Liu Jing, Yuan Yongze, Wu Zhi, Li Na, Chen Yuanlei, Qin Tingting, Geng Hui, Xiong Li, Liu Deli
Hubei Key Laboratory of Genetic Regulation and Integrative Biology, School of Life Science, Central China Normal University, Wuhan, 430079, China.
PLoS One. 2015 Feb 20;10(2):e0117115. doi: 10.1371/journal.pone.0117115. eCollection 2015.
Penicillium digitatum is the most destructive postharvest pathogen of citrus fruits, causing fruit decay and economic loss. Additionally, control of the disease is further complicated by the emergence of drug-resistant strains due to the extensive use of triazole antifungal drugs. In this work, an orthologus gene encoding a putative sterol regulatory element-binding protein (SREBP) was identified in the genome of P. digitatum and named sreA. The putative SreA protein contains a conserved domain of unknown function (DUF2014) at its carboxyl terminus and a helix-loop-helix (HLH) leucine zipper DNA binding domain at its amino terminus, domains that are functionally associated with SREBP transcription factors. The deletion of sreA (ΔsreA) in a prochloraz-resistant strain (PdHS-F6) by Agrobacterium tumefaciens-mediated transformation led to increased susceptibility to prochloraz and a significantly lower EC50 value compared with the HS-F6 wild-type or complementation strain (COsreA). A virulence assay showed that the ΔsreA strain was defective in virulence towards citrus fruits, while the complementation of sreA could restore the virulence to a large extent. Further analysis by quantitative real-time PCR demonstrated that prochloraz-induced expression of cyp51A and cyp51B in PdHS-F6 was completely abolished in the ΔsreA strain. These results demonstrate that sreA is a critical transcription factor gene required for prochloraz resistance and full virulence in P. digitatum and is involved in the regulation of cyp51 expression.
指状青霉是柑橘类水果采后最具破坏性的病原菌,会导致果实腐烂并造成经济损失。此外,由于三唑类抗真菌药物的广泛使用,耐药菌株的出现使该病的防治更加复杂。在这项研究中,在指状青霉基因组中鉴定出一个编码假定的甾醇调节元件结合蛋白(SREBP)的直系同源基因,并将其命名为sreA。假定的SreA蛋白在其羧基末端含有一个功能未知的保守结构域(DUF2014),在其氨基末端含有一个螺旋-环-螺旋(HLH)亮氨酸拉链DNA结合结构域,这些结构域在功能上与SREBP转录因子相关。通过根癌农杆菌介导的转化,在对咪鲜胺耐药的菌株(PdHS-F6)中缺失sreA(ΔsreA),导致其对咪鲜胺的敏感性增加,与HS-F6野生型或互补菌株(COsreA)相比,EC50值显著降低。致病性测定表明,ΔsreA菌株对柑橘类水果的致病性存在缺陷,而sreA的互补可在很大程度上恢复其致病性。通过定量实时PCR进一步分析表明,在ΔsreA菌株中,咪鲜胺诱导的PdHS-F6中cyp51A和cyp51B的表达完全被消除。这些结果表明,sreA是指状青霉对咪鲜胺抗性和完全致病性所需的关键转录因子基因,并参与cyp51表达的调控。