Aphids efficiently ingest dsRNA, which persists in the midgut but is rapidly degraded by aphid nucleases when unprotected. Chitosan and LDH-based formulations improve dsRNA stability, triggering RNAi....

Darwin is an AI research agent for gene regulation from CAIRN Institute — explore regulatory networks, evaluate evidence, predict perturbations, and design experiments.

This study introduces MADAM (Monoclonal Anti-dsRNA Antibody-Based Metagenomics), a novel approach that integrates multiple technical modules previously used independently in other protocols. MADAM combines monoclonal antibody-mediated double-stranded RNA (dsRNA) enrichment, sequence-independent RT-PCR, and Oxford Nanopore Technologies (ONT) sequencing. Applied to Pyricularia oryzae , the causal agent of rice blast disease, MADAM enabled the comprehensive characterization of mycovirus genomes from four fungal isolates collected in Yunnan, China. The approach achieved high viral read recovery rates (46.9–72.7%) and identified 18 P . oryzae -associated RNA viruses spanning seven families: Botourmiaviridae , Deltaormycoviridae , Mymonaviridae , Partitiviridae , Polymycoviridae , Splipalmiviridae , and Ambiguiviridae . Seventeen nearly complete to complete viral genomes (1,226–6,085 nucleotides) were recovered, with sequence coverage ranging from 88% to 100%. Co-infections were detected in three of the four isolates, with notable discoveries including the first deltaormycovirus reported in P. oryzae , a putative novel member of Botourmiaviridae , and an additional genomic segment of a polymycovirus. MADAM successfully detected positive-sense, negative-sense ssRNA, and dsRNA viruses, demonstrating its broad applicability. By uncovering novel viruses and resolving complex co-infections, this method proves invaluable for fungal virology, with potential applications in diagnostics, surveillance, and biological control. Ultimately, MADAM advances our understanding of fungal viral diversity and paves the way for further exploration of mycovirus ecology and evolution. ### Competing Interest Statement The authors have declared no competing interest. University of Montpellier, Key Initiative Clapas CIRAD - Direction générale déléguée à la recherche et à la stratégie, https://ror.org/049x36t95, Cresi

This study introduces MADAM (Monoclonal Anti-dsRNA Antibody-Based Metagenomics), a novel approach that integrates multiple technical modules previously used independently in other protocols. MADAM combines monoclonal antibody-mediated double-stranded RNA (dsRNA) enrichment, sequence-independent RT-PCR, and Oxford Nanopore Technologies (ONT) sequencing. Applied to Pyricularia oryzae , the causal agent of rice blast disease, MADAM enabled the comprehensive characterization of mycovirus genomes from four fungal isolates collected in Yunnan, China. The approach achieved high viral read recovery rates (46.9–72.7%) and identified 18 P . oryzae -associated RNA viruses spanning seven families: Botourmiaviridae , Deltaormycoviridae , Mymonaviridae , Partitiviridae , Polymycoviridae , Splipalmiviridae , and Ambiguiviridae . Seventeen nearly complete to complete viral genomes (1,226–6,085 nucleotides) were recovered, with sequence coverage ranging from 88% to 100%. Co-infections were detected in three of the four isolates, with notable discoveries including the first deltaormycovirus reported in P. oryzae , a putative novel member of Botourmiaviridae , and an additional genomic segment of a polymycovirus. MADAM successfully detected positive-sense, negative-sense ssRNA, and dsRNA viruses, demonstrating its broad applicability. By uncovering novel viruses and resolving complex co-infections, this method proves invaluable for fungal virology, with potential applications in diagnostics, surveillance, and biological control. Ultimately, MADAM advances our understanding of fungal viral diversity and paves the way for further exploration of mycovirus ecology and evolution. ### Competing Interest Statement The authors have declared no competing interest. University of Montpellier, Key Initiative Clapas CIRAD - Direction générale déléguée à la recherche et à la stratégie, https://ror.org/049x36t95, Cresi

This study introduces MADAM (Monoclonal Anti-dsRNA Antibody-Based Metagenomics), a novel approach that integrates multiple technical modules previously used independently in other protocols. MADAM combines monoclonal antibody-mediated double-stranded RNA (dsRNA) enrichment, sequence-independent RT-PCR, and Oxford Nanopore Technologies (ONT) sequencing. Applied to Pyricularia oryzae , the causal agent of rice blast disease, MADAM enabled the comprehensive characterization of mycovirus genomes from four fungal isolates collected in Yunnan, China. The approach achieved high viral read recovery rates (46.9–72.7%) and identified 18 P . oryzae -associated RNA viruses spanning seven families: Botourmiaviridae , Deltaormycoviridae , Mymonaviridae , Partitiviridae , Polymycoviridae , Splipalmiviridae , and Ambiguiviridae . Seventeen nearly complete to complete viral genomes (1,226–6,085 nucleotides) were recovered, with sequence coverage ranging from 88% to 100%. Co-infections were detected in three of the four isolates, with notable discoveries including the first deltaormycovirus reported in P. oryzae , a putative novel member of Botourmiaviridae , and an additional genomic segment of a polymycovirus. MADAM successfully detected positive-sense, negative-sense ssRNA, and dsRNA viruses, demonstrating its broad applicability. By uncovering novel viruses and resolving complex co-infections, this method proves invaluable for fungal virology, with potential applications in diagnostics, surveillance, and biological control. Ultimately, MADAM advances our understanding of fungal viral diversity and paves the way for further exploration of mycovirus ecology and evolution. ### Competing Interest Statement The authors have declared no competing interest. University of Montpellier, Key Initiative Clapas CIRAD - Direction générale déléguée à la recherche et à la stratégie, https://ror.org/049x36t95, Cresi

This study introduces MADAM (Monoclonal Anti-dsRNA Antibody-Based Metagenomics), a novel approach that integrates multiple technical modules previously used independently in other protocols. MADAM combines monoclonal antibody-mediated double-stranded RNA (dsRNA) enrichment, sequence-independent RT-PCR, and Oxford Nanopore Technologies (ONT) sequencing. Applied to Pyricularia oryzae , the causal agent of rice blast disease, MADAM enabled the comprehensive characterization of mycovirus genomes from four fungal isolates collected in Yunnan, China. The approach achieved high viral read recovery rates (46.9–72.7%) and identified 18 P . oryzae -associated RNA viruses spanning seven families: Botourmiaviridae , Deltaormycoviridae , Mymonaviridae , Partitiviridae , Polymycoviridae , Splipalmiviridae , and Ambiguiviridae . Seventeen nearly complete to complete viral genomes (1,226–6,085 nucleotides) were recovered, with sequence coverage ranging from 88% to 100%. Co-infections were detected in three of the four isolates, with notable discoveries including the first deltaormycovirus reported in P. oryzae , a putative novel member of Botourmiaviridae , and an additional genomic segment of a polymycovirus. MADAM successfully detected positive-sense, negative-sense ssRNA, and dsRNA viruses, demonstrating its broad applicability. By uncovering novel viruses and resolving complex co-infections, this method proves invaluable for fungal virology, with potential applications in diagnostics, surveillance, and biological control. Ultimately, MADAM advances our understanding of fungal viral diversity and paves the way for further exploration of mycovirus ecology and evolution. ### Competing Interest Statement The authors have declared no competing interest. University of Montpellier, Key Initiative Clapas CIRAD - Direction générale déléguée à la recherche et à la stratégie, https://ror.org/049x36t95, Cresi

Capsaicinoids are synthesized in the placenta of Capsicum fruit, where vanillylamine aminotransferase (VAMT) catalyzes the formation of vanillylamine, the precursor of the pathway. The modulation of pungency has relied on genetic breeding and transgenic approaches, and this pathway has not been addressed by spray-induced gene silencing. The aim of this study was to evaluate whether a single non-invasive spray of double-stranded RNA (dsRNA) targeting VAMT allows the gene to be silenced and capsaicinoid accumulation to be modified in Capsicum chinense fruit. The molecule was designed in silico and applied at 10 days post-anthesis. Pedicel injection reduced the VAMT transcript in a dose-dependent manner, with three levels of inhibition distinguishable from one another. Spraying with surfactant reduced it by 86.2 %, a magnitude statistically indistinguishable from the 90.6 % obtained by injection, and also reduced the Pun1 transcript, a co-regulation previously described only as a difference between cultivars. Analysis by gas chromatography coupled to mass spectrometry showed reductions of 84.4 % in capsaicin and 68.5 % in dihydrocapsaicin, the loss of nonivamide and one further vanillylamine-derived compound, and the detection of capsiate and a second capsinoid, absent in control fruits. The siRNA was detected in non-treated tissues, and a single topical application is therefore sufficient to silence an endogenous biosynthetic gene and shift the metabolic profile of the fruit without genetic modification.