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This article summarises the activities of the Bacterial Viruses Subcommittee of the International Committee on Taxonomy of Viruses for the period of March 2021−March 2022. We provide an overview of the new taxa proposed in 2021, approved by the Executive Committee, and ratified by vote in 2022. Significant changes to the taxonomy of bacterial viruses were introduced: the paraphyletic morphological families Podoviridae, Siphoviridae, and Myoviridae as well as the order Caudovirales were abolished, and a binomial system of nomenclature for species was established. In addition, one order, 22 families, 30 subfamilies, 321 genera, and 862 species were newly created, promoted, or moved.

Background Characterizing phage–host interactions is critical to understanding the ecological role of both partners and effective isolation of phage therapeuticals. Unfortunately, experimental methods for studying these interactions are markedly slow, low-throughput, and unsuitable for phages or hosts difficult to maintain in laboratory conditions. Therefore, a number of in silico methods emerged to predict prokaryotic hosts based on viral sequences. One of the leading approaches is the application of the BLAST tool that searches for local similarities between viral and microbial genomes. However, this prediction method has three major limitations: (i) top-scoring sequences do not always point to the actual host; (ii) mosaic virus genomes may match to many, typically related, bacteria; and (iii) viral and host sequences may diverge beyond the point where their relationship can be detected by a BLAST alignment. Results We created an extension to BLAST, named Phirbo, that improves host prediction quality beyond what is obtainable from standard BLAST searches. The tool harnesses information concerning sequence similarity and bacteria relatedness to predict phage–host interactions. Phirbo was evaluated on three benchmark sets of known virus–host pairs, and it improved precision and recall by 11–40 percentage points over currently available, state-of-the-art, alignment-based, alignment-free, and machine-learning host prediction tools. Moreover, the discriminatory power of Phirbo for the recognition of virus–host relationships surpassed the results of other tools by at least 10 percentage points (area under the curve = 0.95), yielding a mean host prediction accuracy of 57% and 68% at the genus and family levels, respectively, and drops by 12 percentage points when using only a fraction of viral genome sequences (3 kb). Finally, we provide insights into a repertoire of protein and ncRNA genes that are shared between phages and hosts and may be prone to horizontal transfer during infection. Conclusions Our results suggest that Phirbo is a simple and effective tool for predicting phage–host relationships.

BackgroundBacteriophages from the Bastillevirinae subfamily have proven effective against bacteria from the Bacillus genus including organisms from B. cereus group which causes food poisoning and persistent contamination of industrial installations. However, successful application of these phages in biocontrol depends on understanding of their biology and stability in different environments.MethodsIn this study we isolated a novel virus from the garden soil in Wrocław (Poland) and named it Thurquoise. Genome of the phage was sequenced using Illumina technology and assembled as a single continuous contig that represents consensus result of different assembly algorithms. Its morphology was determined using Cryo-EM imaging while the dynamics of replication by turbidimetric lysis assay. We also determined phage host range in the efficiency of plating (EOP) assay. Finally, the stability of the Thurquoise was tested by long term incubation in buffers containing different ions (Mg2+, Ca2+, Mn2+, Zn2+, Cu2+, K+, Co2+, Fe3+, Cs+) and freezing with various cryoprotectants (glycerol, gelatin, saccharose, trehalose).ResultsWe present a complete, carefully annotated genome of the Thurquoise phage with 226 identified protein genes and 18 tRNAs. The complex virion structure of this phage is typical for the Bastillevirinae family. Confirmed hosts include selected bacteria from the Bacillus cereus group - specifically B. thuringiensis and B. mycoides. Latent and eclipse periods of Thurquoise in the isolation host last ~40 min and ~50 min respectively. The phage remains viable for more than 8 weeks in variants of SM buffer with magnesium, calcium, cesium, manganese or potassium and can withstand numerous freeze-thaw cycles if protected by the addition of 15% glycerol or, to a lesser extent, 2% gelatin.ConclusionThe Thurquoise phage is the exemplar of the new candidate species in Caruleovirus genus in the Bastillevirinae subfamily of the Herelleviridae family with genome, morphology and biology typical for these taxa.With proper buffer formulation, this virus (and likely related phages) can be safely stored in common freezers and refrigerators for a considerable time.

Tailed bacteriophages are the most abundant and diverse viruses in the world, with genome sizes ranging from 10 kbp to over 500 kbp. Yet, due to historical reasons, all this diversity is confined to a single virus order—Caudovirales, composed of just four families: Myoviridae, Siphoviridae, Podoviridae, and the newly created Ackermannviridae family. In recent years, this morphology-based classification scheme has started to crumble under the constant flood of phage sequences, revealing that tailed phages are even more genetically diverse than once thought. This prompted us, the Bacterial and Archaeal Viruses Subcommittee of the International Committee on Taxonomy of Viruses (ICTV), to consider overall reorganization of phage taxonomy. In this study, we used a wide range of complementary methods—including comparative genomics, core genome analysis, and marker gene phylogenetics—to show that the group of Bacillus phage SPO1-related viruses previously classified into the Spounavirinae subfamily, is clearly distinct from other members of the family Myoviridae and its diversity deserves the rank of an autonomous family. Thus, we removed this group from the Myoviridae family and created the family Herelleviridae—a new taxon of the same rank. In the process of the taxon evaluation, we explored the feasibility of different demarcation criteria and critically evaluated the usefulness of our methods for phage classification. The convergence of results, drawing a consistent and comprehensive picture of a new family with associated subfamilies, regardless of method, demonstrates that the tools applied here are particularly useful in phage taxonomy. We are convinced that creation of this novel family is a crucial milestone toward much-needed reclassification in the Caudovirales order., Systematic Biology, 69 (1), ISSN:1063-5157, ISSN:1076-836X

Thaumatin-like proteins (TLPs, osmotins) form a protein family which shares a significant sequence homology to the sweet-tasting thaumatin from the plant Thaumatococcus daniellii. TLPs are not sweet-tasting and are involved in response to biotic stresses and developmental processes. Recently it has been shown using a proteomic approach that the tuber extract from Corydalis cava (Papaveraceae) contains a TLP protein. The aim of this work was to characterize the structure and expression of TLP from C. cava tubers. The results obtained using a PCR approach with degenerate primers demonstrated a coding sequence of a novel protein, named CcTLP1. It consists of 225 aa, has a predicted molecular weight of 24.2 kDa (NCBI GenBank accession no. KJ513303 ) and has 16 strictly conserved cysteine residues, which form 8 disulfide bridges and stabilize the 3D structure. CcTLP1 may be classified into class IX of plant TLPs. The highest CcTLP1 expression levels were shown by qPCR in the stem of the plant compared to other organs and in the medium-size plants compared to other growth phases. The results confirm that CcTLP1 is expressed during plant growth and development until flowering, with a possible defensive function against different stress conditions.

Viruses of prokaryotes are extremely abundant and diverse. Culture-independent approaches have recently shed light on the biodiversity these biological entities1,2. One fundamental question when trying to understand their ecological roles is: which host do they infect? To tackle this issue we developed a machine-learning approach named Random Forest Assignment of Hosts (RaFAH), based on the analysis of nearly 200,000 viral genomes. RaFAH outperformed other methods for virus-host prediction (F1-score = 0.97 at the level of phylum). RaFAH was applied to diverse datasets encompassing genomes of uncultured viruses derived from eight different biomes of medical, biotechnological, and environmental relevance, and was capable of accurately describing these viromes. This led to the discovery of 537 genomic sequences of archaeal viruses. These viruses represent previously unknown lineages and their genomes encode novel auxiliary metabolic genes, which shed light on how these viruses interfere with the host molecular machinery. RaFAH is available at https://sourceforge.net/projects/rafah/.

Members of the family Herelleviridae are bacterial viruses infecting members of the phylum Firmicutes. The virions have myovirus morphology and virus genomes comprise a linear dsDNA of 125–170 kb. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Herelleviridae, which is available at ictv.global/report/herelleviridae.

Bacteriophage therapeutic development will clearly benefit from understanding the fundamental dynamics of in vivo phage-bacteria interactions. Such information can inform animal and human trials, and much can be ascertained from human cell-line work. We have developed a human cell-based system using Clostridium difficile, a pernicious hospital pathogen with limited treatment options, and the phage phiCDHS1 that effectively kills this bacterium in liquid culture. The human colon tumorigenic cell line HT-29 was used because it simulates the colon environment where C. difficile infection occurs. Studies on the dynamics of phage-bacteria interactions revealed novel facets of phage biology, showing that phage can reduce C. difficile numbers more effectively in the presence of HT-29 cells than in vitro. Both planktonic and adhered Clostridial cell numbers were successfully reduced. We hypothesise and demonstrate that this observation is due to strong phage adsorption to the HT-29 cells, which likely promotes phage-bacteria interactions. The data also showed that the phage phiCDHS1 was not toxic to HT-29 cells, and phage-mediated bacterial lysis did not cause toxin release and cytotoxic effects. The use of human cell lines to understand phage-bacterial dynamics offers valuable insights into phage biology in vivo, and can provide informative data for human trials.

In this paper, we describe two independent isolates of a new member of the subfamily Autographivirinae, Pseudomonas phage KNP. The type strain (KNP) has a linear, 40,491-bp-long genome with GC content of 57.3%, and 50 coding DNA sequences (CDSs). The genome of the second strain (WRT) contains one CDS less, encodes a significantly different tail fiber protein and is shorter (40,214 bp; GC content, 57.4%). Phylogenetic analysis indicates that both KNP and WRT belong to the genus T7virus. Together with genetically similar Pseudomonas phages (gh-1, phiPSA2, phiPsa17, PPPL-1, shl2, phi15, PPpW-4, UNO-SLW4, phiIBB-PF7A, Pf-10, and Phi-S1), they form a divergent yet coherent group that stands apart from the T7-like viruses (sensu lato). Analysis of the diversity of this group and its relatedness to other members of the subfamily Autographivirinae led us to the conclusion that this group might be considered as a candidate for a new genus. Electronic supplementary material The online version of this article (doi:10.1007/s00705-017-3419-9) contains supplementary material, which is available to authorized users.

Members of the family Herelleviridae are bacterial viruses infecting members of the phylum Firmicutes. The virions have myovirus morphology and virus genomes comprise a linear dsDNA of 125-170 kb. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Herelleviridae, which is available at ictv.global/report/herelleviridae.

Microcystins produced by several toxic cyanobacterial strains constitute an important problem for public health. Bacterial degradation of these hepatotoxins may play an important role in natural ecosystems, however the nature of the process is very poorly understood. The aim of our study was to investigate the possible interactions between cyanotoxin producers and degraders. Samples collected from 24 water bodies in western Poland were analysed to determine the chemo-physical parameters, phytoplankton content, bacterial community structure and microcystin-biodegradation potency. A redundancy analysis identified a positive correlation between the capacity of a community to degrade microcystin LR (MC-LR) and temperature, pH, chlorophyll a concentration and the abundance of MC-producers. The relative abundance of classes F38, TM7-3 and the order WCHB1-81c (Actinobacteria) was significantly higher in the lakes with MC-biodegradation potency. Some specific bacterial genera belonging to Acidobacteria, Chloroflexi, Gemmatimonadetes, Firmicutes and TM7 were closely correlated with the occurrence of Microcystis spp. Furthermore, the MC biodegradation process was connected with the same bacterial groups. Thus, our approach allowed us to provide a broader picture of some specific relations between microcystin producers and potential microcystin degraders. A more comprehensive analysis of the existing correlations may be helpful in our understanding of natural mechanisms of MC elimination using bacteria such as MC-degraders.

Taxonomic status of endangered peat-bog pine, Pinus uliginosa (Neumann) classified within the Pinus mugo complex, still remains to be elucidated. Here we present a complete chloroplast genome of P. uliginosa, to aid resolve its complex systematical position. The total genome size was 119,877 bp in length and contained a total of 112 genes, including 73 protein-coding genes, 35 tRNAs, and four rRNAs. The most of genes occur as a single copy. Five tRNA genes were duplicated from two to four times. Eighteen genes contain one intron, with a single gene containing two introns. No large inverted repeats were identified. The overall G + C content of P. uliginosa chloroplast genome is 38.5%.

Uncoupling proteins (UCPs) mediate inducible proton conductance in the mitochondrial inner membrane. Herein, we summarize our knowledge regarding UCPs in invertebrates. Since 2001, the presence of UCPs has been demonstrated in nematodes, mollusks, amphioxi, and insects. We discuss the following important issues concerning invertebrate UCPs: their evolutionary relationships, molecular and functional properties, and physiological impact. Evolutionary analysis indicates that the branch of vertebrate and invertebrate UCP4-5 diverged early in the evolutionary process prior to the divergence of the animal groups. Several proposed physiological roles of invertebrate UCPs are energy control, metabolic balance, and preventive action against oxidative stress. © 2016 IUBMB Life, 68(9):691–699, 2016

Main conclusion A novel annotated Chelidonium majus L. transcriptome database composed of 23,004 unique coding sequences allowed to significantly improve the sensitivity of proteomic C. majus assessments, which showed novel defense-related proteins characteristic to its latex. To date, the composition of Chelidonium majus L. milky sap and biosynthesis of its components are poorly characterized. We, therefore, performed de novo sequencing and assembly of C. majus transcriptome using Illumina technology. Approximately, 119 Mb of raw sequence data was obtained. Assembly resulted in 107,088 contigs, with N50 of 1913 bp and N90 of 450 bp. Among 34,965 unique coding sequences (CDS), 23,004 obtained CDS database served as a basis for further proteomic analyses. The database was then used for the identification of proteins from C. majus milky sap, and whole plant extracts analyzed using liquid chromatography–electrospray ionization-tandem mass spectrometry (LC–ESI-MS/MS) approach. Of about 334 different putative proteins were identified in C. majus milky sap and 1155 in C. majus whole plant extract. The quantitative comparative analysis confirmed that C. majus latex contains proteins connected with response to stress conditions and generation of precursor metabolites and energy. Notable proteins characteristic to latex include major latex protein (MLP, presumably belonging to Bet v1-like superfamily), polyphenol oxidase (PPO, which could be responsible for browning of the sap after exposure to air), and enzymes responsible for anthocyanidin, phenylpropanoid, and alkaloid biosynthesis. Electronic supplementary material The online version of this article (doi:10.1007/s00425-016-2566-7) contains supplementary material, which is available to authorized users.

Efficient delivery of heterologous molecules for treatment of cells is a great challenge in modern medicine and pharmacology. Cell-penetrating peptides (CPPs) may improve efficient delivery of a wide range of macromolecular cargos, including plasmid DNA, small interfering RNA, drugs, nanoparticulate pharmaceutical carriers, and anticancer drugs. In this paper, we present the history of CPPs' discovery with special attention drawn to sequences of viral origin. We also describe different CPP families with regard to their physicochemical properties and numerous mechanisms of CPP cell uptake by direct penetration and endocytotic pathways. A detailed description is focused on formation of carrier-cargo complexes, which are needed for practical use of CPPs in medicine and biotechnology. Examples of successful application of CPPs in treatment of human diseases are also presented, including decreased tumor growth and induction of cancer cell death. Finally, we review modern design approaches to novel CPPs and prediction of their activity. To sum up, the current review presents a thorough and up-to-date knowledge of CPPs and may be a valuable source of information for researchers in pharmacology designing new therapeutic agents.

It is almost a cliché that tailed bacteriophages of the order Caudovirales are the most abundant and diverse viruses in the world. Yet, their taxonomy still consists of a single order with just three families: Myoviridae, Siphoviridae, and Podoviridae. Thousands of newly discovered phage genomes have recently challenged this morphology-based classification, revealing that tailed bacteriophages are genomically even more diverse than once thought. Here, we evaluate a range of methods for bacteriophage taxonomy by using a particularly challenging group as an example, the Bacillus phage SPO1-related viruses of the myovirid subfamily Spounavirinae. Exhaustive phylogenetic and phylogenomic analyses indicate that the spounavirins are consistent with the taxonomic rank of family and should be divided into at least five subfamilies. This work is a case study for virus genomic taxonomy and the first step in an impending massive reorganization of the tailed bacteriophage taxonomy.

Bacteriophage therapeutic development will clearly benefit from understanding the fundamental dynamics of in vivo phage-bacteria interactions. Such information can inform animal and human trials, and much can be ascertained from human cell-line work. We have developed a human cell-based system using Clostridium difficile, a pernicious hospital pathogen with limited treatment options, and the phage phiCDHS1 that effectively kills this bacterium in liquid culture. The human colon tumorigenic cell line HT-29 was used because it simulates the colon environment where C. difficile infection occurs. Studies on the dynamics of phage-bacteria interactions revealed novel facets of phage biology, showing that phage can reduce C. difficile numbers more effectively in the presence of HT-29 cells than in vitro. Both planktonic and adhered Clostridial cell numbers were successfully reduced. We hypothesise and demonstrate that this observation is due to strong phage adsorption to the HT-29 cells, which likely promotes phage-bacteria interactions. The data also showed that the phage phiCDHS1 was not toxic to HT-29 cells, and phage-mediated bacterial lysis did not cause toxin release and cytotoxic effects. The use of human cell lines to understand phage-bacterial dynamics offers valuable insights into phage biology in vivo, and can provide informative data for human trials.

Oxyrrhynchium speciosum (Brid.) Warnst. is a relatively uncommon moss that can be found in wetland habitats across Europe and New Zealand. In this paper, we address the morphological, molecular and ecological relationships between the aquatic form of O. speciosum discovered in a deep, Chara-dominated lake (Budzislawskie, NW Poland) and its terrestrial relatives. Morphological assessments revealed a considerable difference between terretrial and deep-water samples of O.speciosum. The aquatic form of O. speciosum was, on average, 12 times larger, with more branched stems (4–30 times) and longer leaves (1–2 times).Investigation of ITS1-5·8S-ITS2-26S region sequences, however, revealed no distinction between the terrestrial and aquatic forms. An analysis of the ecological data suggests that the presence of O. speciosum in deep lake habitats is not incidental (as with many land species developing in aquatic environments). On the contrary, aquatic forms have a distinct ecological niche. Our results conf...

In the course of the nanoLC-nanoESI-MS/MS analysis of Chelidonium majus proteins we detected an extremely abundant 12 aa peptide (sequence: TNAENEFVTIKK). The Mascot search against NCBInr database with Viridiplantae taxonomic restriction revealed its complete identity to Unknown protein 18 from Pseudotsuga menziesii (P85925). The same sequence has also been submitted as Unknown protein 1 (P86104) from Vitis rotundifolia and as a part of the oxygen-evolving enhancer protein 1 (OEE1) from Pinus strobus (P84718). After careful inspection the record P84718 turned out to comprise a set of peptides of unconfirmed origin rather than complete protein sequence. In this paper we present extensive data indicating that the peptide in question may originate from type II cytoskeletal keratin - a common contaminant in protein samples. We found empirical evidence that it can be detected in several types of keratin-contaminated samples and its sequence is identical to one of the proteotypic peptides commonly observed for keratins. Nevertheless, the peptide has been annotated as plant protein and thus leads to data misinterpretation. We advise extreme caution when dealing with sequences of Unknown protein 18 (P. menziesii), Unknown protein 1 (V. rotundifolia) and OEE1 (P. strobus) since they are at best poorly annotated, if not artifactual. Biological significance To our knowledge, this is the first report indicating that the peptide TNAENEFVTIKK, which is identical to Unknown protein 18 from P. menziesii (P85925), Unknown protein 1 (P86104) from V. rotundifolia and a part of the oxygen-evolving enhancer protein 1 (OEE1) from P. strobus (P84718), may originate from type II cytoskeletal keratin - a common contaminant in protein samples. We found empirical evidence that it can be detected in several types of keratin-contaminated samples and its sequence is identical to one of the proteotypic peptides commonly observed for keratins. Nevertheless, the peptide has been annotated as plant protein and thus, without proper quality assessment, leads to biological misinterpretation of proteomic data.

Uncoupling proteins (UCPs) mediate inducible proton conductance in the mitochondrial inner membrane. Herein, we summarize our knowledge regarding UCPs in invertebrates. Since 2001, the presence of UCPs has been demonstrated in nematodes, mollusks, amphioxi, and insects. We discuss the following important issues concerning invertebrate UCPs: their evolutionary relationships, molecular and functional properties, and physiological impact. Evolutionary analysis indicates that the branch of vertebrate and invertebrate UCP4-5 diverged early in the evolutionary process prior to the divergence of the animal groups. Several proposed physiological roles of invertebrate UCPs are energy control, metabolic balance, and preventive action against oxidative stress. © 2016 IUBMB Life, 68(9):691-699, 2016.

sp. (based on com-plete sequences of 16S rDNA as well as genes encoding hsp 60GroELchaperonin, -subunitsofRNApolymerase,andDNAgy-rase).Thegenomewassequencedusingthe454methodinGenomedSA(Warsaw,Poland)andassembledusingNewblerversion2.5.3(454LifeSciences)andGeneiousversion6.1(6)(64.98-foldcov-erage).ProteincodinggeneswerepredictedusingGeneMarkS(7)and Prodigal version 2.60 (8). Their functional annotation wasperformed based on BLASTx hits against the UniprotKB virusesdatabase (9). To search for tRNA genes we used tRNAscan-SEversion 1.21 (10). Mapping of pyrosequencing reads that allowsdetermination of genome ends was performed with the Pausepipeline (https://cpt.tamu.edu/computer-resources/pause/).phD2BisanSP6-likepodophage.Thecompletegenomeofthephageis44,366bpinlengthwithaG Ccontentof51%andendswith direct repeats (262 bp). We identified 49 coding sequences,all on one strand.The arrangement of the modules seems to roughly reflect theorder of expression. The left end of the genome contains a groupof genes involved in the takeover of a host metabolism (

The Bacillus phage phiAGATE is a novel myovirus isolated from the waters of Lake Goreckie (a eutrophic lake in western Poland). The bacteriophage infects Bacillus pumilus, a bacterium commonly observed in the mentioned reservoir. Analysis of the phiAGATE genome (149844 base pairs) resulted in 204 predicted protein-coding sequences (CDSs), of which 53 could be functionally annotated. Further investigation revealed that the bacteriophage is a member of a previously undescribed cluster of phages (for the purposes of this study we refer to it as “Bastille group”) within the Spounavirinae subfamily. Here we demonstrate that these viruses constitute a distinct branch of the Spounavirinae phylogenetic tree, with limited similarity to phages from the Twortlikevirus and Spounalikevirus genera. The classification of phages from the Bastille group into any currently accepted genus proved extremely difficult, prompting concerns about the validity of the present taxonomic arrangement of the subfamily.

Plant antimicrobial peptides (AMPs) are a component of barrier defense system of plants. They have been isolated from roots, seeds, flowers, stems, and leaves of a wide variety of species and have activities towards phytopathogens, as well as against bacteria pathogenic to humans. Thus, plant AMPs are considered as promising antibiotic compounds with important biotechnological applications. Plant AMPs are grouped into several families and share general features such as positive charge, the presence of disulfide bonds (which stabilize the structure), and the mechanism of action targeting outer membrane structures.

The occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) in freshwater reservoirs is a common phenomenon. However, the biodegradation of this toxin in environmental samples has been observed only occasionally. In this work the biodegradation ability of cylindrospermopsin was investigated based on isolates from lakes with previous cyanotoxin history. Bacterial strains were identified based on the 16S rDNA and rpoD gene comparison. CYN biodegradation was monitored using the HPLC method. The R6 strain identified as Aeromonas sp. was documented as being capable of CYN removal. This biodegradation was dependent on the pH and temperature. Additionally, the stimulation of the growth of the R6 strain in the presence of CYN was indicated. Our discovery supports the hypothesis that (in analogy to the well-known phenomenon of microcystin biodegradation) in lakes dominated by potential CYN-producing cyanobacteria, the processes of microbial utilization of this toxin may occur.