Department of Botany and Plant Pathology, Oregon State University, Corvallis, Oregon 97331.
Appl Environ Microbiol. 1987 Nov;53(11):2660-5. doi: 10.1128/aem.53.11.2660-2665.1987.
Serological techniques and restriction enzyme cleavage patterns of total DNA were used to differentiate strains of Agrobacterium spp. Forty-five wild-type and plasmid-cured Agrobacterium strains were tested by immunodiffusion and immunofluorescence against polyclonal antisera to a crude ribosome preparation from Agrobacterium strains K84, U11, B6, A323, NT1, and C58. In immunodiffusion gels, these antisera reacted only with water-phenol extracts of the homologous strain, producing a single, strain-specific precipitin line. In contrast, when the same antisera were used in immunofluorescence staining, cross-reactions occurred with a limited number of heterologous Agrobacterium strains. However, the cross-reacting heterologous cells fluoresced generally less brightly than the homologous cells. When the EcoRI-digested DNA profiles from the same Agrobacterium strains were compared, 34 distinct cleavage patterns were observed. The DNA profiles were the same for all strains sharing a common chromosomal background and correlated with the strain-specific serological reaction. The presence or absence of plasmid DNA did not alter the strain-specific serological reaction or the DNA cleavage patterns. Both the serological reaction and the restriction enzyme digestion of total DNA were complementary to each other. These methods were used successfully to identify A. radiobacter K84 strains which were recovered 6 months after being inoculated to young trees in the field.
血清学技术和限制性内切酶切割总 DNA 的模式被用来区分根癌农杆菌（Agrobacterium）的菌株。用针对根癌农杆菌菌株 K84、U11、B6、A323、NT1 和 C58 的粗核糖体制剂的多克隆抗血清，通过免疫扩散和免疫荧光对 45 株野生型和质粒消除的根癌农杆菌菌株进行了检测。在免疫扩散凝胶中，这些抗血清仅与同源菌株的水-酚提取物发生反应，产生单一的、菌株特异性的沉淀线。相比之下，当相同的抗血清用于免疫荧光染色时，与有限数量的异源根癌农杆菌菌株发生了交叉反应。然而，发生交叉反应的异源细胞荧光通常比同源细胞弱。当比较相同根癌农杆菌菌株的 EcoRI 消化 DNA 图谱时，观察到 34 种不同的切割模式。具有共同染色体背景的所有菌株的 DNA 图谱相同，并与菌株特异性血清反应相关。质粒 DNA 的存在与否不会改变菌株特异性血清反应或 DNA 切割模式。血清学反应和总 DNA 的限制性内切酶消化是互补的。这些方法成功地用于鉴定在野外接种到幼树后 6 个月恢复的 A. radiobacter K84 菌株。
