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By rootrot
Posts:  3
Joined:  Sun Apr 21, 2019 10:03 pm
#332913
Hello,

I'm new to TC, I started with Dionaea leaves, Drosera capensis leaves and Nepenthes seeds.
I sterilized leaves and seeds with 4% PPM for 12-16 h and I plated them, without rinse, inside a glovebox sprayed with 70% ethanol (and left standing for 40 min) on this medium:
- 1/4 MS (+ 1/4 vitamins)
- 30 g/L sucrose
- 5.5 g agar
- 1 mg/L 6-BAP
- 1 mL/L PPM
pH regulated to 5.6.
After 2 weeks only one flask was contaminated but both Dionaea and Drosera leaves turned yellow-brown and Nepenthes seeds color turned from light brown to dark brown/black.
I haven't used vented caps.
What could the problem be?
https://ibb.co/jwmSxRL
https://ibb.co/gjc7BHp
https://ibb.co/HhH7W1P

Thanks
Last edited by rootrot on Sun Apr 21, 2019 10:30 pm, edited 1 time in total.
By Toshi
Posts:  18
Joined:  Sun Apr 21, 2019 3:21 pm
#332915
To say nepenthes seeds,that media might be strong concentration about sugar and hormone.
Normally,nepenthes seeds turns green and germinate in few weeks.
Image
By rootrot
Posts:  3
Joined:  Sun Apr 21, 2019 10:03 pm
#332932
Thank you for the reply.
Have you sterilized those seeds with PPM or NaDCC/NaOCl/H2O2? they looks really "transparent".
Which concentrations do you typically use for Nepenthes?

I read that BigBella used similar concentrations
https://www.terraforums.com/forums/trop ... ro-14.html

Also I was thinking that I used a glovebox sprayed with ethanol, could be alcohol vapors inside the flasks a problem?
I'm still building my flow hood.
By jasonh
Posts:  12
Joined:  Sun Jan 28, 2018 9:11 pm
#332958
rootrot wrote:Thank you for the reply.
Have you sterilized those seeds with PPM or NaDCC/NaOCl/H2O2? they looks really "transparent".
Which concentrations do you typically use for Nepenthes?

I read that BigBella used similar concentrations
https://www.terraforums.com/forums/trop ... ro-14.html

Also I was thinking that I used a glovebox sprayed with ethanol, could be alcohol vapors inside the flasks a problem?
I'm still building my flow hood.
500 ppm (0.5g/L) of NaDCC (I'm using Milton tablets which is about 20% NaDCC, so 2.5g/L) for 24 hours followed by 5-10 minutes in 5000 ppm and 2* rinsing works well for nepenthes seeds (and pretty much all other seeds), except for some lowlander seeds which are *** to clean. 500 ppm NaDCC for 12 hours and 4% PPM for 12 hours worked as well.

Alcohol vapor shouldn't be a huge problem; I work with a glove box too and it doesn't seem to affect the seeds too much.

A good indicator of sufficient sterilization with NaDCC / bleach is that the "wings" of the seeds are transparent while the middle is still light brown.
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