What agar is best to grow plant leaf bacteria - (Jan/04/2006 )

hi everyone,
i am trying to grow some unkown bacteria from the surface and inside of a leaf but i don't know what agar i should grow them on. Does anyone have ANY suggestions because this would be helpful. If someone could just give me some type of feedback that would really be appreciated because i need to purchase some type of agar so that i can try out whether or not the bacteria will grow on that agar. THANKS loulou

I would try several different agars. Probably stay away from minimal media, just in case. How are you planning to avoid fungal contamination?

Hi Captain DNA,
Thanks for replying. What is minimal media? Now about contamination, i just really havent' taken that factor into account though it did cross my mind. I was more worried about what type of media i would need to use to grow ANYTHING on my dishes that i simply overlooked that detail. I am open to any advice though and i would really like to hear your opiniion on this. Thanks a lot,
Loulou

What media do you have available to you?

Any rich media (TSA, LB, BHI, or Nutrient agar, for example) would be worth a shot.

These are "undefined media" -- they contain lots of a lot of things. Minimal media, by comparison, is a "defined media" -- you know everything that's in it, 'cause you start from scratch and make it yourself. It would not be useful in your case, as Captain_DNA pointed out, because your bacteria may have some nutritional need you don't know about (a particular vitiamin, perhaps, or an amino acid), and if you don't put it in, your bacteria will not grow.

Thanks for the tips. I do have those media available to me at my highschool and i plan on using those. but as captain DNA mentioned how would i prevent fungal contamination if spores are floating around and other types of organisms that i don't know about. I am going to steralize everything of course before begining just to be on the safe side but preventing fungal contamination... i am not sure about what i should do there. do you have any suggestions. Thanks again for all your help.
Loulou

you will as likely find fungi on the leaf as in the air

The only thing I can think of is time. fungi usually grow quite a bit more slowly than bacteria. if you pull your bacteria off your early plates quickly for subculture (within 3 days, maybe?), hopefully you can cut down on fungal contaminants

do you have someone who can teach you aseptic technique? the importance of that cannot be overstressed...you don't want to test the effects of your skin commensals on the leaf pathogens

Here is my thought on this. While leafing (pun intended) though the promega catalog, I looked up antibiotics that affect only eukaryotes. G418 apparently binds only to 80S ribosomal units. Maybe adding this in small amounts to the media will act as an anti-fungal agent while keeping your bacteria alive. Anyone have thoughts on this approach, for or against? Aseptic technique or not you are bound to have some fungi on the surface of the leaf. Maybe this can help? Also found hygromycin works only on 80S as well.