The problem that I regularly face is contamination of cultures of R. solani. I usually use PDA media but the other fungi also grow. I am using streptomycin to reduce bacteria, still, some bacterial colonies are appearing. Are there any specific media for the growth of R. solani? or What should I do to avoid contamination?
Hi , you can use Cycloheximide (CHX) to to reduce bacteria ..good luck.
Hi: if you have contamination with bacteria only , use different antibiotic or mix two different antibiotics with the medium, you should add the antibiotic after you cool the medium and prepare it to pour it in the petridish.
Hi i think its better for you to do purification of R.solani on water agar +chloromphnicol after that you can use PDA with antibiotics +3 drop of lactic acid
hi
if you have bacterial contamination then yuo can use chloramphenicol to inhibit bacterial growth
Check your PDA. It should be sterilized for 20-30 minutes at 15 pounds to avoid bacterial contamination.. To avoid fungal contamination do your work under aseptic conditions.
Dear , Aqleem Abbas . first you should sterilize the medium carefully at temperature of 121 C for 30 min. Then you should add suitable antibiotic such as Chloramphenicol in average of 250 mg per letter to prevent bacterial contamination .
Best regards
Dear , Aqleem Abbas . Also you can see the following article .
with my best wishes ..
If just adding chloramphenicol fails you can inoculate a small drop of your fungus on water agar. The fungus will outgrow the bacteria with quick growing running hyphea. Just pick from the edge of the colony.
Alternatively I used 'submarine' culturing. Basically just pour an extra layer of medium (not too hot!) on top of your newly inoculated fungal colony. The fungus will grow out of the second layer leaving the bacteria behind.
PDA if not sterilized properly may have bacteria. Potato agar needs longer sterilization than others.. Be sure PDA IS STERILIZED AT 20-30 MINUTES AT 15 POUNDS before using it.
Hi , you can use Cycloheximide (CHX) to to reduce bacteria ..good luck.
Dear, you can exclude bacterial contamination from R. solani cultures by cultivating small inoculum of the fungus on water ager and after the growth reach the edge of the plate, take small piece of hyphal tips and transfer it to another WA and you can repeat this procedure three time s then yo can get pure culture of R. solani. Dont use any antibiotic for elimination of bacteria because it cant interfere with its pathogenecity.
Thank you and best wishes.
You can do single tip hyphae culture... By growing on water agar... The fungus is very fast growing. In Petri plate usually contamination appears. However If you transfer young culture to slant... Contamination problem is solved to some extent. I am also working on R. solani
Well you can use Chloramphenicol and tip hyphae culture. In addition I will suggest ultra low pH of media. it also helps to solve bacterial problem to some extent.
Dear Aqleem Abbas
To reduce the level of contamination, you have to ensure strict septic conditions in the lab and make sure that all devices you use in lab are appropriately disinfected before starting inoculation process of Rhizoctonia solani or during the transference of this fungus with addition of antibiotics like chloramphenicol. It is necessity to avoid any kind of food and drinks or eating in the lab.. etc.
Good luck
Hyphal tip transfer method::: Most of the time I face bacterial contamination, I usually use CF but failed.
Bacterial contamination! Be sure how your medium is sterilized. Sterilization may be inadequate.
I think bacterial disease is in Potatoes which I use for making PDA media.
If you are autoclaving properly.... No issues. Use antibiotics before culturing. Leave poured plates under laminar for one week. If still contamination comes fumigate your laminar. Also maintain uncultured plates in the incubator for 7 to 10 days... If bacterial growth comes... Fumigate the incubator..... Contamination is handling and maintainence problem
The problem you faced now I faced when I was a student in 1960. Potato agar need longer time of sterilization.
Dear , Aqleem Abbas..
PDA needs longer sterilization period than other media . You should sterilize the medium for 30 minuets at 121 oC and 15 pounds , You should also add Chloramphenicol to avoid contamination with bacteria.
Best regards
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