Small Subunit Research Articles

The First Introduction of an Exogenous 5' Untranslated Region for Control of Plastid Transgene Expression in Chlamydomonas reinhardtii.

The utilization of heterologous 5' untranslated regions (5'UTRs) for expressing foreign proteins in the chloroplast of Chlamydomonas reinhardtii (C. reinhardtii) has posed a persistent challenge over the years. This challenge stems from the lack of a defined and comprehensive set of translational cis-elements responsible for stability, ribosome binding, and translation initiation, which are mediated by trans-acting factors native to C. reinhardtii. In the current study, we aimed to address this bottleneck by employing the 5'UTR from gene 10 of the T7 bacteriophage (T7g10 5'UTR), fused to the promoter of C. reinhardtii small subunit ribosomal RNA (rrnS), to facilitate the translation of a reporter gene, YFP. Using a chimeric construct, the YFP mRNA was efficiently translated utilizing the heterologous T7g10 5'UTR. Furthermore, the accumulation of YFP protein under the control of the T7g10 5'UTR was approximately one third of that observed under the control of the endogenous psaA promoter/5'UTR in the C. reinhardtii chloroplast. The results of computational analyses demonstrated that the T7g10 5'UTR sequence shares common elements with the endogenous 5'UTRs of the chloroplast genes. Moreover, the findings of the current study highlighted the potential of employing bacteriophage 5'UTRs for the foreign protein accumulation from the chloroplast genome of C. reinhardtii.

Impact of Molecular Identification of Palm leaves on the preservation and conservation of Palm Leaves-Tala patra Manuscripts

Palm leaves manuscripts preservation is the main target of this research work. Therefore, this research study aimed at molecular identification of palm leaves by DNA Barcoding to assess the potential level on manuscript preservation. This study aims to amplify the rbcL and matK genes in palm leaves collected from different regions of Bangalore. The DNA was isolated by CTAB method and the amplification was done by PCR analysis. DNA barcoding proved to be an efficient tool for proper discrimination of the related species by generating standers that can be used universally. rbcL: A gene found in the chloroplast genome, encoding a large subunit of the enzyme ribulose-bisphosphate carboxylase. matK: Another chloroplast gene coding for maturase K, involved in splicing group II introns. The discrimination is done by accurately sequencing the standard gene region in a very short time (Hebert et al., 2003). rbcL and matK have been having most recommended and vasty studied DNA barcodes for plants, as they are proved by PWG (Pair wise group) to be potential standardized barcodes for plant DNA barcoding. These barcodes are used to properly identify and discriminate any native or foreign plant species (Maloukh et al., 2017). This work guides us the impact/role of molecular identification-DNA Barcoding of palm leaves on the preservation of Palm leaves manuscripts

First Molecular Detection and Genetic Characterization of Tetratrichomonas buttreyi and Pentatrichomonas hominis in Donkeys in Shanxi Province, China.

Two species of trichomonads, Tetratrichomonas buttreyi and Pentatrichomonas hominis, are common intestinal parasites that can impact animal health and productivity. Severe infection by these parasites can lead to diarrhea and wasting in affected animals. Notably, P. hominis is known to cause diarrhea and has the potential to be transmitted between animals and humans. Donkeys hold significant economic importance in China's agricultural sector. However, whether donkeys are infected with T. buttreyi and P. hominis remains unknown globally. To address this gap in knowledge, 815 fecal samples were collected from donkeys in three representative regions in Shanxi Province, North China. Then, the presence and genetic characteristics of T. buttreyi and P. hominis were examined using species-specific PCR primers amplifying the small subunit ribosomal RNA genes. The overall prevalence was detected to be 25.4% (207/815) for T. buttreyi and 0.7% (6/815) for P. hominis in donkeys in Shanxi Province. All obtained P. hominis sequences were identified as genotype CC1. Genetic analysis revealed that all P. hominis isolates from donkeys were clustered into the same branch with isolates detected in humans, suggesting possible zoonotic transmission. This study is the first to report the occurrence and prevalence of T. buttreyi and P. hominis in donkeys globally. These findings expand the host range of trichomonads and improve our understanding of their genetic diversity and zoonotic potential, providing essential baseline data for the prevention and control of these parasites in donkeys in the region.

COMPARISON OF COMPUTATIONAL METHODS FOR ANALYSING 16S rRNA SEQUENCING DATA PRODUCED WITH THE USE OF SHORT AND LONG READS

This research is driven by the importance of identifying species in microbial community for use in medical testing. Conventional methods including culturing, biochemical assays are often limited to the culturable organisms and do not provide information about relative abundance of different microbial taxa within a community. Comparative analysis of metagenomic programs method was used to identify the most appropriate option for long-read 16S rRNA data. Since the development of high-throughput sequencing technologies, novel methods of identifying species include taxonomic profiling of 16S rRNA. 16S rRNA gene contains variable regions (V1-V9) that are specific to different taxonomic groups. Illumina short-read sequencing technology provides read lengths 2*251 base pairs (bp) for paired-end sequencing, while fraction of 16S rRNA gene is around 1500 bp, thus reads can successfully cover 2 of 9 regions (300-400 bp), but not the whole gene. The Oxford Nanopore Technology (ONT) representing long-read sequencing or so-called 3rd generation sequencing is much more applicable for 16S rRNA profiling, as it can provide reads with length 1500 bp (exactly in line with the length of 16S rRNA gene). Extended read length provides high alignment quality making the downstream taxonomy classification and relative abundance calculation much easier and relevant. The samples from endometrium of uterus were studied in the first part of the research. Endometrial microbiome composition and balance can play an important role in the success of in vitro fertilization (IVF). Taxonomical classification was tested with NCBI, Silva databases. NCBI database consists of full genomes of Bacteria, Archaea and Eukaryote, Silva contains curated and annotated sequences of only small subunit ribosomal RNA of these domains. Among the enormous collection of metagenome classifiers there were selected three particular programs applying completely different algorithms: Emu, PathoScope and Kraken2. The comparative process was divided in three phases. In the first one, pipelines were run with three programs and 11 samples using Silva and NCBI databases, the results were compared both quantitively and visually. In the second phase, the results were compared to the short-read taxonomical classification, the discrepancies were analyzed and questions were put together for further investigation. During the third phase pipelines with best performed programs were run on sample with acknowledged microbial diversity and following metrics were assessed and visualized: True positive, False positive, Precision, Recall, F1-score, L1-norm, L2-norm. Results: After the first phase it was revealed that Kraken2, Emu, PathoScope produce very similar results, while MetaScope discrepancies exceeded substantially 10% threshold for 70% of the reads. Additionally, Kraken2 annotated significantly more genera upon the threshold than other programs indicating its low specificity. Consequently, these two programs were excluded from further analysis. The second phase brought to the conclusion that short-read sequencing reports reveal only one prevailing genus in the community, while long-read establish 3 of them, thus it was assumed that short-read classification is much less sensitive. In the third phase it was determined that Emu and PathoScope have least deviation from reference value of abundances (ground truth) and they successfully annotate all the species introduced in the sample. Therefore, the main outcome of the research is the recommendation to use Emu and PathoScope programs for the purpose of identifying species using long-read data.

Synergistic Effect Mechanism of Binary Sweet Taste Compounds.

In our previous study, phloridzin, sucrose, l-alanine, and dulcitol presented synergistic effects in Camellia nanchuanica black tea (NCBT). This study aims to verify the synergistic effects of the aforementioned sweet taste compounds and the mechanism involved. By conducting σ-τ plot analysis, phloridzin at the recognition threshold concentration (phl) exhibited synergistic effects with different concentrations of sucrose (Lsuc-6suc). Various concentrations of sucrose, phloridzin, and their combinations were selected to investigate the impact on sweet taste receptor cells. The results revealed that sucrose/phloridzin significantly increased the calcium signal compared to phloridzin and sucrose alone, attributed to the greater stability of the sucrose/phloridzin combination when binding to Taste 1 Receptor Member 3 (TAS1R3; one subunit of sweet taste receptor proteins). Ultimately, the sweet taste signal of sucrose/phloridzin was transmitted to the brain, triggering the activation of more brain regions associated with sweet taste perception (right insular, postcentral, and amygdala).

Biological and genomic characterization of the novel bacteriophage vB_VpM-pA2SJ1, which infects Vibrio parahaemolyticus associated with acute hepatopancreatic necrosis disease.

Vibrio parahaemolyticus is a major seafood-borne zoonotic pathogen that causes gastroenteritis in humans and acute hepatopancreatic necrosis disease (AHPND) in shrimp. In this study, we isolated and characterized Vibrio phage vB_VpM-pA2SJ1, which infects clinical and AHPND-associated strains of V. parahaemolyticus. The phage genome is a linear dsDNA 51,054 bp in length with a G + C content of 43.7%, and it contains 89 open reading frames. Genome comparisons revealed basal similarity to other Vibrio phages, particularly Vibrio phage vB_VpP_1, with 84.2% identity and 46% coverage. Phylogenetic analysis based on the whole genome, the terminase large subunit, and the major capsid protein revealed that phage vB_VpM-pA2SJ1 did not cluster with other known phage families, thus indicating its uniqueness.

Quality characteristics and fibrous structure formation mechanism of walnut protein and wheat gluten meat analogues during high-moisture extrusion cooking process

Blending two or more materials to create better high-moisture meat analogues has been actively studied in the food science and technology field. Walnut protein is a high-quality plant-based protein resource, yet its full potential remains underexploited. Thus, this study focused on exploring the quality characteristics and fibrous structure formation mechanism of walnut protein (WP) and wheat gluten (WG) meat analogues during high-moisture extrusion cooking process. Results showed that the optimized WP and WG-blended high-moisture meat analogues exhibited a more pronounced anisotropic and oriented fibrous structure. The blending of WP and WG can protect the molecular chains from the thermal transition, and promote the aggregation of protein molecules mainly by enhancing the interaction between hydrophobic interactions and hydrogen bonds, increasing the apparent viscosity and forming protein subunits with larger molecular weights (>100 kDa) to stabilize the newly formed conformation. Additionally, the content of α-helix was the highest among the secondary structures. This study provides a theoretical basis for the application of WG and WP to produce HMMAs with rich fibrous structures.

Ultrastructural and molecular characterization of Glugea sp. (microsporidia), a parasite of the Red Sea fish Carangoides bajad (Carangidae).

Glugea sp. found infecting the liver of the teleost fish Carangoides bajad from the Red Sea, Egypt, is described based on light microscopy and ultrastructural characteristics combined with phylogenetic analyses. This microsporidium forms whitish xenomas up to ~4 mm in size. Xenomas display numerous parasitophorous vacuoles totally filled by mature spores, no other life cycle stages were observed. Mature spores ellipsoidal and measuring 6.3 × 4.0 μm in size. The polaroplast appears composed of two distinct regions: an electron-dense vesicular region and a densely packed lamellar region. The polar tubule forms approximately 24-27 coils arranged in three layers encircling the posterior vacuole. The small subunit (SSU) rRNA gene and its ITS region were sequenced and showed the highest similarity of 99.4% to other Glugea spp. Bayesian inference and maximum likelihood analyses place the novel isolate within the Glugea clade, more specifically within a subclade that predominantly grouped species described from fish inhabiting the Arabian Gulf or Red Sea. The results validate the parasite's classification in the Glugea genus. Nevertheless, until more detailed ultrastructural and molecular data are obtained, the identification of the current Glugea species is hampered by the absence of some developmental stages and the high degree of genetic similarity.

A new brown rot disease of plum caused by Mucor xinjiangensis sp. nov. and screening of its chemical control.

A novel species of Mucor was identified as the causal agent of a brown rot of Prunus domestica (European plum), widely grown in the south of Xinjiang, China. This disease first appears as red spots after the onset of the fruits. With favorable environmental conditions, fruit with infected spots turn brown, sag, expand, wrinkle, and harden, resulting in fruit falling. Fungal species were isolated from infected fruits. A phylogenetic analysis based on internal transcribed spacer (ITS) regions and the large subunit (LSU) of the nuclear ribosomal RNA (rRNA) gene regions strongly supported that these isolates made a distinct evolutionary lineage in Mucor (Mucoromycetes, Mucoraceae) that represents a new taxonomic species, herein named as Mucor xinjiangensis. Microscopic characters confirmed that these strains were morphologically distinct from known Mucor species. The pathogenicity of M. xinjiangensis was confirmed by attaching an agar disk containing mycelium on fruits and re-isolation of the pathogen from symptomatic tissues. Later, fourteen fungicides were selected to determine the inhibitory effect on the pathogen. Further, results showed that difenoconazole had the best effect on the pathogen and the strongest toxicity with the smallest half maximal effective concentration (EC50) value, followed by a compound fungicide composed of difenoconazole with azoxystrobin, mancozeb, prochloraz with iprodione, pyraclostrobin with tebuconazole, and trifloxystrobin with tebuconazole and ethhylicin. Present study provides the basis for the prevention and control of the novel plum disease and its pathogen.

Development of a PCR assay for detection and identification of Eimeria spp. in cattle

Eimeria spp. are important coccidian parasites causing diarrhea and significant mortality in cattle worldwide. To date, at least 13 Eimeria species with varying pathogenicity have been identified in cattle. Efficient detection and identification of Eimeria spp. is therefore essential for the prevention and control of bovine coccidiosis. However, the commonly used microscopic examination for Eimeria spp. is time-consuming and requires considerable expertise. In this study, we aligned the nucleotide sequences of the small subunit (SSU) rRNA gene of common Eimeria species and developed a nested PCR assay targeting the polymorphic SSU rRNA region of Eimeria spp. from cattle. Initially, the SSU rRNA gene PCR assay was compared with microscopic examination for sensitivity and detection range of Eimeria species using fecal samples from dairy cattle. Of the 193 fecal samples, 131 (67.9 %) and 78 (40.4 %) were positive for Eimeria by PCR and microscopy, respectively. Sequence analysis of the PCR products identified six Eimeria species, including E. cylindrica (n = 76), E. bovis (n = 54), E. auburnensis (n = 30), E. zuernii (n = 25), E. wyomingensis (n = 10), E. canadensis (n = 1), and co-infections of 2–4 species (n = 55). In contrast, only the first four species and co-infections of 2–3 species were identified by microscopy. The PCR assay was able to detect as few as 50 Eimeria oocysts per gram of feces. Thus, the developed SSU rRNA gene PCR assay has a high sensitivity and allowed easy identification of at least six common Eimeria species and their co-infections in cattle. It should be useful in molecular epidemiological studies of bovine coccidiosis.

Genome resequencing reveals genetic status of Macrobrachium rosenbergii populations in China

The giant freshwater prawn (Macrobrachium rosenbergii) has been widely cultured in China since 1990s. In this study, 189 individuals were sampled from 10 distinct populations, including original populations introduced from abroad and domesticated strains in China, namely Florida (FD), "Nantaihu No.2" (NT), Vietnam (YN), Sri Lanka (SK), Bangladesh (ML), Thailand (TG), Taiwan (TW), "Shufeng No.1" (SF), Zhengda (ZD) and New Myanmar (NMD). The genetic diversity and population structure of these populations were analyzed based on a dataset of 300,828 genome-wide SNP loci. The population structure analysis showed that the 10 populations shared 4 ancestors, and the domestic populations (i.e., SF, ZD, TW and NT) shared a common ancestor. The phylogenetic analysis showed that the 10 populations were grouped into 4 branches, with all the domestic populations clustering together. Estimation of fixation index (Fst) values revealed relatively lower Fst values among the domestic populations, in contrast to the more significant differences observed between domestic and foreign-origin populations. Gene flow analysis revealed three significant events of genetic exchange among the 10 populations: (1) from YN to the common ancestor of FD, TG and the domestic populations, (2) from NMD to the common ancestor of TG and FD, and (3) from the common ancestor of NMD, YN, ML and SK to ZD population. In addition, selective sweep analysis, utilizing Fst and nucleotide diversity (Pi) estimations, was conducted on representative populations (i.e., ML and SF), and 7 positively selected genes were identified, including Prospero Homeobox 1 (Prox1), Forkhead Box L1 (Foxl1), Tousled Like Kinase 2 (TLK2), G Protein Subunit Alpha 14 (Gna14), Uronyl 2-sulfotransferase (Ust), MAX Network Transcriptional Repressor (Mnt) and Importin-9 (Ipo9). These findings contribute to elucidating the breeding potential of the introduced populations and provide insights into the genetic status of the domestic breeding strains of M. rosenbergii in China.

Detection of Podosphaera macularis in Air Samples by Quantitative PCR.

Detection and quantification of pathogen propagules in the air or other environmental samples is facilitated by culture-independent assays. We developed a quantitative PCR assay for the hop powdery mildew fungus, Podosphaera macularis, for detection of the organism from air samples. The assay uses primers and a TaqMan probe designed to target species-specific sequences in the 28S large subunit of the nuclear ribosomal DNA. Analytical sensitivity was not affected by the presence of an exogenous internal control or potential PCR inhibitors associated with DNA extracted from soil. The level of quantification of the assay was between 200 and 350 conidia when DNA was extracted from a fixed number of conidia. The assay amplified all isolates of P. macularis tested and had minimal cross-reactivity with other Podosphaera species when assayed with biologically relevant quantities of DNA. Standard curves generated independently in two other laboratories indicated that assay sensitivity was qualitatively similar and reproducible. All laboratories successfully detected eight unknown isolates of P. macularis and correctly discriminated Pseudoperonospora humuli and a water control. The usefulness of the assay for air sampling for late-season inoculum of P. macularis was demonstrated in field studies in 2019 and 2020. In both years, airborne populations of P. macularis in hop yards were detected consistently and increased during bloom and cone development.

Flagellar point mutation causes social aggregation in laboratory-adapted Bacillus subtilis under conditions that promote swimming.

Motility allows microbes to explore and maximize success in their environment; however, many laboratory bacterial strains have a reduced or altered capacity for motility. Swimming motility in Bacillus subtilis depends on peritrichous flagella and is carried out individually as cells move by biased random walks toward attractants. Previously, we adapted Bacillus subtilis strain 3610 to the laboratory for 300 generations in lysogeny broth (LB) batch culture and isolated lab-adapted strains. Strain SH2 is motility-defective and in broth culture forms large, frequently spherical aggregates of cells. A single point mutation in the flagellin gene hag that causes amino acid 259 to switch from A to T is necessary and sufficient to cause these social cell aggregates, and aggregation occurs between flagellated cells bearing this point mutation regardless of the strain background. Cells associate when bearing this mutation, but flagellar rotation is needed to pull associating cells into spherical aggregates. Using electron microscopy, we are able to show that the SH2 flagellar filament has limited polymorphism when compared to other flagellar structures. This limited polymorphism hinders the flagellum's ability to function as a motility apparatus but appears to alter its function to that of cell aggregation/adhesion. We speculate that the genotype-specific aggregation of cells producing HagA259T flagella could have increased representation in a batch-culture experiment by allowing similar cells to go through a transfer together and also that this mutation could serve as an early step to evolve sociality in the natural world.IMPORTANCEThe first life forms on this planet were prokaryotic, and the earliest environments were aquatic, and from these relatively simple starting conditions, complex communities of microbes and ultimately multicellular organisms were able to evolve. Usually, motile cells in aqueous environments swim as individuals but become social by giving up motility and secreting extracellular substances to become a biofilm. Here, we identify a single point mutation in the flagellum that is sufficient to allow cells containing this mutation to specifically form large, suspended groups of cells. The specific change in the flagellar filament protein subunits causes a unique change in the flagellar structure. This could represent a distinct way for closely related cells to associate as an early precursor to sociality.

Comparative Analysis of the Mitochondrial Genome of Eggplant (Solanum melongena L.) to Identify Cytoplasmic Male Sterility Candidate Genes.

Cytoplasmic male sterility (CMS) is important for commercial hybrid seed production. However, it is still not used in eggplant (Solanum melongena L.), and corresponding regulatory genes and mechanisms of action have not been reported. We report CMS line 327A, which was derived from the hybridization between cultivated and wild eggplants. By looking at different stages of anther development under a microscope, we saw that the 327A anther's tapetum layer vacuolized during meiosis, which caused abortion. To investigate the 327A CMS regulatory genes, the mitochondrial genomes of 327A and its maintainer line 327B were assembled de novo. It was found that 15 unique ORFs (Open Reading Frame) were identified in 327A. RT-PCR and RT-QPCAR tests confirmed that orf312a and orf172a, 327A-specific ORFs with a transmembrane domain, were strongly expressed in sterile anthers of 327A. In addition, orf312a has a chimeric structure with the ribosomal protein subunit rpl16. Therefore, orf312a and orf172a can be considered strong candidate genes for CMS. Concurrently, we analyzed the characteristics of CMS to develop a functional molecular marker, CMS312, targeting a future theoretical basis for eggplant CMS three-line molecular breeding.

MiRNA-132/212 encapsulated by adipose tissue-derived exosomes worsen atherosclerosis progression

BackgroundVisceral adipose tissue in individuals with obesity is an independent cardiovascular risk indicator. However, it remains unclear whether adipose tissue influences common cardiovascular diseases, such as atherosclerosis, through its secreted exosomes.MethodsThe exosomes secreted by adipose tissue from diet-induced obesity mice were isolated to examine their impact on the progression of atherosclerosis and the associated mechanism. Endothelial apoptosis and the proliferation and migration of vascular smooth muscle cells (VSMCs) within the atherosclerotic plaque were evaluated. Statistical significance was analyzed using GraphPad Prism 9.0 with appropriate statistical tests.ResultsWe demonstrate that adipose tissue-derived exosomes (AT-EX) exacerbate atherosclerosis progression by promoting endothelial apoptosis, proliferation, and migration of VSMCs within the plaque in vivo. MicroRNA-132/212 (miR-132/212) was detected within AT-EX cargo. Mechanistically, miR-132/212-enriched AT-EX exacerbates palmitate acid-induced endothelial apoptosis via targeting G protein subunit alpha 12 and enhances platelet-derived growth factor type BB-induced VSMC proliferation and migration by targeting phosphatase and tensin homolog in vitro. Importantly, melatonin decreases exosomal miR-132/212 levels, thereby mitigating the pro-atherosclerotic impact of AT-EX.ConclusionThese data uncover the pathological mechanism by which adipose tissue-derived exosomes regulate the progression of atherosclerosis and identify miR-132/212 as potential diagnostic and therapeutic targets for atherosclerosis.Graphical

Genetic variability of Myxobolus nagaraensis (Bivalvulida: Myxobolidae) infecting freshwater gobies Rhinogobius Gill 1859 (Gobiiformes: Oxudercidae) from rivers in Japan

Myxobolus nagaraensis is a myxozoan parasite first reported in freshwater gobies (Rhinogobius spp.) from the Nagara River, Gifu Prefecture, Japan. Myxospores of M. nagaraensis form plasmodia in the visceral cavities of gobies, commonly presenting as distended abdomens. Although Rhinogobius is a common fish genus in Japan, details of M. nagaraensis, including genetic information, remain unknown. We compared the nucleotide sequences of the ribosomal RNA gene (rDNA) of M. nagaraensis from three different host species (R. fluviatilis, R. nagoyae, and R. similis) caught in three different rivers in Japan (Sakai, Sagami, and Kaname). The ITS region (ITS-1, 5.8S rDNA, and ITS-2) and large subunit (LSU) rDNA exhibited 49 and 55 variable sites, respectively. The highest nucleotide diversity was observed in the ITS region (0.00962), whereas that of the LSU rDNA was 0.00187. Differences in host species, rather than rivers, were a significant factor for genetic variation in both the ITS region (62.58%; P < 0.001) and LSU rDNA (55.22%; P < 0.01). Significant genetic variation was observed in M. nagaraensis from R. similis compared to R. fluviatilis (P < 0.001) or R. nagoyae (P < 0.001) from the same river. Such details are valuable for understanding parasite dispersal and its ecological impact on Rhinogobius hosts.

Composition and temporal dynamics of the phytoplankton community in Laizhou Bay revealed by microscopic observation and rbcL gene sequencing

Laizhou Bay, a major breeding ground for economic marine organisms in the northern waters of China, is facing rapid environmental degradation. In this study, field surveys in this area were conducted in the spring, summer, and autumn of 2020. Microscopic observation and RuBisCO large subunit (rbcL) gene analysis were employed to understand the community structure and temporal dynamics of phytoplankton. The phytoplankton community structures detected by the two methods showed significant differences. Microscopic observation revealed the dominance of dinoflagellates in spring that shifted to the dominance of diatoms in summer and autumn. However, rbcL gene sequencing consistently identified diatoms as dominant throughout all three seasons, with their relative abundance showing an increasing trend. Conversely, the relative abundance of the second- and third-most abundant taxa, namely, haptophytes and ochrophytes, decreased as the seasons transitioned. rbcL gene sequencing annotated more species than microscopy. It could detect haptophytes and cryptophytes, which were overlooked by microscopy. In addition, rbcL gene sequencing detected a remarkable amount of Thalassiosira profunda, which was previously unidentified in this sea area. However, it appeared to underestimate the contribution of dinoflagellates considerably, with most taxa being only identified through microscopic identification. The two methods jointly identified 28 harmful algal bloom taxa with similar detection quantities but substantial differences in species composition. Phytoplankton communities were influenced by temperature, salinity, and nutrients. The results of this work suggest that a combination of multiple techniques is necessary for a comprehensive understanding of phytoplankton.

Developing the script "degenerate primer 111" to enhance the coverage of universal primers for the small subunit rRNA gene on target microorganisms.

Amplifying small subunit (SSU) rRNA genes with universal primers in assessing microbial populations diversity, but target microorganisms are sometimes omitted due to inadequate primer coverage. Adding degenerate bases to primers can help, but existing methods are complex and time-consuming. This study introduces a user-friendly tool called "Degenerate primer 111" for adding degenerate bases to existing universal primers. By aligning one universal primer with one uncovered target microorganism's SSU rRNA gene, this tool iteratively generates a new primer, maximizing coverage for the target microorganisms. The tool was used to modify eight pairs of universal primers (515F Parada-806R Apprill, S-D-Bact-0341-b-S-17/S-D-Bact-0785-a-A-21, OP_F114-KP_R013, 27F-1492R, 341F-806R, OP_F066-KP_R013, 515F Parada-926R Quince, 616*F-1132R), and generated 29 new universal primers with increased coverage of specific target microorganisms without increasing coverage of non-target microorganisms. To verify the effectiveness of the improved primers, one set of original and improved primers (BA-515F-806R and BA-515F-806R-M1) was used to amplify DNA from the same sample, and high-throughput sequencing of the amplicons confirmed that the improved primers detected more microbial species compared to the original primers. Future researchers can use this tool to develop more personalized primers to meet their diverse microorganism detection needs.

PRL1 interacts with and stabilizes RPA2A to regulate carbon deprivation-induced senescence in Arabidopsis.

Leaf senescence is a developmental program regulated by both endogenous and environmental cues. Abiotic stresses such as nutrient deprivation can induce premature leaf senescence, which profoundly impacts plant growth and crop yield. However, the molecular mechanisms underlying stress-induced senescence are not fully understood. In this work, employing a carbon deprivation (C-deprivation)-induced senescence assay in Arabidopsis seedlings, we identified PLEIOTROPIC REGULATORY LOCUS 1 (PRL1), a component of the NineTeen Complex, as a negative regulator of C-deprivation-induced senescence. Furthermore, we demonstrated that PRL1 directly interacts with the RPA2A subunit of the single-stranded DNA-binding Replication Protein A (RPA) complex. Consistently, the loss of RPA2A leads to premature senescence, while increased expression of RPA2A inhibits senescence. Moreover, overexpression of RPA2A reverses the accelerated senescence in prl1 mutants, and the interaction with PRL1 stabilizes RPA2A under C-deprivation. In summary, our findings reveal the involvement of the PRL1-RPA2A functional module in C-deprivation-induced plant senescence.

Збудники інфекційних хвороб людини і тварин та їх розповсюдження в Європі іксодовим кліщем Dermacentor reticulatus

Due to climate change, the possibility of human infective diseases caused by pathogens from the genera Borelia, Anaplasma, Rickettsia, Babesia, Bartonella, Coxiella, etc. is increasing in Europe. Generally, these pathogens are directly related to arthropod vectors, in particular, to Ixodes ticks. The aim of the study was to determine the current trends in the spread of tick-borne infections transmitted by Dermacentor reticulatus ticks in some European countries, including Ukraine, based on the analysis of literature. Over the past 10 years, in the vast majority of countries, including Europe, the situation with the defeat of farm and wild animals, as well as humans, by pathogens of severe infectious diseases from the genera Borrelia, Anaplasma, Rickettsia, Babesia, Theileria, Coxiella, Bartonella, Francisella, etc. has been aggravated. Representatives of these genera lead to severe diseases, such as tick-borne borreliosis, which are accompanied by damage to the central nervous system, joints, skin, and cardiovascular system. Members of the Rickettsiales genus cause a variety of feverish conditions in animals and humans. Species of the genus Babesia cause babesiosis, a disease caused by damage to red blood cells and the appearance of anti-erythrocyte antibodies, and representatives of the genus Toxoplasma cause toxoplasmosis, accompanied by damage to the nervous, lymphatic, and cardiovascular systems. Q fever is caused by species of the genus Coxiella. Severe human diseases are caused by tick-borne encephalitis viruses. Various PCR test systems have been developed and used to detect these pathogens. For example, in Poland, PCR for Borrelia burgdorferi s.l. was performed using the B. burgdorferi s.l. PCR kit (GeneProof, Czech Republic) for in vitro diagnosis. Anaplasma PCR kits (Blirt-DNA Gdańsk, Poland) were used to detect Anaplasma phagocytophilum DNA. For the detection of Babesia spp., a fragment of the 18S rDNA gene encoding a small ribosomal subunit localized in the conserved region V4 was used. To detect Bartonella spp., the tests Hum PCR Bartonella, Hum PCR Coxiella burnetii, and Vet PCR Rickettsia kit (BioIngenTech, Chile), Coxiella burnetii and Rickettsia spp. were used. In the Laboratory for the Study of Ticks and Tick-Borne Infections of Ternopil National Medical University (Ukraine), pathogens were identified using several test systems, namely DNA A. phagocytophilum, / DNA E. muris, / DNA E. chaffeensis, DNA B. burgdorferi s.l., DNA B. myamotoi; DNA Babesia sp., and RNA Ticks borne of encephalitis viruses (MolGen, Italy), which allow real-time determination on anamplifier "ROTOR Gene 6000" by Corbett Research. Studies in many European countries have shown infection of the Dermacentor reticulatus tick with the pathogens B. burgdorferi s.l. in the range from 1.6 to 49.8 %; A. phagocytophilum – 0.1–2.2 %; Babesia sрp. – 9.8–43.8 %; Coxiella spр. – 0.1–0.2%; Francisella spp. – 0.7–95%; Toxoplasma spр. – 3.2–16.7 %; and ticks borne of encephalitis viruses 8.5–16.6 %. Some Ukrainian scientists presented data on D. reticulatus infection, in particular B. burgdorferi s. l. – 10.1–22.1 %; A. phagocytophilum – 3–25.36 %; Babesia spр. – 1–41.2 %; Rickettsia sp. – 28 %; Bartonella spp. – 5 %; Coxiella spр. – 5.1 %, and tick-borne f encephalitis viruses – 11.9–71 %. The spread of many infectious agents is largely correlated with the spread of their vectors, including ticks of the D. reticulatus species. According to many scientists, this species is widespread in Europe, in particular, two metapopulations are currently distinguished as the western one, which includes the territories of France, Belgium, Slovakia, the Czech Republic, the Netherlands, and Germany, and the eastern one, which covers Lithuania, Latvia, Belarus, eastern and central Poland, and west of the Vistula River up to the Ural Mountains. In the Czech Republic, this species is found in all regions, more often in the southeastern and northwestern regions. In the UK, ticks have been found in West Wales, Devon, and Essex. In Romania, D. reticulatus was found in 17 counties, and in Moldova – in the Lower Dniester region. In Ukraine, this species is widespread from the Carpathians and Prykarpattia, Polissya, and Forest-Steppe, to the southeastern regions and Crimea.