Exploration

General overview

^[1]

Useful host strains:

• Streptomyces venezuelae ^[1]

This species is currently the most well characterised for exploration.

Abiotic induction protocol:

• Make 33-50mL YP plates
• Inoculate 5μL of Actinomyces spp.. spore stock in the centre of the plate
• Incubate at 30ºC for 14 days
• Image plate

This protocol is more reliable as you are not reliant on the activity of the yeast co-culture

Biotic induction protocol:

• Grow Saccharomyces cerevisiae in an overnight culture of liquid YPD
• Make 33-50mL YPD plates
• Inoculate 5μL of Actinomyces spp. spore stock in the centre of the plates
• Inoculate 5μL of Saccharomyces cerevisiae overnight culture next to the other inoculation and blend gently
• Incubate at 30ºC for 14 days
• Image plate

This protocol can be less reliable due to the nature of the yeast co-culture but allows for reciprocal testing if required.

References: [1] Jones, S., Ho, L., Rees, C., Hill, J., Nodwell, J. and Elliot, M. (2017). Streptomyces exploration is triggered by fungal interactions and volatile signals. eLife, 6.

Protocol adapted for Actinobase by Thomas C. McLean, The University of East Anglia