苏铁在种子植物进化中的位置：分子生物学的证据

用大分子rRNA快速测序法测定了苏铁(Cycas revoluta Thunb.)、水松[Glyptos-trobus pensilis(Staunt.)Koch]和南方红豆杉[Taxus mairei(Lemee et Levl)S.Y.Hu]三种裸子植物Ls-rRNA5′端区108个核苷酸的序列。用这些数据构建的rRNA系统树揭示了苏铁与水松和南方红豆杉构成一个关系密切的姐妹群,而所分析的被子植物都为另一自然类群。另以绿藻作为种子植物群外参照物种,表明苏铁这一位置并不是由于一个特异的进化速度所造成的。这一结果支持了苏铁与松杉类为一自然群类,它们的分歧发生在裸子植物与被子植物分歧之后的假说。     

A taxonomic investigation of variation withinCissus quadrangularis L. (Vitaceae) in Kenya

Using information from literature as well as records of localities on herbarium specimen labels at the East African Herbarium Nairobi (EA), materials ofCissus quadrangularis L.,C. cactiformis Gilg. andC. quinquangularis Chiov. were collected and new localities were sought. On the basis of the nature of their stem angles, the specimens ofC. quadrangularis were sorted into two variants, A and B. Variant A has smooth stem angles while variant B has rough stem angles. The authors have proposed that variant B should be recognized as a new variety withinC. quadrangularis. Variant A is the type variety ofC. quadrangularis. A map showing the distribution of the two variants in Kenya has been prepared. The rough-edged variant was found to be absent from the low-lying semi-arid regions of northern Kenya. The variants were found occurring in distinct, separate populations but without separate geographical patterns. The results of t-test show no significant difference (p=0.05) between the means of measurements of dimensions of various organs in the two variants. The stem angles of specimens of the two variants were studied under scanning electron microscope (SEM). This showed a clear morphological difference between the two variants. The results of the investigation support the hypothesis that the variation withinC. quadrangularis represents distinct kinds that can be recognized as taxa at infra-specific level. Foundation item: Supported by the China Ministry of Education Fund for University Staff Development in Key Subject Areas and Kenyatta University. Biography: Robert Wahiti. Gituru (1967-), male, Kenya, Ph. D. candidate, research direction: plant biosystematics.     

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.     

Molecular phylogenetics of Gymnocypris （Teleostei：Cyprinidae） in Lake Qinghai and adjacent drainages

149 complete mitochondrial DNA (mtDNA) cytochrome b (Cyt b) genes (1140 bp) of Gymnocypris przewaiskii, Gymnocypris eckloni and Gymnocypris scolistomus from the Lake Qinghai, Yellow River and Qaidam Basin were sequenced and analyzed. Consistent dendrogram indicated that the samples collected from the same species do not constitute a separate monophyletic group and all the samples were grouped into three highly divergent lineages (A, B and C). Among them, Lineage A contained all samples of G.przewaiskii from the Lake Qinghai and partial samples of the G. eckloni from the Yellow River. Lineage B contained the remaining samples of G. eckloni from the Yellow River.Lineage C was composed of a monophyletic group by G. eckloni from the Qaidam Basin. Analysis of molecular variance (AMOVA) indicated that most of genetic variations were detected within these three mtDNA lineages (93.12%), suggesting that there are three different lineages of Gymnocypris in this region. Our Cyt b sequence data showed that G.przewaiskii was not a polytypic species, and G. scolistomus was neither an independent species nor a subspecies of G.eckloni. The divergent mtDNA lineages of G. eckloni from theYellow River suggested that gene flow between the different populations was restricted to a certain extent by several gorges on the upper reach of the Yellow River. Lineage B of G. eckloni might be the genetic effect from the ancestor which was incorporated with the endemic schizothoracinefishes when the headward erosion of the Yellow Riverreached to its current headwaters of late. The G. eckloni from Basin Qaldam was a monophyletic group (lineage C) and Fst values within G. eckloni from the Yellow River were higher than 0.98, suggesting that the gene flow has been interrupted for a long time and the G. eckloni from Basin Qaidam might have been evolved into different species by ecology segregation. The correlation between the rakers number of Gymnocypris and population genetic variation was not significant.All Gymnocypris populations exhibited a low nucleotide diversity (n=0.00096-0.00485). Therefore the Gymnocypris populations from Basin Qaidam could have experienced severe bottleneck effect in history. Our result suggested Gymnocypris populations of Basin Qaidam should give a high priority in conservation programs.     

基于COI和D-Loop序列豫北虾虎鱼分子系统分析

虾虎鱼在河南省北部广泛分布,但其系统分类和多样性未有报道.从豫北原阳黄寺、安阳小南海、淇县淇河、长垣孙东闸、林州弓上水库等地采集虾虎鱼样本共计23尾,成都岷江样本6尾,分别扩增了线粒体COI基因和D-Loop序列.联合GenBank下载虾虎鱼的COI序列和D-Loop序列,构建了虾虎鱼的系统进化树.结果显示,采自黄寺、小南海、淇县、岷江地区的子陵吻虾虎鱼(Rhinogobius giurinus)聚为一个分支,而采自孙东闸和弓上水库的褐吻虾虎鱼(Rhinogobius brunneus)聚为一个单系群;这两种虾虎鱼遗传分化主要来自群体间.系统发育结果支持子陵吻虾虎鱼的韩国群体与中国群体构成姐妹群关系,中国不同地理种群间则相互交叉.     

中国主要红树科植物的分子系统发育

测定和分析了红树科（Ｒｈｉｚｏｐｈｏｒａｃｅａｅ）５属６种植物的ｒＤＮＡＩＴＳ区序列，所构建的ＩＴＳ区分子系统树图表明：红树科植物可形成１个单系的类群．但在部分属种ＩＴＳ序列分析的结果与红树科传统的分类系统有所不同，陆生红树科植物竹节树属（Ｃａｒａｌｉａ）的竹节树（Ｃｂｒａｃｈｉａｔａ）和旁杞木（Ｃｐｅｃｔｉｎｉｆｏｌｉａ）之间的分子差异很大，呈复系演化的特征．旁杞木与红树林主要植物秋茄属（Ｋａｎｄｅｌｉａ）的秋茄（Ｋｃａｎｄｅｌ）和红树属（Ｒｈｉｚｏｐｈｏｒａ）的红海榄（Ｒｓｔｙｌｏｓａ）的关系较近，竹节树则与另一红树林的主要种类角果木属（Ｃｅｒｉｏｐｓ）的角果木（Ｃｔａｇａｌ）呈姐妹群关系．     

Expression ofBcl-2 in cells with different telomerase activities

Both telomerase andBcl-2 are important genes in controlling apoptosis. The activation of telomerase and the abnormal regulation ofBcl-2 are also closely related to carcinogenesis. However, little is known about the linkage between telomerase andBcl-2. The effect of activated telomerase on the expression ofBcl-2 has been investigated. It is demonstrated that in tumor and transformed cells with higher telomerase activity,Bcl-2 expression is significantly lower than that in telomerase negative or less telomerose activity cells. Further study showed that in the telomerase gene-transformed 2BS-fibroblasts,Bcl-2 expression is inhibited significantly while the exogenous telomerase catalytic subunit gene is re-expressed in fibroblasts. Results indicated that there might be a certain linkage between the expression of telomerase andBcl-2, and overexpression of exogenous telomerase gene might down regulate the expression ofBcl-2. They contributed equally to this work.     

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     

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.     

Deuterostome phylogeny reveals monophyletic chordates and the new phylum Xenoturbellida

Deuterostomes comprise vertebrates, the related invertebrate chordates (tunicates and cephalochordates) and three other invertebrate taxa: hemichordates, echinoderms and Xenoturbella. The relationships between invertebrate and vertebrate deuterostomes are clearly important for understanding our own distant origins. Recent phylogenetic studies of chordate classes and a sea urchin have indicated that urochordates might be the closest invertebrate sister group of vertebrates, rather than cephalochordates, as traditionally believed. More remarkable is the suggestion that cephalochordates are closer to echinoderms than to vertebrates and urochordates, meaning that chordates are paraphyletic. To study the relationships among all deuterostome groups, we have assembled an alignment of more than 35,000 homologous amino acids, including new data from a hemichordate, starfish and Xenoturbella. We have also sequenced the mitochondrial genome of Xenoturbella. We support the clades Olfactores (urochordates and vertebrates) and Ambulacraria (hemichordates and echinoderms). Analyses using our new data, however, do not support a cephalochordate and echinoderm grouping and we conclude that chordates are monophyletic. Finally, nuclear and mitochondrial data place Xenoturbella as the sister group of the two ambulacrarian phyla. As such, Xenoturbella is shown to be an independent phylum, Xenoturbellida, bringing the number of living deuterostome phyla to four.     

A Note on the Prime Spectrums of Character Rings of Finite Groups

0Introduction LetGbeafinitegroupoforder|G|=g,andletR(G)denotethecharacterringofG,whichisgeneratedbyalltheirreduciblecomplexcharactersofG.LetZbetherationalintegerringandNthesetofnatural numbers,andletZ[ω]betheintegralextensiongeneratedbyaprimitiveg throotωofunity.SupposethatSisasubringof thealgebraicnumberfieldsuchthatZ[ω]S.πisasetofra tionalprimenumbersdefinedasfollowsπ={p｜pisarationalprimenumbersuchthatp-1S}.Definition1WecallthataconjugacyclassCofthefinite groupGisaπregularconju…     

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.     

Alteration of the specificity ofPst I restriction endonuclease

The influence of factors on the substrate-specificity ofPst I restriction endonuclease has been studied with the method of electrophoresis. The results show that, the specificity ofPst I almost can not be influenced by the single alteration of the concentration of Tris·HCl, Mg²⁺ or Na⁺ in the reaction system, but it can be altered by the reduction of any two of them. The specificity can not be altered by the single alteration of pH or the replacement of Mg²⁺ with Mn²⁺. The addition of glycerol or dimethylsulphoxide (DM-SO) to the reaction system results in the relaxation of the substrate-specificity ofPst I, but dimethyl-methylformide, glycol and ethyl alcohol can not bring about the alteration ofPst I specificity. Through the method of cloning and sequencing, the nucleotides of No. 1 and 6 in the recognition sequence ofPst I have changed (1C→A or 6G→T). Used with the enzyme analysis of an artificially synthetic DNA segment containing a special sequence, the nucleotides of No. 1 and 6 have both changed (1C→A and 6G→T). The recognition sequence ofPst I is speculated to be changed from CTGCA→G to TGCA→. Foundation item: Supported by the Research Fund for the Doctoral Program of Higher Education. Biography: Zou Guo-lin (1947-), male, professor. research direction, biochemistry.     

Molecular phylogenetics of <Emphasis Type="Italic">Gymnocypris</Emphasis> (Teleostei: Cyprinidae) in Lake Qinghai and adjacent drainages

149 complete mitochondrial DNA (mtDNA) cytochrome b (Cyt b) genes (1140 bp) of Gymnocypris przewalskii, Gymnocypris eckloni and Gymnocypris scolistomus from the Lake Qinghai, Yellow River and Qaidam Basin were sequenced and analyzed. Consistent dendrogram indicated that the samples collected from the same species do not constitute a separate monophyletic group and all the samples were grouped into three highly divergent lineages (A, B and C). Among them, Lineage A contained all samples of G. przewalskii from the Lake Qinghai and partial samples of the G. eckloni from the Yellow River. Lineage B contained the remaining samples of G. eckloni from the Yellow River. Lineage C was composed of a monophyletic group by G. eckloni from the Qaidam Basin. Analysis of molecular variance (AMOVA) indicated that most of genetic variations were detected within these three mtDNA lineages (93.12%), suggesting that there are three different lineages of Gymnocypris in this region. Our Cyt b sequence data showed that G. przewalskii was not a polytypic species, and G. scolistomus was neither an independent species nor a subspecies of G. eckloni. The divergent mtDNA lineages of G. eckloni from the Yellow River suggested that gene flow between the different populations was restricted to a certain extent by several gorges on the upper reach of the Yellow River. Lineage B of G. eckloni might be the genetic effect from the ancestor which was incorporated with the endemic schizothoracine fishes when the headward erosion of the Yellow River reached to its current headwaters of late. The G. eckloni from Basin Qaidam was a monophyletic group (lineage C) and F_st values within G. eckloni from the Yellow River were higher than 0.98, suggesting that the gene flow has been interrupted for a long time and the G. eckloni from Basin Qaidam might have been evolved into different species by ecology segregation. The correlation between the rakers number of Gymnocypris and population genetic variation was not significant. All Gymnocypris populations exhibited a low nucleotide diversity (π = 0.00096–0.00485). Therefore the Gymnocypris populations from Basin Qaidam could have experienced severe bottleneck effect in history. Our result suggested Gymnocypris populations of Basin Qaidam should give a high priority in conservation programs.     

Aberrant activity of Wnt/Frizzled signaling pathway in renal cancer cell lines

The expression ofWnt, Wntreceptor-Frizzled, and several other key components in Wnt pathway in renal cancer cell lines was studied. The result of semi-quantitative RT-PCR has shown that the expression level ofWnt5A andhFz5 mRNA were higher in renal cancer cell line (GRC-1) than in normal renal cell line (HK-2). This result has been confirmed byin situ hybridization. The expression of β-catenin protein was obviously higher in GRC-1 than in HK-2 (P< 0.01), but there were no different expressions of its mRNA between 3 lines. The reasons of the overexpression of β-catenin has been investigated by means of immunocytochemistry, SSCP and so on, no mutation ofβ-catenin gene and APC were found. That means that the overexpression ofWnt5A/hFz5 might be the reason of overexpression of β-catenin. It was concluded that the aberrant activity of Wnt pathway might play an important role in renal cell carcinoma.     

Phylogeny of fireflies (Coleoptera: Lampyridae) inferred from mitochondrial 16S ribosomal DNA，with references to morphological and ethological traits

We sequenced partial mitochondrial 16S ribosomal DNA (16S rDNA) of 18 firefly species from Southwest of China. Combined with homologous sequences previously reported, phylogenetic trees including Japanese, Korean and Chinese species were reconstructed by neighbor-joining, maximum parsimony and Bayesian methods. All reconstructions agree on most nodes of the trees. Monophyly of Lampyridae is not supported because Rhagophthalmus ohbai in Rhagophthalmidae is included within it. Lamprigera, a genus placed unreliably in Lampyrinae, shows a close relationship to Amydetinae. Monophyly of Luciolinae is not supported because Pristolycus sagulatus (Lampyrinae) is included within it. In the Luciolinae, monophyly of Curtos and Hotaria is well established, respectively, but both morphological and molecular data continue to indicate that Luciola is not monophyletic and its subdivision is indeed necessary. Within the Lampyrinae, both Pyrocoelia and Diaphanes are not monophyletic, but monophyly of Pyrocoelia+Diaphanes is well supported. Phylogeny of Diaphanes is discussed for the first time. Generic placement of a newly discovered species (Diaphanes pectinealis Li et Liang) sharing some characters of Pyrocoelia and Diaphanes challenges the delimitation of these two similar genera. With references to the firefly mating systems, we suggest that more emphases should be placed on those sexually selected characters such as antennal structure in taxonomy of Lampyridae.     

医蛭形亚目常见物种系统发育基因组学研究

本研究基于GenBank中10个医蛭形亚目物种及2个近缘外类群的基因组或转录组数据,提取COI、18 S和蛋白编码核基因的直系同源序列（Orthologs）,开展系统发育分析。比对后的COI、18 S和Orthologs序列长度分别为1533、1832、220095 bp。最大似然法和贝叶斯推断法分析显示,基于Orthologs序列构建的系统发育树在几乎所有节点上的支持率都达到100%,明显优于COI和18 S序列得到的结果。研究表明,医蛭科的欧洲医蛭、东方医蛭和侧纹医蛭组成一个单系群,医蛭科的日本医蛭与黄蛭科的宽体金线蛭和尖细金线蛭组成另一单系群,而黄蛭科的马蛭与上述6个物种形成姐妹群。医蛭科的两种牛蛭（菲牛蛭和棒纹牛蛭）组成一个单系群,此单系群与医蛭科的其他物种的遗传关系比马蛭更疏远;山蛭科的洞穴山蛭在10个医蛭形亚目种类中处于最基部位置。本研究阐明了医蛭形亚目常见物种的系统发育关系,为这些物种的资源开发利用和保护提供科学依据。     

Sauropterygian from Triassic of Guizhou, China

A new sauropterygian,Anshunsaurus huangguoshuensis, is described on the basis of a well-preserved skull from the Late Triassic Wayao Member of Falang Formation, Guizhou Province, southwestern China. It is ascribed to Eusauroptetrygia because it possesses the following characters: the rostrum is mainly composed by the premaxillae; the nasals are small, paired and separated from one another by the premaxillae; the external nares are close to the orbits; and the frontals are fused.Anshunsaurus differs from other Triassic sauropterygians in its large size, long rostrum and the supratemporal fenestrae slightly smaller than orbits. It is further considered to be closely related toPistosaurus on the basis of the nasals contact with the prefrontals; large pineal foramen, lain in the anterior part of the parietal table; the squamosals not in contact posterior to the supratemporal fenestra. The discovery ofAnshunsaurus with the ichthyosaurs in the same locality indicates a new assemblage of Triassic marine tetrapods in China.     

Phylogeny of higher taxa of hexapoda according to 12sRNA sequences

The phylogenetic relationship of Hexapoda has been debated for a long time, which will be resolved mainly depending on the settlement of monophyly, affinities and interrelationships among Protura, Collembola and Diplura. Mitochondrial 12sRNA gene about 355 bp fragments from one proturan species, two collembolan species, two dipluran species and one oribatid species were sequenced. The Kimura 2-parameter distances were calculated and a series of molecular phylogenetic trees were reconstructed by using the N-J method, from which the following points were drawn: (ⅰ) Protura and Collembola compose a monophyletic group representing absent-cerci; (ⅱ) Diplura is not a monophyletic group, in which Campodeoid with filiform cerci belongs to a clade and Japygoid with pincer cerci and Ectognatha compose another clade, that is, Insecta s. str. stemmed from Japygoid. So it would be suggested that the phylogenetic relationship of Hexapoda is [Parainsecta (Collembola + Protura) +Campodeoid +Insecta (Japygoid + Ectognatha)].