Institute of Biosciences and Bioresources, CNR, Research Division Bari, Via Amendola 165/A, 70126, Bari, Italy.
Department of Biotechnology, University of Verona, Strada Le Grazie 15, 37134, Verona, Italy.
Plant J. 2019 May;98(4):590-606. doi: 10.1111/tpj.14275. Epub 2019 Mar 12.
Cyclic AMP plays important roles in different physiological processes, including plant defence responses. However, as little information is known on plant enzymes responsible for cAMP production/degradation, studies of cAMP functions have relied, to date, on non-specific pharmacological approaches. We therefore developed a more reliable approach, producing transgenic Arabidopsis thaliana lines overexpressing the 'cAMP-sponge' (cAS), a genetic tool that specifically buffers cAMP levels. In response to an avirulent strain of Pseudomonas syringae pv. tomato (PstAvrB), cAS plants showed a higher bacterial growth and a reduced hypersensitive cell death in comparison with wild-type (WT) plants. The low cAMP availability after pathogen infection delayed cytosolic calcium elevation, as well as hydrogen peroxide increase and induction of redox systems. The proteomic analysis, performed 24 h post-infection, indicated that a core of 49 proteins was modulated in both genotypes, while 16 and 42 proteins were uniquely modulated in WT and cAS lines, respectively. The involvement of these proteins in the impairment of defence response in cAS plants is discussed in this paper. Moreover, in silico analysis revealed that the promoter regions of the genes coding for proteins uniquely accumulating in WT plants shared the CGCG motif, a target of the calcium-calmodulin-binding transcription factor AtSR1 (Arabidopsis thaliana signal responsive1). Therefore, following pathogen perception, the low free cAMP content, altering timing and levels of defence signals, and likely acting in part through the mis-regulation of AtSR1 activity, affected the speed and strength of the immune response.
环腺苷酸(cAMP)在包括植物防御反应在内的不同生理过程中发挥着重要作用。然而,由于对负责 cAMP 产生/降解的植物酶知之甚少,迄今为止,cAMP 功能的研究依赖于非特异性的药理学方法。因此,我们开发了一种更可靠的方法,生产过表达“cAMP 海绵”(cAS)的转基因拟南芥(Arabidopsis thaliana)系,这是一种专门缓冲 cAMP 水平的遗传工具。与野生型(WT)植物相比,cAS 植物在响应无毒型(avirulent)丁香假单胞菌 pv.番茄(PstAvrB)菌株时,细菌生长更高,过敏性细胞死亡减少。病原体感染后 cAMP 可用性降低,延迟了细胞质钙的升高、过氧化氢的增加以及氧化还原系统的诱导。感染后 24 小时进行的蛋白质组学分析表明,两种基因型中共有 49 种核心蛋白发生了调节,而 WT 和 cAS 系中分别有 16 种和 42 种蛋白独特地发生了调节。本文讨论了这些蛋白在 cAS 植物防御反应受损中的作用。此外,计算机分析表明,WT 植物中唯一积累的蛋白编码基因的启动子区域共享 CGCG 基序,这是钙-钙调蛋白结合转录因子 AtSR1(Arabidopsis thaliana signal responsive1)的靶标。因此,在感知病原体后,低的游离 cAMP 含量改变了防御信号的时间和水平,可能部分通过 AtSR1 活性的失调来影响免疫反应的速度和强度。