Mol Plant Microbe Interact. 2013 Oct;26(10):1200-10. doi: 10.1094/MPMI-01-13-0002-R.
Several EZ-Tn5 insertions in gene locus XALc_0557 (OmpA1) of the sugarcane leaf scald pathogen Xanthomonas albilineans XaFL07-1 were previously found to strongly affect pathogenicity and endophytic stalk colonization. XALc_0557 has a predicted OmpA N-terminal outer membrane channel (OMC) domain and an OmpA C-like domain. Further analysis of mutant M468, with an EZ-Tn5 insertion in the upstream OMC domain coding region, revealed impaired epiphytic and endophytic leaf survival, impaired resistance to sodium dodecyl sulfate (SDS), structural defects in the outer membrane (OM), and hyperproduction of OM vesicles. Cloned full-length XALc_0557 complemented M468 for all phenotypes tested, including pathogenicity, resistance to SDS, and ability to survive both endophytically and epiphytically. Another construct, pCT47.3, which expressed only the C-like domain of XALc_0557, restored resistance to SDS in M468 but failed to complement any other mutant phenotype, indicating that the C-like domain functioned independently of the OMC domain to help maintain OM integrity. pCT47.3 also complemented pathogenicity, resistance to SDS, and stalk colonization in mutant M1152, which carries an EZ-Tn5 insert in the C-like coding region, indicating that both predicted domains are modular and necessary but neither is sufficient for X. albilineans pathogenicity, endophytic survival in, and epiphytic survival on sugarcane.
先前发现,甘蔗叶枯病菌 Xanthomonas albilineans XaFL07-1 基因座 XALc_0557(OmpA1)中的几个 EZ-Tn5 插入强烈影响致病性和内生茎定殖。XALc_0557 具有预测的 OmpA N 端外膜通道(OMC)结构域和 OmpA C 样结构域。对具有 EZ-Tn5 插入上游 OMC 结构域编码区的突变体 M468 的进一步分析表明,其附着和内生叶存活率受损,对十二烷基硫酸钠(SDS)的抗性受损,外膜(OM)结构缺陷,以及 OM 泡囊的过度产生。全长 XALc_0557 的克隆完全互补了 M468 的所有表型,包括致病性、对 SDS 的抗性以及在内生和附着两种环境下的生存能力。另一个构建体 pCT47.3 仅表达 XALc_0557 的 C 样结构域,恢复了 M468 对 SDS 的抗性,但未能互补任何其他突变体表型,表明 C 样结构域独立于 OMC 结构域发挥作用,有助于维持 OM 完整性。pCT47.3 还互补了突变体 M1152 的致病性、对 SDS 的抗性和茎定殖,M1152 携带 C 样编码区的 EZ-Tn5 插入,表明这两个预测结构域是模块化的,是必需的,但都不足以使 X. albilineans 致病,内生在甘蔗上的生存能力,和附着在甘蔗上的生存能力。