Dear Researchers, Greetings!
I am trying to develop in vitro propagation protocol for Dioscorea species and phenolic exudation was found at initiation and subsequent stages, but the major problem was at the initiation stage. I would like to know whether pre-treatment of explants to cold exposure at 4 oC along with antioxidants for few minutes can reduce the level of phenolic exudation in the medium. Suggestions appreciated.
Dear M. Manokari,
Yes, there are ways to use low temperature to minimize phenolic compound exudation in plant tissue culture.
1. Pretreatment of explants with antioxidant solutions such as ascorbic acid and citric acid (each 100-500 mg/l) and store the solution containing the explants for several hours (1-3) at low temperature (4-8 °C), regarding the plant species.
2. Treating the explants with a sterile antioxidant solution (ascorbic and citric acid in 100-250 mg/l) for several hours after the sterilization stage and then culturing the explants on the culture medium.
3. Using antioxidants such as ascorbic acid, citric acid, and phloroglucinol in 100-200 mg/l concentration in the culture medium and keeping the vessels under low temperature (4-8 °C) for few days and then transfer to the optimum growth condition.
Best regards
M. Manokari, yes, by use of antioxidants we can reduce the oxidation of phenolics from the cut ends of the plant tissues. Even low temperatures also help in the reduction of phenolics leach outs.
Yes, we have faced same problem in Leptadenia reticulata leaf and nodal explants.
We have given pretreatment with chilled (4°c) PVP solution prior to surface sterilization and it worked well.
Yes,
Am working on in vitro propagation of different mulberry varieties by taking explants from temperate climatic conditions....which has responded well ...
Mulberry is a tree species and known to release phenolics from the explants under the tropical conditions........in that case cold treatment of explants can reduce the release of phenolics
But in temperate conditions as the explants are already under cold conditions....they have responded well for in vitro cultures in terms of callus induction, somatic embryogenesis and indirect regeneration......we haven't observed any phenolic problems when explants are taken during Feb-March (10-15°C) months of temperate region .....
cold treatment although may temporarily slow down leach of phenolic compounds in medium but explant growth needs optimum temperature which being higher will not stop exudation of phenolics.
Gulab Khan Rohela Sir, Thank you very much for sharing your experience. It seems analysing plant physiological behaviours in altered environments sometimes may solve the problems under in vitro conditions.
Ashutosh Pathak Sir, Thank you, I will try the PVP solution also in the future.
Cold pretreatment have an important efficacy on phenol reduction. You can also place your explants in a liquid medium (MS) with PVP for 20 minutes before culturing in the medium, so that most phenols are secreted. Based on my personal experiences, this method is very effective.
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