Overexpression of AtPAD4 in transgenic Brachypodium distachyon enhances resistance to Puccinia brachypodii.

Brachypodium distachyon (L.) has recently emerged as a model for temperate grasses for investigating the molecular basis of plant-pathogen interactions. Phytoalexin deficient 4 (PAD4) plays a regulatory role in mediating expression of genes involved in plant defence. In this research, we generated transgenic B. distachyon plants constitutively overexpressing AtPAD4. Two transgenic B. distachyon lines were verified using PCR and GUS phenotype. Constitutive expression of AtPAD4 in B. distachyon enhanced resistance to Puccinia brachypodii. P. brachypodii generated less urediniospores on transgenic than on wild-type plants. AtPAD4 overexpression enhanced salicylic acid (SA) levels in B. distachyon-infected tissues. qRT-PCR showed that expression of pathogenesis-related 1 (PR1) and other defence-related genes were up-regulated in transformed B. distachyon following infection with P. brachypodii. Our results indicate that AtPAD4 overexpression in B. distachyon plants led to SA accumulation and induced PR gene expression that reduced the rate of colonisation by P. brachypodii.