true T. lepturus Linnaeus (caught off the Atlantic coast of the United States and Brazil) of the family Trichiuridae using 10, 10, and 15 specimens, respectively. Based on phylogenetic analysis using a neighbor-joining (NJ) algorithm, the haplotypes of Trichiurus sp. 2, T. japonicus, and T. lepturus indicated three distinct monophyletic lineages, being supported by 100% bootstrap values with no haplotypes overlapping or sharing among the lineages. Trichiurus sp. 2, T. japonicus, and T. lepturus are genetically different from each other, suggesting that they are three distinct species.     

Molecular phylogeny of the lionfish genera Dendrochirus and Pterois (Scorpaenidae, Pteroinae) based on mitochondrial DNA sequences

This study investigates the molecular phylogeny of seven lionfishes of the genera Dendrochirus and Pterois. MP, ML, and NJ phylogenetic analysis based on 964 bp of partial mitochondrial DNA sequences (cytochrome b and 16S rDNA) revealed two main clades: (1) “Pterois” clade (Pterois miles and Pterois volitans), and (2) “Pteropterus–Dendrochirus” clade (remainder of the sampled species). The position of Dendrochirus brachypterus either basal to the main clades or in the “Pteropterus–Dendrochirus” clade cannot be resolved. However, the molecular phylogeny did not support the current separation of the genera Pterois and Dendrochirus. The siblings P. miles and P. volitans are clearly separated and our results support the proposed allopatric or parapatric distribution in the Indian and Pacific Ocean. However, the present analysis cannot reveal if P. miles and P. volitans are separate species or two populations of a single species, because the observed separation in different clades can be either explained by speciation or lineage sorting. Molecular clock estimates for the siblings P. miles and P. volitans suggest a divergence time of 2.4–8.3 mya, which coincide with geological events that created vicariance between populations of the Indian and Pacific Ocean.     

Oral Lactobacillus species in type 2 diabetic patients living in southern Thailand

Although lactobacilli are part of normal oral, gastrointestinal and genitourinary flora, they are an uncommon cause of infections in human. Lactobacillus-associated infections have generally occurred in patients with serious underlying conditions e.g. diabetes and cancer that might favour certain microorganisms. The aim of this study was to characterize species and genotypes of lactobacilli isolated from diabetic patients and non-diabetic subjects. One hundred and five type 2 diabetic patients and 103 non-diabetic subjects were recruited in this study. A total of 170 isolates of Lactobacillus were identified using 16S rRNA gene PCR-RFLP and genotyping were performed using AP-PCR by ERIC primers. It was found that type 2 diabetic patients had a significantly higher prevalence (p = 0.008) and level of lactobacilli than non-diabetic controls (p = 0.030). The most frequently isolated Lactobacillus spp. were L. casei/paracasei and L. fermentum in both the diabetic and non-diabetic groups. Strains of L. casei/paracasei and L. fermentum from between and within individuals were genotyped, and the genotyping of Lactobacillus strains showed diversity between individuals. One up to three genotypes of these two species could be found in the same subject. Interestingly, fewer genotypes were found in the diabetic patients than in the non-diabetic subjects.     

Molecular phylogeny and zoogeography of the freshwater crayfish genus Cherax Erichson (Decapoda: Parastacidae) in Australia

The evolutionary history and biogeography of freshwater-dependent taxa in Australia is of intrinsic interest given the present-day aridity of this continent. Cherax is the most widespread and one of the most species-rich of Australia's nine freshwater crayfish genera. The phylogenetic relationships amongst 19 of the 23 Australian Cherax were established from mitochondrial DNA sequences representing the 12S rRNA and 16S rRNA gene regions. The relationships among species support an initial east–west separation, followed by a north–south divergence in eastern Australia. Molecular clock estimations suggest that these divergences date back to the Miocene. The phylogenetic relationships support endemic speciation within geographical regions and indicate that long-distance dispersal has not led to recent speciation as previously hypothesized. This new evolutionary scenario is consistent with the climatic history of Australia and the evolutionary history of other similarly distributed freshwater-dependent organisms in Australia.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 81 , 553–563.     

乌鳢源杀鱼爱德华菌的分离鉴定及药敏特性研究

【背景】江苏省扬州市某乌鳢养殖场发生疾病,给养殖户造成了严重的经济损失。【目的】确定病因并筛选出敏感药物,为乌鳢相关疾病的防治提供参考。【方法】从患病乌鳢体内分离致病菌,并从形态、生理生化特征、16S rRNA和gyrB基因序列及特异性PCR检测等方面对分离菌株进行鉴定,同时开展人工感染试验分析其致病性,通过纸片扩散法进行药敏特性分析。【结果】从患病乌鳢体内分离获得优势菌株SHL,经形态特征、理化特性、16SrRNA和gyrB基因序列及特异性PCR检测鉴定为杀鱼爱德华菌。进一步人工感染试验证实其对乌鳢有较强的致病性,LD50为1.6×105 CFU/g,发病症状与自然发病症状相似。药敏试验结果显示,该菌株对青霉素、氯霉素、四环素等28种抗菌药物高度敏感,对红霉素中度敏感,对苯唑西啉、克拉霉素、万古霉素等6种药物耐药。【结论】引起江苏省扬州市某养殖场的乌鳢体表溃烂及死亡的病原菌为杀鱼爱德华菌,这是我国首次从淡水鱼类中检出致病性杀鱼爱德华菌,表明该菌的感染谱在扩大,需引起水产养殖领域的重视,在养殖过程中可根据药敏实验结果选用合适的国标渔药进行防治。     

南海野生斑节对虾的线粒体16S rRNA基因序列单倍型分析

采用聚合酶链式反应(PCR)技术对我国南海海域5个地点(三亚、深圳、阳江、湛江、北海)的野生斑节对虾100个个体的mtDNA 16S rRNA序列进行扩增,扩增产物经纯化后进行测序.用Clustal_X排序软件对所得的100个mtDNA 16S rRNA序列进行比对.通过ARLEQUIN软件对所得100个mtDNA 16S rRNA序列进行比较分析,共检测出28个变异位点,19种单倍型.19单倍型序列的碱基组成显示出较高的A T比例(69.0%).19种单倍型序列的遗传距离在0.002～0.033.根据构建的单倍型进化关系网,5个地点群体中的单倍型呈现一种混杂的分布格局.此外,根据单倍型在5个地点群体出现的频率和单倍型进化关系网,单倍型7是最为原始的单倍型,其他18种单倍型均起源于单倍型7.     

Hypocrella panamensis sp. nov. (Clavicipitaceae, Hypocreales): a new species infecting scale insects on Piper carrilloanum in Panama

A new species, Hypocrella panamensis, is described from collections and cultures obtained on Barro Colorado Island, Panama. In order to aid in placement of this fungus, phylogenetic analyses were conducted using LSU (rDNA) sequences. Hypocrella panamensis is characterized by possessing pulvinate stromata with a Lecanicillium-like anamorphic state and superficial perithecia. Hypocrella panamensis consistently grouped in a clade containing Hypocrella nectrioides, H. phyllogena, and H. africana (100 % PP). Most species of Hypocrella possess Aschersonia or Hirsutella anamorphs. Hypocrella panamensis is unique in the genus Hypocrella in possession of a Lecanicillium-like anamorphic state. In its biological habit Hypocrella panamensis is similar to other species in Hypocrella in that it infects and degrades the scale insect, then grows superficially on nutrients that emerge to the plant surface through the stylet wound.     

Molecular phylogeny of the Herpestidae (Mammalia, Carnivora) with a special emphasis on the Asian Herpestes

Until now, phylogenetic studies of the mongooses (Carnivora, Herpestidae) have not included an exhaustive sampling of the Asian members of this family. In this study, we used mitochondrial (Cytochrome b and ND2), nuclear (β-fibrinogen intron 7 and Transthyretin intron 1) sequences from almost all of the recognized mongoose species to produce a well-resolved phylogeny of the Herpestidae. We also performed molecular dating analyses to infer divergence dates of the different lineages within the Herpestidae. Our results confirmed the paraphyly of the Herpestes genus and other phylogenetic relationships, which previously had only been moderately supported. The Asian herpestid species were found to form a monophyletic group within the Herpestidae. Within the Asian species, a cyto-nuclear conflict was discovered between the small Indian mongoose (Herpestes auropunctatus), the Indian gray mongoose (Herpestes edwardsii) and the Javan mongoose (Herpestes javanicus), which may have occurred through interspecific hybridization. This study inferred an Early Miocene origin for the Herpestidae and a Middle Miocene origin for the Asian mongooses.     

Revision of some Thecaphora species (Ustilaginomycotina) on Caryophyllaceae

Specimens of Thecaphora saponariae s. lat. from several caryophyllacean host plants belonging to the genera Cerastium, Dianthus, Petrorhagia, Saponaria, Silene, and Stellaria were studied by means of both LM, SEM, and molecular phylogenetic analyses using ITS and LSU rDNA sequences. The data show that T. saponariae s. lat. is not uniform but represents several taxa. Molecular phylogenetic analyses, correlated with morphology of the sori, spore balls, and spores, permitted the recognition of five species. Two new species, Thecaphora italica and T. cerastii are described, and two new combinations, T. alsinearum and T. melandrii are proposed. The anamorph of Thecaphora saponariae is reported for the first time. A lectotype is designated for Sorosporium silenes-inflatae. Evolutionary aspects are discussed.