Tas E, Saano A, Leinonen P, Lindström K
Department of Applied Chemistry and Microbiology, University of Helsinki, Finland.
Appl Environ Microbiol. 1995 May;61(5):1822-7. doi: 10.1128/aem.61.5.1822-1827.1995.
Procedures based on DNA hybridization and PCR were developed for quality control of Rhizobium inoculants. Inoculants for pea and goat's rue were produced by Elomestari Ltd., Juva, Finland, in sterile dry fine peat by the standard procedure used by the company. The inoculants contained Rhizobium galegae HAMBI 1174 and HAMBI 1207 and an R. leguminosarum biovar vicia strain, 16HSA, either solely or in combinations of two or three strains. DNA was isolated from 1-g samples of each peat inoculant and analyzed by nonradioactive DNA-DNA hybridization and by PCR. The hybridization probes were total DNAs from pure cultures of R. galegae HAMBI 1207 and R. leguminosarum biovar viciae 16HSA and a 264-bp strain-specific fragment from the genome of R. galegae HAMBI 1174. The total DNA probes distinguished inoculants containing R. galegae or R. leguminosarum, and the strain-specific probe distinguished inoculants containing R. galegae HAMBI 1174. The hybridization results for R. galegae were verified in a PCR experiment by amplifying an R. galegae species-specific fragment and an R. galegae HAMBI 1174 strain-specific fragment in the same reaction. When suitable probes and primers are available, the methods described here offer promising alternatives for the quality control of peat-based inoculants.
开发了基于DNA杂交和PCR的程序用于根瘤菌接种剂的质量控制。豌豆和山羊豆的接种剂由芬兰尤瓦的Elomestari有限公司按照该公司使用的标准程序在无菌干燥细泥炭中生产。接种剂含有山羊豆根瘤菌HAMBI 1174和HAMBI 1207以及豌豆根瘤菌蚕豆生物型菌株16HSA，单独或两种或三种菌株组合存在。从每个泥炭接种剂的1克样品中分离DNA，并通过非放射性DNA-DNA杂交和PCR进行分析。杂交探针是来自山羊豆根瘤菌HAMBI 12
