CLI: A new protocol for the isolation of Lactic Acid Bacteria from complex plant samples
CLI: A new protocol for the isolation of Lactic Acid Bacteria from complex plant samples
Authors
Chavez, Mariannys
Viscardi, Sharon
Ruiz, Maria Jose
Sans Serramitjana, Eulalia
Duran, Paola
Viscardi, Sharon
Ruiz, Maria Jose
Sans Serramitjana, Eulalia
Duran, Paola
Profesor GuĆa
Authors
Date
Datos de publicaciĆ³n:
10.1016/j.mimet.2024.106937
JOURNAL OF MICROBIOLOGICAL METHODS,Vol.221,2024
JOURNAL OF MICROBIOLOGICAL METHODS,Vol.221,2024
Tipo de recurso
Article
Keywords
Materia geogrƔfica
Collections
Abstract
Lactic Acid Bacteria (LAB) are predominantly probiotic microorganisms and the most are Generally Recognized As Safe (GRAS). LAB inhabit in the human gut ecosystem and are largely found in fermented foods and silage. In the last decades, LAB have also has been found in plant microbiota as a new class of microbes with probiotic activity to plants. For this reason, today the scientific interest in the study and isolation of LAB for agronomic application has increased. However, isolation protocols from complex samples such as plant tissues are scarce and inefficient. In this study, we developed a new protocol (CLI, Complex samples LAB Isolation) which yields purified LAB from plants. The sensitivity of CLI protocol was sufficient to isolate representative microorganisms of LAB genera (i.e. Leuconostoc, Lactococcus and Enterococcus). CLI protocol consists on five steps: i) sample preparation and pre-incubation in 1% sterile peptone at 30 degrees C for 24-48 h; ii) Sample homogenization in vortex by 10 min; iii) sample serial dilution in quarter-strength Ringer solution, iv) incubation in MRS agar plates with 0.2% of sorbic acid, with 1% of CaCO3, O-2 < 15%, at pH 5.8 and 37 degrees C for 48 h.; v) Selection of single colonies with LAB morphology and CaCO3-solubilization halo. Our scientific contribution is that CLI protocol could be used for several complex samples and represents a useful method for further studies involving native LAB.