CLI: A new protocol for the isolation of Lactic Acid Bacteria from complex plant samples
Date
Authors
VISCARDI, SHARON
Chavez, Mariannys
Viscardi, S.
Ruiz, María José
Sans-Serramitjana, Eulália
Durán, Paola C.
Chavez, Mariannys
Viscardi, S.
Ruiz, María José
Sans-Serramitjana, Eulália
Durán, Paola C.
Authors
Date
Datos de publicación:
10.1016/j.mimet.2024.106937
Keywords
Complex Samples - Lactic Acid Bacteria - New Method - Plant Probiotic - Ringer Solution - Sorbic Acid - Lactic Acid - Ringer Solution - Sorbic Acid - Lactic Acid - Probiotic Agent - Article - Bacterium Colony - Bacterium Isolate - Bacterium Isolation - Colony Forming Unit - Dilution - Enterococcus - Lactic Acid Bacterium - Lactococcus - Leuconostoc - Nonhuman - Ph - Plant - Sensitivity Analysis - Solubilization - Classification - Isolation And Purification - Lactobacillales - Metabolism - Microbiology - Lactic Acid - Plants - Probiotics
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 °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 CaCO<inf>3</inf>, O<inf>2</inf> < 15%, at pH 5.8 and 37 °C for 48 h.; v) Selection of single colonies with LAB morphology and CaCO<inf>3</inf>-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. © 2024 Elsevier B.V., All rights reserved.
Description
Keywords
Complex Samples , Lactic Acid Bacteria , New Method , Plant Probiotic , Ringer Solution , Sorbic Acid , Lactic Acid , Ringer Solution , Sorbic Acid , Lactic Acid , Probiotic Agent , Article , Bacterium Colony , Bacterium Isolate , Bacterium Isolation , Colony Forming Unit , Dilution , Enterococcus , Lactic Acid Bacterium , Lactococcus , Leuconostoc , Nonhuman , Ph , Plant , Sensitivity Analysis , Solubilization , Classification , Isolation And Purification , Lactobacillales , Metabolism , Microbiology , Lactic Acid , Plants , Probiotics
Citation
10.1016/j.mimet.2024.106937