Assessment of the roles of SPO11-2 and SPO11-4 in meiosis in rice using CRISPR/Cas9 mutagenesis

Fayos Ian, Meunier Anne Cecile, Vernet Aurore, Navarro Sanz Sergi, Portefaix Murielle, Lartaud Marc, Bastianelli Giacomo, Périn Christophe, Nicolas Alain, Guiderdoni Emmanuel. 2020. Assessment of the roles of SPO11-2 and SPO11-4 in meiosis in rice using CRISPR/Cas9 mutagenesis. Journal of Experimental Botany, 71 (22) : 7046-7058.

https://doi.org/10.1093/jxb/eraa391

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Quartile : Outlier, Sujet : PLANT SCIENCES

Résumé : In Arabidopsis, chromosomal double-strand breaks at meiosis are presumably catalyzed by two distinct SPO11 transesterases, AtSPO11-1 and AtSPO11-2, together with M-TOPVIB. To clarify the roles of the SPO11 paralogs in rice, we used CRISPR/Cas9 mutagenesis to produce null biallelic mutants in OsSPO11-1, OsSPO11-2, and OsSPO11-4. Similar to Osspo11-1, biallelic mutations in the first exon of OsSPO11-2 led to complete panicle sterility. Conversely, all Osspo11-4 biallelic mutants were fertile. To generate segregating Osspo11-2 mutant lines, we developed a strategy based on dual intron targeting. Similar to Osspo11-1, the pollen mother cells of Osspo11-2 progeny plants showed an absence of bivalent formation at metaphase I, aberrant segregation of homologous chromosomes, and formation of non-viable tetrads. In contrast, the chromosome behavior in Osspo11-4 male meiocytes was indistinguishable from that in the wild type. While similar numbers of OsDMC1 foci were revealed by immunostaining in wild-type and Osspo11-4 prophase pollen mother cells (114 and 101, respectively), a surprisingly high number (85) of foci was observed in the sterile Osspo11-2 mutant, indicative of a divergent function between OsSPO11-1 and OsSPO11-2. This study demonstrates that whereas OsSPO11-1 and OsSPO11-2 are the likely orthologs of AtSPO11-1 and AtSPO11-2, OsSPO11-4 has no major role in wild-type rice meiosis.

Mots-clés Agrovoc : méiose, Oryza sativa, chromosome, mutant, aberration chromosomique, Arabidopsis, mutation, riz, tétrade

Mots-clés libres : CRISPR/Cas9 mutagenesis, Meiosis, Recombination, Rice, SPO11-1

Agences de financement hors UE : Agence Nationale de la Recherche

Projets sur financement : (FRA) Développment d'une infrastructure française distribuée coordonnée

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