Laboratory of Plant Microbiology, Intercollegiate Faculty of Biotechnology of University of Gdansk and Medical University of Gdansk, University of Gdansk, Gdansk, Poland.
Sci Rep. 2021 May 26;11(1):10994. doi: 10.1038/s41598-021-90488-w.
Pseudomonas donghuensis P482 is a tomato rhizosphere isolate with the ability to inhibit growth of bacterial and fungal plant pathogens. Herein, we analysed the impact of the carbon source on the antibacterial activity of P482 and expression of the selected genes of three genomic regions in the P482 genome. These regions are involved in the synthesis of pyoverdine, 7-hydroxytropolone (7-HT) and an unknown compound ("cluster 17") and are responsible for the antimicrobial activity of P482. We showed that the P482 mutants, defective in these regions, show variations and contrasting patterns of growth inhibition of the target pathogen under given nutritional conditions (with glucose or glycerol as a carbon source). We also selected and validated the reference genes for gene expression studies in P. donghuensis P482. Amongst ten candidate genes, we found gyrB, rpoD and mrdA the most stably expressed. Using selected reference genes in RT-qPCR, we assessed the expression of the genes of interest under minimal medium conditions with glucose or glycerol as carbon sources. Glycerol was shown to negatively affect the expression of genes necessary for 7-HT synthesis. The significance of this finding in the light of the role of nutrient (carbon) availability in biological plant protection is discussed.
铜绿假单胞菌 P482 是一种番茄根际分离物,具有抑制细菌和真菌植物病原体生长的能力。在此,我们分析了碳源对 P482 抗菌活性和其基因组中三个基因区域选定基因表达的影响。这些区域参与了绿脓菌素、7-羟基色酮(7-HT)和一种未知化合物(“簇 17”)的合成,负责 P482 的抗菌活性。我们表明,这些区域有缺陷的 P482 突变体在特定营养条件下(以葡萄糖或甘油为碳源)对靶病原体的生长抑制表现出变化和对比模式。我们还选择和验证了铜绿假单胞菌 P482 基因表达研究的参考基因。在十个候选基因中,我们发现 gyrB、rpoD 和 mrdA 表达最稳定。使用选定的参考基因进行 RT-qPCR,我们评估了在以葡萄糖或甘油为碳源的最小培养基条件下,感兴趣基因的表达情况。结果表明,甘油会对 7-HT 合成所需基因的表达产生负面影响。鉴于营养(碳)可用性在生物植物保护中的作用,讨论了这一发现的意义。