Germinating Very Old Seed

[deluxestogie]

CR,
Thanks for the reference. It's discouraging to see in that snippet that sensitivity to GA-3 seems to vary by variety. And their study used seed that was, at most, only 9 years old. (ARS-GRIN's Nicotiana Seedbank is renewed at a rate of 10% of the accessions per year. Perhaps their "alternate site" does the same, though with different varieties, which would be a 5 year cycle between the two sites, but to my knowledge, it's a 10 year cycle.)

The Sweet Oronoko seed was completely used in 8 trials, 6 of which are still awaiting their outcome. Only two of the trials used 1000 ppm. Others were exposed to lower concentrations, though the methodology was not adequate to say exactly what concentration, since it was not used as a "soak" with those other trials, but lightly misted onto already moistened germination mixture in various ratios. In some of the trials, the seed had already imbibed water prior to GA-3 exposure. The two trials that molded were performed exclusively on filter paper, with no sign of germination. I would guess that at least one of the trials was at less that 1/4 of the starting concentration of 1000 ppm. With no more of the seed, whatever these remaining 6 trials yields is all she wrote.

A lab setup would be nice. At this point in my growing season, I might still be able to produce some seed from any successful germinations, but add another two or three weeks and it gets down to a pretty tight schedule. If Don still has any of the Sweet Oronoko seed, then maybe next March would be a suitable time to attempt it again with more precise methods. Given that the seed is already likely to be 0% viable (130 years old, and probably stored in a kitchen drawer for a century), I'm not particularly optimistic. It's still a fun thought.

Bob

 

[CoralReefs]

CR,
Thanks for the reference. It's discouraging to see in that snippet that sensitivity to GA-3 seems to vary by variety. And their study used seed that was, at most, only 9 years old. (ARS-GRIN's Nicotiana Seedbank is renewed at a rate of 10% of the accessions per year. Perhaps their "alternate site" does the same, though with different varieties, which would be a 5 year cycle between the two sites, but to my knowledge, it's a 10 year cycle.)

The Sweet Oronoko seed was completely used in 8 trials, 6 of which are still awaiting their outcome. Only two of the trials used 1000 ppm. Others were exposed to lower concentrations, though the methodology was not adequate to say exactly what concentration, since it was not used as a "soak" with those other trials, but lightly misted onto already moistened germination mixture in various ratios. In some of the trials, the seed had already imbibed water prior to GA-3 exposure. The two trials that molded were performed exclusively on filter paper, with no sign of germination. I would guess that at least one of the trials was at less that 1/4 of the starting concentration of 1000 ppm. With no more of the seed, whatever these remaining 6 trials yields is all she wrote.

A lab setup would be nice. At this point in my growing season, I might still be able to produce some seed from any successful germinations, but add another two or three weeks and it gets down to a pretty tight schedule. If Don still has any of the Sweet Oronoko seed, then maybe next March would be a suitable time to attempt it again with more precise methods. Given that the seed is already likely to be 0% viable (130 years old, and probably stored in a kitchen drawer for a century), I'm not particularly optimistic. It's still a fun thought.

Bob

Hmmmm....
I have two thoughts on this:
1) Have you contacted a local university to see if anyone would be interested in trying to get these going? I know at my University, there are people actively doing research on tobacco mosaic virus and as such do experiements on plants. The interest seems to be around. If nothing else, if you have lab experience you might be able to get access to one of the labs. (Since I do volunteer work in my University's plant conservatory managing one of the collections, I have been told I can pretty much have access to just about anything I need- despite having a very meager background in biology).
2) Ever look into plant tissue culture? A technique that I have used, with some success, for some more difficult to germinate species is to grow them in vitro. That is to make a sterile nutrient enriched agar media (kind of like vitamin enriched jello if you are not familiar with it) and attempt to germinate the seedlings there. I have heard of people successfully pretreating seeds with GA3 and then innoculating the media with the pretreated seeds.
Personally, I have not yet done this as most of the plant tissue culturing I have done has been with grown plant material, not seeds, so GA3 has never come up. I have had some success with seed though. It is a very powerful technique that can really work wonders (if you have viable material in the first place of course). Plant TC, for instance, is one of the only ways to get Orchid seeds to germinate and grow since orchid seeds lack stored nutrients and rely on a symbiotic fungus in nature.

I know I have a paper around here somewhere with a TC protocol for TCing tobacco from seed, I will see if I can find it.

 

[deluxestogie]

My Long Red has germinated very poorly, and none of the tiny seedlings had survived. Ten days ago, I started a third batch of them on fully moistened germination mix, and lightly spritzed it with my 1000 ppm GA-3. I now have two apparently viable seedlings from that batch. I suspect they will be my only two Long Red plants, if they survive.

Apart from that benefit, it does serve as an indication that the method I used for applying GA-3 seems to have been more productive than without it. Who knows if a weaker GA-3 solution might have yielded a greater number of happy seedlings. The light spray of 1000 ppm GA-3 was not a death sentence in this instance.

Bob

EDIT: A photo from the delivery room:

 

[wazzappenning]

My Long Red has germinated very poorly, and none of the tiny seedlings had survived. Ten days ago, I started a third batch of them on fully moistened germination mix, and lightly spritzed it with my 1000 ppm GA-3. I now have two apparently viable seedlings from that batch. I suspect they will be my only two Long Red plants, if they survive.

Apart from that benefit, it does serve as an indication that the method I used for applying GA-3 seems to have been more productive than without it. Who knows if a weaker GA-3 solution might have yielded a greater number of happy seedlings. The light spray of 1000 ppm GA-3 was not a death sentence in this instance.

Bob

EDIT: A photo from the delivery room:

if worse comes to worse, you might have one left to germinate seed for next year

 

[CoralReefs]

My Long Red has germinated very poorly, and none of the tiny seedlings had survived. Ten days ago, I started a third batch of them on fully moistened germination mix, and lightly spritzed it with my 1000 ppm GA-3. I now have two apparently viable seedlings from that batch. I suspect they will be my only two Long Red plants, if they survive.

Apart from that benefit, it does serve as an indication that the method I used for applying GA-3 seems to have been more productive than without it. Who knows if a weaker GA-3 solution might have yielded a greater number of happy seedlings. The light spray of 1000 ppm GA-3 was not a death sentence in this instance.

Bob

EDIT: A photo from the delivery room:

Since you sprayed it like that, the concentration was probably much lower than 1000PPM. Particularly if the soil was premoisened.