Day 2 and still no sprouting, but it looks like there is some activity in some samples. Let’s keep an eye on it…
Here is day 3 and there are already some visible sprouting. Once again I’m not sure from what point Crumley considered a seed to be germinating but I definitely see radicle growth in many of the seeds in the ddw, and di water samples, and some sprouting in the 33% d2o sample. As expected, the sample with no seeds shows no sprouting (or abnormal growth of any sort).
I counted anything that had a white tip as the beginning of germination and here are my results (feel free to double check my numbers):
- No seeds – 0/0
- DDW – 19/30
- DI water – 20/30
- 33% D2O – 3/30
- 66% D2O – 0/30
- 99.9% D2O – 0/30
Before counting for germination, I counted to ensure there were 30 seeds in each sample, then I counted the white tips. It should be noted that after I took the picture for the 33% D2O I dropped the sample, a drop of water was lost, but more importantly the bubble moved all the seeds around.
Also to be noted, in the 33% D2O sample there is a white floating thing. I’m not sure what it is, but it could be a premature seedling that shed it’s seed coat. I still counted 30 seeds without the white thing, so we’ll see what happens. This is visible in the picture before I dropped it, so the trauma from dropping isn’t what caused the premie.
Update to Data: I added the original Crumley data to the spreadsheet on it’s own sheet and I made a graph of the current data. So far it matches up pretty well, but it is only day 3. Crumley reported a lot of growth in 33% D2O by day 5 and a lot of growth in 66% D2O by day 8. Around that time is when the first seeds started to sprout in 99.9% D2O.
Two very important things to note:
- There is still nothing growing in the sample with no seeds.
- I’m not sure when to start counting the seeds in 66% D2O, it seems that there are definitely some seeds that are showing initial signs of germination, but nothing obvious (like an extended radicle). I’ll compare this picture to the day 3 images when I considered those seeds to have begun germintion.
- Also note that yesterday I didn’t look at the seeds until around 5pm, and today I’m looking at them at 10am. Obviously much less time has passed between now and yesterday than yesterday and the day before.
Here are today’s results.
And the notes: All seeds showing advanced germination (meaning longer than just a white tip) by my eye exhibit some form of root fro. As per usual it is most prevalent in the deuterium depleted water but this time the di water samples aren’t far behind. While it isn’t noticeable yet in 66% D2O samples, there is some root fro in the 33% samples, but I think it is too soon to tell if the fro compares with that in the ddw/di water samples because the germination isn’t as far along in that sample compared to the others. Right now I’d say the fro’s are just puffs.
It was suggested by Bill Hooker in the comments for Day 5’s results that I point out what I am determining is sufficient germination for the seed to be counted. If you look at the 66% D2O sample there are three seeds towards the bottom with white tips (not close to each other) and I would say that is sufficient germination for sure. That is probably the make or break point. Any less than that and it’s hard to tell if it’s germinating or if the seed coat is just becoming a lighter color, which apparently it does once it starts absorbing water.
Here are the Day 7 results. Looks good so far. There are some troubles though. It looks like I have extensive leakage in the 66% D2O and 99.9% D2O samples, because each day the air bubbles get bigger. How can I fix this without restarting the experiment?
Also in the 33% D2O sample there is that white thing near the center and two unsprouted seeds. I have a feeling the white thing may either be from one of those two seeds. It could also be from a broken seed from the pack that was stuck to the tweezers and which I didn’t notice got added to the lot when I poured the seeds into here. What to do? I won’t know for sure until like Day 15 or so when most of the seeds in this sample would have sprouted. Ideas?
Finally you can definitely see the root hairs in the DDW sample and the DI water samples this time. Not so much in the 33% sample. In all honestly I can’t even tell by the naked eye. Hopefully this will be obvious by day 10. And as usual, nothing to report in the no seed sample.
Bill Hooker suggested (and rightly so!) that I document what I consider to be typical specimens. I found, today that the zoom feature on the webcam I’m using is quite sufficient for getting close enough to demonstrate this and so I snapped a picture and tried to document. Powerpoint messed me up a little bit, but this is good enough.
The orange box is highlighting a seed that is showing no signs of germination. From my studies I’ve found that the the seed coat becomes slightly transparent just before radicle (the pre root) penetration and you can see the precursor to puncture. The seed coat usually gets lighter too and in this case is really dark in color.
The pink box (hehe) is what I would count as germination when I’m counting the seeds. I see the tip of the radicle penetrating the seed coat and so germination is officially underway.
In the very near dead center of the image is a seed that has what I referred to in an earlier post as an extended radicle. Basically I meant that the radicle was pushed through and the root is now forming. The root will continue to grow until the seed coat has come off, which over to the right it has (there is a green leaf that is cropped on the edge of the image).