We aim to produce axenic plants (plants without associated microbes) from field-grown Sporobolus alterniflorus . We then will propagate the plants under sterile conditions, producing many clones of each individual, for experiments re-combining plants with selected, known microbial partners.
S. alterniflorus can be grown in the lab from undifferentiated "callus" tissue (i.e. growing masses of undifferentiated and unorganized cells). Hormones are used to drive production of new shoots and roots. But this approach tends to produce many spontaneous mutations. We are minimizing this risk by developing propagation methods that mimic the way plants naturally propagate themselves in the marsh.
We are pursuing two strategies
We also are gathering seeds and peeling away their protective coverings, leaving the kernal that can be treated with ethanol and bleach. Cleaned seeds sprout in agar in individual test tubes.
One approach is to take advantage of the natural ability of S. alterniflorus to make new shoots and roots rapidly from robust rhizomes (belowground stems). We can grow field-gathered plants hydroponically in the lab, producing far more clean new plant material. Standard approaches such as rinsing with soap and 10% bleach solutions can then be used to reduce, and hopefully eventually completely eliminate, microbes on the tissues.