用核糖体DNA的ITS序列探讨裂叶沙参的系统位置——兼论ITS片段在沙参属系统学研究中的价值

本文采用PCR直接测序方法,对来自沙参属全部2组7亚组的10个种和作为外类群的风铃草属2个种的核糖体DNA　ITS片段进行了序列分析。在重点探讨裂叶沙参分类地位的同时,分析了ITS片段序列在沙参属系统发育重建中的价值。结果表明,在沙参属中,ITS片段在长度、GC含量和位点变异量上均比较一致;长度539bp～541 bp,GC含量57％～60％,信息位点只占总位点的3．9～6％。采用PAUP软件进行的系统发育分析表明,裂叶沙参A．lobophylla与大花盘亚组的A．himalayana组成一支,而不是象以往形态学和杂交试验所推断的与泡沙参A．potaninii或A．stenanthina近缘。可见,ITS序列进一步支持将裂叶沙参移出泡沙参复合体甚至移出有齿亚组的推论,但同时也表明将其作为A．stenanthina的近缘种是不适宜的。尽管本研究所测定的类群已涵盖了整个沙参属,但种间序列的两两比较表明,沙参属种间在ITS片段上的分化很小(0.0～3.9％),相比之下,沙参属类群与风铃草属类群间的分化却很高(17.8％～19.2％)。这大概和沙参属起源较晚、遗传分化较小有关。本文还就ITS片段在沙参属和桔梗科系统学研究中的价值进行了讨论。     

A Reevaluation of the Status of A. lobophylla Based on ITS Sequence,with Reference to the Utility of ITS Sequence in Adenophora

Adenophora Fisch. (Campanulaceae)is a medium-sized and diverse genus, which occurs mainly in eastern Asia, particularly in China. Although several taxonomic systems have been proposed, there is still much debate on the delimitation and systematic arrangement of the species within the genus, due mainly to high variation in morphology, habitats, phenotypic plasticity, and potential hybridization. A. lobophylla Hong is a rare perennial endemic to several counties of Sichuan, China. Since this species was recognized, much work has been done on its taxonomic status, including morphology, crossing experiment and allozymes. However, evidence available is not concordance so far, and its relationship with other species within the genus remains largely uncertain. In the present paper, the internal transcribed spacer(ITS) region of 18s～26s nuclear ribosomal DNA was sequenced in 10 representatives of Adenophora and two outgroup species of Campanula to assess its utility for phylogenetic reconstruction in Adenophora, in addition to reevaluation of the taxonomic status of A. lobophylla. The results indicate that there exist high sequence alignability and length conservation among ITS sequences, with informative sites being 3.9% and length ranging from 539 to 541 bp and G + C content varying from 57% to 60%. Phylogenetic　analyses using Fitch parsimony show that A. lobophylla is closely related to A. himalayana instead of A. potaninii or A. stenanthina as revealed by previous morphological work and crossing experiment. The ITS phylogeny is generally congruent with other evidence in that A. lobophylla should be removed from the A. potaninii complex or even subsect. Microdiscus. Its appropriate placement, however, should be further explored. In pairwise comparisons at unambiguously aligned sites, sequence divergence was relatively low among Adenophora species with the values ranging from 0.0 to 3.9% in spite of the fact that the species surveyed covered all of the sections and subsections of the genus. In contrast, high sequence divergence is found among Adenophora and Campanula species with the values from 17.8 % to 19.2 %. The low divergence among Adenophora species is justified considering its recent origin, and also suggests that ITS sequence would provide a promising source of nuclear phylogenetic markers to reconstruct the phylogeny within Campanulaceae on the tribe and family levels.     

中国特有植物同钟花(桔梗科)的细胞学研究

对分布我国西南的特有属同钟花属的同钟花(Homocodon brevipes)进行细胞学研究,其染色体数目为68,核型为2n=68=56m 10sm 2st,可能是四倍体。通过比较桔梗科已有的大量细胞学资料认为该类群应该与风铃草属和沙参属所代表的类群关系较近。     

中国特有植物同钟花( 桔梗科) 的细胞学研究

对分布我国西南的特有属同钟花属的同钟花( Homocodon brevipes) 进行细胞学研究, 其染色体数目为68 , 核型为2n = 68 = 56m+ 10sm + 2st, 可能是四倍体。通过比较桔梗科已有的大量细胞学资料认为该类群应该与风铃草属和沙参属所代表的类群关系较近。     

Molecular evidence for the phylogenetic position ofHanabusaya asiatica Nakai (Campanulaceae), an endemic species in Korea

The phylogenetic relationship of the Korean endemic genus,Hanabusaya, to other campanulaceous genera has been controversial since it was described by Nakai in 1911. Three genera of Campanuloideae,Symphyandra, Adenophora, andCampanula, have been considered closely related by various taxonomists on the basis of anther shape, gross morphology, and pollen characters, respectively. We have tested these competing taxonomic hypotheses using the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA from 12 taxa representing 7 genera of Campanulaceae. The molecular phylogeny indicates strongly thatHanabusaya is more closely allied toAdenophora than toCampanula orSymphyandra. The phylogenetic affinity ofHanabusaya andAdenophora is supported by a 100% bootstrap value and a high decay index (13). The average sequence divergence value (Kimura’s 2-parameter method) betweenHanabusaya and theAdenophora species is 2.58. The value is significantly (about ten times) lower than the ones observed betweenHanabusaya and the species ofCampanula (average of 23.52) and betweenHanabusaya andSymphyandra (24.95). The ITS sequence phylogeny suggests that some morphological characters, such as fused anthers and corolla shape, are homoplastic in the Campanulaceae genera.     

Phylogenetic relationships of Polyporus and morphologically allied genera

Polyporus accommodates morphologically heterogeneous species and is divided into six infrageneric groups based on macromorphological characters. On the other hand allied genera have macro- and microscopic characters similar to those of Polyporus. The phylogenetic relationships of Polyporus and allied genera were established from sequences of RNA polymerase II second largest subunit (RPB2), nuclear ribosomal large subunit (nucLSU) and mitochondrial ATPase subunit 6 (ATP6). The molecular phylogenetic trees confirmed that Polyporus is a polyphyletic genus and recognized six major clades (1-6) containing species of Polyporus and several allied genera. Among the clades one contained three infrageneric groups of Polyporus and two allied genera, Datronia and Pseudofavolus while one other contained group Polyporellus and Lentinus. Five of the six major clades contained species belonging to a single infrageneric group, Favolus, Melanopus, Polyporellus or Polyporus. This suggests that morphological characters used to define these groups have phylogenetic significance and reveals the need for a taxonomic revision of Polyporus and its allied genera.     

Flowering phenology of Campanula on Mt Olynipos, Greece

We studied the fowering phenology of the genus Campanula (represented by nine species) along the elevation gradient of Mt Olympos (Greece) in order to assess whether there are elevation patterns at the genus level and whether these relate to patterns previously observed along such gradients at the community level. The traits examined were time and duration of the flowering period, flower life span, and duration of male and female flower phases. Floral attributes, such as number of flowers per plant, flower biomass, flower size, were also studied in order to examine whether they change with elevation or influence flower phenology. Flowering of Campanula species started in mid-May and ended in late September. The average duration of flowering of the genus was ca 27 d and the average floral longevity 4 d. In all but one species, the female phase lasted longer than the male. Campanula versicolor differed remarkably from all others in flower phenology and other floral trails. Nearly all Campanula populations studied had right or positively skewed flowering distributions indicating that flowering begins more abruptly than it ends. At the genus level, the time of flowering increased with elevation by 2 3d for every 100 m. Floral longevity also increased with elevation, by 0,2 d for every 100 m. Neither duration of flowering nor duration of the flower phases showed any consistent change with elevation. The same is true of the non–phenological floral traits examined. No trade–off between duration of flowering and flower life span or between structure and maintenance of flowers was apparent. The pattern of increasing floral longevity along the elevation gradient suggests a mechanism of compensation for reduecd pollinator availability.     

Resolving the evolutionary history of Campanula (Campanulaceae) in western North America

Recent phylogenetic works have begun to address long-standing questions regarding the systematics of Campanula (Campanulaceae). Yet, aspects of the evolutionary history, particularly in northwestern North America, remain unresolved. Thus, our primary goal in this study was to infer the phylogenetic positions of northwestern Campanula species within the greater Campanuloideae tree. We combined new sequence data from 5 markers (atpB, rbcL, matK, and trnL-F regions of the chloroplast and the nuclear ITS) representing 12 species of Campanula with previously published datasets for worldwide campanuloids, allowing us to include approximately 75% of North American Campanuleae in a phylogenetic analysis of the Campanuloideae. Because all but one of North American Campanula species are nested within a single campanuloid subclade (the Rapunculus clade), we conducted a separate set of analyses focused specifically on this group. Our findings show that i) the campanuloids have colonized North America at least 6 times, 4 of which led to radiations, ii) all but one North American campanuloid are nested within the Rapunculus clade, iii) in northwestern North America, a C. piperi-C. lasiocarpa ancestor gave rise to a monophyletic Cordilleran clade that is sister to a clade containing C. rotundifolia, iv) within the Cordilleran clade, C. parryi var. parryi and C. parryi var. idahoensis exhibit a deep, species-level genetic divergence, and v) C. rotundifolia is genetically diverse across its range and polyphyletic. Potential causes of diversification and endemism in northwestern North America are discussed.     

A new Campanula (Campanulaceae) from east Anatolia, Turkey

Campanula demirsoyi A. Kandemir (Campanulaceae) is described and illusturated from a localised area of Erzincan province. It is closely allied to C. hedgei Davis and C. coriacea Davis. Diagnostic morphological characters are discussed and compared with those of related taxa.     

我国新记录海链藻属物种的形态学和系统学研究

为了廓清我国海链藻属的物种多样性, 并丰富其分子生物学信息, 为后续的系统学研究提供基础数据, 从我国沿海分离并建立了海链藻的单克隆培养株系, 利用光学显微镜和电镜技术进行了形态学研究, 同时还对其核糖体大亚基的高变区序列进行了扩增和测序分析。结合形态学和分子生物学数据, 鉴定了我国海链藻属的2个新记录种: 狭线形海链藻Thalassiosira anguste-lineata (Schmidt) Fryxell & Hasle和碟形海链藻T. minicosmica Lee & Park。对它们的形态学特征进行了较为详尽的描述, 并与相似种进行了比较研究。此外, 还基于核糖体大亚基高变区的碱基序列信息, 分析了它们的分子系统学位置。在分子系统树上, 狭线形海链藻和碟形海链藻均不与该属模式种——诺氏海链藻T. nordenskioeldii 聚在同一分支上, 显示它们与典型海链藻属物种之间具有较大的遗传差异。分子系统树还显示, 目前基于形态学建立的海链藻属并非自然类群, 而是被骨条藻属Skeletonema、小环藻属Cyclotella、漂流藻属Planktoniella等邻近属种分隔成多个分支, 这预示着现存的海链藻属应该是一个并系类群, 在后续的系统学研究中, 或许会有较大的系统学调整, 但目前有限的分子生物学信息是相关研究推进的重要限制。     

Re-examining the phylogeny of clinically relevant Candida species and allied genera based on multigene analyses

Yeasts of the artificial genus Candida include plant endophytes, insect symbionts, and opportunistic human pathogens. Phylogenies based on rRNA gene and actin sequences confirmed that the genus is not monophyletic, and the relationships among Candida species and allied teleomorph genera are not clearly resolved. Protein-coding genes have been useful to resolve taxonomic positions among a broad range of fungi. Over 70 taxa of the genus Candida and its allied sexually reproducing genera were therefore selected, and their phylogenetic relationships were investigated using nuclear sequences of the largest subunit and second largest subunit of RNA polymerase II gene, actin, the second subunit of the mitochondrial cytochrome oxidase gene, and D1/D2 LSU rRNA gene. The DNA sequences were analysed by maximum parsimony and Bayesian inference, resulting in the recognition of six major phylogenetic groups (A-F). Group A contains six facultative pathogenic Candida species, which seem to have derived from nonpathogenic species, while Group B contains species of Clavispora, Metschnikowia, and Pichia guilliermondii. Species of Debaryomyces form an independent group C that is related to groups A and B. Pichia fermentans and other environmental species are concentrated in Group D. Group E, containing Pichia anomala, may be a sibling to group F, which is represented by the Saccharomyces species complex.     

Using 18S rDNA to resolve diaptomid copepod (Copepoda: Calanoida: Diaptomidae) phylogeny: an example with the North American genera

The phylogenetic relationships among the numerous genera of diaptomid copepods remain elusive due to difficulties in obtaining sufficient numbers of phylogenetically informative morphological characters for cladistic analysis. Molecular phylogenetic techniques offer high potential to resolve phylogenetic relationships in the absence of sufficient morphological characters because of the ease in which many characters can be unambiguously coded. I present the first molecular phylogeny for diaptomid copepod genera using 18S rDNA. Specifically, I test Light’s (1939) hypothesis regarding the interrelationships among the North American diaptomid genera. The 18S phylogeny is remarkably consistent with Light’s hypothesis. The endemic North American genera represent a monophyletic group exclusive of the non-endemic genera. Moreover, his hypothesized basal genus for the North America genera, Hesperodiaptomus, is the basal genus in this analysis. However, his Leptodiaptomus group is not reciprocally monophyletic with his Hesperodiaptomus group, but is rather a derived member of the latter group. Finally, the genus Mastigodiaptomus is found to be more closely allied with the non-endemic genera, as Light suggested. This phylogeny contributes heavily to the understanding of phylogenetic relationships among North American diaptomids and has large implications for the systematics of diaptomids in general. The use of 18S rDNA sequences in phylogenetic analyses of diaptomid copepods can be used to confirm the monophyly of recognized genera, the interrelationships among genera, and subsequent biogeographic interpretation of the family’s diversification. The use of molecular data, such as 18S rDNA sequences, to test phylogenetic hypotheses based on a very limited number of morphological characters will be a particularly useful approach to phylogenetic analysis in this system.     

Tanacetum joharchii sp. nov. (Asteraceae–Anthemideae) from Iran, and its taxonomic position based on molecular data

Tanacetum joharchii Sonboli & Kazempour Osaloo sp. nov. (Asteraceae–Anthemideae) is described and illustrated from the Khorasan province, northeast Iran. Tanacetum joharchii is a suffruticulose species from rocky limestone mountains in Hezarmasjed and Binalud (Khorasan province) at altitudes of 1900–2500 m a.s.l. The diagnostic morphological characteristics that distinguish it from the allied species T. kotschyi are presented. In addition, a distribution map of T. joharchii and the related species is given. In order to provide some hypothesis on its phylogenetic position, a molecular phylogenetic analysis based on nrDNA ITS sequence data of 14 representatives of the genus Tanacetum was performed.     

A Comparison of ITS Nuclear rDNA Sequence Data and AFLP Markers for Phylogenetic Studies in Phyllostachys (Bambusoideae, Poaceae)

Phyllostachys , a large, economically important genus of woody bamboos. DNA sequences of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) were used in a parsimony analysis. Phyllostachys was well supported as monophyletic with Chimonobambusa as its closest allied genus. The 5S spacer region of nrDNA was investigated but found unsuitable for this purpose. The AFLP analysis showed much higher discriminating power between species and was more useful for phylogenetic reconstruction at this taxonomic level. The combined data were used to review the previous infra-generic classifications. Section Heteroclada Wang & Ye is strongly supported and can be further divided into sub-groups. A group within section Phyllostachys is strongly supported, but a further group of taxa previously included in this section is difficult to place. The ability of the methods to help separate species such as P. sulphurea and investigate genetic diversity at the infra-specific level was also assessed. It is argued that AFLPs could often be the method of choice for phylogenetic studies of closely related taxa for which DNA sequence data provide insufficient resolution. Received 18 April 2000/ Accepted in revised form 10 June 2000     

Phylogenetic relationships of Salix (Salicaceae) based on rbcL sequence data

Nucleotide sequences of the chloroplast-encoded rbcL gene were used to examine phylogenetic relationships of the genus Salix together with other allied genera of the family Salicaceae. Phylogenetic analyses of rbcL sequences strongly suggest the monophyly of three commonly recognized genera (Chosenia, Salix, and Toisusu). Two monophyletic groups are recognized within the larger monophyletic group. They do not correspond with any infrageneric taxa proposed so far. With regard to character evolution, it is thought that the reduction of stamen number from more than two stamens to two might occur in at least three lineages and that fused bud scales evolved several times and/or the reverse evolution occurred from fused to free. Some types of pollen surfaces are considered to have evolved independently.     

Systematics of the marine microfilamentous green algae Uronema curvatum and Urospora microscopica (Chlorophyta)

The microfilamentous green alga Uronema curvatum is widely distributed along the western and eastern coasts of the north Atlantic Ocean where it typically grows on crustose red algae and on haptera of kelps in subtidal habitats. The placement of this marine species in a genus of freshwater Chlorophyceae had been questioned. Molecular phylogenetic analysis of nuclear-encoded small and large subunit rDNA sequences reveal that U. curvatum is closely related to the ulvophycean order Cladophorales, with which it shares a number of morphological features, including a siphonocladous level of organization and zoidangial development. The divergent phylogenetic position of U. curvatum, sister to the rest of the Cladophorales, along with a combination of distinctive morphological features, such as the absence of pyrenoids, the diminutive size of the unbranched filaments and the discoid holdfast, warrants the recognition of a separate genus, Okellya, within a new family of Cladophorales, Okellyaceae. The epiphytic Urospora microscopica from Norway, which has been allied with U. curvatum, is revealed as a member of the cladophoralean genus Chaetomorpha and is herein transferred to that genus as C. norvegica nom. nov.     

DIVERSITY AMONG TETRASPORALEAN TAXA: EVIDENCE FROM ULTRASTRUCTURE AND MOLECULES

Recent phylogenetic studies of tetrasporalean exemplars using 18S rDNA revealed the extent of diversity among taxa in this non-monophyletic group (Booton et al. 1998). In particular, two distinct tetrasporalean lineages were identified; one group allied with chlamydomonadalean taxa and the other group comprising taxa now placed in a separate order, the Chaetopeltidales. Using these observations as a framework, a new investigation of diversity among a broader sampling of putative tetrasporalean genera (e.g. Asterococcus , Chloranomala , Chlorophysema , Gloeocystis , Gloeodendron , Palmella , Paulschulzia , Physocytium , Schizochlamys and Tetraspora ) was undertaken. Phylogenetic studies of both 18S and 26S rDNA were compared with ultrastructural investigations of vegetative cells. Molecular phylogenetic analyses corroborate the earlier 18S rDNA results, but also reveal additional diversity. The new data raise doubts regarding the monophyly of two genera, Palmella and Tetraspora. The new data also link two enigmatic green algal genera, Physocytium and Heterochlamydomonas , in a long-branch lineage within the Chlamydomonadales. Another enigmatic genus, Schizochlamys , is allied with Bracteacoccus in the Sphaeropleales. Lastly, Chloranomala is resolved as an ally of Paulschulzia , Tetraspora sp., and the green flagellate, Lobomonas. Comparison of pyrenoid ultrastructure generally supports the molecular phylogenetic analyses, suggesting that this non-molecular character will be a useful marker for broad phylogenetic studies of chlamydomonadalean taxa. (Supported by NSF grant, DEB 9726588)     

The phylogeny of Paralabrax (Perciformes: Serranidae) and allied taxa inferred from partial 16S and 12S mitochondrial ribosomal DNA sequences

Partial sequences of 16S and 12S mitochondrial ribosomal DNA were used to examine the phylogenetic relationships of the primarily eastern Pacific genus Paralabrax (Perciformes: Serranidae) and allied taxa. Paralabrax is considered a basal serranine, which is itself considered the basal subfamily in the Serranidae. Multiple serranines reported closely related to Paralabrax from the genera Serranus, Hypoplectrus, Cratinus, and Centropristis were used as outgroups. Species from the remaining two subfamilies, Epinephilinae and Anthiinae, of the Serranidae were also used in the analyses. The tree of the Serranidae was rooted with the families Polyprionidae and Priacanthidae. Paralabrax, the Serranidae, and the Serraninae were monophyletic in this study. Serranus was found to be paraphyletic. Centropristis, formerly considered the sister taxon to Paralabrax, was not closely related in these analyses. Cratinus agassizii, a monotypic genus from the eastern Pacific, was found to be the sister taxon to Paralabrax. There is greater resolution for intergeneric and subfamily relations than interspecific relationships. A single most parsimonious tree for the interspecific relationships of Paralabrax and allied taxa is proposed. This proposed molecular phylogeny is consistent with known biogeographic processes in the eastern Pacific.     

Andalucinetes hastatus nov. gen., nov. sp., nouveau Chonetacea (Brachiopoda) dévonien du Sud-Ouest de l'Espagne

A new genus and its type-species: Andalucinetes hastatus nov. gen., nov. sp., are described and illustrated. The erection of the new genus is warranted by a comparison with the two more closely allied genera.