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From 2014.igem.org

Contents 1 Sexual Reproduction 1.1 Inducing sexual reproduction 1.2 Fertilising 1.3 Storing sporangia 1.4 Notes

Sexual Reproduction

You will need to do this to collect spores for transformation. See it as 'making a competent stock'. The following protocols apply for working with Marchantia growing in soil, or in sterile culture.

Inducing sexual reproduction

The sexual phase can be induced by incubating Marchantia in far red light (650nm+) for approximately 2 weeks.

Fertilising

1. Pipette ~ 50-100 ul sterile water onto sexual organs

2. Using a sterile brush, brush spermatozoa from archaegoniaphore onto female sexual organ

3. Sporeheads start to be visible in 2 weeks

Storing sporangia

Cut individual sporeheads and store with silica beads at -80 for long term storage, or at -20 for medium term storage. For short term use, sporangia can be left on a bench indefinitely, with little loss of viability.

Notes

Try and be sterile! Fungal contamination is persistent and difficult to get rid of.