Galanthus breeding

[loes]

Martin,
I was joking! and glad you explain about the chromosomes.
In humans people want "normal"and healthy kids,in gardening healthy is also important but 'normal' sounds a bit boring!

[Martin Baxendale]

Martin,
I was joking! and glad you explain about the chromosomes.
In humans people want "normal"and healthy kids,in gardening healthy is also important but 'normal' sounds a bit boring!

I know you were joking. I just thought I'd explain, as an interesting point, why extra complete sets of chromosomes (as in triplopids and tetrapliods) don't neccessarily cause abnormalities, but the splitting of chromosome sets when breeding with triploids can cause abnormalties which may be horticulturally interesting.

[Gerard Oud]

If you do the procedure right there is a good chance that about 5 til 20 % will be polyploid and the rest stays diploid or triploid, they wont die because the mixture is a few milligrams on a bucket full of water.

For comparing the stomata you need references of species that are for sure triploid or tetraploid, polyploids stomata are bigger and more.

Trying the seed route, getting tetraploids is very difficult because you have to be sure you have two instabile diploids to melt together, is easier to win the lottery.

I am going to try to make some triploids, hexaploid with oryzalin, and after this i start breeding with interesting diploids, that will give one set diploid and two sets tetraploids.
For examening the ploidlevel there is a company here in holland called Plant Cytometry Services, you can find details on www.PlantCytometry.nl
It is not expensive 35  euro and 6 samples(2,60 each) will give a average of 8,43 per sample

[Martin Baxendale]

Gerard, thanks for the information. Maybe I'll try the internet and see if I can get hold of some Oryzalin as an alternative route to tetraploids.

For the seed route, I'm intercrossing triploids, on the basis that they should have just a very little fertility rather than being completely sterile (partly through producing unreduced gametes) - so if I do enough crosses with enough flowers, I just might get the occasional one or two seeds set, which may be tetraploid. According to a botanist at Kew Gardens who specialises in this sort of thing, the chances are very, very low; but in theory if you do enough crosses (lots and lots) then you may eventually get a tetraploid (it's how the first tetraploid daffodil was produced, by the crossing of two trilpoid daffodils, which luckily gave a tetraploid seedling). But it might take many years! 

[Martin Baxendale]

Rob, can I assume from your sign-off on your message (ahoy!) that your family roots are in Slovakia or Czech Republic? Sorry, I accidentally deleted your message before I could reply to it. 

[Diane Whitehead]

In Snowdrops, it was written that Dr. Ben Zonneveld of Leiden University began
measuring the amount of DNA in snowdrop species and some cultivars, using
Flow Cytometry, in 2001.

I just googled "Flow Cytometry Galanthus" and found an abstract of "The systematic
value of nuclear DNA content in Galanthus"
by Zonneveld, Grimshaw and Davis, published in 2003 in Plant Systematics
and  Evolution

There are two abstracts of this article:
http://www.cababstractsplus.org/google/abstract.asp?AcNo=20033208109 http://www.springerlink.com/content/gp1r12uh0cb0x33m/

Tetraploids were found in G. transcaucasicus and hexaploids in G. elwesii
and G. lagodechianus.

[Martin Baxendale]

Flow cytometry is the method used by the company Gerard recommended in Holland. Seems quite cheap.

[rob krejzl]

Maybe I'll try the internet and see if I can get hold of some Oryzalin as an alternative route to tetraploids

It seems to me that even if an individual can't get a permit to import this for use as a chemical reagent rather than a herbicide, any suitably qualified laboratory should be able to. Over here in Australia orchid breeding is a fairly widespread hobby. Home hobbyists send their seed pods away to a lab for in-vitro germination, and get back flasked plantlets. Amongst the services on offer is tissue conversion. Even our local orchid nursery doesn't convert it's own tetraploids but sends them to someone else. It would be a more expensive route, but might be worth exploring.

Rob, can I assume from your sign-off on your message (ahoy!) that your family roots are in Slovakia or Czech Republic?

<Sigh> As your wife will probably tell you, mine is a fairly common Czech name - it must be true 'cos everyone in my family has it. Any Krejzl you should meet in the UK will be at least a cousin of mine. Be kind.

[Gerard Oud]

If you cant get some oryzalin Martin, i can try to get some for you.
If you really want to get polyploids you need it, otherwise with the seedroute you will probably have some succes after 10 or 20 years and a awful lot of work,administration etc.
See you in 20 years?

[Martin Baxendale]

Thanks for the offer, Gerard. I might see if I can order some oryzalin from the American company that Rob gets his from. They supply it as a chemical for plant lab culture etc rather than a herbicide, so maybe they'll ship to the UK.

It just seems a waste to have to order 500 ml of the stuff when I'd probably only ever use a few spoonfuls. I'll let you know if I have any success.

[Martin Baxendale]

Rob, can you tell me how you prepare your Oryzalin solution (using the oryzalin from Phytotechlab, which I might try to order) to get the right dilution, what you dissolve it in (distilled water?) and how, and how long you soak your lily scales for. I assume, from skimming through the info I've seen, that you take scales at the normal time, soak, then incubate scale bulbils as normal - so I guess I'd do the same but with snowdrop bulb chips, as normal but with a soak in the oryzalin before putting into incubation bags of vermiculite.

[Gerard Oud]

Martin it is good to know you are going to start, for me its also better to keep motivated and to know there are others with a mission.
Have you heard of the book The electronic comparative plant ecology ?
I will try to find it in the KAVB library, but i dont think it will be there, its published in 1995 Chapman&Hall in London and written by  JG Hodgson, JP Grime,R Hunt and K Thompson.
There should be information in it about polyploidity in galanthus and maybe some species are named in it.

Thanks Diane for the information

[Diane Whitehead]

How about this:  put some colchicum flowers in a small
ziplock bag, close it, rub the bag so the flowers inside
are crushed, then, wearing disposable gloves,  put some
snowdrop bulb sections inside.

From a website that was not identified:  One Colchicum
autumnale flower contains about 12 mg colchicine, a 20g
tuber provides 60mg, single seed provides 3.5mg

The message warned about the extreme toxicity of colchicine,
but it seems to me that keeping oneself protected by plastic
would be safe.

[Martin Baxendale]

Diane, that's an interesting idea. I've certainly got plenty of colchicum bulbs in the garden. The problem with colchicine, apart from the extreme toxicity, is I believe that it also tends to produce other genetic abnormalities than simply polyploidy. Of course that might not be a bad thing in snowdrop circles, where wierd and wonderful is often highly esteemed!

By the way, I wonder why colchicums aren't all polyploid if colchicine induces polyploidy?! Or maybe they are. That would certainly account for those ridiculously massive bulbs!

[rob krejzl]

Martin,

The Art Evans link I posted covers most of your questions pretty well, but I've sent you something by PM. Although it doesn't dissolve easily, you can use hot water to dissolve Oryzalin and still get reasonable results, but if that doesn't work for you try alcohol instead - avoid DMSO since it can deliver whatever's dissolved in it straight into your bloodstream.

I wonder why colchicums aren't all polyploid if colchicine induces polyploidy?!

And why opium poppies aren't all high since they contain opium..

I'd avoid making your own colchicine. It's not just the snowdrops that might show wierd and wonderful abnormalities...