"Harvey Brenneise" <[EMAIL PROTECTED]> queried thusly:

> Does anyone out there know of any work done with orchid seed viability, 
> particularly long-term and/or of seeds stored at very low temps? 

There are a few papers out there; they're all referenced in my first book, and 
I can fetch those up if you really want to go back to the literature.

About 99% of what we store is refrigerated at 4C, maintained over doubled 
calcium chloride (individual containers have calcium chloride slurry, and 
refrigeration units are dried with calcium chloride) to provide the right 
moisture balance. Although some are surprisingly short-lived, we've had good 
experiences with seeds that have been stored for years- even labile genera, 
like Stanhopea. Just had one accession that was 3-4 years old come up quite 
nicely, although that is an exception.

We do maintain stores of seeds in liquid nitrogen for research purposes. The 
great thing about this kind of work is that the data tend to get better with 
age (even if it is a sort of binary response, making it tough to say precisely 
WHEN a given accession is flat-out nongerminable due to age), making it perfect 
for procrastinators like myself.

Orchid seeds can be stored in liquid nitrogen and maintain their germinability; 
of the very few we've tested, it compares well with seeds that were never 
placed into cryogenic storage. However, there may be a difference between LN2 
(at -196C) and deep refrigeration (at -70 or -80), and there's certainly a 
difference between these and domestic freezers (at -20); orchid seeds don't 
really freeze at -20. Instead, the liquid components turn into a biological 
glass. This is important as freeze-fracturing is what normally kills plant 
cells- the expansion of water as it freezes, shattering membranes and so forth.

With orchid seeds, the endotherm (as gauged by differential scanning 
calorimetry- Pritchard and Seaton's work, although I did a very little bit of 
it back when I was an explosives chemist and had access to a DSC) is rather 
lower. I'd have to look up the values to be certain. Anyway- it's well below 
-20C, meaning that frozen storage (in domestic freezers) may be more effective 
than 4C while more convenient than -70 (or lower) without any additional damage.

There are concerns with cycling, however, and this gets into some pretty deep 
issues in physical chemistry and why seeds die; there are also handling issues, 
and problems with moisture cycling (wet/dry from condensation)- even just on 
the packaging, which later translates to the seeds. If Dr. Norman Deno is 
correct, then moisture may play a much larger role in telomere degradation than 
is generally thought and casually cycling seeds through temperatures (as in 
pulling the box out to retrieve seeds, then replacing it) may have a greater 
role in shortening seed life than has been previously thought.

This is one of the nice things about LN2- as the temperature of the boiling 
liquid nitrogen (as it is stored in a Dewar) is very nearly a constant, there 
is no cycling at all. The nitrogen itself is chemically inert, and may interact 
less with the seeds than wonderfully corrosive atmospheric oxygen. Best of all, 
a Dewar and a fill of LN2 is far less expensive than a -70 or -80 freezer, and 
the operating costs (about $10/month for adding more LN2) is probably less than 
the cost of keeping a freezer plugged in.

Of course, the space is smaller- but if there's one kind thing afforded by 
orchid seeds, it's their space requirements. At least I'm not the coconut 
seedbank.

Cheers,

-AJHicks
The Orchid Seedbank
Chandler, AZ



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