Revisiting the origins of the Sobemovirus genus: A case for ancient origins of plant viruses.

The discrepancy between short- and long-term rate estimates, known as the time-dependent rate phenomenon (TDRP), poses a challenge to extrapolating evolutionary rates over time and reconstructing evolutionary history of viruses. The TDRP reveals a decline in evolutionary rate estimates with the measurement timescale, explained empirically by a power-law rate decay, notably observed in animal and human viruses. A mechanistic evolutionary model, the Prisoner of War (PoW) model, has been proposed to address TDRP in viruses. Although TDRP has been studied in animal viruses, its impact on plant virus evolutionary history remains largely unexplored. Here, we investigated the consequences of TDRP in plant viruses by applying the PoW model to reconstruct the evolutionary history of sobemoviruses, plant pathogens with significant importance due to their impact on agriculture and plant health. Our analysis showed that the Sobemovirus genus dates back over four million years, indicating an ancient origin. We found evidence that supports deep host jumps to Poaceae, Fabaceae, and Solanaceae occurring between tens to hundreds of thousand years ago, followed by specialization. Remarkably, the TDRP-corrected evolutionary history of sobemoviruses was extended far beyond previous estimates that had suggested their emergence nearly 9,000 years ago, a time coinciding with the Neolithic period in the Near East. By incorporating sequences collected through metagenomic analyses, the resulting phylogenetic tree showcases increased genetic diversity, reflecting a deep history of sobemovirus species. We identified major radiation events beginning between 4,600 to 2,000 years ago, which aligns with the Neolithic period in various regions, suggesting a period of rapid diversification from then to the present. Our findings make a case for the possibility of deep evolutionary origins of plant viruses.

Sequencing and biological characterization of historical cynosurus mottle virus isolates from Germany.

We report sequencing of four historical cynosurus mottle virus (CnMoV) isolates, originating from different hosts and locations. The CnMoV genome, ranging from 4417 to 4419 nt, encodes five ORFs. It shares 48.1% nucleotide sequence identity with cocksfoot mottle virus and 69.8% with the recently discovered Poaceae Liege sobemovirus. Phylogenetic analysis supports classification within the genus Sobemovirus. Sequenced CnMoV isolates exhibit 96.4-99.9% identity. Nucleotide substitutions leading to amino acid changes showed no host associations. However, amino acid changes in the coat protein appear to be linked to differences in serological properties. Aphid transmission tests confirmed non-transmissibility, consistent with earlier observations for the English isolate.

ALMT-independent guard cell R-type anion currents.

Plant transpiration is controlled by stomata, with S- and R-type anion channels playing key roles in guard cell action. Arabidopsis mutants lacking the ALMT12/QUAC1 R-type anion channel function in guard cells show only a partial reduction in R-type channel currents. The molecular nature of these remaining R-type anion currents is still unclear. To further elucidate this, patch clamp, transcript and gas-exchange measurements were performed with wild-type (WT) and different almt mutant plants. The R-type current fraction in the almt12 mutant exhibited the same voltage dependence, susceptibility to ATP block and lacked a chloride permeability as the WT. Therefore, we asked whether the R-type anion currents in the ALMT12/QUAC1-free mutant are caused by additional ALMT isoforms. In WT guard cells, ALMT12, ALMT13 and ALMT14 transcripts were detected, whereas only ALMT13 was found expressed in the almt12 mutant. Substantial R-type anion currents still remained active in the almt12/13 and almt12/14 double mutants as well as the almt12/13/14 triple mutant. In good agreement, CO2 -triggered stomatal closure required the activity of ALMT12 but not ALMT13 or ALMT14. The results suggest that, with the exception of ALMT12, channel species other than ALMTs carry the guard cell R-type anion currents.

CRISPR/Cas-mediated plant genome editing: outstanding challenges a decade after implementation.

The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding of the plant genome. CRISPR/Cas has been used for over a decade to modify plant genomes for the study of specific genes and biosynthetic pathways as well as to speed up breeding in many plant species, including both model and non-model crops. Although the CRISPR/Cas system is very efficient for genome editing, many bottlenecks and challenges slow down further improvement and applications. In this review we discuss the challenges that can occur during tissue culture, transformation, regeneration, and mutant detection. We also review the opportunities provided by new CRISPR platforms and specific applications related to gene regulation, abiotic and biotic stress response improvement, and de novo domestication of plants.

Improving abiotic stress tolerance of forage grasses - prospects of using genome editing.

Due to an increase in the consumption of food, feed, and fuel and to meet global food security needs for the rapidly growing human population, there is a necessity to obtain high-yielding crops that can adapt to future climate changes. Currently, the main feed source used for ruminant livestock production is forage grasses. In temperate climate zones, perennial grasses grown for feed are widely distributed and tend to suffer under unfavorable environmental conditions. Genome editing has been shown to be an effective tool for the development of abiotic stress-resistant plants. The highly versatile CRISPR-Cas system enables increasingly complex modifications in genomes while maintaining precision and low off-target frequency mutations. In this review, we provide an overview of forage grass species that have been subjected to genome editing. We offer a perspective view on the generation of plants resilient to abiotic stresses. Due to the broad factors contributing to these stresses the review focuses on drought, salt, heat, and cold stresses. The application of new genomic techniques (e.g., CRISPR-Cas) allows addressing several challenges caused by climate change and abiotic stresses for developing forage grass cultivars with improved adaptation to the future climatic conditions. Genome editing will contribute towards developing safe and sustainable food systems.

An Epigenetic Alphabet of Crop Adaptation to Climate Change.

Crop adaptation to climate change is in a part attributed to epigenetic mechanisms which are related to response to abiotic and biotic stresses. Although recent studies increased our knowledge on the nature of these mechanisms, epigenetics remains under-investigated and still poorly understood in many, especially non-model, plants, Epigenetic modifications are traditionally divided into two main groups, DNA methylation and histone modifications that lead to chromatin remodeling and the regulation of genome functioning. In this review, we outline the most recent and interesting findings on crop epigenetic responses to the environmental cues that are most relevant to climate change. In addition, we discuss a speculative point of view, in which we try to decipher the "epigenetic alphabet" that underlies crop adaptation mechanisms to climate change. The understanding of these mechanisms will pave the way to new strategies to design and implement the next generation of cultivars with a broad range of tolerance/resistance to stresses as well as balanced agronomic traits, with a limited loss of (epi)genetic variability.

ICTV Virus Taxonomy Profile: Solemoviridae 2021.

The family Solemoviridae includes viruses with icosahedral particles (26-34 nm in diameter) assembled on T=3 symmetry with a 4-6 kb positive-sense, monopartite, polycistronic RNA genome. Transmission of members of the genera Sobemovirus and Polemovirus occurs via mechanical wounding, vegetative propagation, insect vectors or abiotically through soil; members of the genera Polerovirus and Enamovirus are transmitted by specific aphids. Most solemoviruses have a narrow host range. This is a summary of the International Committee on Taxonomy of Viruses (ICTV) Report on the family Solemoviridae, which is available at ictv.global/report/solemoviridae.

ABCE1 is essential for S phase progression in human cells.

ABCE1 is a highly conserved protein universally present in eukaryotes and archaea, which is crucial for the viability of different organisms. First identified as RNase L inhibitor, ABCE1 is currently recognized as an essential translation factor involved in several stages of eukaryotic translation and ribosome biogenesis. The nature of vital functions of ABCE1, however, remains unexplained. Here, we study the role of ABCE1 in human cell proliferation and its possible connection to translation. We show that ABCE1 depletion by siRNA results in a decreased rate of cell growth due to accumulation of cells in S phase, which is accompanied by inefficient DNA synthesis and reduced histone mRNA and protein levels. We infer that in addition to the role in general translation, ABCE1 is involved in histone biosynthesis and DNA replication and therefore is essential for normal S phase progression. In addition, we analyze whether ABCE1 is implicated in transcript-specific translation via its association with the eIF3 complex subunits known to control the synthesis of cell proliferation-related proteins. The expression levels of a few such targets regulated by eIF3A, however, were not consistently affected by ABCE1 depletion.

Overview on Sobemoviruses and a Proposal for the Creation of the Family Sobemoviridae.

The genus Sobemovirus, unassigned to any family, consists of viruses with single-stranded plus-oriented single-component RNA genomes and small icosahedral particles. Currently, 14 species within the genus have been recognized by the International Committee on Taxonomy of Viruses (ICTV) but several new species are to be recognized in the near future. Sobemovirus genomes are compact with a conserved structure of open reading frames and with short untranslated regions. Several sobemoviruses are important pathogens. Moreover, over the last decade sobemoviruses have become important model systems to study plant virus evolution. In the current review we give an overview of the structure and expression of sobemovirus genomes, processing and functions of individual proteins, particle structure, pathology and phylogenesis of sobemoviruses as well as of satellite RNAs present together with these viruses. Based on a phylogenetic analysis we propose that a new family Sobemoviridae should be recognized including the genera Sobemovirus and Polemovirus. Finally, we outline the future perspectives and needs for the research focusing on sobemoviruses.

ABCE1 is a highly conserved RNA silencing suppressor.

ATP-binding cassette sub-family E member 1 (ABCE1) is a highly conserved protein among eukaryotes and archaea. Recent studies have identified ABCE1 as a ribosome-recycling factor important for translation termination in mammalian cells, yeast and also archaea. Here we report another conserved function of ABCE1. We have previously described AtRLI2, the homolog of ABCE1 in the plant Arabidopsis thaliana, as an endogenous suppressor of RNA silencing. In this study we show that this function is conserved: human ABCE1 is able to suppress RNA silencing in Nicotiana benthamiana plants, in mammalian HEK293 cells and in the worm Caenorhabditis elegans. Using co-immunoprecipitation and mass spectrometry, we found a number of potential ABCE1-interacting proteins that might support its function as an endogenous suppressor of RNA interference. The interactor candidates are associated with epigenetic regulation, transcription, RNA processing and mRNA surveillance. In addition, one of the identified proteins is translin, which together with its binding partner TRAX supports RNA interference.

Cocksfoot mottle virus coat protein is dispensable for the systemic infection.

BACKGROUND: The Sobemovirus genome consists of polycistronic single-stranded positive-sense RNA. The first ORF encodes P1, a suppressor of RNA silencing required for virus movement. The coat protein (CP) is expressed from the 3' proximal ORF3 via subgenomic RNA. In addition to its structural role, the CP of some sobemoviruses has been reported to be required for systemic movement and to interact with P1. The aim of this study was to analyse the role of Cocksfoot mottle virus (CfMV) CP in the suppression of RNA silencing and virus movement. METHODS: Agrobacterium-mediated transient expression method was used for testing CfMV CP capacity to suppress RNA silencing. CP substitution and deletion mutants were generated to examine the role of this protein in CfMV infection, using three host plants (oat, barley and wheat). The viral movement was characterised with CfMV expressing EGFP fused to the C-terminus of CP. RESULTS: In the current study we show that CfMV CP is an additional RNA silencing suppressor. Interestingly, we observed that all CP mutant viruses were able to infect the three tested host plants systemically, although usually with reduced accumulation. CfMV expressing EGFP was detected in epidermal and mesophyll cells of inoculated leaves. Although EGFP fluorescence was not detected in upper leaves, some plants displayed CfMV symptoms. Analysis of the upper leaves revealed that the viruses had lost the EGFP sequence and sometimes also most of the CP gene. CONCLUSIONS: The present study demonstrates that CfMV CP suppresses RNA silencing but, surprisingly, is dispensable for systemic movement. Thus, CfMV does not move as virion in the tested host plants. The composition of the movement RNP complex remains to be elucidated.

Effects of viral silencing suppressors on tobacco ringspot virus infection in two Nicotiana species.

This study investigated the effects of silencing suppressors derived from six different viruses (P1, P19, P25, HcPro, AC2 and 2b), expressed in transgenic Nicotiana tabacum and Nicotiana benthamiana plants, on the infection pattern of tobacco ringspot virus (TRSV) potato calico strain. In N. benthamiana, this virus produced an initial infection with severe systemic symptoms, but the infection was strongly reduced within a few weeks as the plant recovered from the infection. P25 and HcPro silencing suppressors effectively prevented recovery in this host, allowing continuous accumulation of the viral RNA as well as of the virus-specific small interfering RNAs, in the systemically infected leaves. In the P1-, P19-, AC2- or 2b-expressing transgenic N. benthamiana, the recovery was not complete. Susceptibility of N. tabacum to this virus was temperature sensitive. At lower temperatures, up to 25 degrees C, the plants became systemically infected, but at higher temperatures, the infections were limited to the inoculated leaves. In these preventative conditions, all silencing suppressor transgenes (except P25, which was expressed at very low levels) allowed the establishment of systemic infections. Very strong and consistent systemic infections were observed in HcPro- and AC2-expressing plants.

Phenotypes and functional effects caused by various viral RNA silencing suppressors in transgenic Nicotiana benthamiana and N. tabacum.

RNA silencing suppressor genes derived from six virus genera were transformed into Nicotiana benthamiana and N. tabacum plants. These suppressors were P1 of Rice yellow mottle virus (RYMV), P1 of Cocksfoot mottle virus, P19 of Tomato bushy stunt virus, P25 of Potato virus X, HcPro of Potato virus Y (strain N), 2b of Cucumber mosaic virus (strain Kin), and AC2 of African cassava mosaic virus (ACMV). HcPro caused the most severe phenotypes in both Nicotiana spp. AC2 also produced severe effects in N. tabacum but a much milder phenotype in N. benthamiana, although both HcPro and AC2 affected the leaf tissues of the two Nicotiana spp. in similar ways, causing hyperplasia and hypoplasia, respectively. P1-RYMV caused high lethality in the N. benthamiana plants but only mild effects in the N. tabacum plants. Phenotypic alterations produced by the other transgenes were minor in both species. Interestingly, the suppressors had very different effects on crucifer-infecting Tobamovirus (crTMV) infections. AC2 enhanced both spread and brightness of the crTMV-green fluorescent protein (GFP) lesions, whereas 2b and both P1 suppressors enhanced spread but not brightness of these lesions. P19 promoted spread of the infection into new foci within the infiltrated leaf, whereas HcPro and P25 suppressed the spread of crTMV-GFP lesions.

Cocksfoot mottle virus P1 suppresses RNA silencing in Nicotiana benthamiana and Nicotiana tabacum.

The Sobemovirus genome consists of positive sense, single-stranded polycistronic RNA. The 5'-terminal ORF, encoding the protein P1, is its most variable region. Sobemoviral P1 has been described as dispensable for replication but indispensable for systemic infection. The P1 of Rice yellow mottle virus-Nigerian isolate (RYMV-N) is the only RNA silencing suppressor reported for sobemoviruses until now. Using an agrobacterium-mediated transient assay, we demonstrate here that P1 of Cocksfoot mottle virus-Norwegian isolate (CfMV-NO) suppresses RNA silencing in Nicotiana benthamiana and Nicotiana tabacum, two non-host plants. CfMV-NO P1 was able to suppress the initiation and maintenance of silencing. The suppression of systemic silencing was weaker with CfMV-NO P1 than in the case of RYMV-N P1. In the case of suppression at the local level, the reduction in the amount of 25-nucleotide small interfering RNAs (siRNAs) was less pronounced for CfMV-NO P1 than it was when RYMV-N P1 was used. At the same time, we show that CfMV-NO P1 did not bind siRNAs.

AtRLI2 is an endogenous suppressor of RNA silencing.

RNA silencing is a mechanism involved in gene regulation during development and anti-viral defense in plants and animals. Although many viral suppressors of this mechanism have been described up to now, this is not the case for endogenous suppressors. We have identified a novel endogenous suppressor in plants: RNase L inhibitor (RLI) of Arabidopsis thaliana. RLI is a very conserved protein among eukaryotes and archaea. It was first known as component of the interferon-induced mammalian 2'-5' oligoadenylate (2-5A) anti-viral pathway. This protein is in several organisms responsible for essential functions, which are not related to the 2-5A pathway, like ribosome biogenesis and translation initiation. Arabidopsis has two RLI paralogs. We have described in detail the expression pattern of one of these paralogs (AtRLI2), which is ubiquitously expressed in all plant organs during different developmental stages. Infiltrating Nicotiana benthamiana green fluorescent protein (GFP)-transgenic line with Agrobacterium strains harboring GFP and AtRLI2, we proved that AtRLI2 suppresses silencing at the local and at the systemic level, reducing drastically the amount of GFP small interfering RNAs.

Experiencia con retenedores linguales de última generación

RESUMEN: Una de las preocupaciones fundamentales de los profesionales que trabajamos en esta especialidad, cuando se comienza un tratamiento de ortodoncia, es mantener la oclusión y la estética lograda., Los esfuerzos que se hacen en el diagnóstico, plan de tratamiento, tiempo de contención y post-contención, estan dirigidos a evitar el potencial de recidiva de los mismos. Desde hace tiempo se usan variados aparatos de contención o retención al finalizar la etapa activa del tratamiento. La mayoria de ellos son los aparatos rermovibles. Con el uso de los retenedores fijos en el maxilar inferior, comienza una nueva etapa en los tiempos de contención. Se ha comprobado que manteniendo alineados los elementos dentarios del maxilar inferior, éstos sirven como modelo para el desarrollo y funcionalidad del maxilar superior.

Experiencia con retenedores linguales de última generación

RESUMEN: Una de las preocupaciones fundamentales de los profesionales que trabajamos en esta especialidad, cuando se comienza un tratamiento de ortodoncia, es mantener la oclusión y la estética lograda., Los esfuerzos que se hacen en el diagnóstico, plan de tratamiento, tiempo de contención y post-contención, estan dirigidos a evitar el potencial de recidiva de los mismos. Desde hace tiempo se usan variados aparatos de contención o retención al finalizar la etapa activa del tratamiento. La mayoria de ellos son los aparatos rermovibles. Con el uso de los retenedores fijos en el maxilar inferior, comienza una nueva etapa en los tiempos de contención. Se ha comprobado que manteniendo alineados los elementos dentarios del maxilar inferior, éstos sirven como modelo para el desarrollo y funcionalidad del maxilar superior.(AU)

Tratamiento de ortodoncia correctivo en adultos / Orthodontics treatment corrective in adults

RESUMEN: Este caso muestra los cambios periodontales y estéticos logrados en una paciente adulta con biomecánica moderna. Se utilizaron alambre y resortes de titanium en varias etapas del tratamiento favoreciendo las respuestas óseas, periodontal y estética. Los resultados indican las apreciables mejorías y el cumplimiento de las necesidades y objetivos del paciente devolviendole su sonrisa. Los adultos a través de la ortodoncia pueden evitar la perdida de elementos dentarios y mejorar su estado de salud bucal. Actualmente los adultos pueden acceder a tratamientos de ortodoncia con exelentes resultados.

Tratamiento de las mordidas cruzadas con el expansor palatino de niquel de titanio / Treatment of the bilateral cross-bite with the nickel titanium palatine expansor

RESUMEN:Los objetivos de este tratamiento consisten en analizar y evaluar los resultados obtenidos en un paciente adulto con diagnóstico de clase II de Angle, severa mordida cruzada bilateral y marcada contracción de la arcada superior en sentido transversal. Mediante el uso de un aparato expansor palatino de niquel titanium, se agiliza en forma significativa la expansión del paladar. esta se realiza por los movimientos dentoalvolares suaves y continuos. La mordida cruzada bilateral se descruza en pocos meses alcanzando la norma deseada.

Tratamiento de ortodoncia correctivo en adultos / Orthodontics treatment corrective in adults

RESUMEN: Este caso muestra los cambios periodontales y estéticos logrados en una paciente adulta con biomecánica moderna. Se utilizaron alambre y resortes de titanium en varias etapas del tratamiento favoreciendo las respuestas óseas, periodontal y estética. Los resultados indican las apreciables mejorías y el cumplimiento de las necesidades y objetivos del paciente devolviendole su sonrisa. Los adultos a través de la ortodoncia pueden evitar la perdida de elementos dentarios y mejorar su estado de salud bucal. Actualmente los adultos pueden acceder a tratamientos de ortodoncia con exelentes resultados.(AU)