Rice genetic variation enhancement through EMS and 5-azac seed priming

Overview

This innovative approach to rice mutagenesis leverages a combination of pre-soaking, EMS (ethyl methanesulfonate), and 5-Azacytidine (5-AzaC) treatments to enhance genetic and epigenetic diversity. By activating seeds metabolically and opening chromatin regions, the method increases the accessibility of DNA to mutagens. The result is a higher mutation frequency without compromising seed viability, making it a valuable tool for trait discovery in rice varieties such as Nipponbare and IR64.

Technical specifications

Key features:

• Pre-soaking: Seeds are pre-soaked in distilled water at 28°C for 8–12 hours to activate early metabolism and chromatin accessibility.
• Chemical mutagenesis: Utilizes EMS (0.5–1%) and 5-AzaC (5–25 µM) combined with low-concentration DMSO (0.1–1%) to enhance mutagen uptake.
• Recovery process: Post-treatment, seeds are rinsed and allowed to recover before sowing, ensuring germination and survival.
• Evaluation: M1 plants are assessed for germination, growth, vigour, phenotypic variations, and chimaeras over a 12-month period.

Technology readiness level

This technology is currently at TRL 3, indicating that it is in the experimental proof-of-concept stage. It has been validated in controlled conditions with Nipponbare and IR64 rice varieties and is progressing through further optimization and evaluation phases.