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Specific and sensitive detection of Alcaligenes species from an agricultural environment
Author(s) -
Nakano Miyo,
Niwa Masumi,
Nishimura Norihiro
Publication year - 2013
Publication title -
microbiology and immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.664
H-Index - 70
eISSN - 1348-0421
pISSN - 0385-5600
DOI - 10.1111/1348-0421.12026
Subject(s) - alcaligenes faecalis , alcaligenes , biology , genomic dna , genus , detection limit , soil water , agriculture , soil microbiology , dna , microbiology and biotechnology , botany , veterinary medicine , bacteria , chromatography , ecology , genetics , chemistry , pseudomonas , medicine
A quantitative real‐time PCR assay to specifically detect and quantify the genus Alcaligenes in samples from the agricultural environment, such as vegetables and farming soils, was developed. The minimum detection sensitivity was 106 fg of pure culture DNA, corresponding to DNA extracted from two cells of Alcaligenes faecalis . To evaluate the detection limit of A. faecalis , serially diluted genomic DNA from this organism was mixed with DNA extracted from soil and vegetables and then a standard curve was constructed. It was found that Alcaligenes species are present in the plant phytosphere at levels 10 2 –10 4 times lower than those in soil. The approach presented here will be useful for tracking or quantifying species of the genus Alcaligenes in the agricultural environment.