Further observations on the genus Didymosphaenia M. Schmidt—D. Sibirica (Grun.) M. Schm.

Maintenance of Didymosphaenia M. Schmidt as a distinct diatom genus is supported by an examination of D. sibirica (Grun.) M. Schm. with the scanning electron microscope. This investigation of D. sibirica reveals features similar to those of D. geminata (Lyngb.) M. Schm. and substantiates the existence of the genus separate from Gomphonema. D. sibirica is, however, a quite distinct species.     

Phylogeny of the Bangia flora of New Zealand suggests a southern origin for Porphyra and Bangia (Bangiales, Rhodophyta)

Analysis of nuclear small subunit ribosomal RNA gene (18S rDNA) sequence data from 123 samples of the red algal genus Bangia from mainland New Zealand has revealed diversity exceeding that reported for the genus from any other region in the world. Our study resolves two New Zealand Bangia taxa basal to the order Bangiales, and five clades of Bangia, four of which include New Zealand members. The basal taxa are separated from each other by 139 bp and differ from all other Bangia taxa in the New Zealand region by 103-163 bp over approximately 1750 bp 18S rDNA sequence data. Our results reveal a Bangia flora of previously unsuspected richness, and show that the simple morphology of these organisms obscures significant levels of genetic diversity. The presence of high diversity and retention of basal taxa in New Zealand Bangia raises the prospect that the southern hemisphere, and particularly eastern Gondwana, is not only a centre of diversity, but a centre of origin for the modern Bangiales.     

Increasing the transient expression of GUS gene in Porphyra yezoensis by 18S rDNA targeted homologous recombination

In order to test whether 18S rDNA can influence positively GUS gene transient expression in the red alga Porphyra yezoensis, a targeting vector pQD-GUS was constructed containing a portion of the 18S rDNA of P. yezoensis and transformed it into the same strain protoplasts. The results showed that GUS protein activity was increased markedly with pQD-GUS compared to the parent pBS-GUS. It is suggested that this system can be used to enhance the expression of exogenous genes in transgenic P. yezoensis.     

Effect of harvest method and timing on yield and regeneration of Karengo (Porphyra spp.) (Bangiales,Rhodophyta) in New Zealand

Commercial interest in harvesting wild stocks ofPorphyra and concern for this prized resource by the Maori community highlighted the need to investigate the impact of harvest method and timing onPorphyra beds. Harvesting trials were carried out at two locations near Kaikoura (South Island) and one in Wellington (southern North Island) between June 1987 and September 1987. At each of five sampling sites, ten replicate sets of four quadrats were used to test the effects of harvest method and timing on yield and regeneration. The method of harvest had a major effect on the extent of regeneration: in quadrats in which thePorphyra had been cut with basal portions left intact there were harvestable plants within two months, whereas in quadrats which were cleared of allPorphyra there was very little growth after the same period. Harvests in the latter half of thePorphyra growing season gave greater yields at all sites except Wellington. Several species ofPorphyra were found to exist at the Kaikoura sampling sites and a single, different, species at the Wellington site. There were site to site differences in the yields.     

What can 18S rDNA do for bivalve phylogeny?

Molecular characteristics, especially 18S rDNA sequences, may be of great value for the study of bivalve evolution and its numerous morphological convergencies once the reliability of these data can be evaluated. The analysis of 11 published complete molluscan sequences and two new ones,Arca noae andAtrina pectinata, reveals considerable differences in relative substitution rates. The gastropod and eulamellibranch species have the fastest and Atrina species have the slowest rates. Two methods are used to assess the information contents of the dataset in addition to bootstrap analysis, spectral analysis, and the “pattern of resolved nodes” technique. Tree reconstructions by parsimony, neighbor-joining, and maximum-likelihood differ in regard to the position of the eulamellibranch family Mactridae and ofCrassostrea. Although there is a signal for the monophyly of Bivalvia, Mactridae cluster with Gastropoda in most runs, rendering Bivalvia diphyletic. The position ofCrassostrea was extremely variable, probably due to the high substitution rate of this species.Atrina roots deeper thanArca in all trees, although a corresponding signal in spectral analysis is absent. Phylogenetic signals among the three pectinid species are low but sufficient to resolve the branching pattern. The tree inferred from the 18S rDNA and from morphological data has Bivalvia monophyletic with a basal polytomy of Mactridae,Crassostrea, and the remaining Pteriomorphia, whereArca branches off before Atrina and the Pectinidae.Argopecten is sister group to the other two pectinids; 18S sequence data will have great impact on our understanding of bivalve phylogeny, but only when more sequences of similar substitution rates are available. Correspondence to: G. Steiner     

An evaluation of characters obtained from life history studies for distinguishing New Zealand Porphyra species

Since 1992, as part of a study to circumscribe the genus Porphyra in New Zealand, an extensive culture programme has been developed at the Museum of New Zealand Te Papa Tongarewa. This collection currently houses more than 800 cultures and 150strains. We currently recognise at least 20 species in New Zealand, most of which are undescribed. Close observation of reproduction and the development of conchocelis-phase filaments grown in the laboratory, under controlled conditions of temperature, irradiance and photoperiod, has enabled the recognition of characters that can assist in species separation. The comparative taxonomic value of a range of such characters is discussed, including reporting a novel reproductive mode. This revised version was published online in August 2006 with corrections to the Cover Date.     

Phylogenetic relationships ofPolemoniaceae inferred from 18S ribosomal DNA sequences

Cladistic analyses of chloroplast DNA disagree with current classifications by placingPolemoniaceae near sympetalous families with two staminal whorls, includingFouquieriaceae andDiapensiaceae, rather than near sympetalous families with a single staminal whorl, such asHydrophyllaceae andConvolvulaceae. To explore further the affinities ofPolemoniaceae, we sequenced 18S ribosomal DNA for eight genera ofPolemoniaceae and 31 families representing a broadly definedAsteridae. The distribution of variation in these sequences suggest some sites are hypervariable and multiple hits at these sites have obscured much of the hierarchical structure present in the data. Nevertheless, parsimony, least-squares minimum evolution, and maximum likelihood methods all support a monophyleticPolemoniaceae that is placed nearFouquieriaceae, Diapensiaceae and related ericalean families.     

Phylogenetic relationships of Amur sturgeon Acipenser schrenckii Brandt, 1869 based on 18S rDNA sequencing

The phylogenetic relationships of Amur sturgeon A. schrenckii Brandt, 1869 with related species have been analyzed based on sequencing of the 18S rDNA small subunit. The complete sequence (1746 bp) of 18S rDNA has been estimated in seven individual A. schrenckii clones. The results show that the rDNA mutation profile of A. schrenckii 18S is very similar to that of A. fulvescens (Genbank data). Structural-functional and phylogenetic analyses allowed us to identify a presumably expressed variant, as well as taxon-specific mutation (adenine insertion after position 658 bp), for A. schrenckii 18S rDNA. Phylogenetic reconstructions performed with various approaches (NJ, MP, ML and Bayesian) support the monophyly of the genus Acipenser and point (1), based on which, in accordance with the classification based on ecological and morphological data (Artyukhin, 2006), the Amur sturgeon is closer to the sterlet than the Baltic sturgeon and (2) to substantial differentiation between North American (A. fulvescens) and Eurasian (A. schrenckii, A. ruthenus, and A. sturio) species of Acipenseridae.     

Phylogenetic positions of rust fungi parasitic on ferns: Evidence from 18S rDNA sequence analysis

Molecular phylogenetic analyses of fern rusts were carried out based on 18S rDNA sequences. We sequenced the 18S rDNAs of fern rusts (Hyalopsora polypodii andUredinopsis intermedia) and non-fern rusts (Aecidium epimedii, Coleosporium asterum, Ochropsora kraunhiae, Puccinia suzutake andPhysopella ampelopsidis) and analyzed their phylogenetic relationships with other members of the Urediniomycetes. Our bootstrapped neighbor-joining tree obtained from these analyses showed that rust fungi were apparently monophyletic at high confidence level (100% bootstrap confidence). In this molecular phylogenetic tree, the two fern rusts did not occupy the basal position within the rust fungal lineage and did not form a monophyletic lineage. Two species of the Cronartiaceae (Peridermium harknessii, Cronartium ribicola) and one species of the Coleosporiaceae (Coleosporium asterum) grouped with the fern rusts. Therefore, our results suggested that the two fern rusts were not primitive. On the other hand,Mixia osmundae, which is parasitic on the primitive fernOsmunda, was phylogenetically far from the fern rusts.     

Cytogenetic analysis in Polypterus ornatipinnis (Actinopterygii, Cladistia, Polypteridae) and 5S rDNA

Polypteridae is a family of archaic freshwater African fish that constitute an interesting subject for the study of the karyological evolution in vertebrates, on account of their primitive morphological characters and peculiar relationships with lower Osteichthyans. In this paper, a cytogenetic analysis on twenty specimens of both sexes of Polypterus ornatipinnis the ornate "bichir", coming from the Congo River basin, was performed by using both classical and molecular techniques. The karyotypic formula (2n=36; FN=72) was composed of 26 M+10 SM. The Alu I banding, performed to characterize heterochromatin in this species, was mainly centromeric. Both the chromosome location of the ribosomal 5S and 18S rRNA genes were examined by using Ag-NOR, classical C-banding, CMA(3) staining and FISH. CMA(3) marked all centromerical regions and showed the presence of two GC rich regions on the p arm of the chromosome pair n°1 and on the q arm of the pair n°14. Staining with Ag-NOR marked the only telomeric region of the chromosome n°1 p arm. After PCR, the 5S rDNA in this species was cloned, sequenced and analyzed. In the 665bp 5S rDNA sequence of P.ornatipinnis, a conserved 120bp gene region for the 5S rDNA was identified, followed by a non-transcribed variable spacer (NTS) which included simple repeats, microsatellites and a fragment of a non-LTR retrotransposon R-TEX. FISH with 5S rDNA marked the subtelomeric region of the q arm of the chromosome pair n°14, previously marked by CMA(3). FISH with 18S rDNA marked the telomeric region of the p arm of the pair n°1, previously marked both by Ag-NOR and CMA(3). The (GATA)(7) repeats marked the telomeric regions of all chromosome pairs, with the exclusion of the n°1, n°3 and n°14; hybridization with telomeric probes (TTAGGG)(n) showed signals at the end of all chromosomes. Karyotype evolution in Polypterus genus was finally discussed, including the new data obtained.     

Phylogenetic relationships of the hyphomycete generaChaetopsina andKionochaeta based on 18S rDNA sequences

A setiform dematiaceous hyphomycete,Kionochaeta spissa was newly collected and isolated from evergreen broad-leaved forests in the southern parts of Japan. Except for its dematiaceous nature, the species is morphologically similar to a nectriaceous hyphomycete,Chaetopsina fulva. The morphology and cultural properties of the Japanese isolates ofK. spissa were described, and the phylogenetic relationships betweenChaetopsina (C. fulva (type species)) andKionochaeta (K. ramifera (type species),K. spissa andK. ivoriensis) were inferred based on nuclear encoded small subunit (18S) rDNA sequences using the neighbor-joining method.Chaetopsina andKionochaeta were found to be phylogenetically related to the Hypocreales and Sordariales, respectively. Both should be maintained as separate genus for phylogenetic classification. The morphological resemblance especially betweenC. fulva andK. spissa is an example of the convergent evolution.     

棕囊藻北部湾株的18S rDNA分子鉴定

为研究北部湾棕囊藻(Phaeocystis)藻华的成因,采用PCR克隆了棕囊藻北部湾株核糖体18S r DNA序列。结果表明,棕囊藻北部湾株具有游动单细胞与群体结构两种形态;其18S r DNA序列和NCBI基因库中球形棕囊藻(Phaeocystis globosa)的同源性为99%~100%,在系统进化树上与不同海域来源的球形棕囊藻聚在一大分支上,且与球形棕囊藻间的遗传距离均小于其他种。首次从分子生物学上确定棕囊藻北部湾株为球形棕囊藻。     

Bryophyte 5S rDNA was inserted into 45S rDNA repeat units after the divergence from higher land plants

The 5S ribosomal RNA genes (5S rDNA) are located independently from the 45S rDNA repeats containing 18S, 5.8S and 26S ribosomal RNA genes in higher eukaryotes. Southern blot and fluorescence in situ hybridization analyses demonstrated that the 5S rDNAs are encoded in the 45S rDNA repeat unit of a liverwort, Marchantia polymorpha, in contrast to higher plants. Sequencing analyses revealed that a single-repeat unit of the M. polymorpha nuclear rDNA, which is 16103 bp in length, contained a 5S rDNA downstream of 18S, 5.8S and 26S rDNA. To our knowledge, this is the first report on co-localization of the 5S and 45S rDNAs in the rDNA repeat of land plants. Furthermore, we detected a 5S rDNA in the rDNA repeat of a moss, Funaria hygrometrica, by a homology search in a database. These findings suggest that there has been structural re-organization of the rDNAs after divergence of the bryophytes from the other plant species in the course of evolution.     

Phylogenetic Relationships Among Xiphinema and Xiphidorus Nematode Species from Brazil Inferred from 18S rDNA Sequences

Maximum likelihood trees produced from 18S rDNA sequences separated 14 Xiphinema and five Xiphidorus nematode species from Brazil into distinct groups that concurred with their current morphological taxonomic status. Species belonging to the X. americanum group (X. brevicolle, X. diffusum, X. oxycaudatum, and X. peruvianum) formed a single group that was clearly separated from the other Xiphinema species. As with previous taxonomic studies that noted only minor morphological differences between putative X. americanum group species, separation of these species based upon 18S rDNA sequences was inconclusive. Thus it is probable that instead of comprising distinct species, the X. americanum group may in fact represent numerous morphotypes with large inter- and intra- population morphological variability that may be environmentally driven. Within the cluster representing non X. americanum group species, there was little statistical support to clearly separate species. However, three subgroups, comprising (i) the X. setariae/vulgare complex, (ii) X. ifacolum and X. paritaliae, and (iii) X. brasiliense and X. ensiculiferum were well resolved.     

汉江硅藻水华优势种的形态及18S rDNA序列分析

我国汉江下游在1992、1998、2000、2003年初春曾先后多次发生水华。水华发生期间,硅藻大量增殖,形成肉眼可见的褐色颗粒悬浮于江水中,使江水呈现黄褐色,在以往的研究中认为汉江硅藻水华优势种为小环藻^[1,2],但并未从分类上展开研究。传统上硅藻种类的鉴定主要是以硅藻壳的形态及壳表面上的花纹为依据。目前国内外学者利用18S rDNA基因分析对水华赤潮原因种的鉴定方面也作了许多相关的研究。陈丽芬等利用18SrDNA序列分析确定香港海域一株棕囊藻为球形棕囊藻^[3]。     

Mapping of the 18S and 5S ribosomal RNA genes in the fish Prochilodus argenteus Agassiz, 1829 (Characiformes,Prochilodontidae)

A single NOR-bearing chromosome pair was identified by silver nitrate staining in a previous study of the fish Prochilodus argenteus from the S ã o Francisco River (MG, Brazil), with a third metacentric chromosome sporadically bearing active NOR. The present study focused on an analysis of the chromosomal localization of both the major (45S) and the minor (5S) rRNA genes using FISH. The use of the 18S rDNA probe confirmed the previous Ag-NOR sites interstitially located in a large metacentric pair and also identified up to three other sites located in the telomeric regions of distinct chromosomes, characterizing an interindividual variation of these sites. In addition, the 5S rDNA site was revealed adjacent to the major NOR site, identified at the end of the large Ag-NOR bearing metacentric chromosome. In a few metaphases, an additional weak hybridization signal was observed in a third chromosome, possibly indicating the presence of another 5S rDNA cluster. Despite a lower karyotype diversification (2n=54 and FN=108) often observed among species of Prochilodontidae, variations involving both 45S and 5S rRNA genes could play an important role in their chromosome diversification.     

Taxonomic relationships ofThiobacillus halophilus,T. aquaesulis,and other species ofThiobacillus,as determined using 16S rDNA sequencing

Total base sequences of the 16S rRNA genes ofThiobacillus halophilus andThiobacillus aquaesulis show that these bacteria fall into the gamma- and beta-subdivisions, respectively of the Proteobacteria. The closest relative ofT. halophilus isThiobacillus hydrothermalis (with 98.7% similarity), and the closest relative ofT. aquaesulis isThiobacillus thioparus (93.2% similarity). Physiological properties and mol% G+C content of their DNA serve to confirm that these four organisms are all distinct species. It is reiterated that the species currently assigned to the genusThiobacillus are clearly so diverse that they need reclassification into several genera. The type species,T. thioparus, is unequivocally placed in the beta-subdivision of the Proteobacteria, thus requiring that the use of the genus nameThiobacillus be restricted to the chemolithoautotrophic species falling into that group.T. aquaesulis andT. thioparus may thus be regarded as true species ofThiobacillus. The relatively large number of obligately chemolithoautotrophicThiobacillus species falling in the gamma-subdivision of the Proteobacteria need further study in order to assess the case for reclassification into one or more new or different genera.     

垃圾填埋场中厌氧真菌18SrDNA的PCR扩增及鉴定

采用机械破壁法直接从来自7个不同地区的垃圾填埋场滤液样本中提取真菌DNA,应用真菌通用引物NS1和NS8扩增18SrDNA（约1800bp),多聚酶链式反应（PCR）产物的琼脂糖凝胶电泳结果表明所有的样本均得到了扩增;以PCR产物作为模板,采用厌氧真菌Chytridiomycetes科的专用引物Chyt-719和Chyt-1553进行二次PCR扩增（约857bp),该阳性扩增产物克隆和测序结果首次表明在食草动物瘤胃中存在的厌氧真菌Chytridiomycetes也存在于垃圾填埋场中,且为Neocallimastix属。