How do I activate germination in dormant seeds of Arabidopsis Thaliana (Ecotype: Col-0, stored in refrigerator) ? I have to sow the seeds in soil.
I have never dealt with plant seeds before and therefore have no previous hand on experience. Need help as a starter.
Hello,
seed incubation for a few days in distilled water with giberellic acid (1000 ppm) should be sufficient to overcome physiological dormancy.
Hi,
Thank you for the reply.
I have been searching around in literature. Found the GA3 treatment. But do I have to use some media? Can't really get the media part though. Any idea what's all that about?
Honestly, Col-0 doesn't need to be chilled to germinate. You can just sow the seeds directly on sterile soil. Chilling just helps to synchronize the germination.
If you have a large refrigerator or walk-in cold room, you can directly sow the seeds onto the soil, make sure it's very well-watered and stick the pots of soil in the cold for 3-4 days to synchronize the germination.
But it's honestly a bit annoying to put seeds directly onto soil. They are so tiny that it's easy to spill too many. And you can't tell the germination rate since the seeds are so tiny.
What I used to do was sterilize my seeds, sprinkle them onto 0.5X MS agar plates, stick the plates in the refrigerator for 2-3 days, and then put the plates under growth lights. They will germinate in 1-2 days. I found it easiest to transplant just barely germinated seeds (root tip protruding from the seed coat). Make sure your soil is sterile, very wet, and fine-textured. Cover with a plastic dome until the cotyledons emerge, then vent the dome for a day or so, then you can take off the dome.
The easier tool for picking up and transferring very tiny plants is just a cheap plastic micropipette tip. I preferred P200 size.
Good luck!
Hello Katie!
Thank you so much. This is very helpful.
But i need to ask that you grow the seeds on the MS media and then just pick it up on the tips and sow them in the soil?
Also any fertilizer for the soil? So far i have read using Osmocote, any help here?
Hello Khudeja, we look at flowers so we regularly sow directly onto soil without any sterilisation. The compost we use is Levington M3. I sprinkle a few amount of seeds onto a small piece of folded paper, and then gently tap the seeds above the pot. If you do it gently enough you can count the seeds as they fall. We normally do between 1-4 seeds per pot. If you are unsure of the germination rate you could do more and then pull out any extra seedlings once they have germinated.
In regards to chilling, they dont need chilling but as Katie says this synchronises germination. We normally chill the seeds for 2-3 days after collection from the plant and then either store at room temperature or in the fridge. And sow directly from these.
Col-0 seeds are not dormant and do not need any GA or chilling to break the dormancy. Water is the only thing needed to break the dormancy and you can germinate them on a wet piece of paper.
If you do grow the seeds on MS media - use half MS 2.15g/L. Then once they are 1-2 weeks old, you can carefully pick the seedings up by their cotyledon - I normally just use my gloved fingers. Do not use forceps around the stem of the plant as this will kill them. To transfer to soil I put a small hole (1cm) into the compost with my finger, lie the seedlings root into this with the shoot to one side, and then carefully cover the root with compost. These will then need to be covered for 2 days to prevent stressing the plant (as they will have come from a very humid environment on the plate), these can then be uncovered/bags opened after this.
Good luck.
We resuspend our seeds in (distilled) water in a microfuge tube and use a P20 pipettor (cut off the tip at the end to have a wider bore) and dispense the seeds (usually 1, but a few is also OK) directly on soil. That way you can have a more even distribution of seeds in the pot (especially larger pots that we use for transformation experiments, 6" dia) or on flats/trays.
Col-0 seeds have week or no dormancy. I would start with sowing the seeds on wet soil, incubating in dark/4degree C/ 4day and place under light. If it fails, I would start treating with GA3.
can we sow seeds on media, Immediately after collecting seeds?
- Badges
- Science topic
- Similar topics
- Agriculture
- Horticulture
- Seeds