Thermosensitive genic male sterile (TGMS) rice genotypes, which are malesterile at the restrictive temperature (>26°C) but becomes male-fertile at the permissive temperature (< 22°C) are widely used in hybrid rice breeding. Mutation in tms5 gene located on chromosome 2 is responsible for the malesterility trait in rice. Through gene editing, it is possible to create a specific mutation in the tms5 locus of male fertile rice lines and convert them into male sterile lines. The CRISPR/Cas9 editing system was employed to induce specific mutation in tms5 gene of rice variety ASD16. CRISPR/Cas9 vector with guide RNA targeting the second exon of tms5 gene was constructed and used for rice transformation. Four putative transgenic ASD16 events were generated through rice immature embryo transformation and analysed for the presence of hpt and mutation in tms5 locus in T0 generation
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