The research project has empowered the BLASTOFF Team to establish a platform for bioengineering disease resistance genes, combining structural and evolutionary biology with synthetic biology. This effort has culminated in the development of novel blast resistant rice and wheat through genome editing of susceptibility genes, alongside expanding the effector recognition profile of a rice NLR immune receptor via protein engineering.
Our study focused on the multihost pathogen Magnaporthe oryzae, which infects over 50 species of grasses, including key cereal crops like wheat, barley, and rice. We completed a comprehensive survey of M. oryzae effectors that interact with small Heavy Metal-Associated (sHMA) proteins, identifying several new effector-HMA interacting pairs through yeast two-hybrid screens. This work has provided valuable insights into the diversification of M. oryzae effectors and their interactions with host proteins.
The project helped the development of Pikobodies, namely bioengineered immune system proteins that exploits antibodies' uniquely flexible immune systems, enabling plants the ability to fight off emerging pathogens.
Significant progress was made in elucidating the binding properties of multiple M. oryzae effector proteins to HMA domains, revealing multiple binding interfaces and the impact of amino acid polymorphisms on effector specificity. Through structure-guided mutagenesis and protein engineering, we have successfully modified the rice NLR immune receptor Pikp to recognize and respond to previously elusive variants of the AVR-Pik effector, demonstrating the potential of targeted molecular interventions to broaden plant disease resistance.
Concurrently, our work on genome editing in rice and wheat aimed at knocking out susceptibility genes has progressed, employing CRISPR technology to target the sHMA gene family. This has led to the generation of lines with targeted deletions for enhanced resistance to various blast fungus isolates. These combined efforts underscore the potential of integrating structural biology, synthetic biology, and genome editing to create crops with improved resistance to devastating diseases.
