Hello all! I am currently facing a big trouble with fungi contamination in in vitro culture of native forest seeds, because they come from field collection with a lot of endogenous contamination (fungi). The disinfestation with alcohol (1min) + 2% hypochlorite (15 min) does not work, they insist to proliferate ... Would pre-treatment with fungicide or some other solution solve this problem? Any suggestion?
Dear Kathryn De Abreu ,
Thanks so much for your comments and contribution!
I added nystatin (fungistatic) in the media and for leaf explants and worked well...
Do a dosis survey on your system.
As a comment, nystatin is not altocaved, Add in the media when temperature gets below 50 C.
I work with citrics. First I disinfect the fruit and later I extract the seed under sterile conditions. My strategy is to extract the seeds before they become contaminated.If the species you work with allows you, you could try.
Hi,
As an undergrad I used to make calli from tree buds coming from outside and I know contamination can be a huge problem. We used the following method: 1min. 96% ethanol, 5min. HgCl (not sure if it is allowed everywhere although it is not volatile or anything and as long as you don't ingest it , it is fine), 3x5min sterile water. I think for seeds ethanol can be left even longer. As you already know that you have a lot of fungal contamination you could also treat with a fungicide prior to the actual sterilisation.
Lorenzo Moncini , Marcin Olszak , Luis Carlos da Silva Ramos , Lilyana Nacheva
, Sivabalan Karthik , Manuel Robles , Svetlana Boycheva Woltering , thank you very much for your comments and contributions, all very valuable and relevant!
Organic techniques I've used in home-based seed starting include a sprinkle of cinnamon on the surface of the soil, and irrigation with strong but cooled chamomile tea. Both cinnamon and chamomile are natural anti-fungals, but I'm not sure how well they would work in culture. Good luck!
You can treat the seed with using bavistin which is one of the important fungicide at 0.1 per cent concerns for 5-10 min may it helps you
Chemical fungicide like bavistin is used to prevent the contamination in tissue culture which raises the environmental concerns. Thus, use of microbially derived antifungals can help in preventing fungal growth with benefit of positively impacting the plant growth.
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