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Filamentous green chaetophoralean algae are distributed mainly in aquatic ecosystems, less known from terrestrial habitats. Many of them have a complicated thalli and complex life cycles that make it difficult to determine these organisms by morphology. Description of new representatives of the Chaetophorales continues. In this study, we have explored the filamentous green alga IRK–A 341 from soil of the Baikal Region by light and electron microscopy along with molecular phylogeny. Based on the results of morphological, ecological, and molecular phylogenetic analyses (18S–28S rDNA, tufA), we described the studied alga as the new species, Barranca variabilis sp. nov. The study complements the data on the diversity of soils green filamentous algae, and their biogeography. For the first time, the data on the structure of the cell walls and the cell ultrastructure of Barranca were established. The information on the morphology of the reproductive and resting cells is updated.

Chaetophora lobata is the type species of the polyphyletic genus Chaetophora. Clarification of its species identification, phylogenetic relationships, and plastome characteristics is helpful for ta...

Considering the phylogenetic differences in the taxonomic framework of the Chaetophorales as determined by the use of nuclear molecular markers or chloroplast genes, the current study was the first to use phylotranscriptomic analyses comparing the transcriptomes of 12 Chaetophorales algal species. The results showed that a total of 240,133 gene families and 143 single-copy orthogroups were identified. Based on the single-copy orthogroups, supergene analysis and the coalescent-based approach were adopted to perform phylotranscriptomic analysis of the Chaetophorales. The phylogenetic relationships of most species were consistent with those of phylogenetic analyses based on the chloroplast genome data rather than nuclear molecular markers. The Schizomeriaceae and the Aphanochaetaceae clustered into a well-resolved basal clade in the Chaetophorales by either strategy. Evolutionary analyses of divergence time and substitution rate also revealed that the closest relationships existed between the Schizomeriaceae and Aphanochaetaceae. All species in the Chaetophorales exhibited a large number of expanded and contracted gene families, in particular the common ancestor of the Schizomeriaceae and Aphanochaetaceae. The only terrestrial alga, Fritschiella tuberosa, had the greatest number of expanded gene families, which were associated with increased fatty acid biosynthesis. Phylotranscriptomic and evolutionary analyses all robustly identified the unique taxonomic relationship of Chaetophorales consistent with chloroplast genome data, proving the advantages of high-throughput data in phylogeny.

Background: Order Chaetophorales currently includes six families, namely Schizomeridaceae, Aphanochaetaceae, Barrancaceae, Uronemataceae, Fritschiellaceae, and Chaetophoraceae. The phylogenetic relationships of Chaetophorales have been inferred primarily through phylogenetic analysis based on rDNA sequences. Most studies have primarily focused on intergeneric phylogenetic relationships within this order and the phylogenetic relationships with four other Chlorophycean orders (Chaetophorales, Chaetopeltidales and Oedogoniales, and Volvocales). The phylogenetic relationships among families in order Chaetophorales remain unclear. This study aimed to phylogenetically reconstruct order Chaetophorales and determine the taxonomic scheme and to further the current understanding of the evolution of order Chaetophorales . Results: In the present study, seven complete and five fragmentary chloroplast genomes were harvested. Phylogenomic and comparative genomic analysis were performed to determine the taxonomic scheme within Chaetophorales. Consequently, Oedogoniales was found to be a sister to a clade linking Chaetophorales and Chaetopeltidales. Schizomeriaceae, and Aphanochaetaceae clustered into a well-resolved basal clade in Chaetophorales, inconsistent with the results of phylogenetic analysis based on rDNA sequences. Comparative genomic analyses revealed that the chloroplast genomes of Schizomeriaceae and Aphanochaetaceae were highly conserved and homologous, highlighting the closest relationship in this order. Germination types of zoospores precisely correlated with the phylogenetic relationships. Conclusions: chloroplast genome structure analyses, synteny analyses, and zoospore germination analyses were concurrent with phylogenetic analyses based on the chloroplast genome, and all of them robustly determined the unique taxonomic scheme of Chaetophorales and the relationships of Oedogoniales, Chaetophorales, and Chaetopeltidales.

Mitogenome evolution in the Chlorophyceae is characterized by the acquisition of a reduced-derived pattern by the Chlamydomonadales + Sphaeropleales clade. Because no mitogenomes are available for the sister clade Oedogoniales + Chaetophorales + Chaetopeltidales, it remains unclear whether the common ancestor of chlorophycean green algae harbored a reduced-derived or ancestral-type mitogenome. The 70,191 and 46,765-bp mitogenomes reported here for Bulbochaete rectangularis var. hiloensis (Oedogoniales) and Stigeoclonium helveticum (Chaetophorales), respectively, shed light on this question. Both contain the same set of 41 conserved genes, a repertoire lacking numerous protein-coding genes but featuring all 27 tRNA genes typically found in ancestral-type mitogenomes.

A green alga in Burmese amber is described as Electrophycus astroplethus gen. et spec. nov. and tentatively placed in the extant family Chaetophoraceae. The new species is characterised by apical–b...

One green algal specimen from China was identified as Gongrosira fluminensis Fritsch, due to its unique morphology that pseudoparenchymal basal stratum with rounded or polygonal cells grew into dense little-branched upright threads that reached approximately the same height with specific akinete formation and germination and formed strong cushions without calcification. Examination of the ultrastructural characteristics of plasmodesmata and pyrenoid confirmed that Gongrosira fluminensis Fritsch should be excluded from the order Chaetophorales. The phylogenetic evidence based on DNA sequence data from the nucleus (18S rDNA, ITS rDNA) and chloroplast (tufA) sequences clearly revealed that the Gongrosira fluminensis Fritsch should be classified in the Ulvales (Ulvophyceae) as the new combination species of the genus Lithotrichon Darienko et Proschold, instead of the Chaetophorales (Chlorophyceae). More specimens in conjunction with natural morphological investigation and molecular analyses are required to reevaluate the microfilamentous genus Gongrosira Kutzing and reveal hidden diversities among the Ulvophyceae.

Filamentous green algae Chaetophorales present numerous taxonomic problems as many other green algae. Phylogenetic analyses based on nuclear genes have limited solutions. Studies with appropriate chloroplast molecular markers may solve this problems; however, suitable molecular markers for the order Chaetophorales are still unknown. In this study, 50 chloroplast genomes of Chlorophyceae, including 15 of Chaetophorales, were subjected to single protein-coding gene phylogenetic analyses, and substitution rate and evolutionary rate assays, and PCR amplification verification was conducted to screen the suitable molecular markers. Phylogenetic analyses of three chloroplast representative genes (psaB, tufA, and rbcL) amplified from 124 strains of Chaetophorales showed that phylogenetic relationships were not improved by increasing the number of samples, implying that the genes themselves, rather than limited samples, were the reason for the unsupported Topology I. Seven genes (atpF, atpI, ccsA, cemA, chlB, psbB, and rpl2) with robust support were selected to be the most suitable molecular markers for phylogenetic analyses of Chaetophorales, and the concatenated seven genes could replace the time-consuming and labor-intensive phylogenetic analyses based on chloroplast genome to some extent. To further solve the taxonomic problems of Chaetophorales, suitable chloroplast markers combined with more taxon-rich approach could be helpful and efficient.

The Chaetophorales (Chlorophyceae) consist of filamentous green algae that were mostly described directly from natural samples by light microscopy. However, the descriptions were often based on mor...

The original report of Hormotila mucigena Borzi for North America was misidentified (Wujek and Wynne, 2016). It was shown earlier to be a stage of Rufusiella insignis (Hassall) Loeblich III (1967). This branched filamentous green alga, a taxon now placed in the order Chaetophorales and originally described from Sicily, is once again reported for North America from Douglas County, Lawrence, Kansas area and Isabella County, Michigan.

The green branched filamentous alga Spongioplastidium proliferum Vischer is reported for the first time in North America. Spongioplastidium proliferum was collected from ponds in Douglas and Neosho Counties, Kansas. Subsequently it was isolated and placed into culture. Zoospore production is detailed for the first time.

It is generally accepted that land plants evolved from streptophyte algae. However, there are also many chlorophytes (a sister group of streptophyte algae and land plants) that moved to terrestrial habitats and even resemble mosses. This raises the question of why no land plants evolved from chlorophytes. In order to better understand what enabled streptophyte algae to conquer the land, it is necessary to study the chlorophytes as well. This review will introduce the freshwater filamentous chlorophyte alga Draparnaldia sp. (Chaetophorales, Chlorophyceae) as a model for comparative analyses between these two lineages. It will also focus on current knowledge about the evolution of morphological complexity in chlorophytes versus streptophytes and their respective morphological/behavioural adaptations to semi-terrestrial habitats, and will show why Draparnaldia is needed as a new model system.

Background Terrestrial, freshwater and marine green algae constitute the large and morphologically diverse phylum of Chlorophyta, which gave rise to the core chlorophytes. Chlorophyta are abundant and diverse in freshwater environments where sometimes they form nuisance blooms under eutrophication conditions. The phylogenetic relationships among core chlorophyte clades (Chlorodendrophyceae, Ulvophyceae, Trebouxiophyceae and Chlorophyceae), are of particular interest as it is a species-rich phylum with ecological importance worldwide, but are still poorly understood. In the Mediterranean ecoregion, data on molecular characterization of eukaryotic microalgae strains are limited and current knowledge is based on ecological studies of natural populations. In the present study we report the isolation and characterization of 11 green microalgae strains from Greece contributing more information for the taxonomy of Chlorophyta. The study combined morphological and molecular data. Results Phylogenetic analysis based on 18S rRNA, internal transcribed spacer (ITS) region and the large subunit of the ribulose-bisphosphate carboxylase (rbcL) gene revealed eight taxa. Eleven green algae strains were classified in four orders (Sphaeropleales, Chlorellales, Chlamydomonadales and Chaetophorales) and were represented by four genera; one strain was not assigned to any genus. Most strains (six) were classified to the genus Desmodesmus, two strains to genus Chlorella, one to genus Spongiosarcinopsis and one filamentous strain to genus Uronema. One strain is placed in a separate independent branch within the Chlamydomonadales and deserves further research. Conclusions Our study reports, for the first time, the presence of Uronema in an aquatic environment up to 40 °C and reveals new diversity within the Chlamydomonadales. The results from the ITS region and the rbcL gene corroborated those obtained from 18S rRNA without providing further information or resolving the phylogenetic relationships within certain genera, due to the limited number of ITS and rbcL sequences available. The comparison of molecular and morphological data showed that they were congruent. Cosmopolitan genera with high worldwide distribution inhabit Greek freshwaters.

Background Green plants comprise two lineages: (1) the streptophytes that colonised land and (2) the chlorophytes that have adaptations to land but remained mostly aquatic. To better understand what made streptophytes so successful, we are currently establishing the chlorophyte alga Draparnaldia sp. (Chaetophorales, Chlorophyceae) as a model for comparative analyses between these two lineages. However, establishing Draparnaldia as a valuable model requires that it can be transformed. Thus, our goal is to develop a transformation protocol for this alga. Results We have established the first transformation protocol for Draparnaldia. This protocol is based on protoplast transformation by electroporation. It includes instructions on protoplast isolation, regeneration and transient transfection. It also provides a list of the effective selective agents for future Draparnaldia transformations. Conclusions Our protocol opens a way for Draparnaldia functional genomics analyses. Moreover, it also provides an important base for establishment of stable transformation. Electronic supplementary material The online version of this article (10.1186/s13007-019-0460-6) contains supplementary material, which is available to authorized users.

The broadly defined genus Chaetophora consisted of species with minute, uniseriate branching filaments enveloped in soft or firm mucilage forming macroscopic growths that are spherical, hemispherical, and tubercular or arbuscular, growing epiphytically on freshwater aquatic plants and other submerged surfaces in standing or fast-flowing water. Recent molecular analyses clearly showed that this genus was polyphyletic. In this study, eight strains of Chaetophora and three strains of Stigeoclonium were identified and successfully cultured. In combination with the morphological data, a concatenated data set of four markers (18S + 5.8S + ITS2+ partial 28S rDNA) was also used to determine their taxonomic relationships and phylogenetic positions. The molecular analysis resolved the broadly defined Chaetophora to at least two genera. Species with a globose thallus of genus Chaetophora formed a separate monophyletic clade, which clearly separated from, a type of lobe-form Chaetophora species. Therefore, we propose to erect a new genus, Chaetophoropsis, which includes all globose species of the Chaetophora. Chaetophoropsis aershanensis was determined to be a new species, based on its special characteristic of profuse long rhizoids. Stigeoclonium polyrhizum, as the closest relative to Chaetophoropsis, revealed its distant relationships to other species of Stigeoclonium. A globose thallus with a thick, soft mucilage matrix, and special rhizoidal branches lent further support to the placement of S. polyrhizum in the genus Chaetophoropsis and had the closest relationship to C. aershanensis. Taxonomic diversity was proven by distinctive morphological differences and by phylogenetic divergence in the broadly defined Chaetophora identified herein.

Background Chlorophyceae is one of three most species-rich green algal classes and also the only class in core Chlorophyta whose monophyly remains uncontested as gene and taxon sampling improves. However, some key relationships within Chlorophyceae are less clear-cut and warrant further investigation. The present study combined genome-scale chloroplast data and rich sampling in an attempt to resolve the ordinal classification in Chlorophyceae. The traditional division into Sphaeropleales and Volvocales (SV), and a clade containing Oedogoniales, Chaetopeltidales, and Chaetophorales (OCC) was of particular interest with the addition of deeply branching members of these groups, as well as the placement of several incertae sedis taxa. Methods We sequenced 18 chloroplast genomes across Chlorophyceae to compile a data set of 58 protein-coding genes of a total of 68 chlorophycean taxa. We analyzed the concatenated nucleotide and amino acid datasets in the Bayesian and Maximum Likelihood frameworks, supplemented by analyses to examine potential discordant signal among genes. We also examined gene presence and absence data across Chlorophyceae. Results Concatenated analyses yielded at least two well-supported phylogenies: nucleotide data supported the traditional classification with the inclusion of the enigmatic Treubarinia into Sphaeropleales sensu lato. However, amino acid data yielded equally strong support for Sphaeropleaceae as sister to Volvocales, with the rest of the taxa traditionally classified in Sphaeropleales in a separate clade, and Treubarinia as sister to all of the above. Single-gene and other supplementary analyses indicated that the data have low phylogenetic signal at these critical nodes. Major clades were supported by genomic structural features such as gene losses and trans-spliced intron insertions in the plastome. Discussion While the sequence and gene order data support the deep split between the SV and OCC lineages, multiple phylogenetic hypotheses are possible for Sphaeropleales s.l. Given this uncertainty as well as the higher-taxonomic disorder seen in other algal groups, dwelling on well-defined, strongly supported Linnaean orders is not currently practical in Chlorophyceae and a less formal clade system may be more useful in the foreseeable future. For example, we identify two strongly and unequivocally supported clades: Treubarinia and Scenedesminia, as well as other smaller groups that could serve a practical purpose as named clades. This system does not preclude future establishment of new orders, or emendment of the current ordinal classification if new data support such conclusions.

Intensive investigation was made on certain ponds of Chidambaram taluk. The present studies are confined to Chlorophyceae and Bacilariophyceae. Studies revealed that the Chlorophyceae is represented by 6 orders i.e., Chlorococcales (19 genera and 41 species), Volvocales (3 genera and 3 species), Chaetophorales (2 genera), Oedogoniales (Oedogonium; 2 species, Bulbochaete; 1 species), 2 Zygnemales (Spirogyra; 4 species) and Conjugales (6 genera and 18 species), constituting 35 genera and 76 species. Chloroccales is most dominant group, representing 35 genera and 76 species. Chloroccales is most dominant group, representing 40% of the total reported green algal population were more followed by Bacilariophyceae members. It has been observed that the green algal population exhibited remarkable variations and fluctations both qualitatively and quantitatively during different seasons and months of year. The ecological characteristics of habitat water ranged remarkably i.e., pH (9-10), temperature (38-40 i‚°C), dissolved oxygen (5-11mg/l), carbondioxide (6-12mg/l), HCO3– (80-100mg/l), Ca++ (15-25mg/l), Mg++ (4-15mg/l) and Cl– (0.70-1.75mg/l).

During taxonomical study of Coleochaete Breb., an epiphytic alga , three species viz . Coleochaete nitellarum Jost, Coleochaete pulvinata A. Br. ex Kutz., Coleochaete soluta (Breb.) Pringsh. belonging to the order Chaetophorales under the class Chlorophyceae were recorded in the winter season from three sites of Hooghly district in West Bengal, India. The thallus of C . nitellarum grew diffusely on the submerged angiospermic plants and consists of irregularly branched prostrate filaments. In C . soluta , the erect filaments were freely branched while, upper cells of erect filaments were cylindrical and thallus was hemispherical in C . pulvinata . The most important finding of the limnological work was the higher concentration of DO in the lentic aquatic ecosystems where they occurred that might be due to higher rate of photosynthetic activities and lower temperature of water. This study also exhibited somewhat, lower nutrient status in respect of NO 3 -N: PO 4 3- : K ratios. The review of literatures revealed that C . pulvinata was the second time report and other two taxa were reported third timely from this state. Our Nature (2015), 13(1): 19-25

Premise of the study To enhance our knowledge of the diversity of microalgae, a phycological survey of the Canary Islands (Spain) was undertaken. Here we report the discovery of a (semi)terrestrial green filamentous alga isolated from a steep volcanic canyon on La Palma. This alga is continually exposed to changing weather conditions (floods vs. droughts) and thus provides a good opportunity to investigate possible adaptations to a semiterrestrial habitat with large fluctuations of environmental parameters. Methods We used axenic cultures, simulated flood and drought stresses and studied their effect on the life history of the alga using light, confocal laser scanning and scanning electron microscopy including fluorescent staining. Furthermore, phylogenetic analyses using rDNA sequence comparisons were performed. Key results Three specific life-history traits that likely represent adaptations to the fluctuating environment of the canyon were observed: (1) fragmentation through "filament splitting", a unique branching mechanism not reported before in algae and initiated by formation of oblique cross walls, (2) aplanospore formation, and (3) reproduction by multiflagellate zoospores with 4-24 flagella arranged in groups of four. Phylogenetic analyses identified the alga as Barranca multiflagellata gen. et sp. nov. (Barrancaceae fam. nov., Chaetophorales, Chlorophyceae). Moreover, the Chaetophoraceae Greville, 1824 was emended and a new family, Uronemataceae (fam. nov.) erected. Conclusions The discovery of Barrancaceae fam. nov. highlights the importance of investigating nonconventional habitats to explore microalgal diversity. The reproductive versatility demonstrated by Barranca suggests adaptation to a semiterrestrial habitat with large fluctuations in water availability.

The definition of species plays a pivotal role in biology. It has been proposed that Compensatory Base Changes (CBCs) in the fast-evolving Internal Transcribed Spacer 2 (ITS2) correlate with speciation and thus can be used to distinguish species. The applicability of CBC - based species concepts using ITS2, however, rests on the homology of the investigated ITS2 positions. We studied the ITS2 molecule of 147 strains of Chlorophyceae (Chlorophyta, Viridiplantae) including 26 new sequences in the order Chaetophorales, and compared their secondary structures to ITS2 in the sister class Ulvophyceae, represented by the order Ulvales. Using a phylogenetic/comparative approach, it was possible to identify 1) the first consensus structure model of the ITS2 molecule that can be applied to two classes of green algae [Ulvophyceae (Ulvales), Chlorophyceae] and 2) landmarks (the spacer regions separating the ITS2 Helices) for more robust prediction of the secondary structures in green algae. Moreover, we found that CBCs in homologous positions in these 147 strains (representing 115 validly described species) are either completely absent or mostly associated with internal branches representing higher order taxonomic levels (genera, families, orders). As reported for the Ulvales, CBCs are not diagnostic at the species level in the dataset used.

BackgroundThe chloroplast genome sustained extensive changes in architecture during the evolution of the Chlorophyceae, a morphologically and ecologically diverse class of green algae belonging to the Chlorophyta; however, the forces driving these changes are poorly understood. The five orders recognized in the Chlorophyceae form two major clades: the CS clade consisting of the Chlamydomonadales and Sphaeropleales, and the OCC clade consisting of the Oedogoniales, Chaetophorales, and Chaetopeltidales. In the OCC clade, considerable variations in chloroplast DNA (cpDNA) structure, size, gene order, and intron content have been observed. The large inverted repeat (IR), an ancestral feature characteristic of most green plants, is present inOedogonium cardiacum(Oedogoniales) but is lacking in the examined members of the Chaetophorales and Chaetopeltidales. Remarkably, theOedogonium35.5-kb IR houses genes that were putatively acquired through horizontal DNA transfer. To better understand the dynamics of chloroplast genome evolution in the Oedogoniales, we analyzed the cpDNA of a second representative of this order,Oedocladium carolinianum.MethodsTheOedocladiumcpDNA was sequenced and annotated. The evolutionary distances separatingOedocladiumandOedogoniumcpDNAs and two other pairs of chlorophycean cpDNAs were estimated using a 61-gene data set. Phylogenetic analysis of an alignment of group IIA introns from members of the OCC clade was performed. Secondary structures and insertion sites of oedogonialean group IIA introns were analyzed.ResultsThe 204,438-bpOedocladiumgenome is 7.9 kb larger than theOedogoniumgenome, but its repertoire of conserved genes is remarkably similar and gene order differs by only one reversal. Although the 23.7-kb IR is missing the putative foreign genes found inOedogonium, it contains sequences coding for a putative phage or bacterial DNA primase and a hypothetical protein. Intergenic sequences are 1.5-fold longer and dispersed repeats are more abundant, but a smaller fraction of theOedocladiumgenome is occupied by introns. Six additional group II introns are present, five of which lack ORFs and carry highly similar sequences to that of the ORF-less IIA intron shared withOedogonium. Secondary structure analysis of the group IIA introns disclosed marked differences in the exon-binding sites; however, each intron showed perfect or nearly perfect base pairing interactions with its target site.DiscussionOur results suggest that chloroplast genes rearrange more slowly in the Oedogoniales than in the Chaetophorales and raise questions as to what was the nature of the foreign coding sequences in the IR of the common ancestor of the Oedogoniales. They provide the first evidence for intragenomic proliferation of group IIA introns in the Viridiplantae, revealing that intron spread in theOedocladiumlineage likely occurred by retrohoming after sequence divergence of the exon-binding sites.

The green algal genus Cloniophora has been classified in the Chaetophorales (Chlorophyceae) based on morphological characters. This study uses DNA sequence data from the nucleus (SSU) and the chloroplast (tufA) from collections in the Hawaiian Islands and a culture originating from Portugal to test this classification. Taxonomic identities of contemporary collections were confirmed by sequencing small fragments of DNA (rbcL and SSU) from type specimens, including the generitype, Cloniophora willei L. H. Tiffany. These molecular data show that Cloniophora does not have close affinities to the Chaetophorales and belongs instead to the Ulvales (Ulvophyceae). The morphological features of eight or more reproductive products per cell and a pyrenoid with a traversing thylakoid membrane support the molecular data and confirm the placement of this group in the Ulvales. As this genus does not belong to any recognized family in the Ulvales, the family Cloniophoraceae, containing the single genus Cloniophora, is proposed.

In the Chlorophyceae, the chloroplast genome is extraordinarily fluid in architecture and displays unique features relative to other groups of green algae. For the Chaetophorales, 1 of the 5 major lineages of the Chlorophyceae, it has been shown that the distinctive architecture of the 223,902-bp genome of Stigeoclonium helveticum is consistent with bidirectional DNA replication from a single origin. Here, we report the 182,759-bp chloroplast genome sequence of Schizomeris leibleinii, a member of the earliest diverging lineage of the Chaetophorales. Like its Stigeoclonium homolog, the Schizomeris genome lacks a large inverted repeat encoding the rRNA operon and displays a striking bias in coding regions that is associated with a bias in base composition along each strand. Our results support the notion that these two chaetophoralean genomes replicate bidirectionally from a putative origin located in the vicinity of the small subunit ribosomal RNA gene. Their shared structural characteristics were most probably inherited from the common ancestor of all chaetophoralean algae. Short dispersed repeats account for most of the 41-kb size variation between the Schizomeris and Stigeoclonium genomes, and there is no indication that homologous recombination between these repeated elements led to the observed gene rearrangements. A comparison of the extent of variation sustained by the Stigeoclonium and Schizomeris chloroplast DNAs (cpDNAs) with that observed for the cpDNAs of the chlamydomonadalean Chlamydomonas and Volvox suggests that gene rearrangements as well as changes in the abundance of intergenic and intron sequences occurred at a slower pace in the Chaetophorales than in the Chlamydomonadales.

The green alga Printzina (Trentepohlia) lagenifera was found in southern Finland in the sori of the leaves of the indigenous fern Polypodium vulgare. The find is noteworthy because P. lagenifera occurred epiphyllous in a boreal, natural habitat. The subcosmopolitan P. lagenifera is a collective species; it was described from a hothouse in Germany. Two earlier finds from Finland are likewise from hothouses. The present alga is hardly conspecific with these lignicolous hothouse algae that most probably are introductions from the tropics. Another green alga, the common Desmococcus olivaceus (Chaetophorales) is reported to occur in Finland on the upper surfaces of the leaves of Polypodium vulgare.

Previously published molecular phylogenetic analyses of the Chaetophorales (Chlorophyceae) suffered from limited taxon sampling (six genera with only a single species per genus). To test the monophyly of species-rich genera, and to analyze the phylogenetic relationships among families and genera in the Chaetophorales, we determined nuclear-encoded SSU rDNA sequences from 30 strains of Chaetophorales, performed phylogenetic analyses using various methods, and screened clades for support by unique molecular synapomorphies in the SSU rRNA secondary structure. The Schizomeridaceae and the weakly supported Aphanochaetaceae were recovered as basal lineages. The derived family Chaetophoraceae diverged into two clades: the "Uronema clade" containing unbranched filaments, and a sister clade designated as "branched Chaetophoraceae" comprising Chaetophora, Stigeoclonium, Draparnaldia, Caespitella, and Fritschiella. Although some terminal clades corresponded to genera described (e.g., Caespitella and Draparnaldia), other clades were in conflict with traditional taxonomic designations. Especially, the genera Stigeoclonium and Chaetophora were shown to be polyphyletic. The globose species Chaetophora elegans was unrelated to lobate Chaetophora spp. (e.g., Chaetophora lobata). Since the original description of Chaetophora referred to a lobate thallus organization, the latter clade represented Chaetophora sensu stricto. In consequence, C. lobata was designated as lectotype of Chaetophora. Two Stigeoclonium species, Stigeoclonium farctum Berthold and Stigeoclonium'Longipilus', diverged independently from the type species of Stigeoclonium, Stigeoclonium tenue (C. Agardh) Kutz. These results indicated that some commonly used taxonomic characters are either homoplasious or plesiomorphic and call for a reevaluation of the systematics of the Chaetophorales using novel morphological and molecular approaches.

The diplobiontic–haplodiplontic life cycle with alternating isomorphic generations in Stigeoclonium tenue (C. Agardh) Kutz. is described for the first time. Sporophytes (2n = 10) arise from tetraflagellate zoospores that are produced by meiosis. Sporic meiosis might be inferred from the cruciform divisions formed during zoosporogenesis and is confirmed through observations of prophase I substages. Zoospores do not germinate directly but produce a haploid cyst that germinates to give rise to a gametophyte (n = 5). Gametophytes produce biflagellate isogametes, which fuse to produce zygotes that germinate by mitosis into the sporophytic stage. Gametophytes and sporophytes reproduce asexually both via mitotic tetraflagellate zoospores and by thallus fragmentation. Results from this study indicate that both the cosmopolitan distribution and dominance of S. tenue in many periphytic communities might be due to its multiple reproductive strategies.

Four specimens of Stigeoclonium spp., sampled in China between 2015 and 2016, were identified as the species Stigeoclonium polyrhizum (Chaetophoraceae, Chaetophorales) due to their unique morphology. A large part of the main filament and branches was tightly enclosed by numerous rhizoidal branches, which extended downward to the base of the plant to form an expanded holdfast. S. polyrhizum has previously been regarded as a synonym of S. longipilum or other related species by most phycologists. Therefore, a reassessment of S. polyrhizum based on morphological and molecular data was conducted. Rhizoidal branch development of S. polyrhizum was also described, showing the abundant rhizoidal branches present from the juvenile stage to the mature stage under controlled culture conditions. Phylogenetic evidence, using nuclear - encoded SSU rDNA data, clearly revealed that the Chaetophoraceae diverged into two well - supported sister clades: the Chaetophora- clade and the Fritschiella- clade. S. polyrhizum was included in the Fritschiella- clade instead of the Chaetophora- clade where Chaetophora draparnaldioides ( S. longipilum ) was found. The morphological and molecular data unambiguously show that S. polyrhizum is a valid species rather than a synonym of S. longipilum or other related species. Thylakoid bands appressed to the periphery of the pyrenoid matrix of S. polyrhizum was also described.

SUMMARY Four species of Stigeoclonium from Argentina were studied by means of transmission electron microscopy and light microscopy. For species identification, we collected data related to the prostrate system and zoospore germination. We also determined the chromosome number for each species. Stigeoclonium aestivale showed a more developed erect system than the prostrate one, zoospore germination was predominantly of erect type and the chromosome number was 8. Stigeoclonium tenue presented well developed, erect and prostrate systems, zoospore germination was initially of the prostrate type and the chromosome number was 5. In Stigeoclonium variabile the prostrate system predominated over the erect one, zoospore germination was strictly of the prostrate type and the chromosome number was 3. Stigeoclonium farctum presented a more developed prostrate system than the erect one, zoospore germination was strictly of the prostrate type and the chromosome number was 8. The ontogeny of the zoospore germination was related to the final relative development of the prostrate and erect portions of adult thalli.

The Chlorophyceae (sensu Mattox and Stewart) is a morphologically diverse class of the Chlorophyta displaying biflagellate and quadriflagellate motile cells with varying configurations of the flagellar apparatus. Phylogenetic analyses of 18S rDNA data and combined 18S and 26S rDNA data from a broad range of chlorophycean taxa uncovered five major monophyletic groups (Chlamydomonadales, Sphaeropleales, Oedogoniales, Chaetophorales, and Chaetopeltidales) but could not resolve their branching order. To gain insight into the interrelationships of these groups, we analyzed multiple genes encoded by the chloroplast genomes of Chlamydomonas reinhardtii P. A. Dang. and Chlamydomonas moewusii Gerloff (Chlamydomonadales), Scenedesmus obliquus (Turpin) Kutz. (Sphaeropleales), Oedogonium cardiacum Wittr. (Oedogoniales), Stigeoclonium helveticum Vischer (Chaetophorales), and Floydiella terrestris (Groover et Hofstetter) Friedl et O'Kelly (Chaetopeltidales). The C. moewusii, Oedogonium, and Floydiella chloroplast DNAs were partly sequenced using a random strategy. Trees were reconstructed from nucleotide and amino acid data sets derived from 44 protein-coding genes of 11 chlorophytes and nine streptophytes as well as from 57 protein-coding genes of the six chlorophycean taxa. All best trees identified two robustly supported major lineages within the Chlorophyceae: a clade uniting the Chlamydomonadales and Sphaeropleales, and a clade uniting the Oedogoniales, Chaetophorales, and Chaetopeltidales (OCC clade). This dichotomy is independently supported by molecular signatures in chloroplast genes, such as insertions/deletions and the distribution of trans-spliced group II introns. Within the OCC clade, the sister relationship observed for the Chaetophorales and Chaetopeltidales is also strengthened by independent data. Character state reconstruction of basal body orientation allowed us to refine hypotheses regarding the evolution of the flagellar apparatus.

Nonspecific acid phosphatases are a group of enzymes whose activity increases the availability of exogenous and endogenous orthophosphate either through extra- or intracellular hydrolysis of phosphate compounds. Our study demonstrates the activity of acid phosphatases in the filamentous freshwater alga Stigeoclonium tenue. These enzymes were detected following a cerium-based method in which cerium was used as an orthophosphate-capture reagent. In thalli from S. tenue from the natural environment, acid phosphatases were found in the longitudinal cell wall, plasmalemma, and vacuole. In thalli from Bold's Basal Medium culture, these enzymes were found mainly in the plasmalemma; they were scarce in the cell wall. In the thalli grown in phosphate-enriched culture medium, enzymes were found only in the plasmalemma. The low availability of orthophosphate in the medium seems to induce the transport of these enzymes to the cell wall. Its abundance, on the contrary, seems to attenuate this response without affecting the localization of acid phosphatases in the plasmalemma.

The evolutionary affinities of Heterochlamydomonas Cox and Deason and Dictyochloris Vischer ex Sterr were investigated using phylogenetic analyses of a combined data set of 18S and 28S rDNA sequences with those from 38 additional green algae. Previous ultrastructural studies have shown that motile cells of Heterochlamydomonas and Duictyochloris have an unusual flagellar apparatus organization in that the two flagella are of unequal length and the basal bodies are persistently parallel. Because of this similarity these taxa, along with Bracteacoccus Tereg, a third taxon with this same flagellar apparatus arrangement, are hypothesized to be closely related. We show, with maximum parsimony and Bayesian analyses, that the parallel basal bodies are not homologous in the three genera. Rather, Heterochlamydomonas is most closely related to Chlamydomonas baca in the clockwise flagellar apparatus clade, and Dictyochloris and Bracteacooccus are nested within the Sphaeropleales, which has the directly opposite flagellar absolute orientation. Surprisingly, Dictyochloris and Bracteacoccus are not supported as closest relatives. These relationships are supported by morphological features such as the presence or absence of a walled motile cell but not by the orientation of the basal bodies. In addition, our data are derived from multiple isolates of each study genera, and the analyses show that Heterochlamydomonas and Dictyochloris are each monophyletic.

Akinete formation in Stigeoclonium tenue (AG) KUTZ began under unfavourable conditions (exhaustion of nutrients and/or evaporation of the culture medium). Cells from both prostrate and erect portions produced akinetes. A gradual change in the shape of vegetative cells was observed. They lost their typical straight edges and acquired an oval contour with constrictions at the level of the transverse cell walls. At the ultrastructural level, the changes were: 1) accumulation of starch and lipids as reserve substances; 2) disorganization of the chloroplast; 3) reduction of the vacuole until it disappeared in mature akinetes; 4) thickening of the wall resulting from the synthesis of both an additional new own wall next to the parental wall and a mucilage external layer; 5) disorganization of the plasmodesmata in the transverse walls of vegetative cells. All of these drastic cytological changes are essential to the development of akinetes which provide S. tenue with an effective survival mechanism under unfavourable conditions.

In aquaculture centers of the northern region of Japan, "Nami-ita" (waved polycarbonate plates), on which the green alga Ulvella lens Crouan frat. (Chaetophoraceae: Chaetophorales) was cultured, are used to promote larval settlement and metamorphosis of the sea urchin species Strongylocentrotus intermedius (A. Agassiz) and S. nudus (A. Agassiz). We investigated chemical inducer(s) for larval settlement and metamorphosis of these sea urchins with extracts of U. lens. Bioassay-guided separation of the methanol extract using a combination of column and thin-layer chromatography led to the isolation of several active compounds, the chemical structures of which were determined by spectral and chemical methods. These active compounds were identified as glycoglycerolipids, all comprising several molecular species: sulfoquinovosyl monoacylglycerols (SQMGs), sulfoquinovosyl diacylglycerols (SQDGs), monogalactosyl monoacylglycerols (MGMGs), monogalactosyl diacylglycerols (MGDGs), digalactosyl monoacylglycerols (DGMGs) and digalactosyl diacylglycerols (DGDGs). Among these glycolipids, SQMGs, MGMGs, MGDGs and DGMGs induced larval metamorphosis of the sea urchin S. intermedius. SQMGs and MGDGs induced larval metamorphosis at a concentration of 5 µg ml–1, whereas SQDGs and DGDGs only induced larval settlement. These glycoglycerolipids are new congeners of chemical inducers to settlement and metamorphosis of planktonic larvae of sea urchins. The findings would provide a better understanding of larval settlement and metamorphosis in sea urchins.

Ultrastructural analyses of the flagellar apparatus suggested that Sphaeroplea, Atractomorpha, the Hydrodictyaceae, and the Neochloridaceae, all of which produce biflagellate motile cells with directly opposed (DO) basal bodies, are allied in an order Sphaeropleales. Recent studies of 18S rDNA sequence data supported an alliance of the DO group, but no data from Sphaeroplea and its allies were included. This investigation presented a test of the phylogenetic hypothesis suggested by the flagellar apparatus evidence using sequence data from the nuclear-encoded small-subunit rDNA (18S) and large subunit rDNA (26S) genes, combined with additional taxon sampling. Results from phylogenetic analyses weakly supported monophyly of biflagellate DO taxa and indicated that pyrenoids with cytoplasmic invaginations are present in numerous distinct lineages. Analysis of both molecular data sets supported a class Chlorophyceae comprised of at least six major groups that generally correspond to currently recognized orders or families: Chaetophorales, Chae- topeltidales, Chlamydomonadales, Sphaeropleales, Sphaeropleaceae, and Oedogoniales. In addition, Cylindrocapsa, Elakatothrix, Treubaria, and Trochiscia formed a seventh chlorophycean clade that is new to science. This investigation demonstrated that the 26S rDNA gene provides more phylogenetic signal, per unit sequence, than the 18S rDNA gene and that combined analysis yields topologies with more robust support than independent analysis of either data set.

SUMMARY Two populations of Chaetophora elegans (Roth) C, Agardh and two of Stigeoclonium helveticum Vischer were investigated for microhabitat characteristics and morphological variation in streams of Sao Paulo State, southeastern Brazil. Different patterns of microhabitat distribution were found between species investigated. Populations of C, elegans were distributed under relatively narrow microhabitat conditions (high irradiance, low depth, moderate to high current velocity, rocky substrata and lower values of niche width) and showing little morphometric variation (colony diameter, main axis cell size, and apical branch number), Stigeoclonium helveticum occurred under more diverse microhabitat conditions, revealed by lack of significant difference between sampling units with and without the alga and wider niche width, but also exhibited relatively narrow morphometric variation (plant length, main axis cell and lateral branch cell sizes). The narrow microhabitat conditions and smaller niche width of C. elegans can explain its low abundance (percentage cover) in streams from the area studied as well as in other regions of Sao Paulo State, In contrast, the wider variation of microhabitat conditions and the higher niche widths of S. helveticum suggest that this green alga is able to grow in a high number of stream ecosystems in the region investigated, ranging from undisturbed to highly disturbed habitats. Thus, the results suggest that S. helveticum is a generalist species.

The order Chaetophorales includes filamentous green algae whose taxonomic relationships to other chlorophycean orders is uncertain. Chaetophoralean taxa include filamentous species which are both branched and unbranched. Ultrastructural studies of zoospores have revealed similar flagellar apparatuses in a number of genera, including Uronema, Stigeoclonium, and Fritschiella, suggesting a close phylogenetic relationship among these taxa. The order Oedogoniales represents a second group of branched and unbranched filamentous green algae whose relationships to other chlorophycean orders also has been unclear. A possible close relationship between the Chaetophorales and Oedogoniales has been suggested. Using DNA sequences from the small-subunit ribosomal RNA gene (SSU rRNA) of several members of each order, we have examined the monophyly of the Chaetophorales and Oedogoniales, as well as the nature of their relationship to other chlorophycean orders. Our results show that chaetophoralean and oedogonialean taxa form separate monophyletic groups. Results also suggest that the two orders are not closely related to each other.

We study the occurrence of the alien macroalga Uronema marinum in the lagoon of Venice, in the lagoons and ponds of the Po Delta and in Pialassa della Baiona in Emilia-Romagna. It was in summer 2012 that U. marinum was identified for the first time, even though it has been present at least since 2008. This species, originally described from South Australia and Western Australia and probably imported with the Manila clam Tapes philippinarum, is prevalently associated with thalli of another introduced species, Agardhiella subulata, and the invasive Gracilaria vermiculophylla, which also have a Pacific origin and have recently colonized the same lagoon areas. Uronema marinum is currently widespread in the whole lagoon surfaces, but is particularly abundant in stagnant waters, rich in nutrients, where Gracilariaceae and Solieriaceae prevail on Ulvaceae.

SUMMARY To elucidate the phylogeny of the Dunaliellales sensu Ettl. the taxon often thought to be intermediate between primitive green flagellates such as the prasinophytes and the advanced chlorophycean algae, the sequences of the nuclear-encoded small subunit ribosomal RNA gene (18SrDNA) were determined and analyzed for five green algae, including three dunaliellalean algae. Phylogenetic trees based on 18SrDNA suggest that Oltmannsiellopsis viridis (Margraves et Steele) Chihara et Inouye represents an early divergence in the Ulvophyceae/Trebouxio-phyceae/Chlorophyceae clade and has no close relationship to any other green algae, as also suggested from ultrastructural characters. We propose Oltmannsiellopsidates ord. nov. for this genus. Hafniomonas and Polytomella are included in the clade which is characterized by clockwise basal bodies (CW group). The 18SrDNA trees suggest that multiple losses of the cell wall of the flagellate cell occurred in the CW group, and that the Dunaliellales sensu Ettl has a polyphyletic nature. This study also suggests that Planophita terrestns Groover et Hof-stetter (Chaetopeltidales) and Chaetophora incras-sata (Hudson) Hazen (Chaetophorales) are distinct lineages in the Chlorophyceae.

Two microfilamentous green algal specimens from Tibet were identified as Ctenocladus circinnatus Borzi, due to their unique morphology and saline habitat. The phylogenetic evidence based on DNA sequence data from the nucleus (SSU) and chloroplast elongation factor TU (tufA) sequences clearly revealed that the genus Ctenocladus, which has been classified in the Chaetophorales (Chlorophyceae) or Trebouxiophyceae incertae sedis by most phycologists, should be included in the Ulvales (Ulvophyceae) instead, and has a close relationship with the genera Pseudendoclonium and Phaeophila. We speculate that there may be undescribed or cryptic species especially in freshwater and other non-marine habitats. A phylogenetic re-evaluation based on large samples of microfilamentous ulvophycean algae especially freshwater specimens is needed.

Clones of the filamentous green alga Ctenocladus circinnatus Borzi were isolated from algae collected at Abert Lake (Oregon) and Mono Lake (California). Stock cultures were exposed to varied salinities of natural lake water to examine the effects on growth rate, cell form, chlorophyll a, and water content. Growth rates were reduced in both clones with increased salinity over the range 25–100 g·L−1 and were almost completely inhibited at 150 g·L−1. Chlorophyll a increased between salinities of 25 and 100 g·L−1, reflecting slower growth, higher proportions of akinetes, and smaller cell sizes as salinity increased. Tissue water content remained essentially constant from 25 to 100 g·L−1 salinity. Shorter cell dimensions with increased salinity suggest that a lower surface-to-volume ratio may reduce the potential for passive loss of cell water. Prior acclimation of stock cultures to elevated salinity provided no enhancement of growth response at any salinity. The results indicate that environmental salinity can limit the productivity and distribution of Ctenocladus in nature.

Vegetative cells and zoospores of Hormotilopsis gelatinosa Trainor & Bold, H. tetravacuolaris Arce & Bold, Planophila terrestris Groover & Hofstetter, and Phyllogloea fimbriata (Korchikov) Silva were examined by transmission electron microscopy. All cells had pyrenoids traversed by cytoplasmic channels. Zoospores were quadriflagellate and had essentially cruciate flagellar apparatuses. Scales were present on free-swimming zoospores. These features are essentially identical to those of Chaetopeltis sp. and are dissimilar to those of other described green algae. The new order Chaetopeltidales is created to accommodate the genera Chaetopeltis, Hormotilopsis, Planophila sensu Groover & Hofstetter, Phyllogloea, Dicranochaete, and Schizochlamys, organisms previously scattered among the orders Tetrasporales, Chloro-coccales, Chlorosarcinales, and Chaetophorales. Members of the order are closely related to the ancestral chlorophycean flagellate genus Hafniomonas, may be ancestral with respect to other Chlorophyceae, and may also be closely related to the ulvophycean order Ulotrichales.

Molecular phylogenetic analyses have had a major impact on the classification of the green algal class Chlorophyceae, corroborating some previous evolutionary hypotheses, but primarily promoting new interpretations of morphological evolution. One set of morphological traits that feature prominently in green algal systematics is the absolute orientation of the flagellar apparatus in motile cells, which correlates strongly with taxonomic classes and orders. The order Sphaeropleales includes diverse green algae sharing the directly opposite (DO) flagellar apparatus orientation of their biflagellate motile cells. However, algae across sphaeroplealean families differ in specific components of the DO flagellar apparatus, and molecular phylogenetic studies often have failed to provide strong support for the monophyly of the order. To test the monophyly of Sphaeropleales and of taxa with the DO flagellar apparatus, we conducted a molecular phylogenetic study of 16 accessions representing all known families and diverse affiliated lineages within the order, with data from four plastid genes (psaA, psaB, psbC, rbcL) and one nuclear ribosomal gene (18S). Although single-gene analyses varied in topology and support values, analysis of combined data strongly supported a monophyletic Sphaeropleales. Our results also corroborated previous phylogenetic hypotheses that were based on chloroplast genome data from relatively few taxa. Specifically, our data resolved Volvocales, algae possessing predominantly biflagellate motile cells with clockwise (CW) flagellar orientation, as the monophyletic sister lineage to Sphaeropleales, and an alliance of Chaetopeltidales, Chaetophorales, and Oedogoniales, orders having multiflagellate motile cells with distinct flagellar orientations involving the DO and CW forms.

Over the past two decades, molecular phylogenetic data have allowed evaluations of hypotheses on the evolution of green algae based on vegetative morphological and ultrastructural characters. Higher taxa are now generally recognized on the basis of ultrastructural characters. Molecular analyses have mostly employed primarily nuclear small subunit rDNA (18S) and plastid rbcL data, as well as data on intron gain, complete genome sequencing, and mitochondrial sequences. Molecular-based revisions of classification at nearly all levels have occurred, from dismemberment of long-established genera and families into multiple classes, to the circumscription of two major lineages within the green algae. One lineage, the chlorophyte algae or Chlorophyta sensu stricto, comprises most of what are commonly called green algae and includes most members of the grade of putatively ancestral scaly flagellates in Prasinophyceae plus members of Ulvophyceae, Trebouxiophyceae, and Chlorophyceae. The other lineage (charophyte algae and embryophyte land plants), comprises at least five monophyletic groups of green algae, plus embryophytes. A recent multigene analysis corroborates a close relationship between Mesostigma (formerly in the Prasinophyceae) and the charophyte algae, although sequence data of the Mesostigma mitochondrial genome analysis places the genus as sister to charophyte and chlorophyte algae. These studies also support Charales as sister to land plants. The reorganization of taxa stimulated by molecular analyses is expected to continue as more data accumulate and new taxa and habitats are sampled. Twenty years ago, a relatively slim volume with chapters by leading chlorophycologists celebrated the systematics of green algae (Irvine and John, 1984), a field that was undergoing rapid and fascinating changes, both in content and theory. ‘‘The present period may be termed the ‘Age of Ultrastructure’ in green algal systematics,’’ wrote Frank Round (1984, p. 7) in the introductory chapter, which summarized the history and state of the art. Round (1984) argued that light microscopy had laid the foundation in the preceding two centuries, but that the foundation was largely descriptive—alpha taxonomy in the most restricted sense. Ultrastructure, he asserted, had enlarged and presumably would continue to expand our horizons to unify systematics of green algae and overcome the fragmented alpha taxonomy that had dominated the field. Little did Round know that this golden age of green algal systematics was about to go platinum. Molecular systematics, in concert with a rigorous theoretical approach to data analysis and hypothesis testing (Theriot, 1992; Swofford et al., 1996), would at first complement and then transform the age of ultrastructure and usher in the ‘‘Age of Molecules.’’ In this article, we review the major advances in green algal systematics in the past 20 years, with a focus on well-supported, monophyletic taxa and the larger picture of phylogeny and evolution of green algae. We will review the types of data that have fueled these advances. As will become obvious, this perspective entails discussion of some embryophytes as well as their closest green algal relatives. In addition, we will point

Since the human inclination to estimate and trace natural diversity, usable species definitions as well as taxonomical systems are required. As a consequence, the first proposed classification schemes assigned the filamentous and parenchymatous taxa to the green algal order Chaetophorales sensu Wille. The introduction of ultrastructural and molecular methods provided novel insight into algal evolution and generated taxonomic revisions based on phylogenetic inference. However, until now, the number of molecular phylogenetic studies focusing on the Chaetophorales s.s. is surprisingly low. To enhance knowledge about phylogenetic relationships among taxa within the order, the nuclear?encoded SSU rDNA sequences from 30 strains covering all three chaetophoralean families have been investigated. All revealed monophyletic groupings were further screened for molecular non-homoplasious synapomorphies within the Viridiplantae. To address the question of the correspondence between morphological characters traditionally used for taxonomical delimitation of the Chaetophorales and the tree topology favored by molecular data, the list of morphological/ ultrastructural/ecological characters was elaborated and further analyzed. In addition, to obtain a close-up view into the evolution of Compensatory Base Changes (CBCs) of the second internal transcribed spacer (ITS2) which is currently often used to delimit putative biological species, 86 newly obtained/published sequences of ITS2 for five families of the order Ulvales were analyzed. Furthermore, a detailed comparative study of all ITS2 substitutions has been done. Subsequently all revealed CBCs and hemi- CBCs have been mapped upon the ITS2 phylogenetic tree topology. Finally, CBCs/hCBCs taxonomic inference in the Ulvales has been discussed.

The Chlorophyceae, an advanced class of chlorophyte green algae, comprises five lineages that form two major clades (Chlamydomonadales + Sphaeropleales and Oedogoniales + Chaetopeltidales + Chaetophorales). The four complete chloroplast DNA (cpDNA) sequences currently available for chlorophyceans uncovered an extraordinarily fluid genome architecture as well as many structural features distinguishing this group from other green algae. We report here the 521,168-bp cpDNA sequence from a member of the Chaetopeltidales (Floydiella terrestris), the sole chlorophycean lineage not previously sampled for chloroplast genome analysis. This genome, which contains 97 conserved genes and 26 introns (19 group I and 7 group II introns), is the largest chloroplast genome ever sequenced. Intergenic regions account for 77.8% of the genome size and are populated by short repeats. Numerous genomic features are shared with the cpDNA of the chaetophoralean Stigeoclonium helveticum, notably the absence of a large inverted repeat and the presence of unique gene clusters and trans-spliced group II introns. Although only one of the Floydiella group I introns encodes a homing endonuclease gene, our finding of five free-standing reading frames having similarity with such genes suggests that chloroplast group I introns endowed with mobility were once more abundant in the Floydiella lineage. Parsimony analysis of structural genomic features and phylogenetic analysis of chloroplast sequence data unambiguously resolved the Oedogoniales as sister to the Chaetopeltidales and Chaetophorales. An evolutionary scenario of the molecular events that shaped the chloroplast genome in the Chlorophyceae is presented.

Three species of Gongrosira were isolated from freshwater habitats in England and Denmark. Developmental morphology and reproduction of the isolated strains were studied over a wide range of light and temperature regimes by means of a cross-gradient apparatus. Some of the traits traditionally used for defining infraspecific taxa (e.g. comparative development of prostrate and erect branches, position of zoosporangia) were found to be of questionable diagnostic value. New information is provided on previously described species and the isolates compared with cultures of related taxa. Gongrosira scourfieldii was collected from its type locality and culture-based observations have enabled an improved description of this little-known alga. A new species of Gongrosira is described, G. pseudoprostrata L. Johnson nov. sp., characterized by the presence of a well-defined primary prostrate axis and secondary axes. Problems of defining some of the main character traits and the possible ecological implications of the ...

Clonal algal cultures isolated from a stream in south-west England and a lake in northern Denmark correspond to the little-known genus Dilabifilum. The developmental morphology and reproduction of these isolates were compared with cultures (often type cultures) of the three species of Dilabifilum species recognized by Tschermak-Woess. All our isolates closely corresponded with a previously described species, Dilabifilum printzii. Some of the vegetative traits traditionally used for defining species were found to be of questionable diagnostic value. The most useful traits proved to be the branching habit, degree of compactness of the filaments, length to breadth ratio of the cells, form of the cells, and physiological features (e.g. growth rate, readiness to become coccoidal). The genus remains ill-defined, with earlier observations concerning the production of quadriflagellate zoospores by the type culture of D. arthopyreniae still requiring reconfirmation.