Shaw J J, Dane F, Geiger D, Kloepper J W
Department of Botany and Microbiology, Alabama Agricultural Experiment Station, Auburn University, Alabama 36849.
Appl Environ Microbiol. 1992 Jan;58(1):267-73. doi: 10.1128/aem.58.1.267-273.1992.
The persistence and movement of strain JS414 of Xanthomonas campestris pv. campestris, which was genetically engineered to bioluminesce, were monitored during a limited field introduction. Bioluminescence and traditional dilution plate counts were determined. Strain JS414 was applied to cabbage plants and surrounding soil by mist inoculation, by wound inoculation, by scattering infested debris among plants, and by incorporating bacteria into the soil. Bioluminescent X. campestris pv. campestris was detected in plant samples and in the rhizosphere up to 6 weeks after inoculation. Movement to uninoculated plants was detected on one occasion, but movement from the immediate release area was not detected. Strain JS414 was detected in soil samples beneath mist- and wound-inoculated plants only at intentionally infested locations and in aerial samples only on the day of inoculation. Our bioluminescence methods proved to be as sensitive as plating methods for detecting the genetically engineered microorganisms in environmental samples. Our results demonstrate that transgenic incorporation of the luxCDABE operon provides a non-labor-intensive, sensitive detection method for monitoring genetically engineered microorganisms in nature.
对经过基因工程改造能够发出生物荧光的野油菜黄单胞菌野油菜致病变种JS414菌株在有限的田间引入期间的存活和移动情况进行了监测。测定了生物荧光和传统的稀释平板计数。通过喷雾接种、伤口接种、在植株间撒播受侵染的残体以及将细菌混入土壤等方式，将JS414菌株施用于甘蓝植株及其周围土壤。在接种后长达6周的时间里，在植物样本和根际中检测到了能发出生物荧光的野油菜黄单胞菌野油菜致病变种。有一次检测到了向未接种植株的移动，但未检测到从直接释放区域的移动。仅在故意侵染的地点，在喷雾接种和伤口接种植株下方的土壤样本中检测到了JS414菌株，且仅在接种当天的空气样本中检测到了该菌株。我们的生物荧光方法在检测环境样本中的基因工程微生物方面被证明与平板计数方法一样灵敏。我们的结果表明，luxCDABE操纵子的转基因整合为监测自然界中的基因工程微生物提供了一种非劳动密集型的灵敏检测方法。
