Shi YingWu, Li Chun, Yang HongMei, Zhang Tao, Gao Yan, Chu Min, Zeng Jun, Lin Qing, Li YuGuo, Huo Xiangdong, Lou Kai
Institute of Microbiology, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, Xinjiang, P. R. China.
Xinjiang Laboratory of Special Environmental Microbiology, Urumqi 830091, Xinjiang, P. R. China.
J Microbiol. 2017 Apr;55(4):267-272. doi: 10.1007/s12275-017-6371-1. Epub 2017 Jan 26.
This study details the introduction of a gfp marker into an endophytic bacterial strain (Achromobacter marplatensis strain 17, isolated from sugar beet) to monitor its colonization of sugar beet (Beta. vulgaris L.). Stability of the plasmid encoding the gfp was confirmed in vitro for at least 72 h of bacterial growth and after the colonization of tissues, under nonselective conditions. The colonization was observed using fluorescence microscopy and enumeration of culturable endophytes in inoculated sugar beet plants that grew for 10 or 20 days. gfp-Expressing strains were re-isolated from the inner tissues of surface-sterilized roots and stems of inoculated plants, and the survival of the Achromobacter marplatensis 17:gfp strain in plants 20 days after inoculation, even in the absence of selective pressure, suggests that it is good colonizer. These results also suggest that this strain could be a useful tool for the delivery of enzymes or other proteins into plants. In addition, the study highlights that sugar beet plants can be used effectively for detailed in vitro studies on the interactions between A. marplatensis strain 17 and its host, particularly if a gfp-tagged strain of the pathogen is used.
本研究详细介绍了将绿色荧光蛋白（gfp）标记引入一种内生细菌菌株（从甜菜中分离出的马尔普拉滕斯无色杆菌菌株17），以监测其在甜菜（Beta. vulgaris L.）中的定殖情况。在非选择性条件下，编码gfp的质粒在细菌生长至少72小时以及组织定殖后，其稳定性在体外得到了证实。使用荧光显微镜观察定殖情况，并对接种后生长10天或20天的甜菜植株中可培养内生菌进行计数。从接种植物经表面消毒的根和茎的内部组织中重新分离出表达gfp的菌株，接种20天后，即使在没有选择压力的情况下，马尔普拉滕斯无色杆菌17:gfp菌株在植物中的存活表明它是一种良好的定殖菌。这些结果还表明，该菌株可能是将酶或其他蛋白质输送到植物中的有用工具。此外，该研究强调，甜菜植株可有效地用于对马尔普拉滕斯菌株17与其宿主之间相互作用的详细体外研究，特别是如果使用带有gfp标签的病原菌菌株的话。
