Functional analysis of citrus AP2 transcription factors identified CsAP2-09 involved in citrus canker disease response and tolerance

Abstract

Genetic engineering approaches offer an alternative method to the citrus canker resistance breeding. The ethylene response factor (ERF) family is a member of families of transcription factors that are particular to plants and contribute significantly to biotic stress response and to plant growth. CsAP2-09 belongs to the citrus AP2/ERF transcription factor family. Initially, we proved the induction of CsAP2-09 in wild-types by Xcc and some hormones involved in pathogen response. We successfully cloned the CsAP2-09 and proved that CsAP2-09 protein is targeted to the nucleus. The CsAP2-09 was functionally characterized with over-expression and RNAi silencing strategy. In the overexpression lines, the diseased lesions and disease index were significantly decreased while in RNAi lines of CsAP2-09 the diseased lesions and disease index were significantly enhanced. Thus, the over-expression conferred Xcc resistance to transgenic citrus while silencing of CsAP2-09 in sweet orange leads to Xcc susceptibility. When the transcriptomes of WT and overexpression transcriptomes were compared, they revealed that some genes involved in phenylpropanoid biosynthesis, pathogen responses, transcript regulation etc. were modified. Our results provide a possibility for improving citrus canker disease resistance by over-expression of CsAP2s. Furthermore, various functions of CsAP2-09 provide significant information about the role of AP2/ERFs in plant disease resistance and stress tolerance.