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By tokol0sh
Posts:  16
Joined:  Tue Dec 11, 2018 3:37 am
#342898
Thanks for the reply.
My media is 1/3 MS with 3% sucrose.
My sterilization protocol was:
Dip in IPA, 10min soak in 10% bleach and rinsed 3 times in sterile DI water (I didn't note how long each rinse was). I figured I might have over sterilized the seeds and tried an 8min soak in the bleach solution with the same results.

There's no signs of contamination, Ill just leave them alone for a month or so longer and see of anything happens.
By tokol0sh
Posts:  16
Joined:  Tue Dec 11, 2018 3:37 am
#345422
Just a little update:
None of my VFT seeds have sprouted and the gel is now starting to get hard.
I decided to tryout some centrifuge tubes (50mL) as this would mean I can put each sterilized plantlet into its own container, alowing me to try more things at once and if one gets contaminated, its contained.

I have one flower stalk and one flower bud that are doing well. The flower stalk is still green and the flower bud has grown a lot. The flowers are starting to open. There looks like some white mold growing inside too, but its not growing on the plantlet and seems to be sinking into the gel (I didn't add enough agar to this vessel so it still very liquid)

Earlier this week I got a some more flower buds to try out.
Here are some of the photos of the flower bud that is growing (It only seems to be growing flowers tho)
The flower bud was place in the vessel on 13 September

Photos from 3 Oct, it started off as a small bud, maybe 5-8mm diameter (I forgot to to take photos of the start):
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Photo from 19 Oct:
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Photos from 23 Oct:
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I have noticed that the gel has started to go a bit green, not entirely sure whats causing this. I have noticed it with some other leaf cuttings too. I might transfer it to a new vessel soon, depending on how the flowers do. Maybe I will get some seeds out of this...
By tokol0sh
Posts:  16
Joined:  Tue Dec 11, 2018 3:37 am
#368473
After a bit of a break I have tried again.
This time I brought a recently repotted VFT indoors (at the start of spring) and placed it under my grow light for a few weeks, until new leaves started growing. After cutting a few leaves, rinsing under flowing water for about 15min, sterilized with H2O2 for 3min and 10% bleach for 10min, rinsed three times and trimmed about 1mm off the ends.

Now about 2 weeks later, I see no contamination (yet) and none of the leaves have turned black. Instead the leaves have turned a deep red (see photos below). Is this normal?
I have had the TC containers next to the plant I cut the leaves from, under my LED grow light at roughly the same distance. None of the leaves on the potted plant are turning red.

Leaf cuttings have turned a deep red
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The colour balance is a bit off with the light from my grow box leaking out. The VFT has a nice green leaf with deep red traps. (It looks a bit sick with this colour balance)
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TC containers and VFT together under the LED grow light
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Is the colouring normal? I have seen photos of callus formation and it normally goes a little red, but I have never seen a leaf go like this before.

Cheers
By tokol0sh
Posts:  16
Joined:  Tue Dec 11, 2018 3:37 am
#378592
I collected some seeds from the flower in the previous post, and have had success in starting cultures with them. I started off in 50mL test tubes and have just moved them into larger containers.
I will try multiplying these out, and then I need to figure out how to get roots to grow and then potting and acclimatisation.

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Yes

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