Development of a PCR-Based Method for Monitoring the Status of Alcaligenes Species in the Agricultural Environment

作者：Nakano Miyo; Niwa Masumi; Nishimura Norihiro^*

来源：Biocontrol Science, 2014, 19(1): 23-31.

DOI：10.4265/bio.19.23

• 16S rRNA PCR method
• Alcaligenes species
• Soil bacteria
• Plant phylloepiphytic bacteria
• Specific primer

To analyze the status of the genus Alcaligenes in the agricultural environment, we developed a PCR method for detection of these species from vegetables and farming soil. The selected PCR primers amplified a 107-bp fragment of the 16S rRNA gene in a specific PCR assay with a detection limit of 1.06 pg of pure culture DNA, corresponding to DNA extracted from approximately 23 cells of Alcaligenes faecalis. Meanwhile, PCR primers generated a detectable amount of the amplicon from 2.2 x 10(2) CFU/ml cell suspensions from the soil. Analysis of vegetable phylloepiphytic and farming soil microbes showed that bacterial species belonging to the genus Alcaligenes were present in the range from 0.9 x 10 CFU per gram (or cm(2)) (Japanese radish: Raphanus sativus var. longipinnatus) to more than 1.1 X 10(4) CFU/g (broccoli flowers: Brassica oleracea var. italic), while 2.4 x 10(2) to 4.4 x 10(3) CFU/g were detected from all soil samples. These results indicated that Alcaligenes species are present in the phytosphere at levels 10-1000 times lower than those in soil. Our approach may be useful for tracking or quantifying species of the genus Alcaligenes in the agricultural environment.

• 出版日期2014-3