Chandra Harish, Khandelwal Puneet, Khattri Arun, Banerjee Nirupama
International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India.
Environ Microbiol. 2008 May;10(5):1285-95. doi: 10.1111/j.1462-2920.2007.01542.x. Epub 2008 Feb 14.
Xenorhabdus nematophila produces type 1 fimbriae on the surface of Phase I cells. Fimbriae mediate recognition and adhesion of the bacteria to its target cell. To investigate the role of fimbriae in the biology of X. nematophila, we have produced a fimbrial mutant strain by insertional inactivation of the mrxA gene, encoding the structural subunit of type 1 fimbriae. Phenotypic characterization of the mutant revealed loss of fimbriae on the cell surface. Cell surface characteristics like dye absorption, biofilm formation, red blood cell agglutination remained unaltered. The mrxA mutant was defective in swarming on soft agar, although swimming motility was not affected. Flagellar expression was suppressed in the mrxA strain under swarming conditions, but not swimming conditions. Agglutination and cytotoxicity of the mutant to larval haemocytes was also reduced. When the mutant cells were injected in the haemocoel of the fourth instar larvae of Helicoverpa armigera, an increase in the LT(50) of 9-12 h was observed relative to the wild-type strain. The nematode growth was slow on the lawn of the fimbrial mutant. The mrxA negative strain was unable to colonize the nematode gut efficiently. This study demonstrates importance of type 1 fimbriae in establishment of bacteria-nematode symbiosis, a key to successful pest management program.
嗜线虫致病杆菌(Xenorhabdus nematophila)在I期细胞表面产生1型菌毛。菌毛介导细菌与靶细胞的识别和黏附。为了研究菌毛在嗜线虫致病杆菌生物学中的作用,我们通过插入失活编码1型菌毛结构亚基的mrxA基因,构建了一个菌毛突变菌株。突变体的表型特征显示细胞表面菌毛缺失。染料吸收、生物膜形成、红细胞凝集等细胞表面特征未发生改变。mrxA突变体在软琼脂上的群游运动存在缺陷,不过其游泳运动能力未受影响。在群游条件下,mrxA菌株中的鞭毛表达受到抑制,但在游泳条件下不受影响。该突变体对幼虫血细胞的凝集和细胞毒性也有所降低。当将突变体细胞注射到棉铃虫(Helicoverpa armigera)四龄幼虫的血腔中时,相对于野生型菌株,观察到其半数致死时间(LT50)增加了9 - 12小时。在菌毛突变体的菌苔上,线虫生长缓慢。mrxA基因阴性菌株无法有效地定殖于线虫肠道。本研究证明了1型菌毛在建立细菌 - 线虫共生关系中的重要性,而这是成功的害虫管理计划的关键。