Identification of Photobionts from the lichen family Physciaceae using algal-specific ITS rDNA sequencing

2001 ◽  
Vol 33 (1) ◽  
pp. 73-86 ◽  
Author(s):  
Gert Helms ◽  
Thomas Friedl ◽  
Gerhard Rambold ◽  
Helmut Mayrhofer

AbstractThe identity of photobionts from 20 species of the Physciaceae from different habitats and geographical regions has been determined by ITS rDNA sequence comparisons in order to estimate the diversity of photobionts within that lichen group, to detect patterns of specificity of mycobionts towards their photobionts and as a part of an ongoing study to investigate possible parallel cladogenesis of both symbionts. Algal-specific PCR primers have been used to determine the ITS rDNA sequences from DNA extractions of dried lichens that were up to 5 years old. Direct comparisons and phylogenetic analyses allowed the assignment of Physciaceae photobionts to four distinct clades in the photobiont ITS rDNA phylogeny. The results indicate a diversity within the genus Trebouxia Puymaly and Physciaceae photobionts that is higher than expected on the basis morphology alone. Physciaceae photobionts belonged to 12 different ITS lineages of which nine could unambiguously be assigned to six morphospecies of Trebouxia. The identity of the remaining three sequences was not clarified; they may represent new species. Specificity at the generic level was low as a whole range of photobiont species were found within a genus of Physciaceae and different ranges were detected. The photobionts of Physcia (Schreb.) Michaux were closely related and represented one morphospecies of Trebouxia, whereas the algal partners of Buellia De Not and Rinodina (Ach.) Gray were in distant lineages of the ITS phylogeny and from several Trebouxia morphospecies. Photobiont variation within a genus of Physciaceae may be due to phylogeny, geographical distance or because photobionts from neighbouring lichens were taken (“algal sharing“). At the species level Physciaceae mycobionts seem to be rather selective and contained photobionts that were very closely related within one morphospecies of Trebouxia.

Author(s):  
Filip Pniewski ◽  
Thomas Friedl ◽  
Adam Latała

Identification of diatom isolates from the Gulf of Gdańsk: testing of species identifications using morphology, 18S rDNA sequencing and DNA barcodes of strains from the Culture Collection of Baltic Algae (CCBA)Eighteen strains of diatoms isolated from different phytoplankton and microphytobenthic assemblages in the Gulf of Gdańsk (Baltic Sea, Poland) and maintained at the Culture Collection of Baltic Algae (CCBA) were investigated for their species identifications. The latter were tested by phylogenetic analyses of nearly full 18S rDNA sequences as well as through sequence comparisons of 5.8S+ITS2 rDNA fragments for use as DNA barcode. The planktonic species were readily identified as a member of the


Nematology ◽  
2003 ◽  
Vol 5 (1) ◽  
pp. 99-111 ◽  
Author(s):  
Zahra Tanha Maafi ◽  
Sergei Subbotin ◽  
Maurice Moens

Abstract RFLP and sequences of ITS-rDNA of 45 populations of cyst-forming nematodes collected from different parts of Iran were analysed and identified as representatives of 21 species. Eight enzymes generated RFLP for all studied populations. Comparison of RFLP profiles and sequences of the ITS regions with published data confirmed the presence of Heterodera avenae, H. filipjevi, H. glycines, H. hordecalis, H. latipons, H. schachtii and H. trifolii in Iran. RFLP patterns and ITS sequences for H. elachista, H. turcomanica, H. mothi and C. cacti were obtained for the first time in this study. Heterodera humuli, H. goettingiana, H. fici, H. elachista, H. turcomanica and Cactodera cacti are recorded for the first time in Iran. These results correspond with morphological and morphometric identification of the populations. Several populations were not identified at the species level and are attributed to Heterodera sp.; some of these may correspond to new species. Twenty-one new sequences from Iranian cyst-forming nematodes and 36 known sequences were used for the phylogenetic analyses. The cyst-forming nematodes formed several clades corresponding to their morphological features. Heterodera mothi and H. elachista clustered with high support with other Cyperi group species and H. turcomanica formed a moderately to highly supported clade with the Humuli group.


2006 ◽  
Vol 41 (10) ◽  
pp. 1507-1516 ◽  
Author(s):  
Érico Leandro da Silveira ◽  
Rodrigo Matheus Pereira ◽  
Denilson César Scaquitto ◽  
Eliamar Aparecida Nascimbém Pedrinho ◽  
Silvana Pómpeia Val-Moraes ◽  
...  

Studies on the impact of Eucalyptus spp. on Brazilian soils have focused on soil chemical properties and isolating interesting microbial organisms. Few studies have focused on microbial diversity and ecology in Brazil due to limited coverage of traditional cultivation and isolation methods. Molecular microbial ecology methods based on PCR amplified 16S rDNA have enriched the knowledge of soils microbial biodiversity. The objective of this work was to compare and estimate the bacterial diversity of sympatric communities within soils from two areas, a native forest (NFA) and an eucalyptus arboretum (EAA). PCR primers, whose target soil metagenomic 16S rDNA were used to amplify soil DNA, were cloned using pGEM-T and sequenced to determine bacterial diversity. From the NFA soil 134 clones were analyzed, while 116 clones were analyzed from the EAA soil samples. The sequences were compared with those online at the GenBank. Phylogenetic analyses revealed differences between the soil types and high diversity in both communities. Soil from the Eucalyptus spp. arboretum was found to have a greater bacterial diversity than the soil investigated from the native forest area.


2014 ◽  
Vol 90 (1) ◽  
pp. 1-6 ◽  
Author(s):  
F. Li ◽  
T. Hu ◽  
N.C. Duan ◽  
W.Y. Li ◽  
Q. Teng ◽  
...  

AbstractThe present study examined sequence variability in two mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1), and internal transcribed spacer (ITS) of nuclear ribosomal DNA (rDNA) among Oesophagostomum asperum isolates from goats in Hunan Province, China. A portion of the cox1 (pcox1), nad1 (pnad1) genes and the ITS (ITS1+5.8S rDNA+ITS2) rDNA were amplified by polymerase chain reaction (PCR) separately from adult O. asperum individuals and the representative amplicons were subjected to sequencing from both directions. The lengths of pcox1, pnad1 and ITS rDNA were 366 bp, 681 bp and 785 bp, respectively. The A+T contents of gene sequences were 71.5–72% for pcox1, 73.7–74.2% for pnad1 and 58–58.8% for ITS rDNA. Intra-specific sequence variations within O. asperum were 0–1.6% for pcox1, 0–1.9% for pnad1 and 0–1.7% for ITS rDNA, while inter-specific sequence differences among members of the genus Oesophagostomum were significantly higher, being 11.1–12.5%, 13.3–17.7% and 8.5–18.6% for pcox1, pnad1 and ITS rDNA, respectively. Phylogenetic analyses using combined sequences of pcox1 and pnad1, with three different computational algorithms (Bayesian inference, maximum likelihood and maximum parsimony), revealed distinct groups with high statistical support. These findings demonstrated the existence of intra-specific variation in mtDNA and rDNA sequences among O. asperum isolates from goats in Hunan Province, China, and have implications for studying molecular epidemiology and population genetics of O. asperum.


2016 ◽  
Vol 48 (3) ◽  
pp. 161-169 ◽  
Author(s):  
Beata GUZOW-KRZEMIŃSKA ◽  
Paweł CZARNOTA ◽  
Anna ŁUBEK ◽  
Martin KUKWA

AbstractMicarea soralifera sp. nov., a new sorediate species belonging to the M. prasina group, is characterized by soralia developing directly from the endoxylic thallus or small external areoles, as well as the presence of micareic acid. Phylogenetic analyses of mtSSU rDNA sequences have shown that its closest relative is M. subviridescens. ITS rDNA sequence, a marker proposed as the universal barcoding region of fungi, was generated from the holotype.


Author(s):  
Niklas Reichelt ◽  
Dorian Leger ◽  
Mareike Daubert ◽  
Paolo Ruffino ◽  
Thomas Pröschold ◽  
...  

AbstractChromochloris zofingiensis (Dönz) Fucíková & L.A.Lewis, due to its production of highly valuable carotenoids such as astaxanthin, is a model organism in biotechnology. Since the recognition of this physiological property, many biotechnological applications have only used a single strain (SAG 211-14 = CCAP 211/14 = UTEX 32 = ATCC 30412) to produce biomass and carotenoids. However, multiple acquisitions of strains putatively belonging to the same species raised the question of the conspecificity of those strains and their properties. In this study, the conspecificity of the available strains, which are deposited axenically in SAG, was tested using SSU and ITS rDNA sequencing and AFLP (EcoRI/PstI) analyses. The comparison of SSU and ITS rDNA sequences as well as the AFLP patterns revealed that the investigated strains formed two very similar groups, (1) SAG 211-14, SAG 4.80, SAG 31.80, and SAG 34.80 and (2) SAG 221-2. All strains belonged to one species, C. zofingiensis, and represented one monophyletic lineage within the so-called DO-group of the Chlorophyceae. The robustness to cryopreservation and the subsequent epigenetic variability was detected using the methylation-sensitive AFLP (EcoRI/MspI and EcoRI/HpaII) among the five Chromochloris strains. All strains showed a high rate of survival (54.4–98.1%) during cryopreservation. The methylation patterns varied between precryo and postcryo in all strains detected among three time points (before, shortly after, and 8 weeks after cryopreservation), showing that the MS-AFLP technique has the potential to detect epigenetic effects occurring in response to cryopreservation and other stresses. Finally, the potential of these five strains for usage in biotechnological applications was proven by growing them in aerated cultures with and without additional carbon dioxide supply. The comparison showed that all strains produced high amounts of biomass and carotenoids under aeration with additional CO2 and were therefore suitable in biotechnology.


1998 ◽  
Vol 64 (11) ◽  
pp. 4333-4339 ◽  
Author(s):  
Daniel H. Buckley ◽  
Joseph R. Graber ◽  
Thomas M. Schmidt

ABSTRACT Within the last several years, molecular techniques have uncovered numerous 16S rRNA gene (rDNA) sequences which represent a unique and globally distributed lineage of the kingdom Crenarchaeotathat is phylogenetically distinct from currently characterized crenarchaeotal species. rDNA sequences of members of this novel crenarchaeotal group have been recovered from low- to moderate-temperature environments (−1.5 to 32°C), in contrast to the high-temperature environments (temperature, >80°C) required for growth of the currently recognized crenarchaeotal species. We determined the diversity and abundance of the nonthermophilic members of the Crenarchaeota in soil samples taken from cultivated and uncultivated fields located at the Kellogg Biological Station’s Long-Term Ecological Research site (Hickory Corners, Mich.). Clones were generated from 16S rDNA that was amplified by using broad-specificity archaeal PCR primers. Twelve crenarchaeotal sequences were identified, and the phylogenetic relationships between these sequences and previously described crenarchaeotal 16S rDNA sequences were determined. Phylogenetic analyses included nonthermophilic crenarchaeotal sequences found in public databases and revealed that the nonthermophilic Crenarchaeota group is composed of at least four distinct phylogenetic clusters. A 16S rRNA-targeted oligonucleotide probe specific for all known nonthermophilic crenarchaeotal sequences was designed and used to determine their abundance in soil samples. The nonthermophilicCrenarchaeota accounted for as much as 1.42% ± 0.42% of the 16S rRNA in the soils analyzed.


Phytotaxa ◽  
2019 ◽  
Vol 388 (1) ◽  
pp. 47 ◽  
Author(s):  
TATYANA DARIENKO ◽  
THOMAS PRÖSCHOLD

The genus Chlorella (in its traditional sense) is polyphyletic and belongs to at least twelve independent lineages of the Trebouxiophyceae and Chlorophyceae. Most of the aquatic species belong to the Chlorella and Parachlorella clades (within the so-called Chlorella-lineage of the Trebouxiophyceae), or to the genera Scenedesmus and Mychonastes (within the DO-group of the Chlorophyceae) according to phylogenetic analyses of the SSU and ITS rDNA sequences. In contrast to the aquatic species, the terrestrial strains investigated so far form a monophyletic lineage (Watanabea-clade) within the Trebouxia-lineage of the Trebouxiophyceae. Several genera with Chlorella-like morphology (Chloroidium, Heterochlorella, Watanabea, Kalinella, Viridiella and others) belong to the Watanabea clade. We studied 22 strains isolated from soil, bark, and artificial hard substrates, which have been traditionally identified as Chlorella luteoviridis or as unidentified Chlorella. To clarify the taxonomical status and intrageneric diversity of this group, we used an integrated approach (molecular phylogeny of SSU and ITS rDNA sequences, secondary structures, DNA barcoding, and morphology) including the ecological distribution. All investigated strains showed a low phenotypic plasticity, but a high genetic diversity, which could be only resolved in complex phylogenetic analyses based on the secondary structures of the investigated genes. Considering these results, we reestablished the genus Jaagichlorella for Heterochlorella and Heveochlorella, and proposed new combinations (J. luteoviridis, J. hainangensis, J. roystonensis, and J. sphaerica) as well as the new species, J. africana.


Plant Disease ◽  
2011 ◽  
Vol 95 (6) ◽  
pp. 633-639 ◽  
Author(s):  
Jason A. Smith ◽  
Kerry O'Donnell ◽  
Lacey L. Mount ◽  
Keumchul Shin ◽  
Kelly Peacock ◽  
...  

A canker disease of Florida torreya (Torreya taxifolia) has been implicated in the decline of this critically endangered species in its native range of northern Florida and southeastern Georgia. In surveys of eight Florida torreya sites, cankers were present on all dead trees and 71 to 100% of living trees, suggesting that a fungal pathogen might be the causal agent. To identify the causal agent, nuclear ribosomal internal transcribed spacer region (ITS rDNA) sequences were determined for 115 fungi isolated from cankers on 46 symptomatic trees sampled at three sites in northern Florida. BLASTn searches of the GenBank nucleotide database, using the ITS rDNA sequences as the query, indicated that a novel Fusarium species designated Fsp-1 might be the etiological agent. Molecular phylogenetic analyses of partial translation elongation factor 1-alpha (EF-1) and RNA polymerase second largest subunit (RPB2) gene sequences indicate that Fsp-1 represents a novel species representing one of the earliest divergences within the Gibberella clade of Fusarium. Results of pathogenicity experiments established that the four isolates of Fsp-1 tested could induce canker symptoms on cultivated Florida torreya in a growth chamber. Koch's postulates were completed by the recovery and identification of Fsp-1 from cankers of the inoculated plants.


2020 ◽  
Vol 94 ◽  
Author(s):  
R. Petkevičiūtė ◽  
G. Stanevičiūtė ◽  
V. Stunžėnas

Abstract Comparative analysis using complete ITS2 and partial 28S rDNA sequence data revealed that cercariaeum developing in rediae in Lithoglyphus naticoides represent two different lissorchiid species. One morphotype of cercariaeum is conspecific with adult Palaeorchis incognitus from European roach, Rutilus rutilus. The other cercariaeum is attributable to the genus Asymphylodora, but the species identity is not yet determined. We also generate the first rDNA sequences for Asymphylodora progenetica based on new collections from Bithynia tentaculata from Lithuania. Phylogenetic analyses of the newly generated sequences, together with information for other lissorchiids available on GenBank, showed that all representatives of Lissorchiidae form a strongly supported clade. Three monophyletic lineages, Asymphylodora, Palaeorchis and Lissorchis, were recognized at the generic level. Karyological analysis of the chromosome set of larval P. incognitus revealed a diploid number of 2n = 20. Its karyotype with subtelocentric chromosomes prevailing can be regarded as comparatively ‘primitive’, which is consistent with the basal position of P. incognitus in the 28S tree relative to the representatives of the genus Asymphylodora. The present study adds significant new information for establishing species-specific markers for the confident characterization of different developmental stages of lissorchiid species and clarification of their life cycles.


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