our stock plants are highly contaminated with internal none aggressive bacteria that live inside of plants ( mostly Begonias and Dieffenbachia ) but make contamination problem in plant tissue culture (leaf petiole and stem segment explants ) . due to some difficulties i can't use antibiotics in media so i decided to use antibiotic solution (dissolved in DMSO ) bath and spray treatment for stock plants and also antibiotic solution (dissolved in DMSO ) bath for explants after sterilization .
and what is safe dose of DMSO per liter of sterilizing solution ?
You have to screen a healthy, bactrial infection free plants to establish asetic in vtro cultures of Begonias and Dieffenbachia plants. An article is attached herewith for identification and control in commercial greenhouse.
Why did use DMSO? Normally the antibiotics have a high solubility in water.
Did you try Plant Preservative mixture having antibacterial and antifungal property. PPM™ is a proprietary broad-spectrum biocide, which can be used to control contamination in plant cell cultures, either during the sterilization procedure, or as a medium component. PPM™ comes in an acidic liquid solution (pH 3.8). PPM™ is useful when routine surface sterilization is insufficient.
Dear Ricardo Julian Licea-Moreno
I use DMSO to increase antibiotic penetration to explants and increase the efficacy of antibiotics .
Dear Nand Lal PPM is made in USA and they don't accept to send to Iran and i need a solution for this problem .
I'm not sure that this method to promote the penetration of antibiotics with DMSO could be useful to "remove" microorganisms that live inside the plants. Endophytics have evolved to live inside the plant tissues, so their "elimination" (I prefer to use control) might be highly difficult. If you're seeking a deep penetration of antibiotics, you could use vacuum. The control of endophytic contaminants can be approached from different points. In my experience a single action to control endophytics usually don't render the expected reponse. See the works of P. Thomas and B. Reeds. Good luck.
Endophytics are very hard to eliminate. The simplest solution is to eliminate the contaminated cultures and to start from new clean material. However, sometimes you will have to deal with certain plant species that always bring to the lab their associated MOs.
In this case, the applied an integrated management considering good micropropagation practices (high quality media, avoiding long term subcultures) to reduce the plant stress. In this case, the endophytic is not a problem until it causes economical losses into the lab. To keep low levels (and even to eliminate) some endophytics we have tested some antibiotics. Even when you have to make your own tests to chose wich is the best for your situation, I can tell you that Vancomycin is, by far, the most effective in our conditions.
We are preparing a paper about some studies we made using sweet potato as model.
PPM™ url: http://www.plantcelltechnology.com/ppm-protocols/
Endogenous Contamination: (a) For explants: gently and routinely shake / stir 1 cm. long explants (or shorter) for 4-12 hours in 4-5% v/v PPM™ solution supplemented as above with full MS strength basal salts without pH ing and without Tween 20. Without rinsing, insert into a medium supplemented with 0.05 - 0.1% PPM™ for herbaceous plants and 0.2% PPM™ for woody plants.
Amoxycilline could work, you could get it from any pharmacy. For PPM you could try to find the active ingredient.
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