Daylilies forum→Wigged Out / Sterile varieties and protoplast fusion

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Name: Dan
Springfield, MO (Zone 6b)
docpaps
May 11, 2021 11:14 AM CST
Wigged Out is my favorite oddball, but like many I've never seen a bloom and have not had any chance to hybridize with it. Today I started working on a protocol for using protoplast fusion to cross sterile or recalcitrant cultivars. Experiment is crossing Wigged Out with Dr. Strangelove and with Believe via protoplast fusion.

Just wondering if anyone has any other favorites that are sterile? Hoping to play around with this method in the coming seasons.
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Name: Larry
Enterprise, Al. 36330 (Zone 8b)
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Seedfork
May 11, 2021 11:28 AM CST
According to Dan Hansen, Chan Dom Vongthirath is sterile.
[Last edited by Seedfork - May 11, 2021 12:30 PM (+)]
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Name: Maurice
Grey County, Ontario (Zone 4b)
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admmad
May 11, 2021 12:40 PM CST
I would guess that the 'Wigged Out' phenotype will act as at least a partial dominant in your cell fusion.

If your interest was more to getting 'Wigged Out' to be usable in crosses then I might try growing it cool to see how that affected its phenotype (or hot).

If only you could be encouraged to try to convert tetraploid daylilies into diploids....
Maurice
[Last edited by admmad - May 11, 2021 12:45 PM (+)]
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Name: Dan
Springfield, MO (Zone 6b)
docpaps
May 11, 2021 1:36 PM CST
Maurice, that's a great suggestion regarding temp. I could move some fans of Wigged Out to the grow tent I have in my basement so I could keep it relatively cool, but I may need the space for babying some conversions. I can probably streamline my process quite a bit, but the steps required for conducting protoplast fusion are insanely onerous, especially if like me you're trying to save money by mixing up your own solutions. Most likely I'll try with Wigged Out a few more times before trying for a cross between daylilies and another species.

I don't have any blooms to work with yet this season, but am planning on anther culturing some toothies in an attempt to bring that characteristic to the dips. If you have any suggestions on a tet that you'd like to see converted to a dip I'm all ears.

Of course all of these are long odds, but with enough tries... hopefully

Name: Maurice
Grey County, Ontario (Zone 4b)
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admmad
May 11, 2021 2:15 PM CST
I was thinking of wide gold picotee and dark picotee edged tetraploids (no particular cultivars) but toothed would be exciting.

I am going to try Lilium pollen on tetraploid daylilies this year as well as Kniphofia. Dianella pollen might be a good one to try on tetraploid daylilies, but I don't have any of it.
Maurice
Name: Dan
Springfield, MO (Zone 6b)
docpaps
May 11, 2021 2:27 PM CST
Kniphofia is on my short list for future protoplast fusion experiments, along with hosta and a few others.
Name: Jim
northern Illinois, USA (Zone 5b)
Lilies Orchids Hybridizer
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Liliophilic
May 28, 2021 8:04 PM CST
Dan and Maurice,

I think Dan's mention of anther culture is definitely worth pursuing. But I'd suggest trying this with both tet and diploid day lilies. For sure start with obtaining diploid plants from culturing tet plant's pollen. But take this a step further and convert those derived dip plants back to tets via oryzalin or colchicine. Imagine having homozygous tetraploid plants to breed with! You could also do the same with diploid daylilies; obtain haploid plants, induce them to homozygous diploids, and now you have all sorts of homozygous dip and tet breeding lines to play with.

Piece of cake, lol. Good luck!
Name: Maurice
Grey County, Ontario (Zone 4b)
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admmad
May 29, 2021 4:48 AM CST
@Liliophilic,
Jim,
If a haploid (e.g. a1 representing one chromosome of one of the 11 daylily sets) is produced from a diploid's pollen and then its chromosomes are doubled that will produce a diploid with all its genes homozygous (e.g. a1a1 for one chromosome pair). But if a diploid (e.g. a1a3 - one of six possible combinations) plant is obtained by culturing the pollen from a tetraploid (e.g. a1a2a3a4) plant it will not be homozygous. If the diploid's chromosomes are then doubled (e.g. a1a1a3a3) the plant will be duplex rather than quadruplex (e.g. a1a1a1a1 homozygous) for each of the 11 sets of daylily chromosomes.
If the tetraploid (a1a1a3a3) is a converted diploid (a1a3) then it is duplex.

Culturing pollen is not always successful, as the pollen of some species does not have chloroplasts (or presumably their proplastid precursors).
Maurice
Name: Dan
Springfield, MO (Zone 6b)
docpaps
Jul 1, 2021 7:16 AM CST
Hi all, just an update. 1st round of protoplast fusion did not produce any callus. But, at least it looks like my homemade air flow hood worked as there was also no contamination.

Few weeks ago started my first attempt at anther culture of a few toothies - Buddy's Kayla and Papio Shelli Mae. It's looking like the interior of unopened buds is less sterile than I'd hoped, as about half have had to be thrown out due to bacterial/fungal contamination.

Will give an update on progress in another few weeks.


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Name: Marika
Kansas City, MO (Zone 6a)
Daylilies
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Mikamajik08
Jul 19, 2021 7:11 PM CST
The technical expertise & science of this thread fills me with joy.

What are we looking at in the latest pictures?
Name: Dan
Springfield, MO (Zone 6b)
docpaps
Jul 20, 2021 11:26 AM CST
Hi Marika! The last pic is of a test tube that contains liquid growth medium (Nitsch and Nitcsch), plus some BAP, 2,4-D, maltose and sucrose along with some anthers from Papio Shelli Mae (which I mispelled on the tube).

As far as I know, no one has anther cultured daylilies to convert tets to dips, or dips to haploids, so I'm unsure if my hormones, nutrients, etc are optimal, or even if it's possible. Some plants can be anther cultured and some can't.

So far, no growth in the test tubes. And, I've had a dickens of a time with fungal contamination ruining about half of my attempts.
Name: Dan
Springfield, MO (Zone 6b)
docpaps
Sep 3, 2021 7:24 AM CST
Update - one anther culture attempt (Frilly Frazzeld) has produced callus tissue. The tissue developed in a liquid medium and appears extremely fragile.

Just transfered this to a more solid growth medium.


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