Hello everyone,
I'm growing plants in hydroponic (A thaliana), and my issue is that there is a big heterogeneity between my plants especially in the first steps of development. Do you have any advice to reduce it?
Thanks,
Julie
Hi Julie,
Hydroponics are tricky and nearly an art to do well. Random distribution, starting material homogeneity, reduced contamination and stratification may help, see below.
I can suggest to have several boxes with random (and blindly) distributed plants on the boxes and on the growing chamber (if you have enough space). For example, you can number the boxes from 1 to 5 (or 10 or as much as you are using) and decide which line you put in box 1 hole "A" (1A = mutant "Research"), "B" (1B = WT), "C" (1C = mutant "Gate") and so on.
In addition, you can sow three seeds in each hole to make sure you get a healthy plant on each one. After a week you can remove two of them (which may have stop growing or did not germinate or are also healthy). It is also good to know the phenotype of your plants before doing hydroponics, just in case all look "unhealthy".
It is true that mold contamination also can affect the plants. To prevent it I washed well the boxes and the pieces for the holes and let them under UV before filling them with media and seeds. I put Scotch tape on the flat part of the pieces of the holes to prevent contamination while filling the holes with agar+media or water (as your lab prefers). When filled, I put them on the box, I removed the tape and sow. I stratified them for 2-3 days (as your lab prefers) at 4 C to coordinate the germination and then added the media before put in in the chamber. In some labs water instead of media is added during the first 1-3 weeks, when the roots are grown enough.
All the best,
Mila
Dear Julie, probably your problem has started with the seed quality. When seeds emerges from substrata along the time, the plants will start the treatments in different stages of development (number of pair of leaves). Please, see my paper about stages of plant development and you have a uniform bedding plant at beginning of your study. Best regards, Walter.
Thanks everyone for your advices. I will start using them. For my seeds they all came from the same batch (same date of sow and recovering)...and they are recent.
Mila I didn't understand where you put Scotch tape? How much time do you put the stuff under UV please?
Hi Julie,
I used araponic boxes and on the holes, there are "clove-shaped" pieces to insert. I put the tape on their "head". Then I put them upside down to fill them with agar from the tip with a needleless syringe; then I removed the tape so the surface of the "head" remains totally flat.
I put 15-20 min on one side of the boxes and 15-20 min on the other. I also put the "clove-shaped" pieces under UV before filling them with agar.
Hi Julie,
Here is the user guide from Araponics. On page 2 you will find a solution to fill the seedholders with agar. We use this method routinely in the lab. Note that to increase plantlet establishment success, we cover the hydroponic boxes with transparent lids during the first 2-3 weeks.
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