Department of Microbiology, Institute of Biology, Martin-Luther University Halle-Wittenberg, Halle (Saale), Germany.
PLoS One. 2012;7(4):e34941. doi: 10.1371/journal.pone.0034941. Epub 2012 Apr 6.
The aerobic plant pathogenic bacterium Xanthomonas campestris pv. vesicatoria (Xcv) colonizes the intercellular spaces of pepper and tomato. One enzyme that might contribute to the successful proliferation of Xcv in the host is the iron-sulfur protein aconitase, which catalyzes the conversion of citrate to isocitrate in the tricarboxylic acid (TCA) cycle and might also sense reactive oxygen species (ROS) and changes in cellular iron levels. Xcv contains three putative aconitases, two of which, acnA and acnB, are encoded by a single chromosomal locus. The focus of this study is aconitase B (AcnB). acnB is co-transcribed with two genes, XCV1925 and XCV1926, encoding putative nucleic acid-binding proteins. In vitro growth of acnB mutants was like wild type, whereas in planta growth and symptom formation in pepper plants were impaired. While acnA, XCV1925 or XCV1926 mutants showed a wild-type phenotype with respect to bacterial growth and in planta symptom formation, proliferation of the acnB mutant in susceptible pepper plants was significantly impaired. Furthermore, the deletion of acnB led to reduced HR induction in resistant pepper plants and an increased susceptibility to the superoxide-generating compound menadione. As AcnB complemented the growth deficiency of an Escherichia coli aconitase mutant, it is likely to be an active aconitase. We therefore propose that optimal growth and survival of Xcv in pepper plants depends on AcnB, which might be required for the utilization of citrate as carbon source and could also help protect the bacterium against oxidative stress.
需氧植物病原细菌丁香假单胞菌番茄致病变种(Xcv)定植于辣椒和番茄的细胞间隙。可能有助于 Xcv 在宿主中成功增殖的一种酶是铁硫蛋白 aconitase,它催化三羧酸(TCA)循环中柠檬酸向异柠檬酸的转化,并且还可能感应活性氧(ROS)和细胞铁水平的变化。Xcv 包含三个假定的 aconitase,其中两个,acnA 和 acnB,由单个染色体基因座编码。本研究的重点是 aconitase B(AcnB)。acnB 与两个基因 XCV1925 和 XCV1926 共转录,编码假定的核酸结合蛋白。acnB 突变体的体外生长与野生型相似,而在辣椒植物中的生长和症状形成受损。虽然 acnA、XCV1925 或 XCV1926 突变体在细菌生长和植物体内症状形成方面表现出野生型表型,但 acnB 突变体在易感辣椒植物中的增殖受到显著抑制。此外,acnB 的缺失导致抗性辣椒植物中 HR 诱导减少,对超氧化物生成化合物亚甲萘醌的敏感性增加。由于 AcnB 弥补了大肠杆菌 aconitase 突变体的生长缺陷,它很可能是一种活性 aconitase。因此,我们提出 Xcv 在辣椒植物中的最佳生长和存活取决于 AcnB,这可能是利用柠檬酸作为碳源所必需的,并且还可以帮助保护细菌免受氧化应激。