Techniques for The Germination of Older Seeds

Well, these have been in a vial with dilute h2o2 and h2o, and gently shaken a couple times daily.

Today I dumped them into a starter dish and added a tiny (pinhead) amount of gibberelic acid and put them under the lights.

I saw one had cracked, and it’s green inside- so maybe there’s some life left after 15+ years? I took a couple of photos, what do you guys think?

There’s no radicle yet, but the cotyledons (or whatever that tissue is) look pretty green.

Cross your fingers, toes, and eyes for me, folks. Send good vibes to the universe and hopefully these guys will grow. I have waited SO FREAKING LONG to grow one of my babies I could cry :sob:

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Looks like some seeds will open…
Good luck!!!

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So far there’s no change. The one that cracked is still green, but there’s no growth. I put them all in a vial with a tiny bit of gibberelic acid, a teeny pinch of table sugar, h2o2 and water. I think the h2o2 helps, because it causes the seeds to float in a bubble of oxygen.

I’m also going to send off for the murashige and skoog formula after the 1st and give it a try with fresh seeds. I’m starting to run out of seeds, so hopefully this does the trick.

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So here’s the vial with the seeds. The problem I have had is that although some of them crack (and they show green tissue inside), they just don’t grow. I really don’t know if the MS formula will help.

I’ve been gently agitating the vial, because otherwise the seeds just float at the top (conveniently in bubbles of pure oxygen).

I have a tray of coir moistened with h2o2 that I put other similarly treated seeds in, but they have done nothing. I started those on Oct 21, and this new batch on Nov 9.

This is the growroom, btw. I have a couple of 1k equivalent LEDS (plus a 600w equivalent that I added later). I still need to hook up the ventilation system, but I’m not growing anything smelly right now so it’s not urgent.

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Here’s some info about using h2o2 in germination, there is such a thing as too much, or too long fwiw. :sunglasses:

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I’m going to wait until I get the Murashige-Skoog formula before attempting this again. I hate wasting my seeds!

H2O2 is fairly safe if diluted properly. I use it constantly in my hydroponics, and my plants love it. It harmlessly degrades into water and oxygen- and that oxygen is a powerful oxidizer that destroys bad bugs. Too much of it can destroy healthy tissue, though, which is why I agitate it constantly.

I’m clearly not doing something right, though I think I have the right general idea. Maybe adding the M/S and soaking the seeds in it might make a difference.

I really wish I had a proper dissection scope and steady hands, because I think it’s possible to dissect the seed and extract the actual embryo and grow it. I know it’s regularly done for agricultural research and propagation in other species (orchids, especially).

I have the vial snuggled in between my lettuce tubs, and it stays a nice constant temperature there.

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I got in contact with a gentleman from Node Labs https://www.nodelabsca.com/.

They are pretty heavy into tissue culture technology, and they also are working with a strain called Black Diamond…which shares a parent (Black Domina) with Black Mamba.

I’m hoping to send him some, and see what he can do with them. Maybe he’ll be able to work some magic 🪄

It would be super freaking cool if these seeds can be used to improve or even help create a strain.

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I have had very good results with a 1% hydrogen peroxide solution. For older seeds, I take an oz of 3% peroxide from CVS and add two oz of water. Here in NYC, we have good quality tap water. If you don’t, use distilled water. I just germinated eight seeds from 2012 and all eight popped within 24 hours. If I had enough old seeds, I’d do a side by side test to confirm it was the peroxide that made the difference, but I don’t.

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I’ve tried that, and had no luck. I have very few seeds left, and I am waiting to hear back from Node Labs- they actually do this kind of thing, and if anyone can get them to grow, I am hopeful that they can. I just hope that if they do use them in their program they will send me some seeds back later on. That sounds fair, doesn’t it?

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Did you try he tech I posted above ? If you have it, I would try at least one bean and see how it goes for you.

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Sure, I don’t have a whole lot of beans left, but I am willing to sacrifice one for the cause. I’m also putting a few outside- some in the ground, and others in a half barrel planter. Maybe they can wake up naturally in the spring.

I’m thinking about doing it on the Solstice. Who knows what will happen?

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Many people use the moon cycle to decide when to germ and say it helps. Many farmers swear by it as well.
Ive tried it a few times, and thou I couldn’t determine if it was the moons cycle, or just having a good technique, it can’t hurt, so you might as well.

I’m sending good vibes to your beans, keep me updated!
:seedling::herb:🪴 :evergreen_tree::green_heart:

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I did a little special something on Samhain, I figured it would do no harm. They’re all in the Dixie cup.

I really want to contribute more to the community, especially since I owe a huge unpaid debt of gratitude to the people who helped me get going all those years ago.

Hopefully Sunday I can finally get someone over here to help me install the exhaust system and get the plastic laid down in my closet. I’m unable to bend down and it’s hard to walk, so I have to work in little fits and starts when I’m physically able to.

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shure? because according another Study,
3 Percent Hydrogen Perloxideand dilluted by a factor of 100 with water, gave best results.

When they increased the percentage of Hydrogen Perloxide then the germrates sunk…

SO: @HappyTrees23s mentioned he used 2 Tablesppon in 2 Cups Water, that would be raundabout lead to the said ratio, that was advised…

If you dont follow that ratio, in my study i seen the Sucessrate was considerably lower…

Im still so thankfull @happytrees saved an old Line of gold-- could been as old as 50years, we dont know the exact age. Various other people failed sprouting the same batch. great.

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The Debacco video is about a study that shows best results with 1%, not 3%. It is most definitely not correct that the higher the peroxide ratio the better.

From the abstract: “In this protocol, all three steps including seed sterilization, germination, and seedlings development were carried out in an H2O2 solution of different concentrations; 1% H2O2 solution showed the fastest and the most efficient germination. This protocol also exhibited high germination efficiency for very old cannabis seeds with lower viability. Overall, this protocol demonstrates superior germination compared to water control and reduces the risk of contamination, making it suitable for tissue culture and other sensitive applications.”

This is the study: Development and Standardization of Rapid and Efficient Seed Germination Protocol for Cannabis sativa - PMC

BTW, the study is not “from Debacco University” which is not an actual university and Debacco is not one of the study authors. Debacco has a Masters in Agronomy and a PhD in Education; he makes pretty good videos that are mostly aimed at new growers.

Sorry to be nit-picky about this, but we have a lot of incorrect info circulating among cannabis growers, so I try to correct it in the cases where I happen to know the facts and appreciate it when others here do the same!

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I prefer the towel method for very old seeds…

These are seedlings from 6-year-old seeds. They were badly stored. High temperatures, low temperatures… I didn’t expect one to grow.

But there is a lot of deformed growth.

Could the reason be poor seed storage and will further growth be somehow normal?

Or is something else involved?

They all pop up in same time, in same conditions?

Thanks

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They should grow out of the abnormal leaves.

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Gonna have to try some of these techniques. Have some nice crosses from years ago but poor germ rates.

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Found this in my notes.

I am no fan of chimera, but this may be of value anyhow…LOL

Originally Posted by Chimera
The protocol I use is pretty simple.

5% bleach in autoclaved, distilled water with a couple of drops of Tween-20 per liter of solution. I sterilize the water in Duran jars (loose lids) in a pressure cooker @ 20 PSI for 20 minutes, and then allow the jars to cool under my flow hood and tighten the lids when the water has cooled.

This is probably overkill for most purposes, but it’s a variation of the protocol I use for sterilizing explants for tissue culture, and it seems to work with minimal contamination. You can get away with a drop of dish detergent instead of the Tween, I just have the tween-20 in the lab so I use it. If you are going to be doing a lot, you can order a few ml of Tween-20 on Amazon if you like.

First I rinse the seeds in a tight mesh strainer to remove any leaf debris that might be harboring bacteria or spores. These seeds were a little dirtier than most having been field grown, so you definitely want to wash away any superfluous materials. This can be up to a 5 minute rinse in lukewarm water. Cannabis achenes have a lot of little crevices where spores and bacteria can hide, so a thorough rinse is a wise step.

Then I take the seeds and drop them in maybe 100-200 mls of the bleach/Tween-20 solution in an Erlenmeyer flask, but any clean glass vessel will do -the seeds should be fully immersed, maybe 2x the volume of water as seed. You want to keep stirring and swishing the seeds around gently to ensure the current is washing away any contaminants. After a minute of swishing I remove the bleach and strain the seeds from the liquid and start again. No more than 3 minutes total, and if I see bleaching on the small test batch (always do a test batch to seed how the seeds react), sometimes I’ll only do 2 minutes.

The next part is key. Immediately rinse 3-5 times (for 5 minutes per rinse) in the same sterilized distilled water (baby water / distilled water from Walmart or the local pharmacy is fine), again using a tight mesh strainer, or simply rinse the seeds in a different vessel with the same volume of water, at least 3 times to ensure the bleach is removed. I use a labratory orbita shaker, but you can do it by hand it’s just a little tedious.

Once the rinses are completed, soak the seeds in the distilled water as before, for 2-24 hours depending on the age of the seed and the medium you will be growing in. Sometimes I’ll use agar plates, but most often I’ll just sprout them in paper towels and transfer to a light dirt (like Light Warrior for you Americans) or surface sow on a .peat based planting substrate. You can use whatever medium you prefer, it’s not that big of a deal as long as the seeds remain moist.

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