FlytrapCare Carnivorous Plant Forums

Hey Matt!

Thanks for your comment, it's really interesting that you got the same result like me. I don't know why we got these problems while others had great success with it. Anyway i gave it one more try a few days ago with strong, big, fresh tissue and if this also doens't work i think i will try NaClO again.

Last time i used NaClO my protocoll was
washing wish antibacterial soap under running water
then placed in 10% NaClO with few drops of Tween 20 for 5 min
and rinse it 3 times.

Tissue had very strong phenolic bleeding and was contamined.

I also tried the protocoll on this website but this lead me to dead tissue... I have really no idea what i am doing wrong...

Seeds work fine for me with NaDCC or NaClO but explants are such a pain in the a** for me.

I think i will try

1 sec 70% Alcohol
5-6 min 7,5% NaClO with tween 20
3x rinse
hope my tissue will survive this.

Greetings Lukas

Sterilizing explants is definitely challenging and requires some practice and trial and error. It's not something that is easily achieved without a lot of experience.

It sounds like you are making adjustments to your methods, which sound similar to mine and should work just fine, and you should find something that works for you with a bit more time and effort

Yes! Explants are very hard to sterilize at least in my opinion most time they are contaminated or just turn brown.

Do you also use 3% H202 ? What's the point of using NaClO and H2O2 ?

Savior7 wrote:Do you also use 3% H202 ? What's the point of using NaClO and H2O2 ?

Yes, my normal routine is a dip in alcohol, a few minutes in H2O2 and then a few more minutes in NaClO.

I don't know for certain, but I'm guessing that the H2O2 and NaClO might kill different bacteria more effectively, thus improving the success rate for shorter soak times.

Very interesting, i think i should give it a try with H2O2.

My guess would be:

-Dip in 70 Alcohol
3 min 3% H202
3-4min 10% NaClO
Rinse 3x

Hope for the best

That's close to what I do. But the time in NaClO is longer or shorter depending on the size and type of explant.

Okay that's nice to hear that i am on the right track

I'm sterilizing media right now. I will give it a try on Saturday. I got some bigger explants so i think i leave them in as long as possible ~5min for the NaClO. Hope phenolic bleeding won't be that bad.

I leave you guys updated

Edit: do you use the H202 before or after the NaClO ?

Very interesting methods, some will work but not for the others.
@savior7, my questions
1. How do treat the mother plants? Indoor/outdoor?
2. Using laminar flow when transfering?


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Hey Ruruanta!,

It's a mix, some of my plants are greenhouse plants (the matured ones) and the indoor plants are the smaller ones which i wil cultivate warm trough winter for this year for faster growth.

And yes i do use a homemade Laminar Flowhood with a HEPA filter H14

And i don't use PPM in my media

Greetings
Lukas

Savior7 wrote:Edit: do you use the H202 before or after the NaClO ?

H2O2 before NaClO

Oh damn!

So i did it in the wrong row.

I did 6 min NaClO 10% and 3 min H2O2 3% with big and more or less fresh tissues but unfortunately so strong phenolic bleeding that the tissue died.

The other one i did 4 min NaClO and 3 min H2O2 but i think it's contaminated. I will try it the other row maybe tomorrow. Any tips for me ? I did spray them with alcohol

Greetings Lukas

Lukas, whats "phenoloc bleeding"? Any pic how its look like?


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Sure.

See how the media turned yellow ? That's phenolic bleeding. It's waste of the tissue due to harsh sterilization or cutting. Normally something that the plant doesn't like

Greetings Lukas

Uh wow... Is the explant still useable? Looks black already.


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No it isn't. It's already dead.