Zaini Paulo A, Fogaça Andréa C, Lupo Fernanda G N, Nakaya Helder I, Vêncio Ricardo Z N, da Silva Aline M
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof. Lineu Prestes 748, 05508-000, São Paulo, SP, Brazil.
J Bacteriol. 2008 Apr;190(7):2368-78. doi: 10.1128/JB.01495-07. Epub 2008 Jan 25.
Xylella fastidiosa is the etiologic agent of a wide range of plant diseases, including citrus variegated chlorosis (CVC), a major threat to citrus industry. The genomes of several strains of this phytopathogen were completely sequenced, enabling large-scale functional studies. DNA microarrays representing 2,608 (91.6%) coding sequences (CDS) of X. fastidiosa CVC strain 9a5c were used to investigate transcript levels during growth with different iron availabilities. When treated with the iron chelator 2,2'-dipyridyl, 193 CDS were considered up-regulated and 216 were considered down-regulated. Upon incubation with 100 microM ferric pyrophosphate, 218 and 256 CDS were considered up- and down-regulated, respectively. Differential expression for a subset of 44 CDS was further evaluated by reverse transcription-quantitative PCR. Several CDS involved with regulatory functions, pathogenicity, and cell structure were modulated under both conditions assayed, suggesting that major changes in cell architecture and metabolism occur when X. fastidiosa cells are exposed to extreme variations in iron concentration. Interestingly, the modulated CDS include those related to colicin V-like bacteriocin synthesis and secretion and to functions of pili/fimbriae. We also investigated the contribution of the ferric uptake regulator Fur to the iron stimulon of X. fastidiosa. The promoter regions of the strain 9a5c genome were screened for putative Fur boxes, and candidates were analyzed by electrophoretic mobility shift assays. Taken together, our data support the hypothesis that Fur is not solely responsible for the modulation of the iron stimulon of X. fastidiosa, and they present novel evidence for iron regulation of pathogenicity determinants.
木质部难养菌是多种植物病害的病原体,包括柑橘杂色黄化病(CVC),这对柑橘产业构成了重大威胁。对该植物病原体的多个菌株的基因组进行了全序列测定,从而能够开展大规模的功能研究。利用代表木质部难养菌CVC菌株9a5c的2608个(91.6%)编码序列(CDS)的DNA微阵列,研究了在不同铁可用性条件下生长期间的转录水平。用铁螯合剂2,2'-联吡啶处理时,193个CDS被认为是上调的,216个被认为是下调的。与100微摩尔焦磷酸铁孵育后,分别有218个和256个CDS被认为是上调和下调的。通过逆转录定量PCR进一步评估了44个CDS子集的差异表达。在两种测定条件下,几个与调节功能、致病性和细胞结构有关的CDS都受到了调节,这表明当木质部难养菌细胞暴露于铁浓度的极端变化时,细胞结构和代谢会发生重大变化。有趣的是,受调节的CDS包括那些与类大肠杆菌素V细菌素合成和分泌以及菌毛/纤毛功能有关的CDS。我们还研究了铁摄取调节蛋白Fur对木质部难养菌铁刺激子的作用。对菌株9a5c基因组的启动子区域进行了推定的Fur框筛选,并通过电泳迁移率变动分析对候选物进行了分析。综上所述,我们的数据支持这样一种假设,即Fur并非唯一负责调节木质部难养菌的铁刺激子,并且它们为致病性决定因素的铁调节提供了新的证据。