Schmidt E L, Bakole R O, Bohlool B B
J Bacteriol. 1968 Jun;95(6):1987-92. doi: 10.1128/jb.95.6.1987-1992.1968.
Application of fluorescent-antibody (FA) techniques to the study of rhizobia as free-living soil bacteria was explored. Antiserum to a particular strain of Rhizobium japonicum proved specific in both agglutination and FA tests. Within the R. japonicum group, 2 of 12 strains were stained by the conjugate and these fluoresced brightly; all others were entirely negative. FA tests were negative for 7 strains of R. meliloti, 9 strains of R. leguminosarum, 9 strains of R. trifolii, 6 strains of R. phaseoli, and 65 unidentified bacteria isolated from 12 soils. R. japonicum grew in autoclaved soil and was readily detectable by FA examination of contact slides. The FA technique also detected antibody-reacting bacteria in a field soil whose rhizobial content was unknown. Fluorescent cells were probably R. japonicum, since nodules developed on soybean plants grown in the same soil sample and FA preparations of the crushed nodules proved positive. Autofluorescence was not a problem, but nonspecific adsorption of conjugate restricted observations to microscopic fields free from soil particles. Nonspecific adsorption was substantial, irrespective of the soil used.
探索了荧光抗体（FA）技术在研究作为自由生活土壤细菌的根瘤菌方面的应用。针对特定日本根瘤菌菌株的抗血清在凝集试验和FA试验中均具有特异性。在日本根瘤菌组内，12个菌株中有2个被结合物染色并发出明亮荧光；其他所有菌株均为完全阴性。对于7株苜蓿根瘤菌、9株豌豆根瘤菌、9株三叶草根瘤菌、6株菜豆根瘤菌以及从12种土壤中分离出的65株未鉴定细菌，FA试验均为阴性。日本根瘤菌在灭菌土壤中生长，通过对接触玻片进行FA检查很容易检测到。FA技术还在一种根瘤菌含量未知的田间土壤中检测到了与抗体反应的细菌。荧光细胞可能是日本根瘤菌，因为在同一土壤样本中种植的大豆植株上形成了根瘤，且对压碎根瘤的FA制剂检测呈阳性。自发荧光不是问题，但结合物的非特异性吸附限制了观察范围，只能观察没有土壤颗粒的微观视野。无论使用何种土壤，非特异性吸附都很显著。
