Author Topic: Germinating super old seeds  (Read 1271 times)

Joseph Lofthouse

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Germinating super old seeds
« on: 2020-11-11, 08:09:55 PM »
I recently traveled to a seed bank in Colorado, and acquired seeds for the Beautifully Promiscuous and Tasty Tomato Project. A plant breeder named GW Denna had started a project like that in about 1971 to 1974, then he died young. The seeds sat in a warehouse in Colorado until they made a new home in the seed bank a few years ago. I was able to tell the custodian of the seed bank why Denna's project was important, and about the meaning of the labels on the seed packets.

The seeds are:

F1 hybrids between Solanum peruvianum.
F2 hybrids between domestic tomatoes and Solanum peruvianum.
A domestic OP tomato that was highly attractive to bumblebees.
LA128 L hirsutum glabratum from Baņos Equador.  Self fertile.
LA387 L hirsutum from Santa Apolonia Peru. Mixed fertility.
"Wild Tomato Crosses" with large-ish seeds indicating domestic ancestors.
L peruvianum. SI F2 Intercrosses with large-ish seeds indicating domestic ancestors.

The seeds are 45 years old and were stored haphazardly. I have about 500 seeds to work with.

Can you recommend germination protocols for old seeds?

Some ideas that have been suggested include.

  • Watering with kelp fertilizer.
  • Soaking in dilute potassium nitrate before germination.
  • Watering with tea. (Helps replace the solutes that leach out of older seeds)
  • Watering them with 0.15% H2O2. 3 tablespoons 3% solution per pint. (Gives them a boost of oxygen)
  • Sprouting them on paper towels instead of soil. (Reduces damping off.)
  • Surface sterilizing and growing on agar with MS/Gamborg nutrients.
  • Keep them in a warm place.
  • Plant them with bio-dynamic calendar.
  • Sing and dance for them. Love on them.

I'm currently fascinated with the idea of making a tea from tomato seeds, and watering with dilute hydrogen peroxide, on some type of hydroponics mat. The peroxide is also an antibiotic.

Anyone want to recommend your favorite method for germinating old seeds?



« Last Edit: 2020-11-11, 08:46:54 PM by Joseph Lofthouse »

William S.

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Re: Germinating super old seeds
« Reply #1 on: 2020-11-11, 10:18:25 PM »
Rough Surface sterilization protocol:

10% bleach with a drop of free and clear dish soap stir
 
Them 3% H202 stir to rinse of the bleach

Then sterile water rinse.

A product called PPM or Plant Protective Material can reduce contamination rates.

If used media sans sugar to soak germination paper wouldn't really need agar and without glucose less risk of mold etc.

Can also sterilize everything in a pressure cooker or microwave depending.

Carol Stiff wrote up detailed instructions for home tissue culture very helpful.

Kinda want to just load up my plant tissue culture kit and come help but probably can't.

Surface sterilization generally increases germination rates

Liquid smoke generally increases germination rates (choose one without added sugar). One of the neat things about forest and chaparral fires is the wildflower flush after. Precedes massive germination events. The smoke may help replace the solutes that leak out and or suppress damping off organisms. Speculating in part as to how it works, Google probably knows more by now than I do about it.

Western Montana garden, glacial lake Missoula sediment lacustrian parent material and shallow 7" silty clay loam mollisoil topsoil sometimes with added sand in places. Zone 6A with 100 to 130 frost free days

Garrett Schantz

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Re: Germinating super old seeds
« Reply #2 on: 2020-11-11, 10:32:48 PM »
The oxygen boost is pretty important considering what you are working with. So I would incorporate that no matter what.
Nitrogen has been found to increase germination in old seeds, unsure if this applies to tomato seed though.
Letting the seeds soak in diluted sugar could improve germination as well - be sure to sterilize the seeds in diluted hydrogen peroxide to reduce chances of mold if you go this route. Diluted Hydrogen Peroxide can help with oxygen for the seed as well, to revitalize enzymes and such.
Some sources mention diluted Gibberellic Acid as an option as well, unsure though.
I would try all of these and see what works best. 45 years combined with not so good storage conditions, the seeds probably need all the help they can get.
 William posted a lot of good information / ideas.

S.Simonsen

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Re: Germinating super old seeds
« Reply #3 on: 2020-11-12, 12:53:15 AM »
How to manage risk and make best use of time and resources germinating different batches depends a lot on the size of each batch as well, and depends on how much time/resources/attention you have to spare.

Personally I would start out with the largest batches where you have the most chances to get it right, and take out a small (5-10%) sample. A germination test on damp paper would be my first step, just to get a sense of standard levels of viability, especially since you know very little about how the seed has fared during the long storage time. If you get absolutely no sign of germination then repeat with a slightly larger percentage of seed. If you get poor germination with sprouts too weak to establish under normal treatment only then would I consider going down the route of sterilisation and boosting treatments or tissue culture. Then you can figure out a successful protocol on small portions of the larger batches before applying the method to the smaller and more valuable batches.

Ocimum

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Re: Germinating super old seeds
« Reply #4 on: 2020-11-12, 03:17:22 AM »
If they were kept dry, first keep them for a few hours/up to days at high humidity, otherwise they soak up the water too fast and may die. At least this is for some species, no idea if it's the same with tomatoes. I germinated seeds which, according to life expectancy in books, should not have sprouted that way.

Oh, and some use Lactobacillus to improve germination in seeds


Joseph Lofthouse

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Re: Germinating super old seeds
« Reply #5 on: 2020-11-12, 09:30:14 AM »
I looked more carefully at the seeds. There are about ~300 seeds from the wild ancestor. ~300 seeds from the F2 of a cross between domestic tomatoes and Solanum peruvianum. And ~40 elite seeds that are exactly what I'm looking for in my breeding project. (These might be G4). So it's looking like I'll be able to successively experiment to maybe find suitable germination conditions. 

Adrian

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Re: Germinating super old seeds
« Reply #6 on: 2020-11-12, 12:43:11 PM »
I choose a potting soil and i add sand in.
The best is a potting soil with mychoryze muschroom
What would the effect of lactobacillus mixed with the coffee ground ?  I will try a mix with potting soil, ash,sand and coffee ground for create a very good seedling potiing soil! For me the most important is the coating of the seed by the potting soil.
The problem of the potting soil alone is the difficulty to humidifated this and have a bad water penetration.
« Last Edit: 2020-11-12, 02:54:43 PM by Adrian »

Steve1

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Re: Germinating super old seeds
« Reply #7 on: 2020-11-13, 04:17:38 AM »
If it was worth alot to me, I'd be going the tissue culture route. Probably with some gibberellic acid. Seems to me with weak seeds that damping off/fungus get hold before germination occurs. Surface sterilisation at least if done properly halts that process. If you have a clean hepa filtered room, and good sterile technique then you may get months before the plate gets infected which means best chance. The 32,000 year old seeds from the Russian permafrost recently germinated were done in tissue culture. Guessing there would be a paper that might help with details.
https://news.yahoo.com/austrian-scientists-revived-32-000-105145497.html

Adrian

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Re: Germinating super old seeds
« Reply #8 on: 2020-11-13, 07:16:11 AM »
For a weak plant i think that the most dangerous is the fly of the seedling and the pythium muschroom.Its very difficult to save a plant after their attacks. The alone solution is the cutting.
I don't know if its a good idea but i hace try to put gibbeleric acid at the germ position before the germination.
« Last Edit: 2020-11-13, 07:50:09 AM by Adrian »

Adrian

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Re: Germinating super old seeds
« Reply #9 on: 2020-11-28, 08:24:46 AM »
This year i have try the mychorize poting soil N=6 P=7 K=8 with coffe ground above the synergy with the coffe and the mychorize is for me miraculous.I have see of lentil in and i have never see a lentil plant so beautiful!
The result is best with a mulch above the coffe ground for fight against the drought of the germ if she is long to germinate.
« Last Edit: 2020-11-28, 08:28:07 AM by Adrian »

Richard Watson

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Re: Germinating super old seeds
« Reply #10 on: 2020-11-28, 10:01:34 PM »
I had a sack of Mangle beet seed that would have to have been 10years old and was not stored in a ideal location really. Thinking bugger all would germinate I sowed thickly, surprised to find i must have had 90%+ success rate, got a big job now thinning out.
Changeable climate manly during winter & spring - just under 500mm average yearly rainfall. 20 years of soil improvements plus sub soil top soil reversal means my garden beds are about half metre deep. Below that is 100's of metres of alluvial out wash from the Southern
alps

Adrian

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Re: Germinating super old seeds
« Reply #11 on: 2020-12-13, 05:30:47 AM »
Has the coffe ground contain an inhibitor of germination,
 i have been an idea for have a best germination with the coffe ground!
I will did a solution of C6H12O6, glucosis with H2O water in my coffe ground.I may stimulated the cellular growing and feed more easily my muschrooms mychorizes.
 have mixed the cofee ground with of granule manure.
« Last Edit: 2020-12-13, 09:09:55 AM by Adrian »

Joseph Lofthouse

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Re: Germinating super old seeds
« Reply #12 on: 2020-12-13, 06:45:22 PM »
I started some preliminary testing with the old tomato seeds...

Attempting germination in petri dishes with quilt batting and coffee filter paper. Three different sets of conditions. In all cases with 0.15% hydrogen peroxide.

Water only
Tomato seed extract
M/S Gamborg fertilizer and vitamins, commonly used in plant tissue culture.

Using my standard germination conditions: 90F and lighted for 16 hours per day. 60F and dark the rest of the time.

The tomato seed extract is encouraging the growth of microbes.

I'm expecting that surface sterilizing of the seed will become part of the procedure for next time. I autoclaved the petri dishes and solutions. 

« Last Edit: 2020-12-13, 06:50:32 PM by Joseph Lofthouse »

Steve1

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Re: Germinating super old seeds
« Reply #13 on: 2020-12-15, 07:57:51 PM »
Joseph, the tomato seed extract could be interesting / helpful. You could mix that with your medium / GA and then sterilize the plates then sow seed after surface sterilization. We use V8 mixed fruit/vegetable juice in medium when tissue culturing specific fungal plant pathogens. Good luck. 

Steph S

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Re: Germinating super old seeds
« Reply #14 on: 2020-12-30, 11:07:26 AM »
Wow, Joseph, what a find!   Hope you have had some success by now?

I've heard of using dilute N ferts in the seed soak for old tomato seeds.  I like the point of humidifying them before soaking.  I have only used an overnight soak in water, for some 15yr old tomato seeds and I did get seedlings.   But 45 years...  I guess much depends on unknown conditions during that time.
One thing I've noticed with old pepper seeds is that they take much longer to germinate and then can suddenly surprise you, weeks after you gave up.  I always soak pepper seeds overnight, even if they're fresh.  I had some hot pepper seed given me, marked "old/low germination".  I put their soaking cup on the heat mat for the overnight soak.  Ended up with seedlings too many to count in just a couple of days.
Tomato seeds that stubbornly don't germinate also have a tendency to pop up much later, unexpectedly.
So whatever treatment/soak you used, I would treat, plant, and then wait at least a month.  Maybe 45 days for 45 years?

My thought about soaking with ferts or kelp is that I didn't really want something that is decomposing or promoting decomposition in the vicinity of the pre-germination seed.
Old seeds definitely do need ferts as soon as they have germinated though.   Brassica seed a case in point which I find deteriorates after the third year or so - they still germinate but the cots will come out yellow and they don't thrive unless you feed them immediately.  Same was true of the old tomato seeds - some cotyledons less than prime condition, so a bit of a handicap in ability to nourish itself.  This is because the nutrients saved in the seed for the purpose of getting started are no more - used up in seed respiration iirc or simply deteriorated over time.

I know lactobacillus is supposed to discourage pathogens on the ungerminated seed surface.  I used whey to soak some tomato seeds before planting.  They didn't mind it, were healthy.

Here's a thought:  Some time ago I read about the microbiome around seed germination - apparently the exudates of the seed promote a very specific and transitory microbial community which assists in getting to the seedling stage.   Maybe an extract could be made by ? washing germinating seeds to collect the exudates and associated microbes, to soak the seeds in before planting and/or to water them with, once they're in the soil.  If you had a big batch of seeds you don't plan to grow out, you could sprout them for the purpose...

That is, if you don't have luck with the agar.  Hope you did. :)