I have isolated many bacteria from plant rhizosphere but before going further for antimicrobial assay all the bacteria die even though they were stored at 40c.
Manual methods are not effective enough to extract the plant antimicrobial compounds.
Please try out the commercial method in your place.
We found the plant antimicrobial compounds to be of better activity extracted by the commercial method in comparison to the manual method.
Dear Renu
I presume you streaked onto agar plates and stored at 4°C. If so, I would recommend to use glycerol stock (in reps) and then store in freezer (‑20 to -80°C) for long-term usage. Also, please note that the repeated thawing and refreezing will reduce the viability of your bacteria and should be avoided.
For stocking the isolates, freeze drying is the best option and the 2nd option would be stocking in enriched broth + glycerol media at -80°C.
For glycerol stock:
A 15 - 20% final concentration of glycerol (V/V) is adequate to preserve viability for most of the Bacteria. After mixing well with a measured aliquot of well grown culture (late log phase culture) it needs to be stored at minus 80 degree C freezer is used.
The people above have provided you with some good suggestions for storage. However, I think another important issue is that it does not sound like you have found the best media conditionsfor your bacteria. I suggest that you try growing them in a variety of media. You may also find that you get different bacterial populations that way. Many bacteria will be out competed in rich media or may not grow. By trying multiple media, you may not only find conditions to keep you cultures alive longer, but you may also find new bacterial species present that you were unable to grow previously.
thanks Dr. Godfred and velivelli but i have already used the glycerol stock and the stock was then stored at -20Oc .
thanks Dr. Ian Edwin for your suggestion ....my work is to isolate a bacteria that have anti fungal activity against phytopathogens and should have plant growth promotional activity that's why i m using 5 different media and the bacteria obtained from these different media died after a month. even though the bacteria in glycerol stock and stock culture were not active
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