Monitoring Lactobacillus plantarum in grass silages with the aid of 16S rDNA-based quantitative real-time PCR assays.
Ensiling plant material with the aid of lactic acid bacteria (LAB) is a common agricultural practice for conserving forages independently of the time point of harvest. Despite ensiling being a natural process, it can be improved by the treatment of the harvested forage with starter cultures before storage. Within this context, Lactobacillus plantarum (L. plantarum) is the most frequently used LAB in commercially available starter cultures. In order to enable the monitoring of the population dynamics of L. plantarum in silage, methods for species-specific detection based on the 16S ribosomal DNA (rDNA) sequence were developed by applying a quantitative real-time polymerase chain reaction (QRT-PCR) approach. The QRT-PCR assay was also applied to estimate the development of the L. plantarum population within experimental grass silages. In addition, a multiplex QRT-PCR assay was developed to estimate the amount of L. plantarum 16S rDNA in relation to total bacterial 16S rDNA. This multiplex QRT-PCR assay was applied to monitor the influence of different silage additives on the L. plantarum population.
Klocke
Michael
M
Leibnitz-Institute for Agricultural Engineering Potsdam-Bornim (ATB), Department of Bioengineering, Max-Eyth-Allee 100, D-14469 Potsdam, Germany. mklocke@atb-potsdam.de
Mundt
Kerstin
K
Idler
Christine
C
McEniry
Joseph
J
O'Kiely
Padraig
P
Barth
Susanne
S
D019342:Acetic Acid / Q000032:analysis; D000480:Alkynes / Q000032:analysis; D020148:Butyric Acid / Q000032:analysis; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D045782:Hemiterpenes; D048191:Lactiplantibacillus plantarum / Q000235:genetics* / Q000254:growth & development*; D010421:Pentanoic Acids / Q000032:analysis; D043264:Phosphodiesterase I / Q000737:chemistry / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D011422:Propionates / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000737:chemistry / Q000235:genetics; D012820:Silage / Q000032:analysis / Q000382:microbiology*; D013045:Species Specificity
D000480:Alkynes; D017931:DNA Primers; D004269:DNA, Bacterial; D045782:Hemiterpenes; D010421:Pentanoic Acids; D011422:Propionates; D012336:RNA, Ribosomal, 16S; D020148:Butyric Acid; C008216:isovaleric acid; D043264:Phosphodiesterase I; C011537:propiolic acid; D019342:Acetic Acid