School of Life Sciences, The University of Warwick, Coventry, England, United Kingdom.
Warwick Integrative Synthetic Biology Centre, The University of Warwick, Coventry, England, United Kingdom.
PLoS One. 2020 Jun 4;15(6):e0234154. doi: 10.1371/journal.pone.0234154. eCollection 2020.
Phytohormones mediate most diverse processes in plants, ranging from organ development to immune responses. Receptor protein complexes perceive changes in intracellular phytohormone levels and trigger a signaling cascade to effectuate downstream responses. The in planta analysis of elements involved in phytohormone signaling can be achieved through transient expression in mesophyll protoplasts, which are a fast and versatile alternative to generating plant lines that stably express a transgene. While promoter-reporter constructs have been used successfully to identify internal or external factors that change phytohormone signaling, the range of available marker constructs does not meet the potential of the protoplast technique for large scale approaches. The aim of our study was to provide novel markers for phytohormone signaling in the Arabidopsis mesophyll protoplast system. We validated 18 promoter::luciferase constructs towards their phytohormone responsiveness and specificity and suggest an experimental setup for high-throughput analyses. We recommend novel markers for the analysis of auxin, abscisic acid, cytokinin, salicylic acid and jasmonic acid responses that will facilitate future screens for biological elements and environmental stimuli affecting phytohormone signaling.
植物激素调节植物中最多种多样的过程，从器官发育到免疫反应。受体蛋白复合物感知细胞内植物激素水平的变化，并触发信号级联反应以实现下游反应。参与植物激素信号转导的元件的植物内分析可以通过质体原生质体中的瞬时表达来实现，这是一种比稳定表达转基因植物系更快、更通用的替代方法。虽然启动子-报告基因构建体已成功用于鉴定改变植物激素信号转导的内部或外部因素，但可用标记构建体的范围并不能满足原生质体技术在大规模方法中的潜力。我们的研究目的是为拟南芥质体原生质体系统中的植物激素信号转导提供新的标记物。我们针对 18 个启动子::荧光素酶构建体的植物激素反应性和特异性进行了验证，并提出了一种用于高通量分析的实验设置。我们推荐用于分析生长素、脱落酸、细胞分裂素、水杨酸和茉莉酸反应的新标记物，这将有助于未来筛选影响植物激素信号转导的生物元件和环境刺激。
