Chloroplast rbcL Research Articles

Global Diversity and Geographic Distributions of Padina Species (Dictyotales, Phaeophyceae): New Insights Based on Molecular and Morphological Analyses.

The taxonomic status and species diversity of the brown algal genus Padina (Dictyotales, Phaeophyceae) was assessed based on DNA sequences and the morpho-anatomy of specimens collected worldwide, especially from tropical and subtropical western Pacific regions. Phylogenetic analyses using chloroplast rbcL and mitochondrial cox3 gene sequences demonstrated four distinct clades for newly collected samples with high bootstrap support. Each species clade possesses a suite of morphological features that are not shared by any known species of Padina. These are P. imbricata sp. nov., Padina lutea sp. nov., P. moffittianoides sp. nov., and P. nitida sp. nov. The occurrence of these and other species of Padina clearly points to an elevated diversity of the genus in tropical/subtropical waters of the western Pacific. Phylogenetic analyses provided new insights into biogeographic characteristics of the genus, with many species in the Pacific Ocean showing shared/overlapping distributions, whereas species from the Mediterranean/Atlantic and/or the Indian Ocean tend to be confined to particular regions. Consideration has also been given to the evolutionary time frame of the genus Padina based on molecular time trees: a time tree of the concatenated data set (rbcL+cox3) revealed the estimated divergence time in the mid-Cretaceous, whereas those of cox3 and rbcL showed older estimates pointing to the periods of mid-Jurassic and Early Cretaceous, respectively.

Molecular characterization of endangered endemic plant Aloe pseudorubroviolacea using chloroplast matK and plastid rbcL gene

An endangered and rare species Aloe pseudorubroviolacea from the plant family Asphodelaceae which is presently recorded as endangered in Saudi Arabia collected from Al-Baha region of Saudi Arabia its GPS Latitude and Longitude coordinates 19.8345, 41.5481. The chloroplast matK and rbcL gene was considered in this study based on molecular identification the size is about 571 and 664 bp respectively. From the sequence analysis the gene matK and rbcL confirm that this species is very much closely related with A. rubroviolacea and also inter related with the species Astroloba rubriflora, Chrysopogon gryllus, Chortolirion angolense shows about 98.7% sequence homology. The partial matK and rbcL gene sequence discriminate Aloe pseudorubroviolacea from the closely related plant species, A. rubroviolacea. The gene sequence of rbcL discriminates the species from Chrysopogon gryllus and Chortolirion angolense, demonstrates the nucleotide variations in 3 different sites (623C/T; 653C/T; 700C/A). This study showed that matK and rbcL sequence region of chloroplast gene used to authenticate the samples of A. pseudorubroviolacea and which provide to help in correct identification and conservation process of this medicinally valuable endangered plant species.

Identification of morphologically similar species of Tribulus (Zygophyllaceae) by employing DNA barcodes and rRNA secondary structures

Abstract Tribulus terrestris is one of the essential constituents in traditional Indian systems of medicine and commonly used in several herbal preparations. It is adulterated with morphologically similar species, viz., T. subramanyamii, T. lanuginosus and T. alatus. T. terrestris is often misidentified for T. subramanyamii and T. lanuginosus. Hence, the error-free recognition and collection of this medicinal herb are important to ensure the drug’s effectiveness and biosafety. In the present study, DNA barcoding technique was employed to identify and to differentiate T. terrestris from its closely related species. Phylogenetic analysis was carried out for the three species of Tribulus using barcode candidates, such as, nuclear ribosomal DNA region ITS, chloroplast plastid gene psbA-trnH and rbcL. Sequence alignment revealed 7.5% polymorphic sites in ITS, with different specific regions showing the distribution of polymorphic sites as follows: ITS1 1.7%, ITS2 3.6% and the plastid intergenic spacer psbA-trnH 2.33%. The region ITS2 possesses a higher rate of transition/transversion ratio, percentage of variation and mean distance as compared to ITS1 and psbA-trnH. In addition, ITS2 has unique polymorphic loci which aid to differentiate T. terrestris from other species of Tribulus. Additionally we predicted the structure of ITS2 and 5.8S rRNA which distinguishes three phenotypically alike species based on the helical structure. The present study opens up new avenue for the identification of closely related species based on ITS2 region and 5.8S rRNA structure which could be employed for diversity studies

An Integrated Approach for Efficient and Accurate Medicinal Cuscutae Semen Identification.

To guarantee the safety and efficacy of herbal medicines, accurate identification and quality evaluation are crucial. The ripe dried seeds of Cuscuta australis R.Br. and C. chinensis Lam. are known as Cuscutae Semen (CS) and are widely consumed in Northeast Asia; however, the seeds of other species can be misidentified as CS owing to morphological similarities, leading to misuse. In this report, we propose a multilateral strategy combining microscopic techniques with statistical analysis and DNA barcoding using a genus-specific primer to facilitate the identification and authentication of CS. Morphology-based identification using microscopy revealed that the useful diagnostic characteristics included general shape, embryo exudation, hairiness, and testa ornamentation, which were used to develop an effective identification key. In addition, we conducted DNA barcoding-based identification to ensure accurate authentication. A novel DNA barcode primer was produced from the chloroplast rbcL gene by comparative analysis using Cuscuta chloroplast genome sequences, which allowed four Cuscuta species and adulterants to be discriminated completely. Therefore, this investigation overcame the limitations of universal DNA barcodes for Cuscuta species with high variability. We believe that this integrated approach will enable CS to be differentiated from other species, thereby improving its quality control and product safety in medicinal markets.

The macroalgal culture collection in Kobe University (KU-MACC) and a comprehensive molecular phylogeny of macroalgae based on the culture strains

ABSTRACT The Kobe University Macroalgal Culture Collection (KU-MACC), as a section of the Kobe University Research Center for Inland Seas, distributes cultures of macroalgae to the public under the umbrella of the National BioResource Project of Ministry of Education, Culture, Sports, Science and Technology and Japan Agency for Medical Research and Development, in collaboration with the Microbial Culture Collection of the National Institute of Environmental Studies. KU-MACC principally holds macroalgae (Phaeophyceae, Rhodophyta and Ulvophyceae), but also holds some important related taxa of Schizocladiophyceae and Charophyceae. They are unialgal strains maintained as vegetative thalli and by cryo-preservation. Stock cultures are grown in culture chambers under various temperature conditions in transparent polypropylene vials containing Provasoli’s Enriched Seawater (PES) or PESI medium. The collection includes the “genome strains” of Ectocarpus siliculosus, Chondrus crispus, Chara braunii and their related stains. Depending on the phylogenetic group, DNA sequences of the nuclear rDNA ITS region, chloroplast rbcL, and mitochondrial cox3 genes are determined and used for taxonomy and as genetic tags. Based on the genetic data, molecular phylogenies of Ulvophyceae, Phaeophyceae and Rhodophyta are presented. In the rbcL molecular phylogeny of Ulvophyceae, due to rather limited coverage of culture strains, only members of Ulothricales, Ulvales and Bryopsidales are represented by KU-MACC strains, whereas 10 orders are recognized in the class. The molecular phylogeny of Phaeophyceae based on the cox3 gene showed a tree topology similar to the published multigene molecular phylogeny except for the position of Ishige okamurae, although the bootstrap supports of most nodes were low: Ishigeales branched near the base of Phaeophyceae next to Discosporangiales in the multigene molecular phylogeny, but in the cox3 tree, it showed a long branch and was sister to Fucales. In the molecular phylogeny of Rhodophyta, 14 orders are represented by KU-MACC strains covering about half of currently recognized Bangiophycean and Florideophycean orders.

Integrative systematics approaches revealed that the rare red alga Schimmelmannia (Shimmelmanniaceae, Acrosymphytales) from Japan is a new species: The description of S. benzaiteniana sp. nov.

SUMMARYThe rare red alga Schimmelmannia (Schimmelmanniaceae, Acrosymphytales) from Japan was taxonomically re‐examined in this study. Japanese Schimmelmannia has previously been identified as S. plumosa (Setchell) I.A.Abbott. However, Japanese Schimmelmannia and S. plumosa have approximately 5 and 1.9% of genetic distance in mitochondrial cox1 and chloroplast rbcL sequences, respectively. They also differ morphologically in the numbers of periaxial cells and auxiliary cell branches indicating that they are distinct species. Based on molecular phylogenetics and morphology, S. benzaiteniana is distinguishable from other Schimmelmannia species, except for S. bollei Montagne which lacks a detailed description. Since S. bollei has been reported only from the North Atlantic coast of Africa, we assumed that it is distinct from the Japanese species. We, therefore, described the Japanese species as a new species, S. benzaiteniana.

Modifying Plant Photosynthesis and Growth via Simultaneous Chloroplast Transformation of Rubisco Large and Small Subunits.

Engineering improved Rubisco for the enhancement of photosynthesis is challenged by the alternate locations of the chloroplast rbcL gene and nuclear RbcS genes. Here we develop an RNAi-RbcS tobacco (Nicotiana tabacum) master-line, tobRrΔS, for producing homogenous plant Rubisco by rbcL-rbcS operon chloroplast transformation. Four genotypes encoding alternative rbcS genes and adjoining 5'-intergenic sequences revealed that Rubisco production was highest (50% of the wild type) in the lines incorporating a rbcS gene whose codon use and 5' untranslated-region matched rbcL Additional tobacco genotypes produced here incorporated differing potato (Solanum tuberosum) rbcL-rbcS operons that either encoded one of three mesophyll small subunits (pS1, pS2, and pS3) or the potato trichome pST-subunit. The pS3-subunit caused impairment of potato Rubisco production by ∼15% relative to the lines producing pS1, pS2, or pST However, the βA-βB loop Asn-55-His and Lys-57-Ser substitutions in the pS3-subunit improved carboxylation rates by 13% and carboxylation efficiency (CE) by 17%, relative to potato Rubisco incorporating pS1 or pS2-subunits. Tobacco photosynthesis and growth were most impaired in lines producing potato Rubisco incorporating the pST-subunit, which reduced CE and CO2/O2 specificity 40% and 15%, respectively. Returning the rbcS gene to the plant plastome provides an effective bioengineering chassis for introduction and evaluation of novel homogeneous Rubisco complexes in a whole plant context.

Using the chloroplast rbcL gene to clarify the relationship between species of the genus <i>Stephania</i> (Menispermaceae) from Vietnam

The rbcL gene of the chloroplast genome is widely used as an additional data for the study of species origin, molecular evolution and phylogeny. In this study, we used the rbcL gene to separate three species of genus Stephania: S. japonica, S. polygona, S. rotunda and one subspecies S. japonica var. discolor from Vietnam. Molecular analysis was performed on 523 bp segment of the rbcL genes with 4 examined samples of the genus Stephania and 18 other genbank sequences of five genera Pachygone, Antizoma, Cissampelos, Cyclea and Syntriandrium. The dataset consists of 22 sequences used to reconstruct the evolutionary tree using two methods: Bayesian Infer (BI) and Maximum Likelihood (MP). The results showed that S. rotunda was able to distinguish from S. japonica or S. polygona, while S. japonica, S. japonica var. discolor and S. polygona could not distinguished each another. The phylogenetic tree splited three examined species into two groups, representing the two main groups of morphology in genus Stephania: a group with tuberous rootstock and another group with main root.

Morphology and molecular phylogeny of Umbraulva spp. (Ulvales, Ulvophyceae), and proposal of Ryuguphycus gen. nov. and R. kuaweuweu comb. nov.

ABSTRACT The reproductive structures, early development and carotenoid composition of Umbraulva kuaweuweu, which is a deep-water ulvacean species recently reported from Hawaii, are reported for the first time using the specimens collected by dredging at ~30 m depth from Ogasawara Islands and Mageshima in southern Japan. Although Umbraulva kuaweuweu had siphonaxanthin as in U. japonica, a characteristic carotenoid of Umbraulva, U. kuaweuweu lacked α-carotene and β-carotene. U. kuaweuweu differed markedly from U. japonica in thallus morphology and reproductive structures: cell shape, presence/absence of inner rhizoidal filaments in the upper parts of thalli, solid/partly saccate thalli, and developmental processes of zoids. Multigene concatenated DNA sequences of chloroplast rbcL and tufA genes, and nuclear 18S, 5.8S, 28S rDNA and rDNA ITS1 and ITS2 regions indicated separation of U. kuaweuweu from the clade comprised of the generitype U. japonica, U. amamiensis, U. dangeardii and U. kaloakulau. In conclusion, based on the distinctive morphology, life history, carotenoid composition and molecular phylogeny, we propose the establishment of a new genus Ryuguphycus gen. nov. to accommodate U. kuaweuweu, as R. kuaweuweu comb. nov.

Morphological and Molecular Diagnostic Characters Reveal a New Species of Pachyphytum (Crassulaceae)

Pachyphytum viscidum (Crassulaceae: Saxifragales) is described as a new species to science. The specimens analyzed come from the Sierra de Santa Barbara, Guanajuato, Mexico. Some morphological diagnostic characters for the new species are: sticky stems and leaves; dark green leaves; absence of wax on all their organs; and pale pink flowers. The new species is placed in the section Diotostemon of the genus, with possible proximity to P. brevifolium and P. hookeri. DNA sequence analysis from the matK and rbcL chloroplast regions, as well as ITS2 nuclear loci, revealed the presence of molecular diagnostic sites which, in addition to the aforementioned morphological character states, further support the taxonomic description as well as future integrative taxonomy studies in Mexican Crassulaceae.

Species delimitation of Planosiphon gracilis morphospecies (Scytosiphonaceae, Phaeophyceae) from Japan and the description of Pl. nakamurae sp. nov.

ABSTRACTMolecular phylogenetic analyses based on mitochondrial cox1 and cox3 and chloroplast rbcL gene sequences revealed that the morphospecies of the brown alga Planosiphon gracilis (Scytosiphonaceae, Ectocarpales) from Japan is a complex of three distinct lineages (lineages I–III). Specimens from these lineages had the typical morphology of Pl. gracilis (i.e. flattened, linear, hollow, and gregarious erect thalli); thus, morphological discrimination among them was difficult in many cases. We observed large genetic divergences among the three lineages (c. 10% in cox1 and cox3). Moreover, we observed a reproductive isolating barrier between lineages I and II, despite the wide overlap in their geographic distribution. Our data, therefore, suggest that these three lineages are distinct species. Since lineage I included the holotype specimen of Pl. gracilis, this lineage was attributed to Pl. gracilis. Lineage III was phylogenetically close to Pl. complanatus from Canada with a 4.1% divergence in cox1. For lineage II, a new species name, Pl. nakamurae sp. nov., was proposed. This new species was also found in Argentina.

Genomic analysis of chloroplast matK and rbcL gene from Flacourtia inermis Roxb for plant DNA barcoding

Genomic analysis of chloroplast matK and rbcL gene from Flacourtia inermis Roxb for plant DNA barcoding

Genomic analysis of chloroplast matK and rbcL gene from Flacourtia inermis Roxb for plant DNA barcoding

Identification of medicinally significant plant species is helpful for their utilization as medicine. Flacourtia inermis Roxb are important medicinal plant. Aim: To Genotypic characterization of Flacourtia inermis through DNA sequence of matK and rbcL gene. Flacourtia inermis Roxb DNA was isolated using NucleoSpin® Plant II Kit (Macherey-Nagel gene extraction) Kit. Isolated DNA was amplified by using PCR (matK and rbcL gene primers). Amplified matK and rbcL gene was sequenced by ABI 3500 DNA analyzer and the sequence quality was checked using sequence scanner software v1. The results exposed basic variability of the sequences. The matK and rcbL gene phylogenic tree was constructed using MEGA 6 software. Flacourtia inermis Roxb DNA was successfully isolated using NucleoSpin® Plant II Kit (Macherey-Nagel gene extraction) Kit and confirmed by agarose gel electrophoresis. Isolated matK and rbcL genomic DNA was successfully amplified by using PCR. From the matK sequence, BLASTn analysis result Flacourtia inermis species showed 99.20% relationship with Flacourtia jangomas and rbcL BLASTn analysis result of Flacourtia inermis species showed100% relationship with Flacourtia jangomas at NCBI database. From this study, the result concluded the matK and rcbL genomes can be used for DNA identification of Flacourtia inermis. The information obtained from the study would be helpful for further research involving genotypic identification of Flacourtia sp.

Reinstatement of Myelophycus caespitosus (Scytosiphonaceae, Phaeophyceae) from Japan

SUMMARYReinstatement of Myelophycus caespitosus Kjellman (Ectocarpales s.l., Phaeophyceae) described from western Japan is proposed based on the comparisons of DNA sequences of Japanese Myelophycus specimens, the type specimen of Chordaria simplex and lectotypified specimen of Myelophycus caespitosus. In the genetic analyses using mitochondrial cox1 and cox3, chloroplast atpB, psbA and rbcL DNA sequences, the specimens morphologically referable to M. simplex formed two distinct clades (clade‐1 and clade‐2) supported by high statistical values. Clade‐1 was distributed on the western coast of Japan (Honshu, Shikoku and Kyushu) from the Kii Peninsula to Tsushima and the Pacific coast of Aomori, northeastern Honshu, and clade‐2 on the Pacific coast of central Honshu from Sanriku to the Kii Peninsula. Based on the cox3 DNA sequences and the location of the type locality at Shimoda, Izu Peninsula, clade‐2 was concluded to correspond to true M. simplex. There were no recognizable differences in the representative morphological features (height and diameter of sporophytes and gametophytes, number of cells comprising cortical, subcortical and medullary layers, thickness of plurilocular gametangia, or length and diameter of unilocular zoidangia) between the specimens included in the two taxa. Analyses of the specimens of the two taxa, including sympatric populations on the Kii Peninsula, using a nuclear genetic marker ocm3 did not suggest any genetic exchanges between the two taxa. On the other hand, cox3 gene sequence of the voucher specimen of Myelophycus caespitosus collected from Goto, Kyushu, western Japan housed in the UPS herbarium was included in clade‐1. In conclusion, we propose the reinstatement of M. caespitosus and to lectotypify the specimen in UPS. Korean Myelophycus specimens reported from Cheju Island and Wando were considered to belong to M. caespitosus based on the reported rbcL sequences.

New morphological and DNA evidence supports the existence of Calligonum jeminaicum Z. M. Mao (Calligoneae, Polygonaceae) in China.

Calligonum jeminaicum Z. M. Mao, a species regarded as endemic to China, was thought to be nonexistent owing to a lack of scientific records. The similarity of C. jeminaicum to C. mongolicum Turcz. warranted an investigation into the taxonomical relationship between these species. In this study, a naturally occurring population of C. jeminaicum was discovered and the taxonomical relationships of this species with C. mongolicum were resolved. Morphological traits, including fruit and flower characteristics, as well as nuclear (ETS, ITS) and chloroplast (psbA-trnH, ycf6-psbM, rpl32-trnL, rbcL, and trnL-F) DNA sequence data were studied to confirm the taxonomic status of C. jeminaicum. The nrDNA data (ITS1-2 and ETS) from C. jeminaicum reflected variability from the whole C. mongolicum complex, showing distinctive haplotypes in the Calligonum sect. Medusa Sosk. & Alexandr. The cpDNA data supplied similar evidence, showing unique branching in Bayesian and ML tree analyses. The specific status of C. jeminaicum is confirmed based on both morphological and molecular analyses. Here we present a revised description of C. jeminaicum along with its DNA barcode and discuss suggestions for the conservation of this species. Based on current evidence, this species was evaluated as Critically Endangered (CR) according to the IUCN criteria.

Dictyota falklandica sp. nov. (Dictyotales, Phaeophyceae) from the Falkland Islands and southernmost South America

ABSTRACTSurveys of the seaweed flora of the Falkland Islands and of Tierra del Fuego revealed the presence of a new species of brown alga. Dictyota falklandica sp. nov. inhabits the shallow rocky infralittoral in sheltered localities and the lower intertidal in more exposed sites. Dictyota falklandica has a regular to irregular habit of dichotomously branched blades, forming erect thalli composed of a single-layered cortex and medulla, with margins in the apical parts dotted with dormant apical cells. Sporangia occur in irregular groups or longitudinal lines on the thallus surface. Molecular phylogenies based on chloroplast psbA and rbcL and mitochondrial cox1 sequences showed that the species from the Falkland Islands is sister to a clade formed by D. korowai, recently described from New Zealand and D. kunthii known from both the Pacific coast of South America and New Zealand. Temperature tolerance experiments, showing mortality at 25 °C but survival at 20 °C, confirm the cold-temperate affinity of this taxon. Its relationship to other cold-temperate Southern Hemisphere species is discussed, with its closest relatives living in regions with sea surface temperatures of at least 7–10 °C higher.

Dual influence of terrestrial and marine historical processes on the phylogeography of the Brazilian intertidal red alga Gracilaria caudata

In this study, we explored how past terrestrial and marine climate changes have interacted to shape the phylogeographic patterns of the intertidal red seaweed Gracilaria caudata, an economically important species exploited for agar production in the Brazilian north-east. Seven sites were sampled along the north-east tropical and south-east sub-tropical Brazilian coast. The genetic diversity and structure of G.caudata was inferred using a combination of mitochondrial (COI and cox2-3), chloroplast (rbcL) and 15 nuclear microsatellite markers. A remarkable congruence between nuclear, mitochondrial and chloroplast data revealed clear separation between the north-east (from 03°S to 08°S) and the south-east (from 20°S to 23°S) coast of Brazil. These two clades differ in their demographic histories, with signatures of recent demographic expansions in the north-east and divergent populations in the south-east, suggesting the maintenance of several refugia during the last glacial maximum due to sea-level rise and fall. The Bahia region (around 12°S) occupies an intermediate position between both clades. Microsatellites and mtDNA markers showed additional levels of genetic structure within each sampled site located south of Bahia. The separation between the two main groups in G.caudata is likely recent, probably occurring during the Quaternary glacial cycles. The genetic breaks are concordant with (i) those separating terrestrial refugia, (ii) major river outflows and (iii) frontiers between tropical and subtropical regions. Taken together with previously published eco-physiological studies that showed differences in the physiological performance of the strains from distinct locations, these results suggest that the divergent clades in G.caudata correspond to distinct ecotypes in the process of incipient speciation and thus should be considered for the management policy of this commercially important species.

Comparative phylogenetics of some species of Ophioglossum L. (Ophioglossaceae: Pteridophyta) in India with comments on evolutionary significance of high palaeoploidy and rare morphological traits

The genomic constitution of the genus Ophioglossum is complicated because of its being geologically ancient and of long evolutionary history. A lot of plants of the same and different species resemble each other as well as show significant differences among morphological characters among themselves. Many of the new species described have exhibited several unknown features not observed in any species of the genus so far. In this paper, we wish to emphasize that mere observations on shape and size of trophophyll (leaf) and rhizome, should not be taken reliably in delimiting different species of the genus Ophioglossum because of several variations already reported for many species. Preference should be given to exine ornamentation of spores as emphasized by the present study. Detailed observations on exine ornamentation of spores under SEM have offered precious and distinctive features; semi-tuberculate, beaded strings, and stripped exines are a few forms of spore ornamentations offering clear distinction among species. Long chains of circulate-alete (round alete) spores, sometimes linear tetrads as described hereunder in Ophioglossum aletum are novel features exclusively confined to this species. Comparative chromosome profiles of many species described earlier do not offer any significant support to the identity except in O. eliminatum, which has the lowest chromosome count (n = 90). This appears that since almost all species possess intraspecific polyploidy, the intragenomic variability must be loaded with interspecific hybridization contents including chromosomal aberrations. Quite possibly, this may be one of the reasons that homologous chromosomes rarely form trivalents and multivalents during early meiotic stages, despite the fact that palaeoautoploidy must have resulted in copying homologous chromosomes. In recent years, repeated phylogenetic analysis of chloroplast rbcL have revealed that many species in India appear to be offshoots of a vulgatum clade which are likely to be the hybridization products involving O. costatum, O. vulgatum, O. reticulatum, and also including of other interspecific hybridizations. It is remarkable to mention that some of the new taxa published by us, viz. O. eliminatum, O. indicum, O. aletum and O. chalonerii (unpublished), explicitly show distinct phylogenetic relationship maintaining independent genomic identity. Furthermore, each new species shows some rare uniquely confined trait not possessed by any other species of the genus. These morphological traits also suggest that the plants may be expressing extremely rare genic-combinations which might have been brought forth due to autopolyploidy and/or allopolyploidy (hybridizations) during the past few decades. Obviously, we will need a multidisciplinary approach (morphological, anatomical, biochemical) in order to identify different species of Ophioglossum with new and unusual genomic identity and also to understand possible evolutionary mechanisms undergoing within the genome.

Haslea nusantara (Bacillariophyceae), a new blue diatom from the Java Sea, Indonesia: morphology, biometry and molecular characterization

Background and aims – The present study aims to describe a new species of pennate blue diatom from the genus Haslea, H. nusantara sp. nov., collected from Semak Daun Island, the Seribu Archipelago, in Indonesian marine waters.
 Methods – Assessment for species identification was conducted using light microscopy, Scanning Electron Microscopy and molecular techniques. The morphological characteristics of H. nusantara have been described, illustrated and compared to other morphologically similar blue Haslea taxa, distributed worldwide. Additionally, molecular characterization was achieved by sequencing plastidial and mitochondrial genomes.
 Key results – This new species, named Haslea nusantara, cannot be discriminated by its morphology (stria density) but it is characterized by its gene sequences (rbcL chloroplast gene and cox1 mitochondrial gene). Moreover, it differentiates from other blue Haslea species by the presence of a thin central bar, which has been previously reported in non-blue species like H. pseudostrearia. The complete mitochondrion (36,288 basepairs, bp) and plastid (120,448 bp) genomes of H. nusantara were sequenced and the gene arrangements were compared with other diatom genomes. Phylogeny analyses established using rbcL indicated that H. nusantara is included in the blue Haslea cluster and close to a blue Haslea sp. found in Canary Islands (H. silbo sp. ined.).
 Conclusions – All investigations carried out in this study show that H. nusantara is a new blue-pigmented species, which belongs to the blue Haslea clade, with an exceptional geographic distribution in the Southern Hemisphere.

Analysis of a New Strain of Pseudomuriella engadinensis (Sphaeropleales, Chlorophyta) for Possible Use in Biotechnology

The species diversity of green coccoid algae is often difficult to study using light microscopy due to their complex morphology. Algae, which have morphological and ecological similarities with the genera Bracteacoccus Tereg., Dictyococcus Gerneck, and Pseudomuriella Hanagata, are one of the most taxonomically complex examples. Often, only a comparison of nucleotide sequences allows one to carry out a reliable identification and find out the phylogenetic identity of the studied objects. Pseudomuriella belongs to the group of genetically diverse genera with a homogeneous Bracteacoccus-like morphology. The combined use of two genetic markers (18S rDNA nuclear gene and chloroplast rbcL gene), along with light microscopy and comparison of fatty acid profiles, allowed the identification of the green algae strain isolated from the soil of the artificial pine and robinia plantation in the steppe zone of Ukraine as Pseudomuriella engadinensis (Kol et Chodat) Fucikova, Rada et Lewis—the first find of the species in the flora of algae in Ukraine. Analysis of the fatty acid composition of the studied strain’s cells showed that the total lipid content in the stationary growth phase was at the level of 87.9 ± 2.1 mg/g dry cell mass, and the main fatty acids were palmitic, hexadecadienoic, ruganic, oleic, linoleic, and α-linolenic—their share accounted for 82.4% of the total amount of fatty acids.