Creation of male‐sterile lines that can be restored to fertility by exogenous methyl jasmonate for the establishment of a two‐line system for the hybrid production of rice (<i>Oryza sativa</i> L.)

Male sterility is widely used in the production of hybrid seeds in rice, but the use of genic male sterility is limited because of the high labour cost for maintaining male-sterile lines. Previous studies using T-DNA insertional mutagenesis demonstrated that disrupting the expression of oxophytodienoic acid reductase 3 (OPR3), which is involved in the jasmonate biosynthesis pathway, results in a kind of male sterility that can be restored to fertility by exogenous jasmonate in Arabidopsis. Here, we created male-sterile mutations by editing the second and fourth exons of OsOPR7 in rice through clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated system 9. The induced mutagenesis at these exons resulted in 31.8% and 23.9% male-sterile plants in the T₀ generation, respectively. We screened male-sterile lines that can be restored to fertility by exogenous methyl jasmonate in the T_0, T₁ and T2 rice populations and characterized the anther and agronomic traits of the transgenic plants. Results showed the successful generation of male-sterile lines through the silencing of OsOPR7, the orthologous gene of Arabidopsis OPR3, in a field crop, paving the way for the establishment of a two-line system for rice hybrid production. The system consists of a male-sterile line that can be maintained by spraying methyl jasmonate and a restoring line that confers pollen.