In natural conditions, plants are subjected to a combination of biotic stresses and often have to cope with simultaneous pathogen infections. In this report, we aim to understand the global transcriptional response of hybrid poplar NM6 (Populus nigra x P. maximowiczii) to infection by two biotrophic Melampsora fungi, Melampsora larici-populina and M. medusae f. sp. deltoidae. These pathogens triggered different responses after inoculation of poplar leaves. Transcript profiling using the GeneChip Poplar Genome Array revealed a total of 416 differentially expressed transcripts whose expression level was > or = twofold relative to controls. Interestingly, approximately half of the differentially expressed genes in infected leaves showed altered expression following interaction with either of the Melampsora spp. We also infected poplar leaves simultaneously with both Melampsora spp. to investigate potential interaction between the responses to the individual pathogens during a mixed infection. For this mixed inoculation, the number of differentially expressed transcripts increased to 648 and our analysis showed that infection with both fungi also induced a common set of genes. The genes induced after Melampsora spp. infection were mainly related to primary and secondary metabolic processes, cell-wall reinforcement and lignification, defense and stress-related mechanisms, and signal perception and transduction.

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Summary• WRKY transcription factors are key regulators that activate and fine-tune stress responses, including defense responses against pathogens. We isolated a poplar (Populus tremula × Populus alba) cDNA sequence, PtWRKY23, that encodes the ortholog of Arabidopsis WRKY23 and present the functional analysis of WRKY23, with emphasis on its potential role in resistance to rust infection.• To investigate the function of PtWRKY23, we examined PtWRKY23 expression after stress treatments by qRT-PCR and generated PtWRKY23-misexpressing plants. Transgenic plants were assessed for resistance to Melampsora rust and were analyzed using the poplar Affymetrix GeneChip ® and histological techniques to study the consequences of PtWRKY23 misexpression.• PtWRKY23 is rapidly induced by Melampsora infection and elicitor treatments and poplars overexpressing and underexpressing PtWRKY23 were both more susceptible to Melampsora infection than wild type. Transcriptome analysis of PtWRKY23 overexpressors revealed a significant overlap with the Melampsorainfection response. Transcriptome analysis also suggests that PtWRKY23 affects redox homeostasis and cell wall-related metabolism, which was confirmed by analyses that showed that PtWRKY23-misexpressing plants have altered peroxidase activity, apparent H 2 O 2 accumulation and lignin deposition.• Our results show that PtWRKY23 affects resistance to Melampsora infection and that this may be caused by deregulation of genes that disrupt redox homeostasis and cell wall metabolism.

In this study, we compared interactions of two Melampsora foliar rust species with poplar, which resulted in either limited or abundant pathogen proliferation. In the pathosystem exhibiting limited pathogen growth, a defence response was observed after invasion of poplar leaf tissues by the biotroph, with late and clear production of reactive oxygen species (ROS) and other products. Characterisation of the histological, biochemical and transcriptional events occurring in both pathosystems showed striking similarity with components of plant defence reactions observed during qualitative resistance. Key components associated with development of an active defence response, such as up-regulation of pathogenesis-related (PR) genes, were observed during infection. Moreover, the time course and strength of gene induction appear to be critical determinants for the outcome of the tree-pathogen interaction. This work provides basic biochemical characterisation and expression data for the study of so-called partial resistance in the poplar-rust pathosystem, which is also applicable to other plant-pathogen interactions resulting in quantitative disease resistance.