Abstract
Primers and conditions for nested PCR were designed to detect the ammonia monooxygenase gene (amo) of chemolithotrophic ammonia-oxidizers in soil.
By the nested PCR, an amo fragment was produced from 10 or fewer fg of genome DNA of pure-cultured Nitrosomonas europaea and Nitrosolobus multiformis. The practical limit for detecting amo was examined by amplifying DNA extracted from autoclaved and inoculated (N. europaea) soil. Soil DNA was prepared by a standard cell extraction method. Before lyzing, cell suspensions were treated with DNase overnight to destroy extracellular DNA which had formed a positive interference. A PCR product was obtained when 103 cells g-1 of fresh soil (1 cell g-1 in the most successful case) were inoculated. The product from amo of the N. europaea-type was detected by Southern hybridization from DNA of cultivated soil, but that of N. multiformis was not.
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References
Head I M, Hiorns W D, Martin T, McCarthy A J and Saunders J R 1993 J. Gen. Microbiol. 139, 1147–1153.
Hiorns W D, Hastings R C, Head I M, McCarthy A J, Saunders J R, Pickup R W and Hall G H 1995 Microbiol. 141, 2793–2800.
Hyman M R, Russell S A, Ely R L, Williamson K J and Arp D J 1995 Appl. Environ. Microbiol. 61, 1480–1487.
Hynes R K and Knowles R 1984 Can. J. Microbiol. 30, 1397–1404.
Rasche M E, Hyman M R and Arp D J 1991 Appl. Environ. Microbiol. 57, 2986–2994.
Stephen J R, McCaig A E, Smith Z, Prosser J I and Embley T M 1996 Appl. Environ. Microbiol. 62, 4147–4154.
Vannelli T, Logan M, Arciero D M and Hooper A B 1990 Appl. Environ. Microbiol. 56, 1169–1171.
Wood P M 1986 In Nitrification. Ed. J I Prosser. pp 39–62. IRL Press, Oxford, Washington DC.
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© 1997 Kluwer Academic Publishers
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Yokoyama, K., Kimura, H., Shinozaki, H., Tajima, S. (1997). Detection of amo gene of chemolithotrophic ammonia-oxidizers in soil by nested PCR. In: Ando, T., Fujita, K., Mae, T., Matsumoto, H., Mori, S., Sekiya, J. (eds) Plant Nutrition for Sustainable Food Production and Environment. Developments in Plant and Soil Sciences, vol 78. Springer, Dordrecht. https://doi.org/10.1007/978-94-009-0047-9_240
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DOI: https://doi.org/10.1007/978-94-009-0047-9_240
Publisher Name: Springer, Dordrecht
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