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Epilithic green algae play crucial roles in aquatic ecosystems. While traditional morphological approaches have provided valuable insights into these algae, their limitations in resolving cryptic species and fully capturing the extent of diversity have impeded a comprehensive understanding. The diversity and phylogenetic relationships of these algae largely remain unexplored. In the present study, single molecular real-time (SMRT) sequencing of the full-length 18S rDNA was employed to investigate the molecular diversity and phylogenetic relationships of epilithic green algae in biodiversity conservation hotspots. A total of 79 709 algal reads were generated, yielding 169 amplicon sequence variants (ASVs) from 21 epilithic riverine green algal assemblages classified in the Ulvophyceae, Chlorophyceae and Trebouxiophyceae. The 10 families in which the greatest number of unique ASVs (diversity) were observed were the Ulvellaceae, Protosiphonaceae, Chlorococcaceae, Watanabeaceae, Chlorellaceae, Neochloridaceae, Chaetophoraceae, Hazeniaceae, Tupiellaceae and Scenedesmaceae. Unique clades formed by a substantial number of sequences of green algae were detected within the Ulvales/Ulotrichales (Ulvophyceae) and the Chlorococcaceae/Protosiphonaceae (Chlorophyceae). This study has unveiled hidden diversity within epilithic green algae, expanding our knowledge of their evolutionary relationships and providing fundamental data for taxonomic investigations within this group, as well as contributing to the assessment of algal diversity in biodiversity conservation hotspots. This study suggests that these epilithic green algal assemblages may be significantly more diverse than previously recognized, but we acknowledge the limited ecological and temporal scope of our sampling effort and the resulting effects on which taxa are identified, warranting further research. Molecular analysis shows high hidden diversity of epilithic green algae The families Ulvellaceae and Protosiphonaceae had the greatest number of unique ASVs Many unique clades within the Ulvales/Ulotrichales and Chlorococcaceae/Protosiphonaceae [ABSTRACT FROM AUTHOR]

Members of the family Scenedesmaceae are some of the most common algal taxa in inland ecosystems, and they are widely distributed in freshwaters, aerial, and sub-aerial habitats. With the continuous updating of methods, the classic morphological taxonomy of this family needs to be revised. In recent years, many genera of Scenedesmaceae have been established via the use of molecular methods. The phylogenetic relationships within Scenedesmaceae were analyzed using different molecular markers and morphological data, and the new freshwater genus Coccoidesmus Wang, Hou et Liu gen. nov. was described. Two new species in this genus were also described. Phylogenetic analysis based on tufA genes revealed that the new genus formed an independent clade closely related to Comasiella. However, these two genera are characterized by significant morphological differences in colony arrangement and cell shape. The chloroplast genome of the type species was assembled and annotated, and analyses of genome structure and sequences were conducted. More genome data could help clarify the phylogenetic relationships within this family. [ABSTRACT FROM AUTHOR]

Stigeoclonium is a genus of green algae that is widely distributed in freshwater habitats around the world. The genus comprises species with variously developed prostrates and erect systems of uniseriate branched filaments and grows attached to a wide range of different surfaces. It holds significant promise for applications in water quality indicators, sewage treatment, and the development of high-value-added products. Nevertheless, our comprehension of Stigeoclonium remains unclear and perplexing, particularly regarding its fundamental systematic taxonomy. Recent molecular analyses have revealed that the morphologically well-defined genus Stigeoclonium is polyphyletic and requires taxonomic revision. Phylogenetic analysis based on a single molecular marker and limited samples is insufficient to address the polyphyletic nature of Stigeoclonium. In the present study, 34 out of 45 strains of Stigeoclonium were newly acquired from China. Alongside the morphological data, a concatenated dataset of three markers (18S rDNA + ITS2 + tufA) was utilized to determine their molecular phylogeny. The phylogenetic analysis successfully resolved the broadly defined Stigeoclonium into three robustly supported clades (Stigeoclonim tenue clade, S. farctum clade, and S. helveticum clade). The morphological characteristics assessment results showed that the cell type of the main axis-producing branch, considered a crucial morphological characteristic of the Stigeoclonium taxonomy, did not accurately reflect the real phylogeny of the genus. A new taxonomical classification of the genus Stigeoclonium was proposed based on zoospores' germination types, which aligned well with the phylogenetic topologies. Species where zoospores showed erect germination (S. helveticum clade) formed a distinct monophyletic clade, clearly separated from the other two clades, with zoospores showing prostrate germination or pseudo-erect germination. Consequently, a new genus, Pseudostigeoclonium gen. nov., is suggested to include all species in the broadly defined Stigeoclonium with zoospores with erect germination. The taxonomic diversity is supported by distinctive morphological differences and phylogenetic divergence within the broadly defined Stigeoclonium identified in this study. Further evaluation of the genus Stigeoclonium is necessary, especially via examining additional specimens and re-evaluating morphological characters under precisely defined laboratory conditions. [ABSTRACT FROM AUTHOR]

Makinoella Okada is a genus of coccoid green algae belonging to the family Oocystaceae with typical morphological characteristics. Reports to date included only one type species. Since the genus is rarely reported, there is a lack of taxonomic research. In this study, 17 strains of this genus were collected from several different places in China, and their taxonomic studies were conducted based on morphological and molecular phylogenetic analysis. The phylogenetic results based on the analysis of ITS, chloroplast genes rbcL and tufA showed that the genus was divided into two branches. One of the branches comprised newly collected algae. Compensatory base changes (CBCs) and hemi-compensatory base changes (hemi-CBCs) within the secondary structure of the ITS-2 confirmed the branches as two species. Compared with the type species of Makinoella tosaensis, the cell size of this new branch was only about 50%, and included 2–4 colonial cells. Therefore, based on the smaller cell size, simpler colony composition, independent phylogenetic position and CBCs and hemi-CBCs, we suggest that the new clade should be designated as a new taxon of Makinoella, namely Makinoella parva. Among the four molecular markers using in this study, the rbcL and newly introduced ITS are recommended for species separation, which can help further studies to revise the species and generic concepts of the family. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Chaetophora lobata is the type species of the polyphyletic genus Chaetophora. Clarification of its species identification, phylogenetic relationships, and plastome characteristics is helpful for taxonomic revisions and evolutionary studies of this genus. This study presents detailed information on morphology, culture, phylogenetic analysis, phylogenomics and comparative genomics of C. lobata. The complete morphological description and phylogenetic analyses based on nuclear rDNA and chloroplast protein-coding genes were robust for the identification of C. lobata. Moreover, de novo sequencing was used to determine the 222 213 bp chloroplast genome, which lacked inverted repeats as in all other members of the Chaetophorales. The chloroplast DNA (cpDNA) of C. lobata was found to be AT-rich (68.2%), consisting of 67 protein-coding genes, among which 62 genes were encoded on the plus strand and five genes were encoded on the minus strand with significant imbalance in distribution. Additionally, 28 tRNA genes, three rRNA genes, 16 introns and 18 putative open reading frames (ORFs) were annotated. Eight ORFs arranged in a row occupied about 18 kbp between psaA and atpI genes, and 16 introns (14 group I introns and 2 group II introns) were annotated in nine genes. Six genes contained nine intronic ORFs. Synteny and rearrangements analysis clearly showed the closest relationships between Chaetophora sp. and C. lobata. Substitution rate estimation indicated that the cpDNA of Chaetophorales was under purifying selection and most species were under similar evolutionary pressure. These findings can help advance research on the taxonomy and phylogeny of the order Chaetophorales. [ABSTRACT FROM AUTHOR]

Cephaleuros is often known as an algal pathogen with 19 taxonomically valid species, some of which are responsible for red rust and algal spot diseases in vascular plants. No chloroplast genomes have yet been reported in this genus, and the limited genetic information is an obstacle to understanding the evolution of this genus. In this study, we sequenced six new Trentepohliales chloroplast genomes, including four Cephaleuros and two Trentepohlia. The chloroplast genomes of Trentepohliales are large compared to most green algae, ranging from 216 to 408 kbp. They encode between 93 and 98 genes and have a GC content of 26–36%. All new chloroplast genomes were circular-mapping and lacked a quadripartite structure, in contrast to the previously sequenced Trentepohlia odorata , which does have an inverted repeat. The duplicated trnD -GTC , petD , and atpA genes in C. karstenii may be remnants of the IR region and shed light on its reduction. Chloroplast genes of Trentepohliales show elevated rates of evolution, strong rearrangement dynamics and several genes display an alternative genetic code with reassignment of the UGA/UAG codon presumably coding for arginine. Our results present the first whole chloroplast genome of the genus Cephaleuros and enrich the chloroplast genome resources of Trentepohliales. [ABSTRACT FROM AUTHOR]

The Oedogoniales comprises the genera Oedogonium, Oedocladium and Bulbochaete which include more than 600 described species. However, only two chloroplast genome sequences have been reported to date, and little is known about the evolutionary relationships between these genera or their infrageneric classification. This group is economically important in astaxanthin production and the energy sector. We examined four Oedocladium strains from different locations in China. One was identified as O. prescottii, and the morphology and molecular data for the other three strains suggested that they were O. carolinianum. Our chloroplast (cp) genome sequences of O. prescottii (FACHB-2452) and O. carolinianum (FACHB-2453) were compared with previously reported Oedogoniales cp genomes. The cp genome of O. carolinianum FACHB-2453 was of similar size, structure and composition to that of O. carolinianum UTEX LB 1686, apart from two missing group I introns in the genes psbA and psbD. O. prescottii showed more differences with regard to genome size, structure, number of genes and introns, and repeats than the other Oedogoniales cp genomes. The inverted repeats (IRs) region of the O. prescottii cp genome had a 10-kb contraction. Synteny analysis of the four cp genomes showed a high degree of syntenic conservation in general; however, some structural variation at genus level was observed in Oedocladium. In phylogenetic analyses of Oedocladium using sequences of 18S rDNA, ITS and 59 cp protein-coding genes, species were placed on two separate branches within the Oedogonium clade, indicating that Oedocladium is a paraphyletic group. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Cephaleuros is a subaerial green alga widely distributed in tropical and subtropical regions. In this study, we investigated the taxonomy of Cephaleuros in southern China. Eight morphospecies (C. virescens, C. karstenii, C. expansus, C. drouetii, C. diffusus, C. tumidae-setae, C. lagerheimii and C. parasiticus) were identified according to previous descriptions, among which C. karstenii was the most frequently recorded. Subsequently, molecular phylogeny of Cephaleuros was inferred from 18S rDNA, ITS rDNA, 28S rDNA and rbcL sequences. Cephaleuros was subdivided into nine clades based on ITS analysis, which were recovered in phylogenies of two concatenated datasets (18S+ITS+28S and 18S+ITS+28S+rbcL). Molecular analyses revealed that Cephaleuros was composed of two main lineages, with one lineage comprising mostly samples of C. karstenii and C. virescens and the other comprising other species. The molecular phylogenies were not congruent with conventional morphological classifications, although some clades were distinguished by one or more diagnostic features. While the morphological features used to distinguish Cephaleuros spp were not reliable, the rbcL gene proved to be a good marker for identification of Cephaleuros. The ITS2 secondary structure prediction model showed that Cephaleuros spp have one ring structure with four helices, with Helix III being the longest without a branch in clades 3, 5 and 8. The present study represents the most detailed taxonomic assessment of Cephaleuros available to date. Further taxonomic research incorporating samples from the type locality is necessary for the reassessment of C. virescens. [ABSTRACT FROM AUTHOR]

Oocystis-like algae are diverse and found throughout the world. In this study, two new strains of algae with colonies resembling Oocystis were identified and successfully cultured in the laboratory. A polyphasic approach, including light microscopy, transmission electron microscopy and phylogenetic analyses of 18S rDNA and rbcL cpDNA, was used to characterize the strains. Observation of the two strains showed a regular colonial morphology (cells/daughter colony are parallel to each other and positioned near the edge of the mother cell wall) and a characteristic fibrillar network structure on the mother cell wall is obvious in light microscopy, which distinguished them from other Oocystis-like members described previously. Molecular analyses revealed that the two strains form an independent lineage within the subfamily Makinoelloideae (Oocystaceae, Trebouxiophyceae), separated from other genera. Therefore, we described this alga as Reticulocystis yunnanensis gen. et sp. nov. The genera of Makinoelloideae with similar colonial morphotypes were revealed as not monophyletic and the relationships between them remained undetermined. Additional taxon sampling and data from additional multiple genes are necessary to assess the taxonomy of the Makinoelloideae. [ABSTRACT FROM AUTHOR]

Samples of Pithophoraceae, collected in diverse freshwater and damp terrestrial habitats from tropical and subtropical China, were characterized morphologically and ultrastructurally, and their phylogenetic position was determined based on nuclear ribosomal DNA sequences. Our phylogenetic analysis resolved a novel lineage of Pithophoraceae, sister to Aegagropilopsis. Based on our phylogenetic results, morphological observations and comparative rDNA ITS2 secondary structure analysis, we propose Chlorocladiella gen. nov., characterized by a well‐developed system of prostrate filaments, and describe four new species, C. cochlea sp. nov., C. erecta sp. nov., C. medogensis sp. nov., and C. pisformis sp. nov. Two species were found growing on damp soil, which is an unusual habitat for cladophoralean green algae, indicating that the diversity of Cladophorales in terrestrial habitats may be greater than currently recognized. [ABSTRACT FROM AUTHOR]

Members of the Watanabea clade of Trebouxiophyceae are genetically diverse and widely distributed in all kinds of habitats, especially in most terrestrial habitats. Ten new strains of terrestrial algae isolated from the tropical rainforest in China, and four published strains were investigated in this study. Morphological observation and molecular phylogenetic analyses based on the 18S, ITS, rbcL, and tufA genes were used to identify the new strains. Four previously described species were reinvestigated to supplement molecular data and autospores' morphological photographs. The phylogenetic analyses based on 18S only, the concatenated dataset of 18S and ITS, as well as the concatenated dataset of rbcL and tufA, showed the same phylogenetic positions and relationships of these new strains. According to the phylogenetic analysis and morphological comparisons results, we described these 10 strains as four new members within the Watanabea clade, Polulichloris yunnanensis sp. nov., Polulichloris ovale sp. nov., Massjukichlorella orientale sp. nov., and Massjukichlorella minus sp. nov., and two known species, Massjukichlorella epiphytica, and Mysteriochloris nanningensis. Additionally, we provide strong evidence proving that Phyllosiphon, Mysteriochloris, Polulichloris, and Desertella all reproduce through unequal sized autospores. [ABSTRACT FROM AUTHOR]

Two unidentified foliicolous algal strains were isolated from living Ficus leaves in a tropical rainforest of China. Morphological, ultrastructural, and molecular phylogenetic investigations of these strains indicated that they differ from each other but cluster together and represent a new lineage of Trebouxiophyceae in the Choricystis/Botryococcus clade. These strains had similar morphologies with solitary, fusiform, and slightly asymmetrical vegetative cells, a basal stalk, and smooth cell walls. Cells contained a single parietal chloroplast with a pyrenoid surrounded by several starch sheaths and penetrating thylakoids. Asexual reproduction occurred in both strains through the splitting of the mother cell wall into two autospores. Phylogenetic analysis based on 18S, ITS and rbcL showed the two strains as a distinct branch in the Choricystis/Botryococcus clade of the Trebouxiophyceae. Here, we describe the two strains as the new genus, Obliquicauda gen. nov., and two new species, Obliquicauda inflata sp. nov. and Obliquicauda apiculata sp. nov. [ABSTRACT FROM AUTHOR]

The genus Coelastrella was established by Chodat (Bull. Soc. Bot. Geneve, 13 [1922] 66), and was characterized as being unicellular or in few‐celled aggregations with many longitudinal ribs on the cell wall. Many species of this genus showed strong ability to accumulate carotenoids and oils, so they have recently attracted much attention from researchers due to its potential applicability in the energy and food industries. In this study, a total of 23 strains of Coelastrella were sampled from China, and three new species and two new varieties were described: C. thermophila sp. nov., C. yingshanensis sp. nov., C. tenuitheca sp. nov., C. thermophila var. globulina var. nov., C. rubescens var. oocystiformis var. nov. Besides 18S rDNA and ITS2 sequences, we have newly sequenced the tufA gene marker for this taxon. Phylogenetic analysis combined with morphological studies revealed four morphotypes within the Coelastrella sensu lato clade, which contained the morphotype of original Coelastrella, original Scotiellopsis, Asterarcys, and morphotype of C. vacuolata and C. tenuitheca sp. nov. The relationships between morphological differences and phylogenic diversity based on different markers were discussed. Our results support that 18S rDNA was too conserved to be used a species‐specific or even a genus‐specific marker in this clade. The topology of tufA gene‐based phylogenetic tree had a better match with the morphological findings. [ABSTRACT FROM AUTHOR]

The colonial volvocine algae span the full range of organizational complexity, from four-celled species to multicellular species, and this group of algae is often used for the study of evolution. In recent years, many organelle genomes have been sequenced using the application of next generation sequencing technology; however, only a few organelle genomes have been reported in colonial volvocine algae. In this study, we determined the organelle genomes of Eudorina elegans and Eudorina cylindrica and analysed the organelle genome size, structure and gene content between these volvocine species. This provided useful information to help us understand the composition of colonial volvocine organelle genomes. Based on the chloroplast genome protein-coding genes, we conducted a phylogenomics analysis of the volvocine algae. The result revealed an unexpected phylogenetic relationship, namely, E. elegans is more closely related to Pleodorina starrii than to E. cylindrica. The substitution rate of volvocine algae was then calculated based on organelle genome protein-coding genes; our analysis suggested the possibility that the two Eudorina species may be under similar evolutionary pressure. Lastly, the synteny analysis of the mitochondrial genome showed that gene arrangements and contents are highly conserved in the family Volvocaceae, and the synteny analysis of the chloroplast genome indicated that the genus Eudorina may have experienced genomic changes. [ABSTRACT FROM AUTHOR]

Many species belonging to the coccoid green algae genus Coelastrella are considered potential candidates for the large-scale production of natural pigments and biofuels. However, little is known about the structural, functional and molecular aspects of the chloroplast genomes (cpDNAs) of this genus. In the present study, the complete sequence of the cpDNA of strain FACHB-2138, which was further identified as Coelastrella saipanensis Hanagata based on morphological and molecular analyses, was elucidated. The 196 140 bp cpDNA sequence that was assembled as a circular map was found to possess the typical quadripartite structure. The two identical copies of 11 897 bp inverted repeat (IR) sequences were separated from one another by single copy regions. The large single copy region (LSC) was 104 949 bp, whereas the small single copy region (SSC) was 67 397 bp. The cpDNA encoded a total of 96 unique genes, which included 67 protein-coding genes, three rRNA genes and 26 tRNA genes. A total of 19 group I introns were annotated in this genome. Comparative analyses with three species from the family Scenedesmaceae showed C. saipanensis had a slightly expanded genome, higher GC content and less skewed distribution of its genes between the two DNA strands than that of the other three species. The cpDNA data deduced from the present study helps to expand our present understanding of plant systematics and phylogenetic reconstruction, and identify the possible biotechnological applications of the species belonging to the studied taxa. [ABSTRACT FROM AUTHOR]

Abstract: The Parachlorella clade was put forward as a group within the family Chlorellaceae in 2004. Recent molecular analyses have revealed that Dictyosphaerium morphotype algae form several independent lineages within the Parachlorella clade, and new genera and species have been established. In this study, we focus on the diversity of Dictyosphaerium morphotype algae within the Parachlorella clade, based on 42 strains from China. We used combined analyses of morphology and molecular data based on SSU and internal transcribed spacer region (ITS) rDNA sequences to characterize these algae. In addition, the secondary structure of ITS2 was compared to delineate new lineages. Our results revealed high phylogenic diversity of Dictyosphaerium morphotype algae, and we describe five distinct lineages. We examined the morphological features of these five lineages, and morphological differences are difficult to find compared with other Dictyosphaerium morphotype algae. The five distinct lineages were not described as new genera currently. We lastly discuss the taxonomic problems regarding the Dictyosphaerium morphotype within the Parachlorella clade, and possible solutions are considered. [ABSTRACT FROM AUTHOR]

Backgroud: Foliicolous algae are a common occurrence in tropical forests. They are referable to a few simple morphotypes (unicellular, sarcinoid-like or filamentous), which makes their morphology of limited usefulness for taxonomic studies and species diversity assessments. The relationship between algal community and their host phyllosphere was not clear. In order to obtain a more accurate assessment, we used single molecule real-time sequencing of the 18S rDNA gene to characterize the eukaryotic algal community in an area of South-western China. Result: We annotated 2922 OTUs belonging to five classes, Ulvophyceae, Trebouxiophyceae, Chlorophyceae, Dinophyceae and Eustigmatophyceae. Novel clades formed by large numbers sequences of green algae were detected in the order Trentepohliales (Ulvophyceae) and the Watanabea clade (Trebouxiophyceae), suggesting that these foliicolous communities may be substantially more diverse than so far appreciated and require further research. Species in Trentepohliales, Watanabea clade and Apatococcus clade were detected as the core members in the phyllosphere community studied. Communities from different host trees and sampling sites were not significantly different in terms of OTUs composition. However, the communities of Musa and Ravenala differed from other host plants significantly at the genus level, since they were dominated by Trebouxiophycean epiphytes. Conclusion: The cryptic diversity of eukaryotic algae especially Chlorophytes in tropical phyllosphere is very high. The community structure at species-level has no significant relationship either with host phyllosphere or locations. The core algal community in tropical phyllopshere is consisted of members from Trentepohliales, Watanabea clade and Apatococcus clade. Our study provided a large amount of novel 18S rDNA sequences that will be useful to unravel the cryptic diversity of phyllosphere eukaryotic algae and for comparisons with similar future studies on this type of communities. [ABSTRACT FROM AUTHOR]

Abstract: The Oocystaceae family is generally considered to contain common freshwater eukaryotic microalgae, and few are reported living in semi‐saline habitats. Our latest ecological survey in Qinghai Lake and Angzicuo Lake, both large, closed, high‐altitude, semi‐saline lakes located on the Qinghai‐Tibet plateau in China, revealed Oocystaceae species as a dominant group among plankton. Since limited knowledge exists about semi‐saline species in the Oocystaceae family, a taxonomical study was carried out using morphological and phylogenetic methods. Using this approach, four new strains of Oocystaceae were identified and successfully cultured in the lab. Molecular results correlated with morphological characters and resolved these species into at least three genera. A new genus, Euchlorocystis, with type species Euchlorocystis subsalina, is described here as having the distinctive morphology of multiple pyrenoids per chloroplast among Oocystaceae, and an independent phylogenetic position at the base of the Oocystaceae. Similarly, the genus Densicystis, with type species Densicystis glomerata, is newly proposed here as having a unique colony morphology of dozens or hundreds of little cells tightly embedded in ellipsoid to round mucilage masses. Oocystis marina, originally described from the Baltic Sea, was also identified in Qinghai Lake and Angzicuo Lake and phylogenetically positioned in the semi‐saline clade of the Oocystaceae. The result that a marine species was detected in the closed inland lakes implies a further need to reevaluate the origins of these species. [ABSTRACT FROM AUTHOR]

Phylogenetic relationships within the green algal phylum Chlorophyta have proven difficult to resolve. The core Chlorophyta include Chlorophyceae, Ulvophyceae, Trebouxiophyceae, Pedinophyceae and Chlorodendrophyceae, but the relationships among these classes remain unresolved and the monophyly of Ulvophyceae and Trebouxiophyceae are highly controversial. We analyzed a dataset of 101 green algal species and 73 protein-coding genes sampled from complete and partial chloroplast genomes, including six newly sequenced ulvophyte genomes ( Blidingia minima NIES-1837, Ulothrix zonata , Halochlorococcum sp. NIES-1838, Scotinosphaera sp. NIES-154, Caulerpa brownii and Cephaleuros sp. HZ-2017). We applied the Tree Certainty (TC) score to quantify the level of incongruence between phylogenetic trees in chloroplast genomic datasets, and show that the conflicting phylogenetic trees of core Chlorophyta stem from the most GC-heterogeneous sites. With removing the most GC-heterogeneous sites, our chloroplast phylogenomic analyses using heterogeneous models consistently support monophyly of the Chlorophyceae and of the Trebouxiophyceae, but the Ulvophyceae was resolved as polyphyletic. Our analytical framework provides an efficient approach to reconstruct the optimal phylogenetic relationships by minimizing conflicting signals. [ABSTRACT FROM AUTHOR]