Estimating divergence times among subfamilies in Nymphalidae

The mitochondrial cytochrome oxidase subunit Ⅰ （COⅠ） gene and the nuclear elongation factor 1α （EF-1α） gene were sequenced from 13 species of Nymphalidae. Phylogenetic trees of Nymphalidae, which is the largest family in butterflies, were constructed based on the sequences determined from 13 species sequenced in our laboratory and an additional 43 species obtained from GenBank using the maximum likelihood （ML） and Bayesian methods. Relative-rate tests between lineages in these phylogenetic trees were performed. On the basis of the results of the relative-rate tests and fossil information of Satyrinae, Nymphalinae and Biblidinae, the average divergence times among the subfamilies are estimated as 44.2-87.1 million years ago （Ma）. These results will be helpful for better understanding of the origin and evolution of this family, as well as the divergence time of butterflies and other complex taxa.     

Phylogenetic relationships and divergence times of the family Araucariaceae based on the DNA sequences of eight genes

Araucariaceae is one of the most primitive families of the living conifers, and its phylogenetic relationships and divergence times are critically important issues. The DNA sequences of 8 genes, i.e., nuclear ribosomal 18S and 26S rRNA, chloroplast 16S rRNA, rbcL, matK and rps4, and mitochondrial coxl and atpl, obtained from this study and GenBank were used for constructing the molecular phylogenetic trees of Araucariaceae, indicating that the phyiogenetic relationships among the three genera of this family should be （（Wollemia, Agathis）, Araucaria）. On the basis of the fossil calibrations of Wollemia and the two tribes Araucaria and Eutacta of the genus Araucaria, the divergence time of Araucariaceae was estimated to be （308± 53） million years ago, that is, the origin of the family was in the Late Carboniferous rather than Triassic as a traditional view. With the same gene combination, the diver- gence times of the genera Araucaria and Agathis were （246 ± 47） and （61 ±15） Ma, respectively. Statis- tical analyses on the phylogenetic trees generated by using different genes and comparisons of the divergence times estimated by using those genes suggested that the chloroplast matK and rps4 genes are most suitable for investigating the phylogenetic relationships and divergence times of the family Araucariaceae.     

Asian origin for Polystichum （Dryopteridaceae） based on rbcL sequences

Chloroplast rbcL sequences of 60 species of Polystichum sensu lato (s.l.), including 23 new sequences from southwest China, were used to assess the phylogenetic relationships within the genus. On the basis of estimated evolution rate of rbcL gene and the genetic distance data that passed relative-rate tests, we further estimated the divergence times between some clades of the genus. The phylogenetic relationships were inferred using the neighbor-joining and maximum-parsimony methods, both methods producing trees with completely congruent topology. These trees reveal that all species of Polystichum s.l. in this study (including Cyrtomium and Cyrtomidictyum) form a monophyletic group. The basal split in Polystichum s.l. separates a clade with all Asian members from a clade containing other species from all over the world. The phylogenetic and divergence time estimation results lead us to suggest that Polystichum s.l. originated in Asia in the late Late Cretacous (≈76 Ma) and migrated into other places in the world in early Eocene(≈46 Ma).     

Phylogeny and divergence of Chinese Angiopteridaceae based on chloroplast DNA sequence data （rbcL and trnL-F）

Marattioid ferns are an ancient lineage of primitive vascular plants that first appeared in the middle Carboniferous. Extant members are almost exclusively restricted to tropical regions, and the spe- cies-rich family Angiopteridaceae are limited in their distribution to the eastern hemisphere; relation- ships within the group are currently vague. Here the phylogenetic relationship between Angiopteris Hoffm. and Archangiopteris Christ et Gies. was evaluated based on the sequence analysis of chloro- plast rbcL gene and trnL-F intergenic spacer with MEGA2 and MrBayes v3.0b4. On the basis of the phy- logenetic pattern and fossil record, we further estimated the divergence time for the two genera. The phy- logenetic trees revealed that all species of Angiopteris and Archangiopteris in this study formed a monophyletic group with strong statistical support, but the relationship between the two genera re- mained unresolved based on individual sequence analysis. On the other hand, the sequence analyses of combined data set revealed that Archangiopteris species diverged first, indicating that Archangiop- teris may not be a direct derivative as traditionally assumed. The clade of Angiopteris and Archan- giopteris appears to have diversified in the late Oligocene (≈26 Ma) based on the molecular estimate. Thus, the evolutionary history of extant Angiopteris and Archangiopteris has been characterized by ancient origin and recent diversification, and these groups are not relic and endangered lineages as traditionally considered.     

Investisations into the perplexing interrelationship of the Genus Takifugu Abe, 1949 （Tetraodontiformes,Tetraodontidae）

The phylogenetic relationships within the genus Takifugu Abe, 1949 （Tetraodontiformes, Tetraodontidae） remain unresolved. Because of the use of Takifugu as model organisms, the resolution of these relationships is crucial for the interpretation of evolutionary trends in biology. Pufferfishes of this genus are comprised of a comparatively small number of species and are mainly distributed along the coastal region of the western part of the Sea of Japan and the coastline of China. Mitochondrial gene sequences were employed to test the phylogenetic hypotheses within the genus. Seventeen species of the genus were examined. Molecular phylogenetic trees were constructed using the maximum parsi- mony, neighbor-joining, maximum likelihood and Bayesian methods. Our hypothesis of internal relationships within the genus differs from previous hypotheses. Our results indicate that （1） the genus Takifugu is a monophyletic assemblage; （2） the genus is divided into 6 subgroups based on the moecular data; and （3） there is low genetic diversity among the species within this genus. In addition, speciation within Takifugu appears to be driven by hybridization and isolation by distribution. Our results also suggested that the taxonomy in the genus should be clarified based on both molecular and morphological data.     

Bayesian mixed models and divergence time estimation of Chinese cavefishes （Cyprinidae： Sinocyclocheilus）

The genus Sinocyclocheilus is distributed in Yun-Gui Plateau and its surrounding region only, within more than 10 cave species showing different degrees of degeneration of eyes and pigmentaUon with wonderful adaptations. To present, published morphological and molecular phylogenetic hypotheses of Slnocyclocheilus from prior works are very different and the relationships within the genus are still far from clear. We obtained the sequences of cytochrome b （cyt b） and NADH dehydrogenase subunlt 4 （ND4） of 34 species within Sinocyclocheilus, which represent the most dense taxon sampling to date. We performed Bayesian mixed models analyses with this data set. Under this phylogenetic framework, we estimated the divergence times of recovered clades using different methods under relaxed molecular clock. Our phyloegentic results supported the monophyly of Sinocyclocheilus and showed that this genus could be subdivided into 6 major clades. In addition, an earlier finding demonstrating the polyphyletlc of cave species and the most basal position of S. jii was corroborated. Relaxed divergence-time estimation suggested that Sinocyclocheilus originated at the late Miocene, about 11 million years ago （Ma）, which is older than what have been assumed.     

The complete mitochondrial DNA sequence and the phylogenetic position of Achalinus meiguensis (Reptilia: Squamata)

The mitochondrial genome complete sequence of Achalinus meiguensis was reported for the first time in the present study. The complete mitochondrial genome of A. meiguensis is 17239 bp in length and contains 13 protein-coding genes, 22 tRNA, 2 rRNA, and 2 non-coding regions (Control regions). On the basis of comparison with the other complete mitochondrial sequences reported, we explored the characteristic of structure and evolution. For example, duplication control regions independently occurred in the evolutionary history of reptiles; the pseudo-tRNA of snakes occurred in the Caenophidia; snake is shorter than other vertebrates in the length of tRNA because of the truncations of TψC arm (less than 5 bp) and “DHU“ arm. The phylogenic analysis by MP and BI analysis showed that the phylogenetic position of A. meiguensis was placed in Caenophidia as a sister group to other advanced snakes with the exclusion of Acrochordus granulatus which was rooted in the Caenophidia. Therefore we suggested that the subfamily Xenodermatinae, which contains A. meiguensis, should be raised to a family rank or higher rank. At the same time, based on the phylogenic statistic test, the tree of Bayesian was used for estimating the divergence time. The results showed that the divergence time between Henophidia and Caenophidia was 109.50 Mya; 106.18 Mya for divergence between Acrochordus granulatus and the other snakes of the Caenophidia; the divergence time of A. meiguensis was 103 Mya, and Viperidae diverged from the unilateral of Elapidae and Colubridae was 96.06 Mya.     

Mitochondria1 cytochromeb sequences variation of Protura and molecular systematics of Apterygota

Mitochondrial cytochromeb genes of about 450 bp fragments from 3 proturan species, 5 collembolan species and 2 dipluran species have been sequenced. The number of nucleotide substitutions and Kimura 2-parameter distances have also been calculated, and a series of molecular phylogenetic trees reconstructed by using parsimony and distance methods. The proturan, collembolan and dipluran species have evolved monophyletic groups. The results suggest that Protura and Collembola are sister groups, while Diplura is more or less demonstrating a closer phylogenetic relationship to the pterygotan insects. The phylogeny and their systematic position of protura and other groups are also discussed.     

牛大力内生细菌的多样性及其促生特性研究

为研究牛大力Callerya speciose内生细菌多样性组成及其促生特性,从经表面消毒处理的牛大力根茎组织中分离内生细菌,采用16SrDNA序列分析法对内生细菌的多样性进行系统发育分析,并以固氮、溶磷性、产铁载体、产IAA能力等为指标,对内生细菌进行促生特性检测。从牛大力根和茎中共分离到59株内生细菌,包括16属20种,芽孢杆菌属是牛大力的优势菌群;在20种分离菌株中,分别有11,17,7株具有潜在的固氮、溶磷性、产铁载体能力,有13株菌可以合成IAA。其中RH17-2、RH17-5、GR2-1 3株菌同时具备固氮、溶磷性、产铁载体、产IAA 4种促生特性。从牛大力中分离得到的内生细菌具有丰富的物种多样性和促生特性,在未来的农业生产上具有潜在的应用价值。     

Mitochondrial 16S rRNA sequence variations and phylogeny of the Chinese sisorid catfishes

Partial sequences of mitochondrial 16S rRNA gene were obtained by PCR amplification for comparisons among nine species of glyptosternoid fishes and six species of non-glyptosternoids representing 10 sisorid genera. There are compositional biases in the A-rich unpaired regions and G-rich paired regions. A-G transitions are primarily responsible for the Ts/Tv bias in unpaired regions. The overall substitution rate in unpaired regions is almost two times higher than that in the paired regions. Saturation plots at comparable levels of sequence divergence demonstrate no saturation effects. Phylogenetic analyses using both maximum likelihood and Bayesian methods support the monophyly of Sisoridae. Chinese sisorid catfishes are composed of two majorlineages, one represented by (Gagata (Bagarius, Glyptothorax)) and the other by “glyptosternoids Pseudecheneis“. The glyptosternoids may not be a monophyletic group. A previous hypothesis referring to Pseudecheneis as the sister group of monophyletic glyptosternoids, based on morphological evidence, is not supported by the molecular data. Pseudecheneis is shown to be a sister taxon of Glaridoglanis. Pareuchiloglanis might be paraphyletic with Pseudexostoma and Euchiloglanis. Our results also support the hypothesis that Pareuchiloglanis anteanalis might be considered as the synonyms of Pareuchiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi.     

Molecular phylogeny and biogeography of the highly specialized grade schizothoracine fishes （Teleostei： Cyprinidae） inferred from cytochrome b sequences

We recovered the phylogenetic relationships among 23 species and subspecies of the highly special-ized grade schizothoracine fishes distributing at 36 geographical sites in the Tibetan Plateau and its surrounding regions by analyzing sequences of cytochrome b genes. Furthermore, we estimated the possible divergent times among lineages based on a historical geological isolation event in the Tibetan Plateau. The molecular data revealed that the highly specialized grade schizothoracine fishes were not a monophyletic group, but were the same as genera Gymnocypris and Schizogypsis. Our results indi-cated that the molecular phylogenetic relationships apparently reflected their geographical and historical associations with drainages, namely species from the same and adjacent drainages clustered together and had close relationships. The divergence times of different lineages were well consistent with the rapid uplift phases of the Tibetan Plateau in the late Cenozoic, suggesting that the origin and evolution of schizothoracine fishes were strongly influenced by environment changes resulting from the upheaval of the Tibetan Plateau.     

基于线粒体蛋白编码基因探讨飞虱科Delphacidae13属系统发育关系

飞虱科昆虫隶属于半翅目蜡蝉总科,是世界上最重要的农业害虫之一.昆虫的线粒体基因是昆虫分子与进化研究中常用且有效的分子标记.本研究基于线粒体蛋白编码基因探讨了飞虱科13属18种的系统发育关系,分别采用距离法、最大似然法和贝叶斯法构建的系统发育树得到的结论是一致的,即在飞虱科13属中,Ugyops属较为原始,处于系统树的基...     

Phylogenetic positions of some genera and species of the family Buccinidae (Gastropoda: Mollusca) from China based on ribosomal RNA and COI sequences

A phylogenetic analysis of members of the family Buccinidae was conducted using 18S rRNA gene, 28S rRNA gene and the mitochondrial cytochrome oxidase Ⅰ gene. We studied 18 species of Buccinidae that belong to eight different genera and inhabit the China coastal seas. We analyzed the patterns of divergence between an outgroup and basal ingroup taxa, the monophyly of the genus Neptunea, and the position of one unnamed species within the Buccinidae. A phylogenetic tree (neighbor-joining (NJ) method) was reconstructed based on the sequences of 18S rRNA, 28S rRNA and COI, with Rapana venosa as outgroup. The NJ tree indicated that the 18 species could be divided into five groups. The genus Buccinum was monophyletic, whereas Neptunea was shown to be paraphyletic since it included Siphonalia subdilatata and Neptunea sp., a new species. This novel species otherwise clustered consistently with Neptunea cumingi in three other phylogenetic trees, showing a low genetic distance and divergence percentage of sequences belonging to the genus Neptunea. A smaller genetic distance and a smaller difference of 18S rRNA, 28S rRNA and COI sequences between Neptunea cumingii and Neptunea arthritica cumingii confirmed them to be the same species.     

金花茶回归地植物群落调查研究

【目的】了解野生金花茶(Camellia nitidissima)种群中植物种类、植物类型和植物群落的特征,为开展金花茶回归提供参考。【方法】在广西防城金花茶国家级自然保护区内选取2个金花茶回归样地,开展回归试验,实地调查样地的植物群落,对样地的土壤性质、植物种类、乔木层结构特征、灌木层结构特征、植物生态特性等进行分析。【结果】2个金花茶回归样地的物种数均超过80种,胸径大于1cm的树种个体数超过250株;所有种类中木本植物比草本植物的种类多,两者的比例为87∶32,其中木本植物中常绿树种和落叶树种的比例为73∶14;植物种类中银柴(Aporosa chinensis)和桃金娘(Rhodomyrtus tomentosa)个体数量占绝对优势,分别为43株和40株。【结论】2个回归地的植物物种多样性较丰富,其中乔灌层的种类数和个体数均比草本层多,且多为常绿树种,因乔灌层的树冠可为金花茶植物提供遮阴环境,这正好符合金花茶作为阴生树种的特性。     

PCR amplification of Sox genes inMisgurnus anguillicaudatus andParamisgurnus dabryanus (Cypriniforms; Cobitidae)

Using the primers which specially amplify the conservative motif of Human SRY gene, we studied the PCR amplification of Sox genes in genomic DNA of two species of mud loach:Misgurnus anguillicaudatus andParamisgurnus dabryanus. Four bands with the length of 200,550,940 and 1000 bp respectively, were presented in the PCR products ofMisgurnus anguillicaudatus. Three bands with the length of 200,550 and 900 bp were presented in that ofParamisgurnus dabryanus. Southern blotting results indicated that the 200 and 550 bp bands are specially positive. There is no difference between male and female individuals as well as between these two species. Chang zhongjie: Born in Nov. 1965, Ph. D. graduate student     

Phylogenetic position of Ephedra rhytidosperma, a species endemic to China： Evidence from chloroplast and ribosomal DNA sequences

The chloroplast genes matK and rbcL, ribosomal gene 18S and ITS regions of nuclear ribosomal DNA from Ephedra rhytidosperma, a species endemic to China, were sequenced and its phylogenetic position was investigated. Independent and combined phylogenetic analyses for the DNA sequences from 16 taxa representing 15 species of the genus Ephedra were performed using the maximum parsimony （MP）, neighbor-joining （N J）, minimum evolution （ME） and maximum likelihood （ML） methods. The results indicate that E. rhytidosperma is closely related to E. equisetina. The divergence time between them is estimated to be 10.85±2.44 Ma based on the results of the relative-rate tests and the evolutionary rate of rbcL gene.     

上海吴淞口浮游植物优势种类调查

自上海吴淞口采集的浮游植物样品,通过显微观察和分子生物学技术,共发现浮游植物6门31属32种,其中绿藻门种类最多,硅藻门次之,鉴定出优势种类17种(属).不同季节水体中出现的优势种类及其优势度都存在差异,土生伪鱼腥藻(Pseudanabaena mucicola)和梅尼小环藻(Cyclotella meneghiniana)为全年优势种.确定了该区域藻类爆发的潜在种类,为后续优势种属的竞争机制研究提供种类纯化培养筛选依据.     

井冈山国家自然保护区毛竹林害虫初步调查

2011~2014年对井冈山国家自然保护区毛竹林内害虫进行初步调查,发现有毛竹害虫8目48科108属127种。按多样性来看,鳞翅目最高(包含15科39属52种),占全部种类的40.94%,其次是半翅目(包含9科30属32种)和鞘翅目(包含7科17属20种)。从危害部位来看,叶、杆类害虫为主,占害虫总数的88.19%。调查发现绝大部分害虫危害较小,仅黄脊竹蝗(Ceracris kiangsu)、青脊竹蝗(Ceracris nigricornis)、小绿叶蝉(Empoasca flavesce)、刚竹毒蛾(Pantana phyllostachysae)等危害较为严重。     

The syntheses and properties of poly(L-lactide)

Poly (L-lactide) was prepared by bulk polymerization ofL-lactide at 140°C in the presence of stannous octoate. The polymer products were characterized by number average molecular weight Mn, weight average molecular weight Mw and molecular weight distribution MWD (M _w/M _n) respectively. The results showed that stannous octoate was a high effective catalyst, and coordinated insertion mechanism of the reaction was proposed. Foundation itemL Supported by the key Project of the Ministry of Science and Technology (96-920-21) Biography: Chen lian-xi (1964-), male, Associate Professor, research direction: organic and polymeric materials.     

Genetic diversity of rhizobial populations recovered from three Lotus species cultivated in the infra-arid Tunisian soils

Eighty-three bacterial strains isolated from root nodules of Lotus creticus, L. pusillus, and L. arabicus grown in infra-arid Tunisian soils were characterized using a polyphasic approach including phenotypic analysis, rep-PCR and PCR-RFLP analyses of the 16S rRNA gene. Phenotypically, all isolates are fast growers the majority of which grow at a pH of between 5.5 and 9. Most of the tested isolates tolerate NaCl concentrations from 1.39% to 3.48%. By rep-PCR fingerprinting, the genomic similarity varied from 30% to 98%. All tested isolates were clustered into 32 rep-PCR clusters at the similarity level of 80%. The genomic divergence of strains revealed by rep/PCR analysis appeared to be very important since a molecular polymorphism delimiting symbionts for each species of Lotus was identified. With the high-resolution of rep-PCR profiles of the isolates obtained using Pearson’s/UPGMA analysis, the isolates were resolved into 60 different profiling groups to undergo 16S ARDRA analyses. The analysis of all restriction fragments from each strain based on the UPGMA algorithm from the combined patterns showed that Lotus isolates are very diverse and that they were affiliated to Sinorhizobium, Rhizobium, and Mesorhizobium genera.