Modification of the Stripe Rust Resistance Gene Yr10 in Triticum aestivum
Stripe rust is a disease in Triticum aestivum (bread wheat) that is caused by the fungal pathogen Puccinia striiformis. The pathogen has evolved to defeat an R gene in T. aestivum known as the Yr10 gene. The Yr10 gene was found to encode an evolutionary-conserved sequence known as the CC-NBS-LRR. This conserved sequence was found to be involved in producing resistance to various pathogens. Within this sequence the coiled coil (CC) and leucine rich repeat (LRR) domains are thought to be important to the protein’s function. This project made use of PCR overlap-extension mutagenesis to mutagenize the CC and LRR domains in an attempt to create modified constructs of the Yr10 gene. PCR reactions gave fragments of the expected sizes which were then assembled into the pANIC6D vector to be used in transformation. Transformation of the vector with inserted sequences into Escherichia coli will be done to confirm the successful insertion of the fragments. There is hope that in the future the modified constructs could be transformed into wheat.