Attempting to germinate 50 year old Seeds from korea

here is another method to consider [MadScientist] posted in aug 2016)
Here is the original inspiration - I saved this post from the original Cannabis World forums years ago:

Through the university, I have been exposed to a ‘germination bomb’ that coincidentally was the same university Steve Tuck attended and learned the same technique (contrary to what he would tell you). Here’s what he stated about it @ OG during his stint and simply, it is as follows:

"Also here’s a free bone for all you old schoolers, while in collage me and a buddy developed a pressure bomb to open/germinate really old seeds. I have taught this trick to a few friends who were amazed at how well it works but neccesity is the mother of invention, here’s how it works at home.

Take an old mayonaise jar and punch a hole in the top a little bit bigger than an aquarium bubbler hose, and run one through it, silicone all around hose on both sides and allow to dry overnight. Now put a little bubbler air blower in it with a stone on end. Now fill with water and 10 drops of superthrive or similar concentrated b1 solution. Next use 10 drops of DMSO per 8 oz.'s of water, float old seeds on top and screw lid on tight, run motor [aquarium air pump] for 24-48 hours to build a little pressure to imbibe fluid in seeds then place on 90 degree F wet dirt and they will usually get a good percentage of those with a spark left in them, let stay at that temp for 3-4 weeks in dirt as some may be slow to respond. You should be able to get DMSO from a pharmacy. And personally I like to add a little sugar water as old seed loses it’s carbohydrates over time. If you cannot find B1, a kelp based mixture will work as well."

The nutrient solution he stated can, obviously, be replaced by the natural banquet of hormones in kelp (like 3LB & VC stated). This ‘germination bomb’ essentially covers each mode of seed scarification in heat, pressure, and water. The air pump provides constant agitation which in turns creates oxygen which is the most abundant element needing in root formation. I have improved my germination by easily 80% since using this technique. I grow solely landrace and heirloom cultivars so needless to say most seeds I posses are old and require special attention.

What’s great about it is, that if the seeds sink - they’re viable. And as I stated, this germination bomb covers all forms of scarification. In my mind, it is the ONLY way to germinate seeds.

And oh yea, DONT USE PAPER TOWELS! Yes, they may work and get the job done(for freshly produced hybrids). But it is an artifical medium and devoid of the microbes necessary to break down (tough, old) seed coats.

For a germination medium I use worm castings and mychorrizal innoculated perlite.

GERMINATION BOMB EXPERIMENTS

Initially, I was primarily interested in whether the seeds would germinate. Since the long-term goal is to pop some of the elite seeds in my collection, I will be germinating the seeds in medium. I will keep track of how many seeds actually grow as well as how many germinate.

I use pint size mason jars with metal lids. Since the air pump I am using has 4 ports, I run 4 jars in each experiment. One jar is a control, the other three jars have different strengths of whatever additive I am testing.

SUPPLIES NEEDED

Glass jars with tight screw-on lids. I use wide-mouth 1 pint Mason jars since replacement lids & rings are easy to get (the lids are going to rust, probably sooner rather than later)
Aquarium air pump
Small airstone for each jar
Air tubing
Silicone sealer
Drill
Drill bit the same size as the air hose
Wood block (scrap)

BASIC SETUP

To prepare the mason jar:

Set the lid on top of the piece of scrap wood to drill the hole in the lid. This greatly reduces the chances of getting sharp shards around the edge of the hole as you drill it. The hole in the lid needs to be just large enough to pass the air hose through. Put a small air stone on the end of the hose coming out the bottom of the lid. Set the lid on top of the jar & adjust the length of the hose inside the jar so that the airstone just touches the bottom of the jar. Seal around the hose on the top & bottom of the lid with silicone; allow to cure.

The jars are filled halfway with water (8 oz). In general, I use reverse osmosis filtered water. The experiment with GA3 was done with distilled water because the instructions specified distilled.

Put in each jar (this amount to 8 oz water):
10 drops Superthrive
1cc food-grade molasses (from the grocery store, a previous experiment using horticultural molasses resulted in a problem with mold)
1 tsp 3% hydrogen peroxide

The jars are set on top of a seedling heat mat to keep them warm with a towel draped over the top to block light. Attach the air line to the air pump and plug the pump in/turn it on. .

EXPERIMENT 1 - DMSO STRENGTH

A word of warning - DMSO is not something to handle casually. It can be dangerous. Before opening the bottle, wash your hands and any exposed skin thoroughly & put on rubber or surgical gloves; long sleeves are also a good idea. Be VERY careful not to get any on your skin. DMSO will take anything it comes in contact with (including anything that is on your skin) and help it to penetrate into your cells - this includes oil, gasoline, and everything else. The system below works reasonably well without the DMSO (unless your seeds are really old or were not stored well), but I suggest scuffing the seeds before putting them in the mix if you don’t use it.

The first experiment I did was to see how various strengths of DMSO affected germination. DMSO can be found easily at Tractor Supply Co or most feed stores. DMSO facilitates penetration of the nutrients & amendments through cell walls/membranes into the seed - no scuffing required. This experiment was done using reverse osmosis (RO) filtered water.

Each mason jar has a different amount of DMSO per 8 oz of water:
Jar 1 - 5 drops DMSO
Jar 2 - 8 drops DMSO
Jar 3 - 10 drops DMSO
Jar 4 - 13 drops DMSO

The jar with 13 drops of DMSO had the best rate of root development. Of course, since the seeds are 18 years old & weren’t stored well at the start, most of the seeds didn’t develop beyond popping roots. Here are the results:
Jar 1 - 16 of 20 seeds cracked. 14 developed visible roots about 1/16 - 1/8" long
Jar 2 - 17 seeds cracked & all developed roots average 1/8" long
Jar 3 - 19 seeds cracked & developed roots average 3/16" long
Jar 4 - all 20 seeds cracked & developed roots average 1/4" long

Once the seeds had roots at about 1/4" long, I transferred them to seed starting trays with purchased sterile seed starting medium. I watered the seedlings with a dilute mix of nutes, Superthrive, & a bit of molasses. Most of them died, but 1 seedling from the jar with 10 drops DMSO grew & 2 seedlings from the jar with 13 drops DMSO grew. They were not the strongest seedlings, of course.

After running this experiment, I decided I wanted to test other mixes, so I will not be using as many seeds for future experiments. I want to use the same seeds for all experiments to eliminate that potential variable. If I keep using 20 seeds in each jar, I won’t have enough seeds to complete all of my experiments. I am also going to germinate using my normal mix of perlite with a bit of vermiculite rather than using a purchased seed starting mix. I grow in an Ebb & Flow system, so it’s easier for me to start the seeds in the same medium I will use for growing. I put the seeds into cups of perlite with a pinch of vermiculite around the seed.

Since the jar with 13 drops DMSO worked best, that will be included in my standard mix. The new mix is (this amount per 8 oz of water):
10 drops Superthrive
1cc molasses (from the grocery store, a previous experiment using horticultural molasses resulted in a problem with mold)
1 tsp 3% hydrogen peroxide
13 drops DMSO

EXPERIMENT 2 - GA3 STRENGTH

Using 90% GA3 powder, I created a 150ppm batch of GA3 by dissolving 1 scoop (1/32 tsp, 0.8 gram) GA3 in 0.5cc 91% rubbing alcohol in a shot glass. The alcohol evaporated as I was working on dissolving the powder, so I added more alcohol as needed (I used close to 1cc total). I then added the dissolved powder/alcohol mix to 16 oz distilled water. I used distilled water (purchased) for this entire experiment since the GA3 instructions specified it.

In addition to the base mix shown previously, I used the following:
Jar 1 - control, no GA3 (8 oz distilled water)
Jar 2 - 50ppm GA3 (5.333 oz distilled water & 2.666 oz of the 150ppm base mix GA3)
Jar 3 - 100ppm GA3 (2.666 oz distilled water & 5.333 oz of the 150ppm base mix GA3)
Jar 4 - 150ppm GA3 (8 oz 150ppm base mix GA3)

I put the seeds in a 1:20 mix of H2O2 & water (1/4 tsp H2O2, 5 tsp RO water) for 1/2 hour to kill pathogens on the shell before putting them in the pressure jars.

After 36 hours in the pressure jar, I put the seeds into cups to germinate:
Jar 1 - 4 of 5 seeds popped open, one not popped. One seed the shell was completely off and the cotelydon leaves were opening. The one that did not pop is in position 4 (bottom left); the one with leaves is in position 3 (center)
Jar 2 - All 5 seeds popped open with roots growing. One seed the shell was completely off and the cotelydon leaves were opening. The one with leaves is in position 3 (center)
Jar 3 - 4 of 5 seeds popped open, one not popped. One seed the shell was completely off and the cotelydon leaves were opening. The one that did not pop is in position 4 (bottom left); the one with leaves is in position 3 (center)
Jar 4 - All 5 seeds popped open, one just barely. One seed the shell was completely off and the cotelydon leaves were opening. The one that did not pop well is in position 4 (bottom left); the one with leaves is in position 3 (center)

Long term survival:
Jar 1 - 2 seedlings grew successfully. The seed that hadn’t popped open never grew.
Jar 2 - 1 seedling grew beyond 1st set of leaves; very stretched - survived a couple of weeks but died. The other 4 grew but did not survive to grow 2nd set of leaves. All somewhat stretched.
Jar 3 - The others made it above ground, but stretched & ended up dying - from the center out, oddly. These died out second quickest. The seed that hadn’t popped open never grew.
Jar 4 - The others made it above ground, but stretched a lot & ended up dying - from the center out. These died out quickest.

The GA3 did not seem to make a significant difference in the number of seeds that developed at least a root tip. The more GA3 in the mix, the shorter the eventual life of the seedling & the more stretch it experienced. The germination bomb without the GA3 seems to be the best long-term germination method.

Future Experiments:

I want to do another experiment with DMSO using 15, 18, and 20 drops per 8 oz of water. Since the strongest mix I tested at the beginning was the best, I want to find out if a stronger mix is better.

I want to try an experiment using IBA in the mix.

I would also like to test using different sources of nutrient for the older seed - molasses, sugar, kelp, mild mix of standard hydro nutrients

Hope you read it, and helps you in some way. Will try it myself.

HppHrvst :robot:

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