Nucleotide differences in the mbf1 gene of the lichenized fungus Umbilicaria decussata collected in polar and non-polar regions

Multiprotein bridging factor 1 (MBF1) is a transcriptional co-activator related to stress tolerance in various organisms. We investigated the nucleotide differences in the mbf1 gene in the lichen-forming fungus Umbilicaria decussata collected from polar (i.e., Antarctica and the Arctic) and non-polar (i.e., Armenia) regions. The 552-bp Udmbf1 genes isolated from eight samples contained numerous sequence variations, including single nucleotide polymorphisms as well as insertions and deletions. The frequency of nucleotide changes was higher in the intron than in the coding sequence. The nucleotide polymorphism levels (π=0.01792, θ=0.01792) and haplotype diversity (Hd=1) in the Udmbf1 gene from Antarctic samples were relatively high. Additionally, of the 19 detected nucleotide sequence variation sites, 15 were observed only in Antarctic samples. The resulting amino acid changes occurred in the N-terminal, whose function remains unknown. Although these DNA polymorphisms and amino acid changes have been verified in Antarctic samples of U. decussata, there is still little evidence indicating that different environmental conditions affected the functional evolution of Udmbf1. Additional studies involving more U. decussata samples from representative ecotypes will be necessary to uncover the relationships among DNA polymorphisms, functional gene evolution, and lichen habitats

Limitations of Species Delimitation Based on Phylogenetic Analyses: A Case Study in the Hypogymnia hypotrypa Group (Parmeliaceae, Ascomycota)

Delimiting species boundaries among closely related lineages often requires a range of independent data sets and analytical approaches. Similar to other organismal groups, robust species circumscriptions in fungi are increasingly investigated within an empirical framework. Here we attempt to delimit species boundaries in a closely related Glade of lichen-forming fungi endemic to Asia, the Hypogymnia hypotrypa group (Parmeliaceae). In the current classification, the Hypogymnia hypotrypa group includes two species: H. hypotrypa and H. flavida, which are separated based on distinctive reproductive modes, the former producing soredia but absent in the latter. We reexamined the relationship between these two species using phenotypic characters and molecular sequence data (ITS, GPD, and MCM7sequences) to address species boundaries in this group. In addition to morphological investigations, we used Bayesian clustering to identify potential genetic groups in the H. hypotrypa/H. flavida Glade. We also used a variety of empirical, sequence-based species delimitation approaches, including: the "Automatic Barcode Gap Discovery" (ABGD), the Poisson tree process model (PTP), the General Mixed Yule Coalescent (GMYC), and the multispecies coalescent approach BPP. Different species delimitation scenarios were compared using Bayes factors delimitation analysis, in addition to comparisons of pairwise genetic distances, pairwise fixation indices (FST). The majority of the species delimitation analyses implemented in this study failed to support H. hypotrypa and H. flavida as distinct lineages, as did the Bayesian clustering analysis. However, strong support for the evolutionary independence of H. hypotrypa and H. flavida was inferred using BPP and further supported by Bayes factor delimitation. In spite of rigorous morphological comparisons and a wide range of sequence-based approaches to delimit species, species boundaries in the H. hypotrypa group remain uncertain. This study reveals the potential limitations of relying on distinct reproductive strategies as diagnostic taxonomic characters for Hypogymnia and also the challenges of using popular sequence-based species delimitation methods in groups with recent diversification histories

Low-frequency electrical stimulation combined with preventative pelvic floor muscle exercises and knowledge-attitude-practice model in women with urinary retention after a pelvic surgery

This study was to explore the efficacy of low-frequency electrical stimulation (LFES) combined with preventative pelvic floor muscle exercises (PPFME) and knowledge-attitude-practice model (KAP model) in women with urinary retention (UR) after a pelvic surgery and the effect on quality of life (QOL). The clinical data of 153 women hospitalized with UR after a pelvic surgery from January 2015 to June 2019 were retrospectively analyzed and divided into the Control Group (CG, n=45, LFES+PPFME) and the Study Group (SG, n=108, LFES+PPFME+KAP model) according to the different treatment methods. Following indicators were compared between the two groups: clinical efficacy, time to first urination, urine output, rate of extubation in 3D, rate of repeated intubation, urodynamic indicators before and after treatment, postvoid residual (PVR), bladder compliance (BC), maximal flow rate (Q-max), pressure of detrusor at Pdet-Q-max (Pdet-Q-max), changes in QOL, incidence of urinary tract infection, and length of stay (LOS). Compared to CG, SG showed higher overall response rate (ORR) and rate of extubation in 3D, shorter time to first urination, higher urine output, lower rate of repeated intubation and incidence of urinary tract infection, and shorter LOS (P<0.05); both groups achieved improvements in urodynamic indicators and QOL score after treatment (P<0.05); the urodynamic indicators and QOL score in SG were better than the CG’s (P<0.05). LFES combined with PPFME and KAP model is effective in treating women with UR after a pelvic surgery by efficaciously improving patients’ urination and QOL

Lichen-associated Fungal Community in Hypogymnia hypotrypa (Parmeliaceae, Ascomycota) Affected by Geographic Distribution and Altitude

Lichen-associated fungal species have already been investigated in almost all the main growth forms of lichens, however whether or not they are homogeneous and constant within each lichen species are still inconclusive. Moreover, the related ecological factors to affect and structure the fungal composition have been poorly studied. In order to answer these questions, we took Hypogymnia hypotrypa as a model to study the relationship between the lichen-associated fungal composition and two ecological factors, i.e. site and altitude, using the method of IlluminaMiSeq sequencing. Four different sites and two levels of altitude were included in this study, and the effects of site and altitude on fungal community composition were assessed at three levels, i.e. OTU, class and phylum. The results showed that a total of 50 operational taxonomic units (OTUs) were identified and distributed in 4 phyla, 13 classes and 20 orders. The lichen-associated fungal composition within H. hypotrypa were significantly affected by both site and altitude at OTU and class levels, while at the phylum level, it was only affected by altitude. While the lichen associated fungal communities were reported to be similar with endophytic fungi of the moss, our results indicated the opposite results in some degree. But whether there exist specific OTUs within this lichen species corresponding to different sites and altitudes is still open. More lichen species and ecological factors would be taken into the integrated analyses to address these knowledge gaps in the near future

Regulation of symbiotic interactions and primitive lichen differentiation by UMP1 MAP kinase in Umbilicaria muhlenbergii

Abstract Lichens are of great ecological importance but mechanisms regulating lichen symbiosis are not clear. Umbilicaria muhlenbergii is a lichen-forming fungus amenable to molecular manipulations and dimorphic. Here, we established conditions conducive to symbiotic interactions and lichen differentiation and showed the importance of UMP1 MAP kinase in lichen development. In the initial biofilm-like symbiotic complexes, algal cells were interwoven with pseudohyphae covered with extracellular matrix. After longer incubation, fungal-algal complexes further differentiated into primitive lichen thalli with a melanized cortex-like and pseudoparenchyma-like tissues containing photoactive algal cells. Mutants deleted of UMP1 were blocked in pseudohyphal growth and development of biofilm-like complexes and primitive lichens. Invasion of dividing mother cells that contributes to algal layer organization in lichens was not observed in the ump1 mutant. Overall, these results showed regulatory roles of UMP1 in symbiotic interactions and lichen development and suitability of U. muhlenbergii as a model for studying lichen symbiosis

3genelocusalignment

DNA sequences alignmen

Comparative Analysis of Chemical Composition, Anti-Inflammatory Activity and Antitumor Activity in Essential Oils from Siegesbeckia orientalis, S. glabrescens and S. pubescens with an ITS Sequence Analysis

Herba Siegesbeckiae (HS), derived from the aerial parts of three plants, Siegesbeckia orientalis (SO), S. glabrescens (SG), and S. pubescens (SP), has been used for the treatment of inflammatory diseases in China for centuries. In the present study, hydrodistillation was applied to extract essential oils from dried SO, SG, and SP aerial parts, and chemical composition analysis by gas chromatography&ndash;mass spectrometry (GC-MS) led to the identification of a total of 148 compounds (56 in SO, 62 in SG, and 59 in SP). The main components in the essential oils of SO, SG, and SP differed significantly. In vitro anti-inflammatory activity assays showed that SP essential oils (IC50, 0.97 &mu;g/mL) significantly reduced the ability of lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages to release NO, and the SO essential oil (IC50, 14.99 &mu;g/mL) was better than the others at inhibiting the LPS-induced release of cytokine IL-6. Furthermore, the essential oils exhibited antitumor activities (IC50, 37.72&ndash;123.16 &mu;g/mL) against Hep3B (liver) and Hela (cervical) cells. Linear regression analysis showed that, caryophyllene oxide peak area percentages showed remarkably high negative correlation coefficients with IC50 values of Hep3B and Hela cytotoxicity, which suggested the contribution of this compound on the cancer cell cytotoxicity of three essential oils. Finally, the ITS1-5.8S-ITS2 region was amplified and sequenced in order to generate genomic reference sequences for each plant. These can be used to identify the origins of the plants, and will assist other research studies related to these three plants