Bacterial culture isolation and maintenance from plant phyllosphere (Pseudomonadaceae)

Knowledge graph centered on Bacterial culture isolation and maintenance from plant phyllosphere (Pseudomonadaceae) with 14 nodes and 29 connections. Top connected: Bacteria, Cardamine cordifolia, Pseudomonas, genome polishing, phyllosphere microbiome.

Method synopsis

Protocol for isolating and maintaining pure bacterial cultures from plant leaf surfaces through surface sterilization, plating on selective media, and cryopreservation in glycerol stocks.

Synthesized from method descriptions across 1 paper using this protocol.

Procedure from a recent peer-reviewed implementation

Steps below were extracted from the most recent peer-reviewed implementation of this protocol in the corpus — Complete Genome Sequences for <i>Pseudomonas sp.</i> Strains 29A and 43A (2020), Microbiology Resource Announcements DOI: 10.1128/MRA.00845-20. Implementations in other papers (listed below) may differ.

1. Culture maintenance and single colony isolation
   Pure cultures of strains 29A and 43A were originally isolated from within surface-sterilized leaves from two individual plants by Humphrey et al. and acquired by the Baltrus lab after a limited (<4) number of passages in rich medium on agar plates. The isolates were streaked to single colonies by the Baltrus lab onto King's B (KB) agar plates.
   Quantities: Two strains (29A and 43A), fewer than 4 passagesDuration: Not specifiedConditions: Rich medium on agar plates
   Equipment: King's B (KB) agar plates
2. Stock culture preparation
   A single colony arising from the original platings was picked to 2 ml KB broth and grown overnight at 27°C, at which point a subset was frozen in −80°C in 40% glycerol to create stock cultures.
   Quantities: 2 ml KB broth, 40% glycerol concentrationDuration: Overnight growthConditions: 27°C temperature, −80°C freezer storage
   Equipment: KB broth, shaking incubator, freezer

Typical Equipment

• King's B (KB) agar plates
• KB broth
• shaking incubator
• freezer

Output Measurements

• pure bacterial cultures
• frozen stock cultures

Papers Using This Protocol (1)

Species Studied With This Protocol