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Development of a 16S rDNA‐targeted PCR assay for monitoring of Lactobacillus plantarum and Lact. rhamnosus during co‐cultivation for production of inoculants for silages
Author(s) -
Klocke M.,
Mundt K.
Publication year - 2004
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2004.01575.x
Subject(s) - microbial inoculant , lactobacillus rhamnosus , biology , lactobacillus plantarum , silage , microbiology and biotechnology , food science , bacteria , agar , multiplex , 16s ribosomal rna , multiplex polymerase chain reaction , polymerase chain reaction , lactobacillus , fermentation , lactic acid , biochemistry , gene , genetics
Aims: This paper reports a simple, rapid approach for the parallel detection of Lactobacillus plantarum and Lact. rhamnosus in co‐culture in order to produce an inoculant mixture for silage purposes. Methods and Results: The 16S rDNA‐targeted PCR primers were established for parallel detection of Lact. plantarum and Lact. rhamnosus in a single multiplex PCR. A protocol for application of these primers in direct PCR as well as colony‐direct (CD) PCR was developed. These primers were also applicable for the estimation of the relative amount of each DNA type in mixed probes (semi‐quantitative PCR). Conclusions: The PCR assay presented in this study is a robust, fast and semi‐quantitative approach for detection of Lact. plantarum and Lact. rhamnosus in liquid cultures as well as on agar plates. Significance and Impact of the study: This method provides an effective tool for the establishment of a regime for co‐cultivation of Lact. plantarum and Lact. rhamnosus . This would enable faster and thus cost‐reduced production of ensiling inoculants.