Key Laboratory of Plant Hormones and Development Regulation of Chongqing, School of Life Sciences, Chongqing University, 401331 Chongqing, PR China.
Citrus Research Institute, Southwest University/Chinese Academy of Agricultural Sciences, Chongqing 400712, PR China.
Gene. 2019 Jul 30;707:178-188. doi: 10.1016/j.gene.2019.04.021. Epub 2019 Apr 13.
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.
遗传工程方法为柑橘溃疡病抗性育种提供了一种替代方法。乙烯响应因子(ERF)家族是转录因子家族的一个成员,该家族是植物特有的,对生物胁迫反应和植物生长有重要贡献。CsAP2-09 属于柑橘 AP2/ERF 转录因子家族。最初,我们证明了 Xcc 和一些参与病原体反应的激素诱导野生型中的 CsAP2-09 表达。我们成功克隆了 CsAP2-09,并证明 CsAP2-09 蛋白靶向细胞核。我们通过过表达和 RNAi 沉默策略对 CsAP2-09 进行了功能表征。在过表达系中,病变和病指明显降低,而在 CsAP2-09 的 RNAi 系中,病变和病指明显增加。因此,过表达赋予转基因柑橘对 Xcc 的抗性,而 CsAP2-09 的沉默导致甜橙对 Xcc 的敏感性。当比较 WT 和过表达转录组时,发现一些参与苯丙烷生物合成、病原体反应、转录调控等的基因发生了修饰。我们的结果为通过过表达 CsAP2s 提高柑橘溃疡病抗性提供了可能性。此外,CsAP2-09 的各种功能为 AP2/ERFs 在植物抗病性和耐胁迫性中的作用提供了重要信息。