Rooted Tissue Culture Acclimatization
Step 1- Prepare Substrate
A) Mix nutrient solution (Clonex is preferred) at 1.5-2.0 EC
B) Wet substrate according to recommended moisture content for propagation. It should be moist but not soaked.
C) Loosen substrate or cut substrate so that the plantlets easily nests without breaking delicate roots
D) Loose or aggregate substrates are best for this type of plantlet such as peat or coir based. Plugs and rockwool are less effective, but can be also be used as long as they are cut to fit the plantlet.
Step 2- Remove agar from the plantlet
A) Using clean forceps, remove plantlet from test tube gently and slowly. The roots at this stage can be brittle.
B) Gently remove chunks of agar using the forceps. A spray bottle or wash bottle with tap water can also be used to wash the roots. It is important to remove the agar since it can attract fungus and mold if too much is left on. A small amount is fine- just remove chunks larger than 2mm.
Step 3- Place plantlet into substrate
A) Using forceps, place plantlets into preferred substrate. Loose or aggregate substrates are preferred. The roots are very delicate and brittle at this stage. It is not necessary to place the plant too firmly or deeply into the substrate.
B) Cuttings with large root masses may be difficult to place. Simply cut or break up substrate to fit the cutting and gently sprinkle additional substrate on top. The plant will root into the rest of the substrate over the next couple days.
Step 4- Grow in propagation grow conditions
A) Place in tray and place dome on top. 100-200 ppfd, 78-83F (25-28C), 65-80% RH
B) Vent as much as possible as long as they aren't wilted. Burp daily. After 3-5 days, remove the dome if no wilting is observed and you should see roots penetrating the substrate. You may need to lightly moisten the cubes during this time, but do not soak or saturate them.
C) After 7-10 days, once roots are fully penetrating the cube, transplant into standard veg conditions.
*D) If you only have a few plantlets to acclimatize, it may be good to place empty substrate or some standing water into the tray to provide additional humidity. Plants acclimatize best when the tray is mostly full.
