Mitochondrial cytochrome b sequences variation of Protura and molecular systematics of Apterygota

Mitochondrial cytochrome b 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.     

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)].     

Testing relative evolutionary rates and estimating divergence times among six genera of Rhizophoraceae using cpDNA and nrDNA sequences

The molecular phylogenetic trees of 10 species representing 6 genera of the family Rhizophoraceae have been constructed using the sequences of chloroplast genesmatK andrbcL as well as the ITS regions of nuclear ribosomal DNA. Relative-rate tests between lineages in these phylogenetic trees have been performed. On the basis of the results of the relative-rate tests and related molecular evolutionary rate data, the divergence times between the lineages are estimated as follows: (i) the first divergence time in these genera is 132.25 million years ago (mya); (ii) the average divergence time between two tribes, i.e. inland Legnotideae (exceptCarallia brachiata) and mangrove Rhizophoreae, is 64.13 mya; and (iii) the average divergence time between two inland species,C. garciniaefolia andC. pectinifolia, is 19.92 mya.     

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.     

Isolation of candidateR disease resistance genes from rice

Using a polymerase chain reaction (PCR) based method six distinct candidate disease resistant gene (R) homologs from rice have been isolated. The rice sequences are organized into two phylogenetic groups with contrasting genomic organization patterns. The first group, represented by a single sequence, Osh359-1, is more similar to non-riceR sequences than to rice ones and has a simple genomic organization. The second group, represented by Osh359-3, contains the remaining five rice sequences. Osh359-3 consists of a multi-gene family. The members of Osh359-3 family are further found to be clustered together in the genome.     

Morphological evolution from aquatic to terrestrial in the genus Oreolalax （Amphibia, Anura, Megophryidae）

A phylogeny of 17 species in the genus Oreolalax is reconstructed based on 21 morphological characters from adult specimens, skeleton specimens, tadpoles and eggs. Four species groups are recognized, of which the O. rugosus species group is the most primitive, the O. weigoldi species group is the second, the O. omeimontis species group is the third and the O. pingii species group is the most recently diversified. Based on the evolutional tendency of the morphological characters on the phylogenetic tree, it is proposed that the evolution of tympanum, tympanic annulus, columella, spoon-like cartilage and the web between toes reflect the habit changes from aquatic to terrestrial. Thus, Oreolalax is regarded as one important representative genus to study further the evolution of morphological characters from aquatic to terrestrial.     

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).     

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。     

3种蓑藓属(Macromitrium)植物DNA C-值测定——兼论苔藓DNA C-值研究的生物学意义

分别以小立碗藓(Physcomitrella patens)和水稻(日本晴亚种)(Oryza sativa spp.japonica)为标样,用流式细胞术分别测定了中华蓑藓(Macromirium cavaleriei)、钝叶蓑藓(M.japonicum)和缺齿蓑藓(M.gymnostomum)3种藓类植物的DNA C-值,由此建立了应用流式细胞术测定蓑藓属植物DNA C-值的方法,讨论了影响藓类植物DNA C-值测定结果的因素.最后讨论了苔藓植物DNA C-值变异式样、系统发育和分类学意义,以及今后有关藓类植物DNA C-值研究中值得关注的几个方向.     

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.     

滇西二高温温泉中Thermus高温菌的16S rRNA

 通过对滇西二温泉纯培养和免培养法获得的Thermus菌株序列进行16SrDNA系统发育及16SrRNA二级结构比较,发现二温泉中至少存在Thermus属的2个潜在新种,它们在系统发育树上形成1个独立的地理簇群分支,它们的16SrRNAhelix10二级结构也与其它地区的Thermus属菌株不同,形成明显的地域进化特征.     

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。     

Phylogenetic position of three Condylostoma species （Protozoa,Ciliophora, Heterotrichea） inferred from the small subunit rRNA gene sequence

The systematically poorly known ciliate genus Condylostoma was erected by Vincent in 1826. About 10 morphotypes have been reported, but any molecular investigations concerning this group so far are lacking. In this work, the small subunit ribosomal RNA （SS rRNA） gene of three marine Condylostoma species was sequenced, by which the phylogenetic trees were constructed by distancematrix, maximum parsimony and Bayesian inference methods. The results show that （1） all the trees have similar topologies with high supports; （2） Condylostoma is mostly related to the genus Condylostentor; and （3） three Condylostoma species as well as Condylostentor auriculatus cluster together and form a sister group with other heterotrichs. This is moderately consistent with the assessment of phylogenetic relationships of Condylostoma-related heterotrichs from the morphological information. The phylogenetic relationship of some other related heterotrichs, Peritromus, Folliculina, Stentor and Blepharisma, has been also discussed.     

Phylogenetic position of three Condylostoma species(Protozoa,Ciliophora,Heterotricheal inferred from the small subunit rRNA gene sequence

The systematically poorly known ciliate genus Conadylostoma was erected by Vincent in 1826.About 10 morphotypes have been reported,but any molecular investigations concerning this group SO far are lacking.In this work,the small subunit ribosomal RNA (SS rRNA)gene of three marine Conaylostoma species was sequenced,by which the phylogenetic trees were constructed by distance-matrix,maximum parsimony and Bayesian inference methods.The results show that(1)all the trees have similar topologies with high supports;(2)Condylostoma is mostly related to the genus Condylostentor;and(3)three Condylostoma species as well as Conadylostentor auriculatus cluster together and form a sister group with other heterotrichs.This is moderately consistent with the assessment of phylo-genetic relationships of Conaylostoma-related heterotrichs from the morphological information.The phylogenetic relationship of some other related heterotrichs,Peritromus,Fotlictllina,Stentor and Blepharisma,has been also discussed.     

Genetic study on the behavior of honeybees influenced by polyandry system

Within a honeybee population, due to polyandry, there are supersister and halfsister relations, thus many subfamilies exist. For the Chinese honeybee (Apis cerana cerana F.) a phylogenetic dendrogram has been constructed in which 4 subfamilies are clustered based on DNA fingerprint patterns. And it has been observed that each kind of workers is distributed to several dierent sub-families.     

基于RAPD标记的15种嵩草属(Kobresia Willd.)植物遗传关系

嵩草属(Kobresia Willd.)植物由于其形态结构的多样性及种内变异式样的复杂性,至今仍被认为是分类学上的"困难属".因此,对于该属系统学其他方面的证据,尤其分子方面的研究则显得极为重要和迫切.本文选用21个RAPD随机引物调查了15种嵩草属植物的遗传关系.结果表明:15种嵩草属植物聚为3类,认为嵩草组和异穗嵩草组的建立是比较自然的,而单穗嵩草组的划分并不自然.此外,分子结论并不支持前人发表的K.maquensis为独立新种,建议降为K.setschwanensis的异名.     

山口红树林土壤芽孢杆菌多样性及纤维素酶活性筛选研究

为探究广西山口红树林保护区内土壤中芽孢杆菌的多样性,挖掘具有稳定性好、耐高温的纤维素酶活性菌株,本研究利用可培养技术和基于16S rRNA基因序列的系统发育树分析研究红树林土壤中可培养芽孢杆菌的多样性,并以羧甲基纤维素钠为唯一碳源,结合刚果红纤维素培养法、滤纸条崩解试验、纤维素酶活力测定法,对分离出的芽孢杆菌开展纤维素降解活性研究。结果表明,从红树林土壤中共分离出171株芽孢杆菌,隶属于4科12属40种,其中芽孢杆菌属为优势菌属;从中筛选出9株具有显著纤维素酶活性的芽孢杆菌,其中4株具有显著的热稳定性。广西山口红树林自然保护区内土壤中蕴含着十分丰富的可培养芽孢杆菌种类,且部分菌株呈现出显著的纤维素酶活性,具有较大的挖掘潜力。     

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.     

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...