Recent changes in the seaweed community of Western Prince Edward Island: implications for the seaweed industry

Prior to 1980, seaweed beds of western Prince Edward Island, Canada, were dominated by Chondrus crispus Stackhouse, associated with 37 other species of common algae. Furcellaria lumbricalis (Huds.) Lamour. occurred in 4.6% of the samples from the Pleasant View bed but never reached measurable levels of cover. By 1991 F. lumbricalis was reported in 67% of the samples from this bed. Furcellaria lumbricalis biomass was negatively correlated with C. crispus biomass and positively correlated with Phyllophora Grev. spp. biomass. F. lumbricalis biomass and cover peaked in subtidal areas below 4 m at 359 ± 299 SD fresh g 0.25 m^–2. Harvesters avoid concentrations of F. lumbricalis, resulting in a 5% content of this species in the commercial harvest. Systematic harvesting located concentrations of F. lumbricalis in seven of 10 commercial beds. Proposed causes for change in species dominance from C. crispus to F. lumbricalis range from general environmental change, natural succession to excessive human perturbation.     

Phylogeny of the Madagascan endemic family Didiereaceae

A molecular phylogeny of the Didiereaceae was produced through parsimony analysis of chloroplastrpl16 intron andtrnL-trnF andtrnT-trnL intergenic spacer sequences of all eleven species of the Didiereaceae and several outgroup taxa from the Portulacaceae. Results indicated that: 1) the Didiereaceae were embedded within the Portulacaceae, withCalyptrotheca as the sister group of the family; 2) present generic limits were supported; 3)Alluaudiopsis was the most basal lineage; 4) at least two separate episodes of polyploidization within the genusAlluaudia had occurred, and 5) unusually low amounts of variation were present in rapidly evolving noncoding plastid sequences.     

Relationships between species ofLeymus,Psathyrostachys, andHordeum (Poaceae,Triticeae) inferred from Southern hybridization of genomic and cloned DNA probes

We have used total genomic DNA as a probe to size-fractionated restriction enzyme digests of genomic DNA from a range ofTriticeae species from the generaLeymus Hochst.,Psathyrostachys Nevski, andHordeum L., and hybrids betweenHordeum andLeymus to investigate their taxonomic relationships. Genomic Southern hybridization was found to be an effective and simple way to assess the distribution and diversity of essentially species-specific and common, repetitive DNA sequences, and is hence especially useful in evolutionary studies. The DNA sequences ofH. vulgare seem to diverge substantially from those ofH. brachyantherum, H. lechleri, H. procerum, andH. depressum. The genome ofThinopyron bessarabicum shows little homology to those of theLeymus species investigated, confirming thatT. bessarabicum is not an ancestral genome inLeymus. Although the genomes ofLeymus andPsathyrostachys share substantial proportions of DNA sequences, they include divergent repeated sequences as well. Hybridization with a ribosomal DNA probe (pTa 71) showed that the coding regions containing structural genes encoding the 18 S, 5.8 S, and 26 S ribosomal RNA were conserved among the species investigated, whereas the intergenic spacer region was more variable, presenting different sizes of restriction fragments and enabling a classification of the species. The rye heterochromatin probe pSc 119.2 hybridized to DNA fromH. lechleri andT. bessarabicum, but not to DNA from the other species investigated.     

Further flavonoid studies onAttalea species and some related cocosoid palms

In a flavonoid survey of direct and hydrolysed leaf extracts of sixteenAttalea, sevenScheelea and fourOrbignya species free tricin, tricin 7-glycosides, tricin 5-glucoside and flavone C-glycosides were the most frequent constituents; present in 100, 89, 70, and 81% of species, respectively. Luteolin, quercetin and isorhamnetin were each found in only 15% of the sample. The present results confirm the findings of a previous survey thatAttalea, Scheelea andOrbignya are chemically heterogeneous with as much variation between species as between genera. Furthermore, threeAttalea species,A. allenii, A. guaranitica andA. victoriana showed some infraspecific variation. On the other hand all three accessions ofA. ferruginea and six ofA. geraensis examined gave identical flavonoid profiles. The results support the view thatA. geraensis andA. guaranitica are closely related but do not support the suggested close relationship based on morphology betweenA. oleifera, A. burretiana andA. piassabossu.     

Molecular phylogeny ofSalicaceae and closely relatedFlacourtiaceae: Evidence from 5.8 S, ITS 1 and ITS 2 of the rDNA

A ribosomal DNA region, including the entire 5.8S RNA gene and the internal transcribed spacers ITS 1 and ITS 2, was used for studying the phylogeny ofSalicaceae and the relationship betweenSalicaceae andFlacourtiaceae. The length of the ITS regions withinSalicaceae andFlacourtiaceae was similar to that found in other angiosperms. The GC content of both ITS regions was high, varying 62.7-72.2%. The most parsimonious tree clusters the wind-pollinatedChosenia bracteosa among theSalix species, suggesting that it should be included in the genusSalix. The grouping withinSalix leaves subg.Salix as paraphyletic, for which reason the subgeneric division is questionable.Populus was monophyletic and formed a sister group toSalix. The interspecific variation of the ITS sequences was very small inSalicaceae, which is in contradiction to the age of the group according to the evidence from fossil data.Idesia polycarpa fromFlacourtiaceae shows great sequence similarity withSalicaceae, but the analysis of 5.8S rDNA supports monophyly of the four species ofFlacourtiaceae sampled for this study.     

Tandem duplication of alkaline phosphatase genes and polymorphism in the intergenic sequence in<Emphasis Type="Italic"> Bombyx mori</Emphasis>

Alkaline phosphatases are ubiquitous in organisms from bacteria to human. Two alkaline phosphatase genes, Alp-m and Alp-s, were independently cloned from the silkworm Bombyx mori. They were mapped to a small DNA region and shown to be organized in tandem. Exon-intron structures of the two genes were highly conserved, with the exception of the second intron in Alp-m, which has no counterpart in Alp-s. The similarity between the nucleotide sequences of the exons of the two genes was strikingly high (60–79%), suggesting that Alp-m and Alp-s originated from a duplication of their common ancestor gene. The intergenic sequence between the two Alp genes shows length polymorphism in different B. mori strains, which can be explained by presence/absence of two putative insertion sequences. This structural variation suggests a possible scenario for the divergence of the two Alp genes after the duplication event.Communicated by G. Reuter     

Recent developments inRhizobium taxonomy

Recent developments inRhizobium taxonomy are presented from a molecular and evolutionary point of view. Analyses of ribosomal RNA gene sequences provide a solid basis to infer phylogenies in the Rhizobiaceae family. These studies confirmed thatRhizobium andBradyrhizobium are only distantly related and showed thatRhizobium andBradyrhizobium are related to other groups of bacteria that are not plant symbionts.Rhizobium andAgrobacterium species are intermixed. Differences in plasmid content may explain to a good extent the different behavior ofRhizobium andAgrobacterium as symbionts or pathogens. Other approaches to identify and classify bacteria such as DNA-DNA hybridization, fatty acid analysis, RFLP and RPD-PCR techniques and phylogenies derived from other genes are in general agreement to the groupings derived by ribosomal sequences. Only a small proportion of nodulated legumes have been sampled for their symbionts and more knowledge is required on the systematics and taxonomy ofRhizobium andBradyrhizobium species.     

Phylogenetic evaluation of 5S ribosomal RNA gene and spacer in theCallistachys group (Fabaceae:Mirbelieae)

Sequences of 5S rDNA from an Australian group of papilionoid legumes were evaluated for phylogenetic informativeness. Twenty four sequences were sampled from ten species in five closely related genera:Brachysema, Callistachys, Jansonia, Nemcia andPodolobium. These sequences fell into two size classes, 200bp and 600bp, which appear to represent paralogous copies of the 5S unit at different loci. As in previous studies, the 5S rRNA gene and both ends of the inter-gene spacer are found to be conserved and almost uninformative about phylogeny at this taxonomic level. By contrast, the middle part of the spacer is highly variable, both in substitutions at individual sites and in length. The short sequences contain virtually no duplications. The long sequences contain numerous short repeats (c.20 bp) which form a regular pattern in the middle section. Phylogenies estimated from these data support monophyly of the study group, and of species. Relationships among species and genera are less clear, perhaps because divergence of the 5S spacer between species is too great. However, non-monophyly ofNemcia andBrachysema, indicated by other data, is corroborated.     

Molecular phylogeny of species in the generaAmylostereum andEchinodontium

Analyses of DNA sequences from the internal transcribed spacer (ITS) region of the nuclear rDNA and from a portion of a manganese-dependent peroxidase gene were used to assess the species inAmylostereum, including isolates from the mycangia of horntails, decay, and basidiomes. Four species are recognized:A. areolatum, A. chailletii, A. laevigatum, andA. ferreum. An unidentifiedAmylosterum isolate from the mycangium ofXoanon matsumurae had an ITS sequence identical to that ofA. areolatum. Another unidentifiedAmylostereum isolate from the mycangium ofSirex areolatus was nearA. laevigatum, which appears to be the mycangial symbiont for those horntails attacking cedar-like trees. The other horntail isolates, primarily from Pinaceae, proved to be eitherA. areolatum orA. chailletii. The DNA sequences ofEchinodontium tinctorium, E. tsugicola andE. japonicum were similar to those of theAmylostereum species, andAmylostereum species are now recognized as members of the family Echinodontiaceae rather than the family Stereaceae.Echinodontium taxodii was found to be distinct from the Echinodontiacaea andStereum, andE. taxodii is recognized as aLaurilia species.     

Molecular phylogenetic analysis ofChrysosplenium (Saxifragaceae) in Japan

The phylogeny of Japanese species ofChrysosplenium (Saxifragaceae) was examined using variation in DNA sequences. Sequences ofrbcL andmatK genes were compared for their feasibility for reconstructing the phylogeny ofChrysosplenium, and thematK sequences was found to give greater resolution. All but one of the 17 Japanese species have been examined formatK gene sequences and phylogenetic analysis of these data resulted in eight most parsimonious trees of 390 steps and a consistency index (Cl) of 0.823. The molecular phylogeny obtained was generally in agreement with Hara's (1957) classification based upon phenotypic similarity, although a conclusion needs extensive examination of the genus on a world-wide level. Using the phylogenetic data, character evolution was examined, especially in the characters traditionally used for grouping infrageneric taxa. Differentiation of opposite and alternate phyllotaxis appears to have occurred only once in the course of evolution ofChrysosplenium.     

Numerical morphological analysis of Lettuce cultivars and species (Lactuca sect.Lactuca,Asteraceae)

L. sativa L. cultivars were compared morphologically with accessions ofL. saligna L.,L. serriola L., andL. virosa L. in order to define the infra- and interspecific variation. Multivariate analysis (principal component analysis) distinguished four groups corresponding with the four species. Infraspecific variation (e.g., colour, leaf shape) withinL. sativa is the largest, because breeding has caused a large variation in vegetative features; infraspecific variation withinL. serriola is also large. Three plants of one accession were placed betweenL. sativa andL. serriola because they showed morphological characteristics of both species. They are still classified asL. sativa. The morphological differences betweenL. sativa andL. serriola are too large to consider these two as one species. The descent of the cultivar groups and the four species is described.     

Studies on the diversity of the distinct phylogenetic lineage encompassing Glomus claroideum and Glomus etunicatum

Morphological and molecular characters were analysed to investigate diversity within isolates of the Glomus claroideum/Glomus etunicatum species group in the genus Glomus. The inter- and intra-isolate sequence diversity of the large subunit (LSU) rRNA gene D2 region of eight isolates of G. claroideum and G. etunicatum was studied using PCR-single strand conformational polymorphism (SSCP)-sequencing. In addition, two isolates recently obtained from Southern China were included in the analysis to allow for a wider geographic screening. Single spore DNA isolation confirmed the magnitude of gene diversity found in multispore DNA extractions. An apparent overlap of spore morphological characters was found between G. claroideum and G. etunicatum in some isolates. Analysis of the sequence frequencies in all G. etunicatum and G. claroideum isolates (ten) showed that four LSU D2 sequences, representing 32.1% of the clones analysed for multispore extraction (564) were found to be common to both species, and those sequences were the most abundant in four of the ten isolates analysed. The frequency of these sequences ranged between 23.2% and 87.5% of the clones analysed in each isolate. The implications for the use of phenotypic characters to define species in arbuscular mycorrhizal fungi are discussed. The current position of G. claroideum/G.etunicatum in the taxonomy of the Glomeromycota is also discussed.     

Phylogenetic relationships in theCampanulales based onrbcL sequences

Phylogenetic relationships within the angiosperm orderCampanulales were investigated by comparative sequencing of the chloroplast generbcL. CompleterbcL sequences were obtained for ten species in six families within the order. These data were analyzed along with previously publishedrbcL sequences from other taxa (for a total of 117 species) within the subclassAsteridae and outgroups, producing 32 equally parsimonious trees. A subset consisting of 44 of these taxa was then chosen and more rigorous analyses performed, resulting in four equally parsimonious trees. Results indicate that two major clades roughly corresponding to traditionally circumscribedAsterales andCampanulales exist as sister taxa. In particular, therbcL trees indicate thatSphenoclea is not a member ofCampanulales orAsterales, thatPentaphragma is more closely allied toAsterales thanCampanulales, that theCyphiaceae are not monophyletic, thatCampanulaceae andLobeliaceae are not sister taxa, and thatStylidiaceae are correctly placed withinCampanulales.     

A phylogenetic study ofPolygonum sect.Tovara (Polygonaceae) based on ITS sequences of nuclear ribosomal DNA

Polygonum sect.Tovara comprises three morphologically very similar species;P. virginianum,P. filiforme, andP. neofiliforme. Sequences of internal transcribed spacers (ITSs) of nuclear ribosomal DNA of these were determined to examine phylogenetic relationships and the levels of differentiation among them. The size of ITS 1 was 241 bp inP. filiforme andP. neofiliforme, and 242 bp inP. virginianum. The size of ITS 2 was 243 bp, and that of the 5.8S rRNA coding region was 163 bp. The ITS sequences clearly separate North AmericanP. virginianum from the eastern Asian species. Nucleotide divergence between them ranges from 3.3% to 3.8% for ITS 1 and from 9.3% to 10.7% for ITS 2. The molecular data also revealed that two eastern Asian species are closely related but should be treated as distinct species.     

Gypsy homologous sequences inDrosophila subobscura (gypsyDS)

Summary Characterization of sequences homologous to theDrosophila melanogaster gypsy transposable element was carried out inDrosophila subobscura (gypsyDS). They were found to be widely distributed among natural populations of this species. From Southern blot and in situ analyses, these sequences appear to be mobile in this species.GypsyDS sequences are located in both euchromatic and heterochromatic regions. A completegypsyDS sequence was isolated from aD. subobscura genomic library, and a 1.3-kb fragment which aligns with the ORF2 of theD. melanogaster gypsy element was sequenced. Comparisons of this sequence in three species (D. subobscura, D. melanogaster, and D. virilis) indicate that there is greater similarity between theD. subobscura-D. virilis sequences than betweenD. subobscura andD. melanogaster. Molecular divergence ofgypsy sequences betweenD. virilis andD. subobscura is estimated at 16 MY, whereas the most likely divergence time of these two species is more than 60 MY. These data strongly suggest thatgypsy sequences have been horizontally transferred between these species.Offprint requests to: T.M. Alberola     

Allozyme variation within and between populations inLolium (Poaceae)

Isozyme analysis was used to determine genetic variation within and between populations of sevenLolium species. All populations from the inbreeding species (L. temulentum, L. remotum, L. loliaceum, andL. persicum) were completely fixed for all enzymes scored. They also contained, for four of the five enzyme systems studied, exactly the same allelic variant. The three cross-breeding species showed large within-population variation and much less between-population variation. The great similarity of the allozymic variants found in all species, made the division of the genusLolium into species on basis of allozymic data difficult. It was not possible to separate the different inbreeding species from each other. Within the cross-breeding groupL. multiflorum andL. rigidum could be distinguished fromL. perenne. L. multiflorum, andL. rigidum could, with more difficulty, also be separated from each other. Allelic variation could have more relation with the provenance of the populations than with taxonomic classification.Genetic variation inLolium spp. II. For first part see Pl. Syst. Evol. 188: 87–99.     

On the conspecificity ofAnopheles fluviatilis species S withAnopheles minimus species C

Anopheles fluviatilis andAn. minimus complexes, each comprising of at least three sibling species, are closely related and important malaria vectors in Oriental Region. RecentlyAn. fluviatilis species S, which is a highly efficient malaria vector in India, has been made conspecific withAn. minimus species C (senior synonym) on the basis of homology in 335 base pair nucleotide sequence of D3 domain of 28S ribosomal DNA(rDNA). We examined the conspecificity of these two nominal species by obtaining and analysing the DNA sequences of nuclear ribosomal loci internal transcribed spacer 2 (ITS2) and D2-D3 domain of 28S rDNA (28S-D2/D3) from those ofAn. fluviatilis S andAn. minimus C. We found that the sequences ofAn. fluviatilis S are appreciably different from those ofAn. minimus C with pair-wise distance (Kimura-2-parametre model) of 3.6 and 0.7% for loci ITS2 and 28S-D2/D3, respectively. Pair-wise distance and phylogenetic analyses using ITS2 sequences of members of Minimus and Fluviatilis Complexes revealedthat An. fluviatilis S is distantly related toAn. minimus C as compared to any other members of the Fluviatilis Complex. These findings suggest that the two nominal species,An. fluviatilis S andAn. minimus C, do not merit synonymy. The study also confirms that the reported speciesAn. fluviatilis X is synonym with species S.     

Arrangement and size distribution of repeat and single copy DNA sequences in four species ofCucurbitaceae

DNA sequence organization patterns have been studied in fourCucurbitaceae plant species, namely,Luffa cylindrica (sponge gourd),L. acutangula (ridge gourd),Benincasa hispida (ash gourd) andCoccinia indica (ivy gourd). Extensive interspersion of repeat and single copy sequences has been observed in sponge gourd and ridge gourd. In ash gourd and ivy gourd, however, there is a limited interspersion of these sequences and a large portion of the single copy DNA remains uninterspersed. The interspersed repetitive sequences are composed of a major class (75–80%) of short repeats (300 base pairs long) and a minor class (15–20%) of long repeats (2 000–4 000 base pairs) in all the four species. The average length of single copy sequences dispersed among repeats is 1 800–2 900 base pairs. In spite of these gross similarities in the genome organization in the four species, the fraction of repeats and single copy sequences involved in short and long period interspersion patterns, and fraction of single copy sequences remaining uninterrupted by repeats are vastly different. The probable implications of these differences with respect to speciation events and rates of genome evolution are discussed.Molecular Analysis ofCucurbitaceae Genomes, III. — NCL Communication No.: 3595.     

Nucleotide sequence ofatpB,rbcL,trnR,dedB andpsaI chloroplast genes from a fernAngiopteris lygodiifolia: a possible emergence of Spermatophyta lineage before the separation of Bryophyta and Pteridophyta

To elucidate the evolutionary relationship between the Spermatophyta, Pteridophyta and Bryophyta, we cloned a fragment of chloroplast DNA from the fernAngiopteris lygodiifolia (Pteridophyta) and determined its nucleotide sequence. The fragment contained theatpB,rbcL,trnR-CCG,dedB andpsaI genes. Comparisons of the deduced amino acid and nucleotide sequences of these genes from the three plant groups indicate thatAngiopteris sequences are more closely related to those of Bryophyta species (85% identity on average) than to those of seed plants (76% identity on average), supporting a hypothesis that the Bryophyta and Pteridophyta diverged more recently from one another than their common progenitor diverged from that of the Spermatophyta.     

Three new species of anamorphic yeasts phenotypically and phylogenetically related toCandida mesenterica: The description ofCandida fungicola sp. nov.,Candida sagamina sp. nov., andCandida fukazawae sp. nov. isolated from fruit bodies of mushrooms

Four strains of anamorphic yeasts isolated from fruit bodies of mushrooms collected in Japan were found to represent three new species of the genusCandida. These species resembleCandida mesenterica in characteristics commonly employed in the classification of yeasts. On the basis of DNA-DNA reassociation, however, they were clearly distinguished fromC. mesenterica and from one another. Three new species,Candida fungicola, Candida sagamina, andCandida fukazawae, are proposed for these yeasts. The analysis of SSU rDNA sequences suggested that these three species were closely related to each other and toC. mesenterica andC. suecica.