Rice trait discovery using pollen mutagenesis and doubled haploid enrichment

Overview

This innovative solution leverages a two-part strategy for rice trait discovery by combining chemical mutagenesis and doubled haploid enrichment. The approach uses EMS to induce mutations in Japonica rice pollen, followed by caffeine treatment to inhibit DNA repair, ensuring permanent mutations. This method specifically targets male gametes, creating heritable and non-chimeric mutations. The subsequent use of anther culture to generate doubled haploid plants rapidly produces homozygous lines, enabling immediate assessment of recessive traits and the creation of stable genetic resources.

Technical specifications

• Mutagenesis method: EMS (ethyl methanesulfonate) treatment of rice pollen induces high mutation rates.
• DNA repair inhibition: Caffeine treatment prevents natural repair processes, increasing mutation retention.
• Homozygous lines: Anther culture generates doubled haploid plants, revealing recessive traits instantly.
• Validation techniques: Phenotypic scoring and molecular validation via high-throughput screening, using Oxford Nanopore technology for precise mutation analysis.

Technology readiness level

The project is at TRL 3, indicating experimental proof of concept. Further validation includes phenotypic scoring of doubled haploid seedlings and molecular validation through high-throughput gene analysis.