As it was demonstrated in earlyer posts in this same blog establishing olive tree explants in vitro is a challenging task.
For more then a year i tried without success to establish Olea europaea cv madural untill i finally got some results. The desinfection procedure itself is cultivar dependent .This means that research with olive starts on the first step for establishment, the desinfection procedure.
I am also convict that some of the contaminations that i had to face in the maintenance of galega vulgar clones were there almost from te beguining. Process in wich i didn't particiapte.
The multiplication protocol on the other hand was so efective and the plants so vigorous that it was possible to keep cultures actively growing without loosing them definetly. -Even from highly contaminated material i was able manny times to save and recycle clean meristems into new clean cultures.
From the earlyer work with Madural there were many discoverys. This was a time when i was searching undreds of synthetic and organic compounds and i found some really efective antibiotics. Curiously one of them was alycin from garlic, wich method of usage, i have also explained in this blog before as well.
Another one of these incredibly efective substances is PPM that stands for plant preservative mixture. A mix of 2 isothizolones one of them chlorinated.
Finally i had discovered as well, a way to convert a fitossanitary antifungal available in a comercial formula into a powerfull antifungal readilly aditionable to the culture medium post autoclaving. The process consists in extracting the active substances of the antifungal switch, fludioxonil and cyprodinil, with a few mililiters of acetone wich is a compatible solvent for both. After this depending on the species you are working with, you will probably have to make trials and see how sensitive your explants are to both molecules as well as to the acetone.
Cefotaxime sodium, gave promissing results in a few of my trials but the explants would not develop after a certain point of the initiation process and a latter experiment with ceftriaxone confirmed toxicity of cefalosporins making the galega explants stop all growth and very slowly become chlorotic. This is in agreement with some of the bibliography available wich says cefalosporins are toxic and have a narrow spectre of action when used alone.
For more then a year i tried without success to establish Olea europaea cv madural untill i finally got some results. The desinfection procedure itself is cultivar dependent .This means that research with olive starts on the first step for establishment, the desinfection procedure.
I am also convict that some of the contaminations that i had to face in the maintenance of galega vulgar clones were there almost from te beguining. Process in wich i didn't particiapte.
The multiplication protocol on the other hand was so efective and the plants so vigorous that it was possible to keep cultures actively growing without loosing them definetly. -Even from highly contaminated material i was able manny times to save and recycle clean meristems into new clean cultures.
From the earlyer work with Madural there were many discoverys. This was a time when i was searching undreds of synthetic and organic compounds and i found some really efective antibiotics. Curiously one of them was alycin from garlic, wich method of usage, i have also explained in this blog before as well.
Another one of these incredibly efective substances is PPM that stands for plant preservative mixture. A mix of 2 isothizolones one of them chlorinated.
Finally i had discovered as well, a way to convert a fitossanitary antifungal available in a comercial formula into a powerfull antifungal readilly aditionable to the culture medium post autoclaving. The process consists in extracting the active substances of the antifungal switch, fludioxonil and cyprodinil, with a few mililiters of acetone wich is a compatible solvent for both. After this depending on the species you are working with, you will probably have to make trials and see how sensitive your explants are to both molecules as well as to the acetone.
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