Tag Archives: tobacco seeds

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 7

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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.

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 6

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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.

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 5

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Two very important things to note:

  1. There is still nothing growing in the sample with no seeds.
  2. 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.
  3. 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.

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 3

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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):

  1. No seeds – 0/0
  2. DDW – 19/30
  3. DI water – 20/30
  4. 33% D2O – 3/30
  5. 66% D2O – 0/30
  6. 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.

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 2

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Day 2 and still no sprouting, but it looks like there is some activity in some samples. Let’s keep an eye on it…

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RC1, Water Type Effects on Organism Growth

The Crumley Spreadsheet

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Spreadsheet

If you don’t want to look at pictures of gradual change every day then you can just come back to this spreadsheet to see the numbers change and then you can look to see what days something new happened and find the corresponding images through the blog. If I remember to, I may link blog posts with activity to the day number in the spreadsheet, which would be a great idea (but like I said, if I remember).

Update: slide all the way over to the right to see links to the image notes from this notebook corresponding to that day.

Update 2: I added the original Crumley Data on a separate sheet, and I began plotting the percent germination in real time. Currently I’m not sure how to label each line in Google Spreadsheets. I can’t find a way to do it, but I’ll mention it here:

  • Blue – DI water
  • Red – 33% D2O
  • Orange – 66% D2O
  • Green – 99.9% D2O
  • Purple – DDW
  • Not shown – di water, no seeds

BTW: It took me a while to figure out how to do this because Google Docs changed their appearance and formatting yet again. In order to embed a Google Doc in the new format there is a “Collaborate” menu along with the usual “File, Edit, etc” menus. No longer can you go to the “Share” button and select “Publish as web page” because that button has been removed (it is not just the original share button that let’s you set documents as public and determine who you want to share with).

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day I Part 2

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These pictures are setup as in picture 1 (with a computer attached to the webcam). The webcam is a Logitech HD Pro Webcam C910 with the capability to take 10MP images and record in full 1080p.

I took one picture of each sample and then discovered that I could increase the image resolution and manually control the focus and some other properties, so I took one more (the second picture of the 66% D2O sample). From the looks of it, the image quality is more than enough for this experiment. And also between the identical pictures (with different settings) the resolution difference doesn’t make that much of a difference.

Opinions, comments, questions? Drop ’em like their hot…

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RC1, Water Type Effects on Organism Growth

Tobacco Seed Growth in Water Day 1

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Here is the first of many posts of the results of this experiment (LIVE!!!).

Outside of the bubbles in the DDW sample, there is no germination almost 24 hours after initial setup.

I’m still experimenting with the setup. These images were taken with a Nikon D40 in the setup I showed yesterday. I aligned everything and put the image in focus with the aid of a webcam. I’m considering just taking pictures with the webcam and will do a trial run this afternoon. More to come…

This post is open to read and review on The Winnower.

RC1, Water Type Effects on Organism Growth

Repeating Crumley: Day 1

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As demonstrated in the setup of the previous post, I have setup 5 samples of Dark Virginia tobacco seeds with each sample being in a different water type: DI water, 33% D2O, 66% D2O, 99.9% D2O, and for poops and giggles 99.9% deuterium depleted water (DDW).

The percentages are by volume (for example 33% D2O is 2mL of D2O and 4mL of DI H2O). Each sample contains 6mL of its respective water type. I’ve determined that the volume of each cell is a little less than 7mL and 6 comfortably fits with no spillage (there is a small air bubble in the chamber). Each sample also has 30 seeds.

The sample cells were setup as follows: Seeds were counted and placed on one of four prefolded sheets of weigh paper. Airflow was a problem so I placed relatively heavy objects on the edges of the paper to prevent a seed catastrophe. After seed sorting, I prepared the water samples in centrifuge tubes for temporary holding and mixing (in the case of the 33% and 66% D2O samples). 6mL of each water type was then poured into prelabeled Analyslides. The seeds were then added with the lid of the analyslide placed on top to “seal” it. Slides were then placed on the pre-assembled photography station.

Today marks day 1 of the experiment and I am proud to report that in the 5 minutes it’s been, 0% germination is seen across all 5 samples. So far results are as expected.

This post is open to read and review on The Winnower.