To begin, weigh five grams of dry super absorbent polymer or SAP, and mix it with 500 milliliters of MMS1 nutrient solution. Approximately one week before assembly, germinate plantago lanceolata seeds on blotting paper moistened with MMS1 nutrient solution in a Petri dish. Then incubate the Petri dish in the dark at room temperature.
To start assembling the SAP-AS system, place the seedling stem in the notch on the side of the Petri dish with the root inward and the cotyledons or leaves and stems outward. Cover the roots with approximately 1.5 to two grams of vermiculite. Afterward, hydrate the root compartment with eight milliliters of MMS1 nutrient solution.
Clear a vermiculite-free space along the nylon mesh filter on the side of the root compartment, and add five grams of hydrated SAP. Then add 15 grams of hydrated SAP to the highfill compartment. Now align the side notches of the lid and the Petri dish bottom to avoid damaging the stem and close the lid.
Then seal the Petri dish with a strip of parafilm, joining the base and lid. Weigh the Petri dish and record its average weight to determine when the SAP-AS needs rehydration. Then stack the Petri dishes to optimize space.
Use an opaque cover with an opening on the side to keep the seedlings in the dark. Using a flame candle or a Bunsen burner, heat the glass pipette and stretch the glass tube as it melts to separate it. Under a stereo microscope, break the tip of the glass pipette to reduce its internal diameter.
Now open the Petri dish and locate where the spore is to be placed. Clear a space among the five grams of hydrated sap along the nylon mesh filter membrane to expose a section of a root. Under the dissecting microscope, pick up a spore in a liquid suspension with the extruded pipette, while pipetting some liquid before collecting the spore.
After that, carefully deposit the spore on the root. Then replace the sap over the root fragment to maintain a moist microenvironment for the root and spore. After 24 hours, if needed, irrigate the root compartment, taking care not to wash off the spore.
Align the side notches of the lid with the bottom of the Petri dish to avoid damaging the seedling stem and close the Petri dish with the lid.
