Germinating Very Old Seed

[deluxestogie]

Since I am looking at the prospect of attempting to germinate some very old tobacco seed (many decades+), I explored some material on the subject.

The following represent some interesting source material on the subject of germinating old seed. None is specific to tobacco.

How Gibberelic Acid and Light affect seed germination:
http://plantphys.info/seedg/seed.html
"The DNA must first be photoactivated before GA can have its effect."
"If we shine red (or it turns out even white) light on imbibing seeds, then virtually all of the phytochrome is converted to the Pfr form and this photoactivates the genes in the DNA. Almost all of the seeds will germinate in this light!"
[After the first long page, subsequent pages are very short, with a diagram. Continue through the discussion of lettuce seed germination.]

Germination of 2000 year old date palm seed from the palace of Herod the Great:
http://en.wikipedia.org/wiki/Judean_date_palm#Germination_of_2000_year-old_seed

Pre-germinating old seeds, best method? [Forum from Seedsavers.org]
http://forums.seedsavers.org/showthread.php?t=1135
[Discusses use of seaweed extract and high nitrogen pre-soaking.]

Seed Germination Testing from Ohio State University:
http://ohioline.osu.edu/b639/b639_2.html


Bob

 

[BarG]

Bob, you may find this interesting, click on the sample kindle book and see how Santa Fe Natural Tobacco company was started.
http://www.amazon.com/Cultivators-Handbook-Natural-Tobacco-ebook/dp/B003GSM2YK#reader_B003GSM2YK

 

[rustycase]

Tnx Bob,
I've got some older veggie seeds and would like to apply 'best practices' to getting them germinated.
I'll need to review those sites you listed to see what encouragement they have.
Best
rc

 

[BigBonner]

Look who finally made it here , Rustycase , Stick around and try to win some tobacco . Be sure to search for contest .

 

[deluxestogie]

rc,
I could have sworn you've been posting here for months. Welcome.

Bob, you may find this interesting, click on the sample kindle book and see how Santa Fe Natural Tobacco company was started.
http://www.amazon.com/Cultivators-Handbook-Natural-Tobacco-ebook/dp/B003GSM2YK#reader_B003GSM2YK

BarG,
Unfortunately, the seed from the Indian "Archaeological Dig" said to be many hundreds of years old was never radio-carbon dated. To my knowledge, no truly ancient tobacco seed has been successfully germinated.

Sacred Cornplanter seed was passed to a museum in the 1910s by the chief of one of the tribes in the NY Fingerlakes region. It was said to be the very strain once grown by Chief Cornplanter. A decade or two later, an ethnologist acquired some of the seed propagated by the museum. So, even in this case, the "very old" seed was probably refreshed every decade or so. And we know that 10 years is a reasonable storage life for N. tabacum seed, since ARS-GRIN has been refreshing (growing-out) their seed at a rate of 10% of the accessions each year, in rotation.

 

[BigBonner]

Isn't there a X -ray they can do to see if seeds are still good . Seems like I have read this some where at one time . Im not for sure .

 

[BarG]

Well that is interesting, Don't believe anything you hear and only half of what you see, We used to play devils advocate me and an old partner and I learned to be prepared to back up what you say. I won't be quoting that particular claim regarding ancient seeds. What I should have said was [Hey Bob, check this site out ,Is there any truth to it you think?]

 

[BigBonner]

I did find a little about x ray on seeds but it would be hard to x ray a small tobacco seed .

http://www.forestry.gov.uk/website/forestresearch.nsf/ByUnique/INFD-8FGBQ4

 

[Jitterbugdude]

I did find a little about x ray on seeds but it would be hard to x ray a small tobacco seed .

I don't know about that. I'm going for my dental check up next month.. send me some and I'll pop them in my mouth before I get my x-rays

 

[rustycase]

An article on web news described Russians who germinated seed found frozen in rodent burrows in Siberia.
They produced plants with further viable seed.
The seeds were thirty thousand years old.
rc

 

[BarG]

Seed banks flash freeze seeds to maintain a certain below freezing temperature and are continualy monitored to maintain a viable seed bank.[research world seed banks] They just took a clue from mother nature. I suspect the freezing temps kept the seeds viable. Possibly along with a heaping helping hand of modern technology. If I lived in alaska I'd be looking to try some fresh mammoth meat,they find every so often.

 

[deluxestogie]

In a personal communication from the (until recently) curator of the ARS-GRIN Nicotiana Collection:

"Bob,
They have been stored at about 8 degrees C [~46.4ºF] for the working part of the collection. For longer-term storage, they have been put in a deep freeze. I have no information to say that this is the best."

So, I would say that refrigerator temp is good, and that a cool, dark pantry or closet would be next best. They should, of course, be enclosed within a tightly sealed container.

Bob

 

[deluxestogie]

Using Gibberellic Acid 3

FmGrowit provided me with a 100mg packet of gibberellic acid 3 (GA-3). When added to 100 cc of distilled water, this makes up a 1000 ppm (parts per million) solution of GA-3.

GA-3 is derived from a fungus, and causes a fatal seedling abnormality in which the seedling goes into overdrive and grows out too long to survive. Good for the fungus, not so good for the seed. While this is a problem for normal seed, and should not be used on healthy, fresh seed, it can be helpful for seed that is otherwise difficult to germinate (like the 160 year old "Sweet Oronoko"). I decided to try it on "Sweet Oronoko" and "Big White Burley KY16," the latter being only a tender 60 years old.

Rather than undertake careful laboratory procedures to expose the selected seed to an exact concentration of GA-3, I added 3.5 ounces of distilled water to a $1 spray bottle (from the Dollar store), and dumped in the 100 mg packet of GA-3. I stirred it, and allowed it to fully dissolve overnight. This comes out to roughly a shaver more dilute than 1000 ppm.

I cut from a #1 coffee filter three rectangular sections, each hinged at one end. These were sized to fit into a tiny Ziplock bag.

I lightly misted pure water onto one opened section of filter paper, sprinkled a few seeds onto one half of it, then folded it closed again, and inserted it into a labeled Ziplock. Using the GA-3 spray, I gave it a couple of sprays into the Ziplock. Since the seed will imbibe at the same rate, regardless of the quantity of liquid it is exposed to, the quantity of GA-3 solution does not matter. Only its concentration (~1000 ppm) matters here. The Ziplock is then closed, and allowed to rest in sunlight for about 8 hours, with the ambient temperature at about 80ºF.

Following this, the seed was rinsed from the filter paper into a prepared jar of moist germination soil using a spray bottle of distilled water. The soil in this case was a coir-based organic seed-starting mix.

I set this jar onto a cheap metal cookie sheet which was placed on an inverted 5 gallon bucket near a baseboard heater in a somewhat darkened room. The baseboard heater seems to keep the temp at a stable 80ºF.

For "normal" seed, I am using 4 oz. jelly jars. The "normal" seed is sprinkled onto the moist soil mix, and lightly tamped onto the surface, to assure good contact. The lid is closed, and the jars added to the aforementioned cheap cookie sheet.

As for healthy seed, I typically see fairly subtle signs of germination by about 60 hours. Shown below is fresh Harrow Velvet seed at 60 hours in the jar. No light treatment; just the warmth. Most of the 12 varieties started 60 hours ago show signs of radicle emergence. I'll be starting 2 more waves of varieties at weekly intervals.

Notice the white tip, which is the emergence of the radicle, clearly visible in the closeup.

While the two ancient seed varieties have swelled with the moisture, they have yet to show signs of life. But I'll give them several weeks before I give up on them. The normal seeds will come out of the jars and into cell trays as soon as enough seeds of a variety are beginning to germinate. My worry with the old seed is the excessive moisture in the jar over a lengthy period, and the increasing risk of fungal growth. After the initial 72 hours, I'll slowly dry-down the soil in those jars to just barely damp.

Bob

 

[Tom_in_TN]

Looking for any Cherry Red Free seeds

Choose "Cherry Red Free" for my FREE seed pick. Then was informed that the last time a batch went out there was a problem with none of them germinating, at all. Still have not received them yet but I plan to pre-soak a few seeds in a weak saltpeter solution for about 6 hours and then some for 10 hours. Then transfer to either potting soil or the paper towel method, maybe both. Wonder if rinsing seeds off the towel into a little cup of distilled water and then scooping them out one a time to place on potting soil would work?

Since the "Cherry Red Free" seed is surely not be as old as the "Sweet Oronoko" and "Big White Burley KY16" there is hope that the nitrogen soak I'll try 1st should work. And there are other methods that I just don't know about, yet. Who knows what will work best? I guess using Gibberellic Acid 3 would be the next option. Seedman.com sells small amounts of it in liquid or powder form.

deluxe, when I read through their instructions, advice and the 2 authors they cite as experts on germinating old seed, it seems there is a chance some seeds will do better at lower concentrations. So, if the 1000mg concentrations fails, try some @ 750 mg, 500mg, or 350 mg. I would say it is worth a try, I mean if you've got enough seeds, why not?

 

[CoralReefs]

Bob,
How is your experiment with the GA3 going? I have tried using GA3 numerous times in the past on some difficult species of plants. I have never had the best of luck- often times seems to encourage fungal growth. Others I know say it works miracles! I have a bunch of notoriously difficult seeds from South Africa I am going to try hitting with GA3 and/or smoke disks here pretty soon here. If you have any insights I would appreciate hearing them.

 

[deluxestogie]

I am not an expert at GA-3 or the germination of old seed. So far, my efforts with GA-3 use with tobacco seed has been pretty much confined to an attempt to germinate what might be considered impossibly old seed (130 year old Sweet Oronoko and 60 year old Big White Burley KY16)--8 different trials of each. Neither of these varieties has shown any convincing sign of life, though the tests are still running.

Bob

 

[Tom_in_TN]

Bob, what is the maximum amount of time are you allowing for each trial run? I still have some of the Yayladag seeds running with the hope that another sprout will show its root. Started 1st trial 03/27 and the 2nd run on 04/03.

Tom TN

 

[deluxestogie]

I essentially let it run until it germinates or molds. After one week, I allow the humidity to decrease, but never allow the soil to dry.

My oldest that are still going were started 2/26.

In some of my standard starts, I'm seeing occasional new sprouts 3 weeks after starting, so waiting a month or more for the stubborn ones seems reasonable.

Bob

 

[CoralReefs]

I essentially let it run until it germinates or molds. After one week, I allow the humidity to decrease, but never allow the soil to dry.

My oldest that are still going were started 2/26.

In some of my standard starts, I'm seeing occasional new sprouts 3 weeks after starting, so waiting a month or more for the stubborn ones seems reasonable.

Bob

I see that you are using 1000ppm for your GA3 concentration. I have never hit baccy seeds with GA3 but some of the plants I work with would be killed by 1000ppm. I usually start with much lower (a few seeds around 250ppm, some at 500 ppm- these are the concentrations I start with), if those concentrations do not work I try tirating upward. I have not had the best of luck in this department though, but then again I am working with seeds that are notoriously impossible to germinate unless sowed immediately upon seed ripening (google search 'genlisea' or 'byblis' for two such examples).

Ok, I just got curious and did a google search- here is an article abstract I found on GA3 and tobacco. Might provide some insight. I will see if I can download it next week when I have access to an institutional journal subscription.

http://aob.oxfordjournals.org/content/51/1/157.extract

 

[CoralReefs]

Re: Looking for any Cherry Red Free seeds

Choose "Cherry Red Free" for my FREE seed pick. Then was informed that the last time a batch went out there was a problem with none of them germinating, at all. Still have not received them yet but I plan to pre-soak a few seeds in a weak saltpeter solution for about 6 hours and then some for 10 hours. Then transfer to either potting soil or the paper towel method, maybe both. Wonder if rinsing seeds off the towel into a little cup of distilled water and then scooping them out one a time to place on potting soil would work?

Since the "Cherry Red Free" seed is surely not be as old as the "Sweet Oronoko" and "Big White Burley KY16" there is hope that the nitrogen soak I'll try 1st should work. And there are other methods that I just don't know about, yet. Who knows what will work best? I guess using Gibberellic Acid 3 would be the next option. Seedman.com sells small amounts of it in liquid or powder form.

deluxe, when I read through their instructions, advice and the 2 authors they cite as experts on germinating old seed, it seems there is a chance some seeds will do better at lower concentrations. So, if the 1000mg concentrations fails, try some @ 750 mg, 500mg, or 350 mg. I would say it is worth a try, I mean if you've got enough seeds, why not?

Oh oops, I did not see this post before I submitted that last one. Sorry for the repeat!