Journal Article Research Support, Non-U.S. Gov't Ppublish 39 citations
PubMed: 32842152 DOI: 10.1093/jxb/eraa391
Assessment of the roles of SPO11-2 and SPO11-4 in meiosis in rice using CRISPR/Cas9 mutagenesis.
J Exp Bot 71(22):7046-7058
Authors: Ian Fayos, Anne Cécile Meunier, Aurore Vernet, Sergi Navarro-Sanz, Murielle Portefaix, Marc Lartaud, Giacomo Bastianelli, Christophe Périn, Alain Nicolas, Emmanuel Guiderdoni
Affiliations
- CIRAD, UMR AGAP, Montpellier Cedex, France.
- Université de Montpellier, CIRAD-INRAe-Institut Agro, Montpellier, France.
- Meiogenix, Paris, France.
- Institut Curie, CNRS UMR 3244, PSL University, Paris Cedex, France.
Abstract
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.
MeSH
Cells, Recombination, Genetic, Mothers, Clustered Regularly Interspaced Short Palindromic Repeats, Mutagenesis, Stem Cells, Chromosomes, Botany, Meiosis
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