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Hebrard, E.

Publications and source records attributed to Hebrard, E..

6 recordsLinked to original sources

eIFiso4G editing confers durable resistance against rice yellow mottle virus

Rice yellow mottle virus (RYMV) is one of the most devastating viral pathogens affecting rice cultivation in Africa. The eukaryotic translation initiation factor 4G isoform eIFiso4G1 plays a pivotal role in rice susceptibility to the virus. Naturally occuring resistance alleles impair infection but are predominantly found in the African cultivated species Oryza glaberrima, whose use in breeding programs is limited by interspecific sterility barriers with the widely grown O. sativa. Here, we used CRISPR-Cas9 to generate knockout (KO) mutations in eIFiso4G1 as well as insertion-deletion (Indel) variants within the region involved in interaction with the viral protein VPg. KO mutations conferred high resistance to RYMV. No resistance-breaking events were observed, suggesting that this resistance may be more durable than that conferred by naturally occuring alleles. However, complete gene KO slightly affected plant growth. Lines carrying Indel mutations in the VPg-interacting region displayed variable resistance levels, with some variants conferring high resistance without compromising plant development. Structural modeling of the eIFiso4G1 variants and their complexes with VPg provided mechanistic insights into how specific Indels modulate the resistance phenotype.

plant biology↗

Symptom observation underestimates co-infections: insight from viral and bacterial diseases in rice fields in Burkina Faso

Co-occurrence of multiple diseases and co-infection of individual plants by various pathogens have potential epidemiological and evolutionary implications. Based on previous information on the co-occurrence of the rice yellow mottle disease (caused by the rice yellow mottle virus, RYMV) and bacterial leaf streak (BLS, due to Xanthomonas oryzae pv. oryzicola, Xoc) in Burkina Faso, and experimental evidence of interactions between the pathogens causing these two diseases, we aimed to monitor the two pathogens more intensively in farmers rice fields. To this purpose, we selected fields showing both types of symptoms to maximize the chance of observing co-infections at the plant scale. We performed observations and sampling in two sites over two consecutive years. Over a global dataset of 1666 samples, 1341 were symptomatic. Although the sampling design aimed to observe co-infections, only 37 of these samples (2.8%) were annotated as presenting both yellow mottle and BLS symptoms. The samples were then subjected to a newly designed molecular detection test that specifically amplifies both the virus (RYMV) and the bacteria (Xo). This revealed that 166 samples, i.e. 12.4% of symptomatic samples, were co-infected by RYMV and Xo, hence showing that symptom observation in the field greatly underestimates co-infection levels. Combining these data with a previously published dataset, we estimated that up to 1-4% of all plants in disease hotspots are simultaneously infected by the two pathogens. Further research on multiple infections would benefit from longitudinal surveys over the crop growing season rather than such a cross-sectional study.

plant biology↗

Understanding evolutionary and functional relationships of RNA polymerases in plant and fungal viruses through structural modeling and divergence date estimations

RNA-dependent RNA polymerases (RdRps) are crucial for RNA virus replication and serve as key marker genes for defining deep taxonomic ranks and for understanding viral evolutionary history. Despite their conserved functions and motifs, the high genetic diversity of RdRps complicates precise sequence comparisons across viral families, hindering accurate taxonomic classification of new species. Three-dimensional (3D) RdRp structures can help overcome these challenges through structure-guided alignments. However, such data are rare for myco- and phytoviruses, limiting investigation of their ecological and evolutionary links. In this study, we focused on the highly divergent order Sobelivirales, which includes sobemoviruses infecting plants -- a taxon known for its ancient origin -- and barnaviruses infecting fungi, which remain relatively unknown. Using deep-learning structural modeling, we generated reliable 3D models for 44 sobemoviral and sobelivirad species. Structure-guided alignments, together with new barnaviral and relevant outgroup sequences, enabled robust phylogenetic reconstruction, allowing us to propose revisions of existing viral families and suggest new evolutionary scenarios. Divergence dates were estimated for the first time at this taxonomical rank using the Prisoner of War model, which revealed a divergence of plant and fungal sobelivirads 27.0 {+/-} 10 million years ago -- much more recent than the separation of their respective hosts. This result suggests that cross-kingdom host shifts have contributed more likely to the evolutionary history of Sobelivirales than strict virus-host codivergence. Based on an extended dataset of 127 species, structure and sequence conservation analyses identified molecular signatures of sobeliviral families. These conserved and extented RdRp motifs will facilitate future taxonomic assignments and the development of diagnostic tools. Our interdisciplinary approach, integrating structure modeling and divergence dating, offers new insights into the evolutionary divergence of plant and fungal viruses, with potential applications to other viral orders and families. Author summaryRNA-dependent RNA polymerases (RdRps) are essential for RNA virus replication and serve as important markers for classifying viruses and understanding their evolution. However, with metagenomic studies that rapidly expand the known diversity of RNA viruses, it is increasingly difficult to compare highly divergent RdRps and accurately classify new species. When available, 3D structures of RdRps can help overcome this challenge through structure-guided alignments. Here, we focused on the highly divergent order Sobelivirales, which groups phytoviruses and mycoviruses. Deep-learning models were used to generate reliable 3D structures for 44 representative viral species. Structure-guided alignments combined with the identification of new barnaviruses and outgroups allowed us to build a more accurate viral phylogeny. Based on this finding, we proposed updates to existing viral families and genera within the order Sobelivirales. We estimated divergence dates using a model that previously uncovered the ancient origins of sobemovirus. Notably, we provided the first estimate of when these plant and fungal viruses diverged [~]27.0 {+/-} 10 million years ago, suggesting cross-kingdom host shifts rather than strict virus-host codivergence. We also identified molecular signatures that are useful for future virus classification and diagnosis, with potential applications to other viral groups.

evolutionary biology↗

Grains, trade and war in the multimodal transmission of Rice yellow mottle virus: an historical and phylogeographical retrospective

Rice yellow mottle virus (RYMV) is a major pathogen of rice in Africa. RYMV has a narrow host range limited to rice and a few related poaceae species. We explore the links between the spread of RYMV in East Africa and rice history since the second half of the 19th century. The phylogeography of RYMV in East Africa was reconstructed from coat protein gene sequences (ORF4) of 335 isolates sampled over two million square kilometers between 1966 and 2020. Dispersal patterns obtained from ORF2a and ORF2b, and full-length sequences converged to the same scenario. The following imprints of rice cultivation on RYMV epidemiology were unveiled. RYMV emerged in the middle of the 19th century in the Eastern Arc Mountains where slash-and-burn rice cultivation was practiced. Several spillovers from wild hosts to cultivated rice occurred. RYMV was then rapidly introduced into the adjacent large rice growing Kilombero valley. Harvested seeds are contaminated by debris of virus infected plants that subsist after threshing and winnowing. Long-distance dispersal of RYMV is consistent (i) with rice introduction along the caravan routes from the Indian Ocean Coast to Lake Victoria in the second half of the 19th century, (ii) seed movement from East Africa to West Africa at the end of the 19th century, from Lake Victoria to the north of Ethiopia in the second half of the 20th century and to Madagascar at the end of the 20th century, (iii) and, unexpectedly, with rice transport at the end of the First World War as a troop staple food from the Kilombero valley towards the South of Lake Malawi. Overall, RYMV dispersal was associated to a broad range of human activities, some unsuspected. Consequently, RYMV has a wide dispersal capacity, its dispersal metrics estimated from phylogeographic reconstructions are similar to those of highly mobile zoonotic viruses. Author summaryRice yellow mottle virus (RYMV) poses a major threat to rice production in Africa. We explored through a multidisciplinary approach the links between the history of rice in East Africa since the second half of the 19th century and the spread of RYMV. The results illuminate the causes of RYMV diffusion. We show the role of long-distance caravan trade, the impact of the First World War and the consequences of seed exchange in the dispersal of RYMV. The paradoxical role of seeds in the spread of RYMV - which is vector transmitted and not seed transmitted - is explained in the light of rice biology and agronomy. Overall, this study reveals the wide range of transmission ways, some unexpected, in the dispersal of plant viruses. It also highlights the role of human transmission of pathogens, even vector-borne, and sheds light on the risk of transmission of RYMV and of other plant viruses from Africa to other continents.

evolutionary biology↗

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 during the Neolithic period. By incorporating sequences collected through metagenomic analyses, the resulting phylogenetic tree showcases increased genetic diversity, reflecting a deep history of sobemovirus species with major radiation events taking place during the Neolithic period, suggesting rapid diversification in that period. Our findings make a case for the possibility of deep evolutionary origins of plant viruses.

evolutionary biology↗

Dynamics of the rice yellow mottle disease in western Burkina Faso: epidemic monitoring, spatio-temporal variation of viral diversity and pathogenicity in a disease hotspot

The rice yellow mottle virus (RYMV) is a model in plant virus molecular epidemiology and phylogeography, with the reconstruction of historical introduction routes at the scale of the African continent. However, information on patterns of viral prevalence and viral diversity over multiple years at local scale remain scarce, in spite of potential implications for crop protection. Here we describe a five-years monitoring of RYMV prevalence in six sites from western Burkina Faso. This study confirmed one irrigated site as a disease hotspot, and found two rainfed lowland sites with occasional high prevalence levels. Within studied field, a pattern of disease aggregation was evidenced at a five-meter distance, as expected for a mechanically transmitted virus. Next, we monitored RYMV genetic diversity in the irrigated disease hotspot site, revealing a high viral diversity, with the current coexistence of various distinct genetic groups at the site-scale (irrigated perimeter of ca. 520 ha), and also within various specific fields (25 meters side). One genetic lineage, named S1bzn, is the most recently introduced group and increased in frequency over the studied period. Its genome results from a recombination between two other lineages. Finally, experimental work evidenced no differences between three rice varieties cultivated in Burkina Faso in terms of resistance level, and no statistical effect of RYMV genetic group on symptom expression and viral load. We found however, that infection outcome depended on the specific RYMV isolate, with various isolates from the lineage S1bzn found to be particularly aggressive, including one accumulating at highest level. Overall, this study documents a case of high viral prevalence and high viral diversity, with the co-occurrence of divergent genetic lineages at small geographic scale. A recently introduced lineage, that includes viral isolates with high symptoms and accumulation in controlled conditions, could be recently rising though natural selection. Following up the monitoring of RYMV genetic and pathogenic diversity in the area is required to confirm this trend and further understand the factors driving the maintenance of viral diversity at local scale.

evolutionary biology↗