Construction and characterization of a knock-down RNA interference line of OsNRPD1 in rice (Oryza sativa ssp japonica cv Nipponbare)

Debladis Emilie, Lee Tzuu-Fen, Huang Yan-Jiun, Lu Jui-Hsien, Mathioni Sandra M., Carpentier Marie-Christine, Llauro Christel, Pierron Davy, Mieulet Delphine, Guiderdoni Emmanuel, Chen Pao-Yang, Meyers Blake C., Panaud Olivier, Lasserre Eric. 2020. Construction and characterization of a knock-down RNA interference line of OsNRPD1 in rice (Oryza sativa ssp japonica cv Nipponbare). Philosophical Transactions B. Biological Sciences, 375 (1795):20190338, 10 p.

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Abstract : In plants, RNA-directed DNA methylation (RdDM) is a silencing mechanism relying on the production of 24-nt small interfering RNAs (siRNAs) by RNA POLYMERASE IV (Pol IV) to trigger methylation and inactivation of transposable elements (TEs). We present the construction and characterization of osnrpd1, a knock-down RNA interference line of OsNRPD1 gene that encodes the largest subunit of Pol IV in rice (Oryza sativa ssp japonica cv Nipponbare). We show that osnrpd1 displays a lower accumulation of OsNRPD1 transcripts, associated with an overall reduction of 24-nt siRNAs and DNA methylation level in all three contexts, CG, CHG and CHH. We uncovered new insertions of known active TEs, the LTR retrotransposons Tos17 and Lullaby and the long interspersed nuclear element-type retrotransposon Karma. However, we did not observe any clear developmental phenotype, contrary to what was expected for a mutant severely affected in RdDM. In addition, despite the presence of many putatively functional TEs in the rice genome, we found no evidence of in planta global reactivation of transposition. This knock-down of OsNRPD1 likely led to a weakly affected line, with no effect on development and a limited effect on transposition. We discuss the possibility that a knock-out mutation of OsNRPD1 would cause sterility in rice.

Mots-clés Agrovoc : Oryza sativa, Génétique moléculaire, Arn messager, Code génétique, Méthylation, ADN

Mots-clés complémentaires : Oryza sativa japonica, interférence par ARN

Mots-clés libres : RNA interference, Polymerase IV, Transposable elements, RNA-dependent DNA methylation

Classification Agris : F30 - Plant genetics and breeding

Champ stratégique Cirad : CTS 1 (2019-) - Biodiversité

Auteurs et affiliations

  • Debladis Emilie, Université de Perpignan (FRA)
  • Lee Tzuu-Fen, Donald Danforth Plant Science Center (USA)
  • Huang Yan-Jiun, Academia Sinica (TWN)
  • Lu Jui-Hsien, Academia Sinica (TWN)
  • Mathioni Sandra M., Donald Danforth Plant Science Center (USA)
  • Carpentier Marie-Christine, Université de Perpignan (FRA)
  • Llauro Christel, CNRS (FRA)
  • Pierron Davy, Université de Perpignan (FRA)
  • Mieulet Delphine, CIRAD-BIOS-UMR AGAP (FRA) ORCID: 0000-0001-6220-0372
  • Guiderdoni Emmanuel, CIRAD-BIOS-UMR AGAP (FRA)
  • Chen Pao-Yang, Academia Sinica (TWN)
  • Meyers Blake C., Donald Danforth Plant Science Center (USA)
  • Panaud Olivier, CNRS (FRA)
  • Lasserre Eric, CNRS (FRA) - auteur correspondant

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