Hi, all
I thought it would be nice to share the outcome of my experiments with supplemented MS medium on some drosera species in vitro. It is well known that the presence of aminoacids in tissue culture media can promote the growth of the plants in a very significant manner (e.g. glycine being part of traditional MS medium). The suplements are sometimes used, such as well known casein hydrolysate and others. The problem with these is that the ratio and amount of aminoacids produced by acidic hydrolysis of caseine is not defined and only aproximate (analysis results may vary considerably). Since I have access to pure L-aminoacids, I did this experiment with various ratios and mixtures of the following: L-Asparagine, L-Tyrosine, L-Arginine, L-Cysteine, L-Glutamine and Glycine. Of course other AA such as Tryptophan, Proline,...can be used as well.
The important benefit of using AAs in medium is buffering activity due to zwitterionic nature of AAs. Thus, the pH of medium becomes much more stabile during the growth of the plants.
The amounts of AA exceeded the figures from literature sources in some cases, generally in a ratio of 10-100mg per L. It is known that too high concentration of AA can thwart or stop the development of plants. But given appropiate concentration, the results are obvious, such as in this case of drosera capensis 'Giant' (supplemented medium on the left):
Some more sensitive species grow on this medium much better than on regular MS.
d.arcturi
d.hilaris
d.anglica "Hawaii" started flowering after replating
d.ascendens 'Palmeira, Parana, Brasil' - the massive germination. The seedlings are less vulnerable to burn by ammonium nitrate. The reduced form of nitrogen (-NH2 group in AA) is more easily available for plants.
d. glanduligera
,,,I have used this medium on tuberous crosses (thankx for the seeds Iggy)
They were treated with GA3. BTW, according my HPLC tests, the original 86% GA3 decomposes to 23% purity after autoclaving (20mins, 120C). I dont think it is as much as 99% as sometimes is stated.
I hope this info will help some TC enthusiasts to nehance the performance or their work
I thought it would be nice to share the outcome of my experiments with supplemented MS medium on some drosera species in vitro. It is well known that the presence of aminoacids in tissue culture media can promote the growth of the plants in a very significant manner (e.g. glycine being part of traditional MS medium). The suplements are sometimes used, such as well known casein hydrolysate and others. The problem with these is that the ratio and amount of aminoacids produced by acidic hydrolysis of caseine is not defined and only aproximate (analysis results may vary considerably). Since I have access to pure L-aminoacids, I did this experiment with various ratios and mixtures of the following: L-Asparagine, L-Tyrosine, L-Arginine, L-Cysteine, L-Glutamine and Glycine. Of course other AA such as Tryptophan, Proline,...can be used as well.
The important benefit of using AAs in medium is buffering activity due to zwitterionic nature of AAs. Thus, the pH of medium becomes much more stabile during the growth of the plants.
The amounts of AA exceeded the figures from literature sources in some cases, generally in a ratio of 10-100mg per L. It is known that too high concentration of AA can thwart or stop the development of plants. But given appropiate concentration, the results are obvious, such as in this case of drosera capensis 'Giant' (supplemented medium on the left):
Some more sensitive species grow on this medium much better than on regular MS.
d.arcturi
d.hilaris
d.anglica "Hawaii" started flowering after replating
d.ascendens 'Palmeira, Parana, Brasil' - the massive germination. The seedlings are less vulnerable to burn by ammonium nitrate. The reduced form of nitrogen (-NH2 group in AA) is more easily available for plants.
d. glanduligera
,,,I have used this medium on tuberous crosses (thankx for the seeds Iggy)
They were treated with GA3. BTW, according my HPLC tests, the original 86% GA3 decomposes to 23% purity after autoclaving (20mins, 120C). I dont think it is as much as 99% as sometimes is stated.
I hope this info will help some TC enthusiasts to nehance the performance or their work
