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Ernesto

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Coming up on five months post flasking of this seedpod, Paph. Jennifer Stage frma. alba (godefroyae x philippinense) x leucochilum frma. album. I’m expecting albino flowers from this weird backcross of an intersectional Paph hybrid (Polyantha x Brachypetalum) crossed back to Brachypetalum. Backcrosses like this aren’t unheard of (think Paph. Frank Hughes, or Hadley Cash’s Paph. Dragon Lord) but as far as I know I don’t think these have been attempted with album plants. These have been exciting to watch grow!

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Pod parent: Paph. Jennifer Stage frma. alba
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Pollen parent: Paph. godefroyae var. leucochilum frma. album
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Proud parents^
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Plantlets have begun differentiating leaves and roots!
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Close up of the first roots to appear on these guys.
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Pollination and sowing dates listed on my tag.
 
Congrats to this cross! It is good, that Jennifer Stage took the pollen and it is even better that you got germination!
Am I right that you did a green seed sowing just four month after pollination???
 
Congrats to this cross! It is good, that Jennifer Stage took the pollen and it is even better that you got germination!
Am I right that you did a green seed sowing just four month after pollination???
Thank you! And correct. I sowed the seeds as soon as I noticed a change in the ovary’s color— it went from green to slight yellow, and was flasked before it could begin splitting.
 
10 month update! These seedlings are MUCH smaller than my other homegrown Paph flask (multifloral x parvi) but they seem to be growing steadily. I’ve noticed some slight browning of old leaves, as well as what seems to be callus tissue that isn’t differentiating into full plants. I was planning on keeping them in their flask for now, but happy to hear any second opinions
 

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i treated the extra plantlets from my replate with 50uM colchicine for five days based off a paper published using Paph. villosum plantlets dosed 50uM for six days. hoping the gentler exposure time makes for higher survival (>65%) and maybe similar conversion rate (~20%)
 

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i treated the extra plantlets from my replate with 50uM colchicine for five days based off a paper published using Paph. villosum plantlets dosed 50uM for six days. hoping the gentler exposure time makes for higher survival (>65%) and maybe similar conversion rate (~20%)

The published paper like several others is unfortunately completely bogus. They never did it, and never got any result... I know how it works in Vietnamese Universities so far, stay at a table, make a paper, find a couple photos ( including in my lab 2 times...), and give results of '2-3 years of trials). Only the sterilization protocol for the shoots of villosum would be extraordinary, but the use of colchicine on differentiated tissues, especially Paphiopedilum, can never work... It needs to reach actively dividing cells, hence meristems or eventually PLB.

The only times suitable to treat Paphiopedilum are the white or green protocorm stage. When they have even a leaf forming, the mortality is extremely high for small plants, for bigger plants, it stunts them a bit, but has no consequence on the formation of polyploids...

A better, proven, reading is Watrous and Wimber, Artificial induction of polyploids in Paphiopedilum, a classic, that works really well...
 
A better, proven, reading is Watrous and Wimber, Artificial induction of polyploids in Paphiopedilum, a classic, that works really well...
hi Roth, lots of people seem to cite this paper, saying the same thing referring to 0.05% colchicine treatment for 3-10 days yielding 50% tetraploid conversion. yet, i can’t seem to find the pdf itself. do you know where i might find it? curious to see what other doses and exposure times they used
 
How did you treat them?
prepared 50uM colchicine in DI water and added enough volume to the flask to cover the seedlings. kept them in the dark for five days, and on day five put them under bright light to get the colchicine to break down as it’s photosensitive. when i find time i’ll pipette out the excess liquid in the flask, grow out the seedlings, and see how they compare to their untreated siblings
 
prepared 50uM colchicine in DI water and added enough volume to the flask to cover the seedlings. kept them in the dark for five days, and on day five put them under bright light to get the colchicine to break down as it’s photosensitive. when i find time i’ll pipette out the excess liquid in the flask, grow out the seedlings, and see how they compare to their untreated siblings
I will check if I still have a scan of it, I have the paper version for certain...

colchicine is not really photosensitive actually... it is UV sensitive, plus anyway it will percolate the agar media, as well colchicine is very toxic to the roots and leaves of the plants in general, especially Paphiopedilum and Phragmipedium. At a concentration of 20mg/L about, I do not expect it to have much effect either...
 
Toxic to humans too.

Not that much actually, by ingestion, and eventually inhalation of the powdered form in the vial, sniffing it is not recommended... The lethal dose would be way over a liter of the solution used to induce polyploidy...

People with gout or Mediterranean Fever can take up to 3mg, sometimes 5mg a day of colchicine orally...

That's another kind of fairy tale that it is highly carcinogenic, etc... in the plant tissue culture laboratories. On the contrary it is widely use for decades in human medicine, without any carcinogenic effect ever reported. It is not classified as a carcinogenic agent anywhere, actually, only by plant tissue culture people...

I would be way more careful with Oryzalin or Trifluralin actually... if I was using them. It is said by plant tissue culture people that it is much safer, yet, they are considered as probable carcinogen. You won't die of drinking it, but you may get cancer. Colchicine, you may die by drinking (a lot of) it, but you won't get cancer. Your call...

One other point I discovered during my long journey in the plant tissue culture and horticulture world, that shocked me, is the absolute lack of knowledge of medicine, or basic mineral or organic chemistry.
 
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