Genome editing of Sal1 to improve drought tolerance in African durum wheat

Overview

This research project applies CRISPR-based genome editing to African durum wheat to improve drought tolerance without sacrificing yield. The approach targets the Sal1 gene, a key regulator of retrograde signaling that connects organelles to the nucleus. By knocking out Sal1, the team aims to enhance ABA-mediated stress responses, increasing water use efficiency and relative water content under drought while preserving productivity. The project addresses a critical challenge: climate change is making traditional high-yield crop varieties increasingly unsustainable, while wild varieties tolerate stress but sacrifice productivity. The goal is a durum wheat variety that achieves a better trade-off between drought tolerance and yield.

Technical specifications

• Target gene: Sal1, a negative regulator of ABA signaling and retrograde communication
• Editing approach: CRISPR/Cas9-mediated knockout using guide RNAs targeting both Sal1 homeologs in wheat
• Delivery method: Agrobacterium-mediated transformation of durum wheat
• Expected outcomes: Reduced stomatal conductance, increased water use efficiency, improved relative water content, and more tillers under limiting water conditions
• Selection strategy: Screening transformed lines for drought tolerance and yield potential, followed by molecular characterization of mutations
• Validation basis: Proof-of-concept demonstrated in Arabidopsis and bread wheat through gene silencing studies showing improved drought tolerance without adverse effects under non-stressed conditions

Technology readiness level

The project is at an early-to-mid stage of development. Guide RNAs targeting both Sal1 homeologs have been designed and subcloned into a wheat-specific plant vector. Agrobacterium-mediated transformation of durum wheat has been performed, and transformants are currently under selection. The next steps include phenotypic evaluation for drought tolerance and yield potential, followed by molecular analysis to confirm and characterize the Sal1 mutations. The team is seeking a partner to contribute to the costs of molecular analysis, cloning, wheat transformation, and phenotypic selection.