Department of Plant Molecular Biology, Leiden University, Nonnensteeg 3, 2311VJ Leiden, The Netherlands; Center for Environmental Biotechnology, Department of Microbiology and the Graduate Program in Ecology, The University of Tennessee, Knoxville, Tennessee 37932 ; and Department of Plant Pathology and Alabama Agriculture Experimental Station, Auburn University, Auburn, Alabama 36859-5409.
Appl Environ Microbiol. 1991 Dec;57(12):3641-4. doi: 10.1128/aem.57.12.3641-3644.1991.
The use of bioluminescence as a sensitive marker for detection of Pseudomonas spp. in the rhizosphere was investigated. Continuous expression of the luxCDABE genes, required for bioluminescence, was not detectable in the rhizosphere. However, when either a naphthalene-inducible luxCDABE construct or a constitutive luxAB construct (coding only for the luciferase) was introduced into the Pseudomonas cells, light emission could be initiated just prior to measurement by the addition of naphthalene or the substrate for luciferase, n-decyl aldehyde, respectively. These Pseudomonas cells could successfully be detected in the rhizosphere by using autophotography or optical fiber light measurement techniques. Detection required the presence of 10 to 10 CFU/cm of root, showing that the bioluminescence technique is at least 1,000-fold more sensitive than beta-galactosidase-based systems.
研究了利用生物发光作为检测根际假单胞菌的敏感标记物。在根际中未检测到生物发光所需的 luxCDABE 基因的连续表达。然而,当将萘诱导型 luxCDABE 构建体或组成型 luxAB 构建体(仅编码荧光素酶)引入假单胞菌细胞中时,通过分别添加萘或荧光素酶的底物正癸醛,可以在测量之前启动发光。可以使用自动摄影术或光纤光测量技术成功地在根际中检测到这些假单胞菌细胞。检测需要根的 10 到 10 CFU/cm 的存在,表明生物发光技术比基于β-半乳糖苷酶的系统至少灵敏 1000 倍。