Inonotus hispidus (Bull.) P. Karst from the Inner Western Anatolia part of Türkiye with a molecular analysis, phenotypic characters study, genus of host determination, and geo-hydrographic distribution factors analysis

As a result of conventional characterization of yeasts isolated from various plant leaves collected in Yunnan, China, six ballistoconidium-forming strains with orange-colored colonies were grouped together. Molecular phylogenetic analysis based on 18S rDNA sequencing showed that two representative strains of this group of yeasts, CH 2.068 and CH 2.497, were closely related to the species in the genus Dioszegia and had signature sequences typical of this genus. However, the six strains from Yunnan differed from the described Dioszegia species remarkably (14.5-17.7% nucleotide divergences) in the ITS (internal transcribed spacer) region sequences, which indicated that they represent a distinct species. Furthermore, among the six strains studied, the ITS region sequence comparison allowed the recognition of two subgroups represented by CH 2.068 and CH 2.497, which differ from each other in three bases in the ITS 2 region. DNA-DNA relatedness revealed that the two subgroups represent two varieties of a new species in the genus Dioszegia, for which Dioszegia zsoltii sp. nov. var. zsoltii and Dioszegia zsoltii var. yunnanensis var. nov. are proposed.

Wheat (Triticum aestivum L.) fields in Zhengzhou, Henan Province that exhibited reduced height and number of tillers were surveyed in 2008, and samples were collected for analyses. Fifteen of thirty-five samples contained a nematode suspected of causing small, brown lesions in roots. Both lateral roots and root hairs were reduced in infected plants. Nematode presence was associated with enlarging lesions and necrotic roots. We also observed lesion nematode infestation in samples with no symptoms in the roots. During the growing season, wheat yield increased by 14% with nematocide treatments in Zhengzhou suburb fields. The morphological and molecular analyses of the nematodes isolated from soil samples established the identity of the species as the lesion nematode, Pratylenchus agilis (2,3). Morphological characteristics that were used for identification included female body, lip annules, tail terminus, number of lines in the lateral field, stylet knobs, stylet length, and vulva location in relation to body length. Females were cylindrical, measured 452 to 811 μm long, and contained two lip annules, a smooth tail terminus, four lateral lines, a vulva at 72.64 to 79.97%, a stylet of 16.20 to 17.55 μm, basal esophageal lobes elongated less than twice the body width, and stylet knobs rounded posteriorly. Males were rare and the spermatheca was empty. Molecular analysis was conducted by amplifying the internal transcribed spacer (ITS) regions ITS1 and 2. Sequence of ITS regions from this population (GenBank Accession No. JQ039330) showed highest sequence homology to P. agilis isolate PagKL5 (GenBank Accession No. FJ71289.1) with the identity of 90%. High variability at species level has been found for Pratylenchus species (1). On the basis of the morphological traits and molecular analyses, the nematodes were identified as P. agilis. To our knowledge, this is the first report of P. agilis parasitizing wheat.

Lipase inhibitors have generated a great interest because they could help in the prevention or the therapy of lipase‐related diseases. Therefore, the aim of this work was to evaluate the inhibitory effect of secondary metabolites extracts such as phenolic compounds and saponins of three Algerian medicinal plants: Achillea santolina, Inonotus hispidus and Zizyphus lotus, indeed their antiradicalaire activity using DPPH• (1, 1‐diphenyl‐2‐picryl‐hydrazyl). The phenolic extracts have shown a strong antiradicalaire activity than the saponin extracts with EC50 values ranged from 6 to 11 μg/ml and from 51 to 82 μg/ml, respectively. The enzymatic inhibition produced by these plant extracts is described here for the first time. The results have shown that the phenolic extracts are more potent than the saponin extracts with Ki values ranged from 0.011 mg/ml to 0.027 mg/ml for phenolic extracts, and ranged from 0.071 mg/ml to 0.69 mg/ml for saponin extracts. The nature, mechanism and possible physiological relevance of lipase inhibition by extract components are discussed. Keywords: Achillea santolina, Inonotus hispidus, Zizyphus lotus, Candida rugosa lipase, inhibition, saponins extracts, phenolic extracts, DPPH•.

Compounds from spices and herbs extracts are being explored as natural antibacterial additives. A plant extract used in traditional folk medicine is Hibiscus sabdariffa L., also known as Roselle. Therefore, the potential use of a phenolic hibiscus extract as antibacterial or natural food preservative was analyzed in vitro and in situ. A phenolic extract was obtained from hibiscus calyces and fractionated, and then the fractions were tested against foodborne pathogen bacteria. Liquid–liquid extraction and solid-phase extraction were used to fractionate the hibiscus extract, and HPLC was employed to analyze the fractions’ phenolic composition. Minimum bactericidal concentration (MBC) and minimal inhibitory concentration (MIC) were calculated for brute hibiscus phenolic extract, each of the fractions and pure commercial phenolic compounds. Bacteria tested were Escherichia coli, Salmonella enterica serovar Typhimurium, Staphylococcus aureus, Listeria monocytogenes and Bacillus cereus. The fraction obtained after liquid–liquid extraction presented the best performance of MBC and MIC against the bacteria tested. Furthermore, a hibiscus ethanolic extract was employed as a natural preservative to extend the shelf-life of beef. Microbiological, color and sensory analyses were performed to the meat during the shelf-life test. The application of the phenolic hibiscus extract also showed an increase of the duration of the meat`s shelf life.

Powdery mildews are obligate biotrophic phytopathogenic fungi with a worldwide distribution. Advances in sequencing and molecular analyses have enabled the identification of cryptic species that were previously indistinguishable through morphological methods alone. Consequently, the discovery of new species continues to increase. This study brings together a comprehensive molecular phylogenetic analysis based on the amplification of ITS and IGS regions and a morphological analysis conducted on a broad spectrum of powdery mildew samples collected on various representatives of the genus Lactuca. Our dataset consisted of 200 accessions of herbaria leaves of Lactuca spp. infected with powdery mildew from different habitats worldwide. Microscopic examination revealed the presence of asexual states observable as conidiophores with two types of conidia, with fibrosin bodies (Podosphaera xanthii), and without fibrosin bodies (Golovinomyces bolayi). In some cases, chasmothecia were also recorded. Molecular identifications based on the ITS region confirmed this conclusion. Among 148 concatenated sequences of ITS and IGS regions, we discovered 12 ribotypes in Lactuca samples infected by G. bolayi. A single ribotype was observed for samples with the presence of P. xanthii. The principal component analysis revealed the pathogen’s host-specificity as an important factor determining host-dependent morphological variability in the case of samples from L. sativa and L. serriola.

The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at 30°C. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

This experiment was undertaken to depict the favourable condition for mycelial growth, molecular identification and phylogenetic relationship of the selected strains of Pleurotus salmoneostramineus. Suitable temperature and pH were obtained at 25ºC and 6, respectively. Mushroom complete, glucose peptone and yeast malt extract culture media were favorable, while Hennerberg and Hoppkins were unfavorable. Dextrin was the best and xylose was the less effective carbon sources. Inorganic nitrogen sources were less effective for the mycelial growth of P. salmoneostramineus. The sequences of internal transcribed spacer (ITS) region of selected strains revealed that the total length ranged from 614 to 663 bp. The size of the ITS1 and ITS2 regions varied among the strains. Sequence analysis showed that 5.8S of rDNA sequences were identical. Phylogenetic tree of the ITS region sequences indicated that strains of P. salmoneostramineus belong to same cluster. The reciprocal homologies of the ITS region sequences ranged from 98 to 100%. The strains of P. salmoneostramineus were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. RAPD results suggested that tested strains of P. salmoneostramineus were genetically similar with some variations, thus it could be concluded that RAPD and ITS techniques were well competent for detecting the genetic diversity of all tested strains of P. salmoneostramineus.

The molecular phylogeny in nine different commercial cultivated strains of Pleurotus nebrodensis was studied based on their internal transcribed spacer (ITS) region and RAPD. In the sequence of ITS region of selected strains, it was revealed that the total length ranged from 592 to 614 bp. The size of ITS1 and ITS2 regions varied among the strains from 219 to 228 bp and 211 to 229 bp, respectively. The sequence of ITS2 was more variable than ITS1 and the region of 5.8S sequences were identical. Phylogenetic tree of the ITS region sequences indicated that selected strains were classified into five clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by RAPD with 20 arbitrary primers. Twelve primers were efficient to applying amplification of the genomic DNA. The sizes of the polymorphic fragments obtained were in the range of 200 to 2000 bp. RAPD and ITS analysis techniques were able to detect genetic variation among the tested strains. Experimental results suggested that IUM-1381, IUM-3914, IUM-1495 and AY-581431 strains were genetically very similar. Therefore, all IUM and NCBI gene bank strains of P. nebrodensis were genetically same with some variations.

Background: In this study, we investigate the inhibitory effects of phenolic and saponins extracts of Cupressus sempervirens cones and seeds. Objective: The ethyl acetate and butanolic extracts of Cupressus sempervirens (L) cones and seeds, collected from Laghouat (Algeria) were analyzed for their saponins and phenolic content. We also studied the in vitro antidiabetic activity of C. sempervirens using α-amylase digestion enzyme. Methods: The amount of total phenolics and saponins in the samples was determined with the Folin- Ciocalteu reagent and with vanillin, respectively. To identify cones and seeds extracts with inhibitory capacities, we have studied the effects of ours extracts on the kinetics catalyzed of α-amylase an enzymes belonging to the class of hydrolase responsible for the digestion and we have subjected our extracts to inhibition assay to determine the inhibition percentage for each extract. Results: The fruits and seeds contained total free phenolic content of 1.96 mg/gGAE and 2.25 mg/gGAE, respectively. The saponin content determined with vanillin reagent shows a good yeild of 119.85 and 131.46 mg/gDE in ethyl acetate and butanolic extracts, respectively. In addition, phenolic and saponins extracts were found to inhibit enzymatic activity of α-amylase under in vitro starch digestion bioassay and the values of the IC50 constants have been determined for both seeds and cones extracts. The values ranged from 0.49 to 1.12 mg/ml. This paper is the first report on antidiabetic activity of saponins and phenolic extracts of cones and seeds from Cupressus sempervirens (L). Keywords: Cupressus sempervirens, diabetes, enzyme inhibition, phenolic compound, α-amylase, vanillin.

Baccharis trimera is a plant popularly used as a tea and to treat gastrointestinal diseases and inflammatory processes as well. The total phenolic content was determined and the antioxidant and anti-inflammatory activities of six extracts (dichloromethane, ethyl acetate, butanol, aqueous, saponin and phenolic) from B. trimera were evaluated. Using carrageenan-induced pleurisy as a model of acute inflammation, the phenolic extract at 15 mg/kg decreased significantly the analyzed parameters when compared to the carrageenan group (p < 0.05), thus showing potential anti-inflammatory activity. The total phenolic content and antioxidant activity were evaluated by the Folin-Ciocalteau and DPPH methods, respectively. Phenolic and ethyl acetate extracts presented higher antioxidant activity (p < 0.05) than ascorbic acid. The phenolic extract also showed the highest antioxidant potential in relation to the other extracts, thus suggesting that the antioxidant and anti-inflammatory activities were due to the presence of phenolic compounds.

Internal transcribed spacer region (ITS) sequencing is the most extensively used technology for accurate molecular identification of fungal pathogens in clinical microbiology laboratories. Intra-genomic ITS sequence heterogeneity, which makes fungal identification based on direct sequencing of PCR products difficult, has rarely been reported in pathogenic fungi. During the process of performing ITS sequencing on 71 yeast strains isolated from various clinical specimens, direct sequencing of the PCR products showed ambiguous sequences in six of them. After cloning the PCR products into plasmids for sequencing, interpretable sequencing electropherograms could be obtained. For each of the six isolates, 10–49 clones were selected for sequencing and two to seven intra-genomic ITS copies were detected. The identities of these six isolates were confirmed to be Candida glabrata (n = 2), Pichia (Candida) norvegensis (n = 2), Candida tropicalis (n = 1) and Saccharomyces cerevisiae (n = 1). Multiple sequence alignment revealed that one to four intra-genomic ITS polymorphic sites were present in the six isolates, and all these polymorphic sites were located in the ITS1 and/or ITS2 regions. We report and describe the first evidence of intra-genomic ITS sequence heterogeneity in four different pathogenic yeasts, which occurred exclusively in the ITS1 and ITS2 spacer regions for the six isolates in this study.

Inonotus hispidus mushroom is a traditional medicinal fungus with anti-cancer, antioxidation, and immunomodulatory activities, and it is used in folk medicine as a treatment for indigestion, cancer, diabetes, and gastric illnesses. Although I. hispidus is recognized as a rare edible medicinal macrofungi, its genomic sequence and biosynthesis potential of secondary metabolites have not been investigated. In this study, using Illumina NovaSeq combined with the PacBio platform, we sequenced and de novo assembled the whole genome of NPCB_001, a wild I. hispidus isolate from the Aksu area of Xinjiang Province, China. Comparative genomic and phylogenomic analyses reveal interspecific differences and evolutionary traits in the genus Inonotus. Bioinformatics analysis identified candidate genes associated with mating type, polysaccharide synthesis, carbohydrate-active enzymes, and secondary metabolite biosynthesis. Additionally, molecular networks of metabolites exhibit differences in chemical composition and content between fruiting bodies and mycelium, as well as association clusters of related compounds. The deciphering of the genome of I. hispidus will deepen the understanding of the biosynthesis of bioactive components, open the path for future biosynthesis research, and promote the application of Inonotus in the fields of drug research and functional food manufacturing.

Background and Purpose:Catheter-related blood circulation infection is the most dangerous and serious side-effects of vascular catheters, which leads to the enhancement of the costs, mortality, and hospital stay duration, especially in the Intensive Care Unit. Regarding this, the aim of the current study was to identify the prevalence of catheter-induced candidemia in the Tehran Heart Center, a heart hospital in Tehran, Iran.Materials and Methods:This study was conducted on patients admitted to Tehran Heart Center for a minimum of 7 days during 18 months. To detect the fungal elements, blood culture and catheter culture were performed in the patients receiving central or peripheral venous catheter. Then, the polymerase chain reaction (PCR) was applied to determine the possible diagnosis.Results:The investigation of 223 samples led to the identification of a total of 15 (6.7%) yeast isolates obtained from 9 (60%), 4 (26.6 %), and 2 (13.4%) catheter, blood, and skin (of the catheter insertion areas) cultures, respectively. Out of nine Candida isolates obtained from the catheter samples, 1 (11.1%), 1 (11.1%), 2 (22.2%), and 5 (55.6%) cases were identified as C. tropicalis, C. membranifaciens, C. glabrata, and C. albicans, respectively, using the internal transcribed spacer region sequencing. Furthermore, the four yeasts isolated from the blood culture included C. tropicalis, C. carpophila, C. membranifaciens, and Cryptococcus albidus. Additionally, one case of C. glabrata and one case of C. albicans were isolated from the skin culture of the catheter insertion areas in patients with positive catheter culture. We reported two cases of catheter-related candidemia caused by C. membranifaciens and C. tropicalis on the basis of the genetic similarity of the species isolated from the blood and catheter. These cases were treated successfully with intravenous fluconazole and catheter removal.Conclusion: There is some evidence indicating the growing prevalence of non-albicans Candida infections. Many risk factors, including prior antibiotic therapy, use of a central venous catheter, surgery, and parenteral nutrition, are considered to be associated with candidemia in hospitalized heart failure patients. The identification of the route of infection in candidemia is difficult. In the current study, the positive blood and catheter cultures for Candida isolates and the similarity of the ITS region of ribosomal DNA sequence of Candida isolated from two patients confirmed the diagnosis of intravenous catheter-related candidemia.

Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a <TEX>$^{60}Co$</TEX> gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

Species of Phaeoacremonium (especially Phaeoacremonium aleophilum) are associated with two severe diseases in grapevines, Petri disease in young plants and Esca disease in adult plants. Phaeoacremonium species grow slowly on culture medium, and it is difficult to identify these species on the basis of morphological characteristics. Primers Pm1 and Pm2 were designed in the ribosomal DNA internal transcribed spacer (ITS) regions ITS1 and ITS2, respectively. They yielded a single amplicon of 415 bp for nine species of Phaeoacremonium that may occur in grapevines. A nested PCR (using general fungal primers ITS1F/ITS4 in the primary reaction) was developed to detect Phaeoacremonium directly in grapevine wood. Molecular detection was more sensitive than the traditional method of culturing in growth medium was. Identification of Phaeoacremonium species was achieved by digesting the PCR-amplified fragment with the restriction enzymes BssKI, EcoO109I, and HhaI. It was possible to distinguish these species by their restriction fragment length polymorphism patterns, except for Phaeoacremonium viticola and Phaeoacremonium angustius, which had 100% similarity in their ITS region sequences. A species-specific PCR amplification of the partial beta-tubulin gene using the primer pair Pbr4_1/T1 and Pbr8/T1 was necessary to differentiate P. angustius from P. viticola, respectively. An easy and fast protocol was developed to detect and identify species of Phaeoacremonium in a few hours. Primers defined here can be used in a plant nursery sanitation program to produce plants free of Phaeoacremonium spp. Use of healthy grapevine plants in new plantations is the most effective measure to manage Petri disease.