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Chironex
12-26-2008, 03:54 PM
After reading some research about "thermodormancy" and seed germination inhibition, it is time to determine what effect, if any, the benefits of cytokinin treatment would have on banana/ensete seeds. Being largely recalcitrant, perhaps due to inhibition by abscisic acid (ABA) and thermodormancy, there appears to be a method of releasing these dormancies by stimulating ethylene production. These studies have shown positive results on breaking seed dormancy on clover, peanut and lettuce seeds.

As soon as I have secured the correct cytokinins, I plan to duplicate the experiment with banana seeds. It is hoped that these results may further improve banana seed germination responses.

Bob
12-26-2008, 04:41 PM
Thanks (I think) Scott I really didn't understand all you said though. I kind of got thermodormancy , but what about the ethylene thing? Should I put my unsprouted seeds in a brown bag with an apple?

Chironex
12-26-2008, 05:38 PM
Hmmm, why not? Try it and see what happens. Worst case scenario, they get mold on the outside of the seeds. I would suggest adding some powdered cinnamon to the bag. I would also try a paper bag and a plastic bag. Set it up by putting the seeds in a dampened (not soaked) coffee filter. Add some cinnamon powder, then fold it twice. Place this in a bag with your apple. See if they germinate quicker than a control group without the apple. Ideal temperature is around 25 C. Alternatively, you can fluctuate the temps from 17 - 25 C for 7-8 hours at the high end and 16-17 hrs at the low end.

It will be interesting to see what if anything happens. I will be using cytokinins to induce ethylene production. I may also use gibberelic acid to see if it makes any difference in germination in darkness.

damaclese
12-27-2008, 08:16 AM
Thanks (I think) Scott I really didn't understand all you said though. I kind of got thermodormancy , but what about the ethylene thing? Should I put my unsprouted seeds in a brown bag with an apple?

Bob not to make fun of you but i was thinking the sam thing and it just made me laff LOL

Wouldn't it be a kick in the pants if after all the thousand of hr and millions of dollars people have spent in labs all over the world we find that just putting them with and apple in some brown paper bag did the trick i would just lafe my %#@^% off lol

O and my spellchecker said that thermodormancy is two words
thermo dormancy

Bob
12-27-2008, 08:18 AM
Thanks for putting this in simpler terms. The fluctuating temps did work with my Darjeeling Giant seeds. No effect with "reg" sikkimensis though as of yet. I'll try the cinnamin /apple recipe though on some Velutina seeds and see what happens. Anything with this combo can't be bad right?

Chironex
12-27-2008, 02:15 PM
Pauly,the spell checker is incorrect.Thermodormancy is correct. It is a combinative, single word.

jmoore
12-28-2008, 05:08 AM
I wasn't aware that apples produced ethylene (or ethene given its IUPAC name), at what point do they produce ethene? I always thought that bananas produce ethene when ripening, which stimulates the ripening of other bananas and indeed other fruit.

Wouldn't it be more logical to put a few banana seeds in a bag with a ripening banana?

As for sikkimensis, bloomin things, can't get them to germinate for love nor money. I'll try the ethene root, worth a go.

jmoore
12-28-2008, 05:16 AM
Well I was wrong, apples do indeed produce ethene. I stand by my ripening banana idea, however.:nanadrink:

damaclese
12-28-2008, 12:16 PM
Pauly,the spell checker is incorrect.Thermodormancy is correct. It is a combinative, single word. thanks for clearing that up i really need to go get a spell checker that has some sort of downloadabal area specific language data bases dose any one know of some good free-ware

Chironex
12-28-2008, 04:17 PM
I just wonder whether sufficient ethylene is produced to have an effect on breaking seed dormancy. But with an open mind, I move forward. I would like to ascertain whether anyone notices any difference in germination rates using this method. Note that ethylene alone will not be an end-all to breaking dormancy. There is a synergistic relationship in the presence of ethrel. This is where I will be focusing my experimentation.
Ethylene plays a hormonal-type role in directing growth, inhibiting growth, fruiting, blossoming, leaf dropping and other functions and is present in most plants. It is one of the most prevalent gases in nature. I see your logic about using a banana as an ethylene source, but I don't think it will make a difference what the source may be as ethylene is ethylene.
If you plan to try this, please set-up a companion test as a control group. Do everything the same, except do not add the apple, banana (or whatever ethylene source) to the control group.

Chironex
01-08-2009, 01:34 AM
OK, all of you chemists out there, I need your help, please. I want to end up with a dilution of 350 micro moles/liter of ethrel (Ethephon - 2-chlorethyl phosphonic acid) in final solution. I am starting with 1 pint containing .04 pounds per 473.16 ml, or (.33 pounds per gallon) from a 3.9% solution containing 96.1% inert ingredients. The molecular weight of this compound is 144.49401 g/mole. I need the final solution to contain .00035 moles of ethephon/liter.

How much would I need to dilute this to achieve the desired amount (350 micro moles) in my final solution?

By my calculations I would need to add 1.93757246 ml to 1 liter of water to achieve 350 micromoles dilution. Can someone please help me to verify this? (I would round it to 1.94 ml/L)


Thanks in advnce for your assistance!

jmoore
01-08-2009, 01:23 PM
OK here goes. The fact you have used two types of unit ie metric and imperial (US imperial at that) and sometimes together is confusing and you lose something in the conversion, so here is how I have interpreted it.

You start with 3.9% active ingredient, which is equal to 39 g / l (grams per litre) stock solution.

You need 350 micromoles in your final solution or 0.05 g / l ( based on the relative molecular mass you supplied of 144.something)

To reach that figure you divide your final mass of 0.05 grams ( as we are talking about 1 litre) by your starting concentration of 39 g / l which gives a figure of 1.28 ml / litre.

I'm sorry, but I don't understand pounds per US gallon and get especially confused when talking about pints. I think you can ignore the inert material as it's inert. On the bottle does it give a concentration as a percentage of active ingredient or a concentration in ounces per gallon?

Hope this helps.

Bob
01-08-2009, 02:09 PM
jmoore, I get confused as well after a few pints!

jmoore
01-08-2009, 02:18 PM
I hear that, in fact going to have one right now. Cheers :bananas_b

buzzwinder
01-08-2009, 03:38 PM
:bananas_b

Richard
01-09-2009, 12:11 AM
...
You start with 3.9% active ingredient, which is equal to 39 g / l (grams per litre) stock solution.

You need 350 micromoles in your final solution or 0.05 g / l ( based on the relative molecular mass you supplied of 144.something)

To reach that figure you divide your final mass of 0.05 grams ( as we are talking about 1 litre) by your starting concentration of 39 g / l which gives a figure of 1.28 ml / litre.
...


Correct. You win this weeks proportionality quiz!

:woohoonaner:

griphuz
01-09-2009, 08:36 AM
I'm still a little confused though about what breacks the dormancy, the cytokinnin or the ethylene?
So there are two possible experiment pathways, treatment with ethylene, and/or treatment with cytokinnin...
Regards,
Remko.

jmoore
01-09-2009, 09:11 AM
That is the underlying issue with biology experiments, they have a lot of variables to control.

I'm sure, though that Scot will control those varibles and come out with a valid answer.

To answer your question, I suppose it could be one or the other or a mixture of the two, or something completely unrelated.

Michael_Andrew
01-09-2009, 03:56 PM
I was kinda wondering what the effect if any there would be subjecting the seeds to the various enzymes, chemicals and the thermo dynamics of the digestive system. Anybody got a pet monkey?

Chironex
01-09-2009, 05:49 PM
James, as you mentioned, it is the two in combination creating a synergy that trigger ethylene production and the inhibition of abscisic acid (ABA). Each of these plays a role in releasing dormancy.
Cytokinens and ethrel stimulate ethylene production in seeds. Further, the synergistic effects of ethrel and kinetin in stimulating polyribosome formation.
Significant variables are temperature, (at 35 C germination is completely inhibited) 25C is ideal; and Light (ethrel is promoted by kinetin in releasing seeds from photodormancy in light, or GA3 induced dark germination). Kinetin is capable of overcoming the inhibition of germination by high temperature, as is ethrel to a lesser extent, but the two combined have been shown to increase both the rate and percentage of germination.
Both kinetin and ethrel appear to be related to the expansion of cotyledons. They work together to assist in ATP synthesis during dormancy release in some seeds.
I should also include that these experiments were initially conducted on lettuce and Xanthium seeds. The trials I will be conducting may be the first efforts on Musa and Ensete seeds. Hopefully similar results will be realized.

Chironex
01-09-2009, 06:30 PM
As a follow-up to these posts, here is a good description of the role ABA (abscisic acid) plays in seed germination and plants in general. Also, from the same website, a description of ethylene and its functions.

Abscisic Acid (http://www.plant-hormones.info/abscisicacid.htm)

Ethylene (http://www.plant-hormones.info/ethylene.htm)

damaclese
01-09-2009, 09:11 PM
Scot this may be a dumb question but as i know almost nothing on this subject all ask any way so when you do a embryo rescue are there still dormancies present in the embryo? and if so could you expose the embryo invetro to thees chemicals or would that be a bad idea?

Chironex
01-09-2009, 09:46 PM
Scot this may be a dumb question but as i know almost nothing on this subject all ask any way so when you do a embryo rescue are there still dormancies present in the embryo? and if so could you expose the embryo invetro to thees chemicals or would that be a bad idea?

Yes, the dormancy still persists even in the embryos, but I must thank you for bringing up this question.

Why? Because in making the more robust embryo rescue medium I was planning to use only kinetin, but now will also make some of the medium including both kinetin and ethrel to see if the synergism applies to embryos as well. This will highlight any advantages of having both chemicals in the medium versus only one.

Great input!

damaclese
01-09-2009, 09:50 PM
Your welcom thanks for all the hard work you are doing on behafe of the Banana world with famon being the nuber 2 consern for the world at larg any and all information pertaining to Muses increast production is a good thing

Richard
01-10-2009, 12:04 AM
...
Both kinetin and ethrel appear to be related to the expansion of cotyledons. They work together to assist in ATP synthesis during dormancy release in some seeds.
I should also include that these experiments were initially conducted on lettuce and Xanthium seeds. The trials I will be conducting may be the first efforts on Musa and Ensete seeds. Hopefully similar results will be realized.

Just a heads' up for the casual reader: Scot is an ace biologist whose thesis subject was the worlds deadliest animal, the Chironex.

Now Scot, you've peaked my interest here because you're discussing cotyledons and vegetables (lettuce) in the same thread. Lettuce seed is usually quick and easy to propagate. Did the research involve a cumbersome variety or was the lettuce being used as a model for "natural" germination?

Looking toward the future, I can see you as a hired gun for propagating some the unusual seeds I obtain from time to time.

Chironex
01-10-2009, 02:43 AM
Thank you Richard, but I must include that botany was only a few hours of elective credit (and that was 35+ years ago.)

Yes, it was on lettuce seeds, however, it is my belief that the process of germination and dormancy have similar characteristics in many seeds. So, while this is a shot in the dark, if it works we will have gained important knowledge to further our efforts.

As to the matter of propagating other unusual seeds, all we can do is try what works. Keep your fingers crossed, seat belts fastened and hope for the best.....

Chironex
01-16-2009, 04:40 PM
Update: Kinetin received today. Will prepare the medium this weekend for the more robust embryo rescue formula, AND will be setting up the soaks using ethrel/kinetin combination for recalcitrant seed germination testing. Will post updates as significant development occur. Thanks again to my chemist buddies out there, Remko (griphuz) and James (jmoore). I was correct on one of the formulas, but slightly off on the other. Your input helps scrape the rust off of this old brain.

Michael_Andrew
01-17-2009, 12:55 PM
I was looking over some experiments online and I think they were using ethylene with CO2.

Chironex
01-17-2009, 04:07 PM
I was looking over some experiments online and I think they were using ethylene with CO2.

That is another experiment that I have read. It is a means of acclimatization without using sugar in the medium. It develops stronger plants sooner and with a higher state of vegetative development. I will be using this in further studies later on.

But, just in case you are reading something different than what I think it is, please email the article to me and I will read it.

Michael_Andrew
01-17-2009, 05:02 PM
Interrelations between Carbon Dioxide and Ethylene on the Stimulation of Cocklebur Seed Germination -- Esashi et al. 86 (1): 39 -- PLANT PHYSIOLOGY (http://www.plantphysiol.org/cgi/content/abstract/86/1/39)
Interaction of Carbon Dioxide and Ethylene in Overcoming Thermodormancy of Lettuce Seeds -- Negm et al. 49 (6): 869 -- PLANT PHYSIOLOGY (http://www.plantphysiol.org/cgi/content/abstract/49/6/869)
www.apsnet.org/phyto/PDFS/1980/Phyto70n12_1158.PDF
www.scielo.br/pdf/sa/v61n2/19356.pdf
None of these look like the one I was reading but they do talk about it. I'll keep looking. The one I read gave charts and percentage of germination and days. I think the best rate and time was a combination of the ethylene and CO2. There were other factors and chemicals involved.

Michael_Andrew
01-17-2009, 05:28 PM
Here is the one I was reading.

http://www.plantphysiol.org/cgi/reprint/56/6/826.pdf

Chironex
01-17-2009, 07:06 PM
Interesting, thanks for bringing it to my attention. So, from what I gather, the synergy is formed when kinetin, ethylene, GA3 and CO2 are combined. Yet each shows little effect when used alone, with the exception of kinetin.

Therefore from least effective to most effective upon germination:

GA3 - no effect

Water + ethylene + CO2 - a light effect

GA3 + kinetin - maximum effect in the dark, but due to presence of kinetin

GA3 + ethylene + CO2 - improved germination rate under any light levels - needed for gemination

kinetin + ethylene + CO2 - improved germination rate under any light levels - not required to add the gases, but interaction improved germination rates

When GA3 and/or kinetin were combined with added ethylene, the need for added CO2 was eliminated. By the same token, when GA3 and/or kinetin was combined with added CO2, the need for added ethylene was eliminated.

These last two observations make it clear that the endogenous ethylene and CO2 gases present in the seeds/atmosphere are sufficient to maximize germination (overcome thermodormancy) in the presence of kinetin, or kinetin and GA3. In the presence of GA3 and kinetin, the need for supplemental CO2 and/or ethylene is reduced, but the 4 appear to work synergistically well to maximize germination rates.

damaclese
01-18-2009, 12:37 PM
wow it sounds like you are getting close to a good stable and reproducible conclusion Scot congrats. I'm just so excited to think that we mite be able to germinate any Musa seed that we want to!!!!! :woohoonaner:

Chironex
01-18-2009, 09:49 PM
Ok, today the modified medium was prepared and next the soaking medium will be prepared. I don't have a cheap source for CO2, so maybe I will just open a can of soda and put it in a zipper bag with the seeds and the soaking medium. (just kidding, but one never knows)
The modified embryo rescue medium will be described further under its own thread.

Michael_Andrew
01-18-2009, 09:55 PM
You can make your own CO2 with some water sugar and yeast.
DIY Yeast CO2 - The Planted Tank (http://www.plantedtank.net/articles/DIY-Yeast-CO2/7/)

Better yet start some wine fermenting!

Chironex
01-18-2009, 10:01 PM
Interesting thought, I might have to use it.

Chironex
01-19-2009, 03:32 AM
This morning (12:00 a.m.) I initiated seeds into the kinetin/ethrel medium. While they probably will not germinate as quickly as the lettuce seed that was used in the research, it is hoped that this method will reduce the period of time and increase the germination percentages of banana seeds.
I set up 6 plates, each containing 8-12 seeds.
These are the seeds initiated:
M. ingens
E. perrieri
M. acuminata ssp. banksii 'Higa'
M. balbisiana
M. aurantiaca
M. paradisiaca var seminifera 'New Bhutan'

I will continue to monitor these plates and advise if there are any significant findings.

griphuz
01-19-2009, 04:12 AM
I take it you also have a control-group right Scot? ;)
Kind regards,
Remko.

Chironex
01-19-2009, 04:30 AM
I take it you also have a control-group right Scot? ;)
Kind regards,
Remko.

I have many control groups! lol Actually, yes I also set-up the seeds in another plate with water only.

Wouldn't it be great if this actually does what it suppose to do? I think I will set-up a CO2 generator and try to adapt the petri dishes to allow some CO2 into them. I need to find some very small tubing, perhaps aquarium sized tubes and fittings could somehow be installed. Anyone have any ideas how to do this? The petri dishes are polycarbonate, so they will easily fracture. I think I will have to melt a hole for a fitting. Then connect it to a gang valve coming from the CO2 generator. Like an octopus rig.

Chironex
01-19-2009, 05:45 AM
Scot this may be a dumb question but as i know almost nothing on this subject all ask any way so when you do a embryo rescue are there still dormancies present in the embryo? and if so could you expose the embryo invetro to thees chemicals or would that be a bad idea?

Update, I read some research articles that indicate some have a dormancy factor within the embryo and others do not. So, I guess it wouldn't hurt to try this on embryos that do not respond otherwise, just to see if it has any effect.

I also read some research that indicated successful pollination of triploid bananas by diploid pollen! The one that was used as an example where it worked was Yangambi KM-5. Most of the positive results were with plantain types, however. But this is perhaps some good news that I stumbled upon by accident.

damaclese
01-19-2009, 08:44 AM
I have many control groups! lol Actually, yes I also set-up the seeds in another plate with water only.

Wouldn't it be great if this actually does what it suppose to do? I think I will set-up a CO2 generator and try to adapt the petri dishes to allow some CO2 into them. I need to find some very small tubing, perhaps aquarium sized tubes and fittings could somehow be installed. Anyone have any ideas how to do this? The petri dishes are polycarbonate, so they will easily fracture. I think I will have to melt a hole for a fitting. Then connect it to a gang valve coming from the CO2 generator. Like an octopus rig.

Scot this sounds like an ideal situation to use a Small aquarium maybe that way the dishes could be set in it and you could pump the hole thing full of CO2 and you wouldn't have to fuss with cutting the dishes and with the hold thing sealed theirs not much risk for contamination how are you going to filter the Co2 id recommend A UV chamber and you will get the added benefit of some small amounts of O3 which is a nice Little anti bacterial gas if its in controlled amounts but i don't know what the optimum would be of if that would retard growth

Michael_Andrew
01-19-2009, 11:06 AM
CO2 Experiment - Video (http://www.metacafe.com/watch/976046/co2_experiment/)

Richard
01-19-2009, 11:08 AM
...
I also read some research that indicated successful pollination of triploid bananas by diploid pollen! The one that was used as an example where it worked was Yangambi KM-5. Most of the positive results were with plantain types, however. But this is perhaps some good news that I stumbled upon by accident.

Yes, there is more than one paper that discusses this in the context of the genetics and heritage of true quadriploids and triploids. Basically, if LMN was produced by LM x LN, then some combination of factors from L, M, N will be needed for successful pollination of LMN.

Chironex
01-19-2009, 03:23 PM
Scot this sounds like an ideal situation to use a Small aquarium maybe that way the dishes could be set in it and you could pump the hole thing full of CO2 and you wouldn't have to fuss with cutting the dishes and with the hold thing sealed theirs not much risk for contamination how are you going to filter the Co2 id recommend A UV chamber and you will get the added benefit of some small amounts of O3 which is a nice Little anti bacterial gas if its in controlled amounts but i don't know what the optimum would be of if that would retard growth

Now why didn't I think of that? I could actually just use a huge zip lock bag! As a matter of fact, I have some. God, I am such a dork sometimes, it amazes me. Thanks Pauly! I am not concerned with contamination as these are seeds, therefore no filtration is needed.
Do you have any tubing? I figure that I will need to buy a stopper and a hard plastic tube a few inches long, plus about 2-3 ft of 1/4 inch tubing. Then, I will set up a CO2 generator using yeast in a pyrex or nalgene bottle. Genius!

Chironex
01-19-2009, 03:24 PM
Is anyone else having trouble playing this video? It starts and i can hear her, but shortly after it begins, the screen goes black.

damaclese
01-19-2009, 03:54 PM
Now why didn't I think of that? I could actually just use a huge zip lock bag! As a matter of fact, I have some. God, I am such a dork sometimes, it amazes me. Thanks Pauly! I am not concerned with contamination as these are seeds, therefore no filtration is needed.
Do you have any tubing? I figure that I will need to buy a stopper and a hard plastic tube a few inches long, plus about 2-3 ft of 1/4 inch tubing. Then, I will set up a CO2 generator using yeast in a pyrex or nalgene bottle. Genius!

yes i hapin to have some tubing its drip line i was going to ues for the garden but im sure i can spair a few ft of it and since im going to the high presure spray systom its probley not even going to get used got drip emiters to if you need any LOL

damaclese
01-19-2009, 04:02 PM
I can only imagine what Scots laboratory is going to look like with 50 zip lock balloons and a bunch of tubing and bottles of yest percolating away good thing the police cant see in God only knows what they will think of it LOL

Um Ya Osifer its Banana seeds ya Yest can you believe that LOL

Scot spreed out across the hood of a police car Better keep some extra money around Scoty LOL

Michael_Andrew
01-19-2009, 06:23 PM
Same experiment different video.
YouTube - Kid's Science Experiment To Show CO2 Is Heavier Than Air (http://www.youtube.com/watch?v=AvpenpBXAmM)

Michael_Andrew
01-19-2009, 09:43 PM
Ok Scot I made that Co2 Generator. Took about 3 minutes with stuff I had laying around. Just drilled a hole in the lid just a bit smaller than some tubing I had. I shaved down the end of the tubing and held it under hot water to get it soft and pushed into the hole. It fit snug so I don't think I need any kind of sealant. Dumped in some Big Chief and some warm water and mixed up. Poured in some regular baking yeast and added some yeast nutrient just cause I had it laying around. In just a few minutes it putting out a bubble every three seconds. I'm gonna let this flow over one of my seedlings and see what happens. According to this site I should double the growth rate.


The Heartland Institute - Environment & Climate News Article (http://www.heartland.org/publications/environment%20climate/article.html?articleid=9831)

<a href=http://www.bananas.org/gallery/showphoto.php?photo=15275&ppuser=3593><img src=http://www.bananas.org/gallery/watermark.php?file=15275&size=1 border=0></a>

Chironex
01-19-2009, 11:17 PM
Michael, I bet that yours was a whole lot cheaper than the one on eBay. Yikes!
Co2 GENERATOR GROW HYDROPONIC LED HPS CARBON DIOXIDE - eBay (item 330301219057 end time Jan-23-09 09:25:56 PST) (http://cgi.ebay.com/Co2-GENERATOR-GROW-HYDROPONIC-LED-HPS-CARBON-DIOXIDE_W0QQitemZ330301219057QQcmdZViewItemQQptZHydroponics_Seed _Starting?hash=item330301219057&_trksid=p3286.c0.m14&_trkparms=66%3A2%7C65%3A1%7C39%3A2%7C240%3A1318)

jmoore
01-20-2009, 06:19 AM
Don't forget to change the yeast and sugar as the CO2 production drops off because the yeast will become denatured by the alcohol it will produce. Also too high a sugar concentration will kill the yeast. Aim for less than 50%.

griphuz
01-20-2009, 06:54 AM
Ehm, just another idea maybe,...
The biological way to make CO2 is good, but not really constant.
There are also chemical ways to make CO@, just by adding an acid to Soda (Sodium carbonate) or Calcium carbonate.
Preferrably use a non gassious acid (so not hydrochloric acid, but rather citric acid e.g.) to avoid traces in your mixture.

Or you could just use those ampul-thins they put in the 'make-your-own-soda-drink' machines,...
Kind regards,
Remko.

jmoore
01-20-2009, 11:14 AM
Adding acid to a carbonate/bicarbonate is sound, but you would have to keep adding acid and carbonate. The soda stream cannisters are also good, but you would need a regulator to release the gas at a constant flow. I think on balance the yeast is good providing it is working at its optimum.

Oh unless it's a one shot system - fill up the baggies with CO2 and away you go. In which case the acid to a carbonate/bicarbonate would be best.

Chironex
01-20-2009, 01:31 PM
There are some considerations:
1. Too much CO@ will probably be inhibitive to growth
2. The acid vapors should not enter the bag.

I just want a light, gradual, continuous flow for several days.

griphuz
01-20-2009, 04:54 PM
Another option would be a completely controlled set of CO2 inlet as used in many planted aquariums.
I know my brother has a system like that, 'feeding' the plants the dissolved CO2 as it were. This is just a small gastank with CO2 (available @ the petstore) with a controlled outlet.

Should you work with either the bio-way or the chemical way, it would be wise to bubble the produced CO2 through some cold water, and next through a piece of cotton to prevent other things than CO2 comming through, and maintain a sterile environment.
It would look a little like this;
http://www.bananas.org/gallery/watermark.php?file=15301 (http://www.bananas.org/gallery/showphoto.php?photo=10222)
CO2 produced in A (either chemically of biologically through fermentation) is washed with cold water in B and filtered (optionally dried with e.g. CaCl2) in C.
From here on it will be passed through to the set-up.


The CO2 can be bubbled through a liquid medium, or just let free in the air part of the set-up, I don't know what the intention is here?
Kind regards,
Remko.

Chironex
01-20-2009, 05:40 PM
This is more like what I was thinking. So how much yeast and how much sugar? Does anyone have a good feel for this? I am not seeking a CO2 generator that emits huge amounts - more like a trickle of gas, slow and steady. The bags are about 2-3 gallons (16-20 liters) in size. So not much gas is needed, just enough to raise the ambient CO2 levels inside the bags to approximately 15% v/v.

damaclese
01-20-2009, 07:53 PM
This is more like what I was thinking. So how much yeast and how much sugar? Does anyone have a good feel for this? I am not seeking a CO2 generator that emits huge amounts - more like a trickle of gas, slow and steady. The bags are about 2-3 gallons (16-20 liters) in size. So not much gas is needed, just enough to raise the ambient CO2 levels inside the bags to approximately 15% v/v. Scot being a chef and doing a grate deal of baking i use yeast all the time typically to a packet which is approx .5oz of dry active yeast you add 1tsp of sugar and this yeast typically will feed for up to 24hr of that amount of sugar although if I'm creating a sour dour which takes up to 7 days i add !tsp of sugar ever 2 days to keep the yeast active a vary small amount of yeast can make a vary large amount of gas this probably doesn't relate all that well to what you are trying to do but its a start and as some one pointed out all ready to much sugar will kill the yeast and after a few days the alcohol will start to build up but all you have to do is strain it throw a coffee filter and scrape whats left back in to the jar and add more luke warm water and some sugar and you could be ready to go again one thing i know about yeast is its every were and it grows at the drop of a hat you will have to keep it warm about 86 deg is a pretty good rate you can go higher but do you really need to you just need a smal amout of gas

Chironex
01-20-2009, 08:15 PM
Perhaps simply exhaling into the bag a few times a day would do the trick. If I hold my breath for several seconds, I can increase the CO2 and simply blow into the bag and seal it quickly.
Any thoughts?

griphuz
01-21-2009, 03:50 AM
No offence Scot, but we do waht to keep thing sterile-ish,....right?
;)

Chironex
01-21-2009, 04:57 AM
No offence Scot, but we do waht to keep thing sterile-ish,....right?
;)
It's not really necessary as these are intact seeds. Actually, they are in more sterile medium than if in regular soil. They are also inside of enclosed petri dishes within the bag, so there really isn't much that can happen. To put your mind at ease, I did not exhale into the bags. I simply opened a fresh can of cola and sealed the open can inside of the plastic bag. (If this were in embryo rescue or tissue culture, then yes, it would be necessary to keep things as sterile as possible.)

I may devise a way to containerize TC jars and keep them in a controlled atmosphere supplemented with CO2 at the proper amounts. But that will have to wait for more funds. I would need to regulate the CO2 to maintain the proper parts per million within the chamber. That sounds like expensive equipment and gauges.

damaclese
01-21-2009, 08:32 AM
i would think the Co2 from soda would work fine its still Co2 interesting i would never have thought that up thanks Scot i wander if i set cans of coke by my orchids if they would grow faster LOL J/K you have got me wondering if with all the plants in my house if theirs a good mix of gases going but i do have three dogs and they respirate a a pretty high volume i think I'm deteriorating in to winter madness now LOL ya know with Bananas its always something A.

Michael_Andrew
01-22-2009, 06:34 PM
I'm beginning to think we should water our plants with club soda!

Michael_Andrew
01-22-2009, 06:37 PM
Well look at this! lol
The Effect of Carbonated Water on Green Plants (http://spot.colorado.edu/~basey/ldanzell.html)

Michael_Andrew
01-22-2009, 06:48 PM
But you would have to choose one without sodium.
Soda-Club | Counting Carbs & Calories (http://www.sodaclubusa.com/dietary_comparison.htm)

Chironex
01-22-2009, 07:00 PM
Well look at this! lol
The Effect of Carbonated Water on Green Plants (http://spot.colorado.edu/%7Ebasey/ldanzell.html)

I wondered about that. Thanks for finding something to back up my thinking. The sugar probably helps too, so I would avoid any diet varieties.

I might try this on established plants when rooting and hardening off. Only one reservation, I wonder if the carbonic acid would cause any problems with soil pH balance? Apparently, whatever it does, the plants seem to like it. It would have been better to see long term results rather than only 10 day results.

Chironex
01-29-2009, 06:50 PM
Update: Day 10
Other than some mold on the outside of 2 seeds, not much change. Some of the seeds do appear to be larger, indicating imbibition of water perhaps. I am monitoring for the ejection of the micropylar plug and/or emergence of plant material.
From conversations with another member, some seeds seem to exude a viscous, non-water soluble sap-like substance just prior to emergence of sprouts. I am trying to identify this exudate. Anyone have any ideas? I am guessing that it may be phenolics or an unidentified gelatinous substance from the vacuoles near the chalazal body.

Chironex
02-03-2009, 08:16 PM
Still nothing significant to report. The seeds are still in the original medium. I hope something germinates soon.

Michael_Andrew
02-03-2009, 09:53 PM
Darn seeds are hard to figure out. I soaked some hibiscus seed for a day and and put them in a zip lock with moist perlite on 2/1/9. Today 2/3/9 one had germinated and had a root about an inch long. I have musa and ensette seeds just sitting there for a couple months antagonizing me! lol

Thanks for the update Scot.

Richard
02-03-2009, 11:11 PM
Now where is inkcube when we need his input?