I have read some methods to start endophyte fermentation. Some using starter, and some are not. I wanna ask three question
1. how important is starter for small scale fermentation (100 mL) ?
2. are there any problem if i start the fermentation culture aged 2 months ?
3. if i want to do mix culture, should i make a starter for each fungus and mix them, or can I just mix them directly in liquid medium ?
Thank you for answering my question :)
Hi, for endophyte fermentation and for any type of fermentation starter is decreasing the risk of contamination. If your cultures are fungi so 2 mounts not so long period for spore forming cultures you can use and you can see how it grows on stater.
If you want mix cultures be sure that our cultures are not antagonists, because you can lose some of them. if your cultures not antagonists, so you can use or not use stater but use same methodology for both with or without stater.
Hi, for endophyte fermentation and for any type of fermentation starter is decreasing the risk of contamination. If your cultures are fungi so 2 mounts not so long period for spore forming cultures you can use and you can see how it grows on stater.
If you want mix cultures be sure that our cultures are not antagonists, because you can lose some of them. if your cultures not antagonists, so you can use or not use stater but use same methodology for both with or without stater.
1. Starter cultures are always a good idea. It will ensure experimental reproduciblity.
2. As long as the culture is still viable it can be used but it is probably a good idea to refresh the starter prior to any experiment.
3. Mixed cultures should be started separately but I agree with Karen that it is a good idea to check the compatibility
Hi Karen and Lucy :)
Thank you very very much to answering the questions :)
I dont know they're antagonist or not, im doing that experiment because I got the inspiration from my isolate. The endophytes that i cultivated, form barrier between others (like a line colored black in the isolate petri dish). and i want to know, is there any difference in HPLC profile of their scondary metabolite when i do mix culture.
i have 2 questions again if you still want to answer,
1. I have pre culture aged 7 days, and then i transfer it to the liquid media, so how i count their age ? 7 days, or start from 1 day again ?
2. have you ever heard or experienced in lost of activity because doing several subculture ?
and do you have suggestion for me in doing this research ? :)
Thank you very much :)
1. Hi it doesn't matter how old the culture, if you transfer them in new nutrient media you can start from 1, but also in grow stage there are several populations of cells you cannot receive by cultivation cultures in same age. During grow, duplication or dividing of cells not in same stage so cells will be in different ages and different populations.
2. During the conservation of microorganisms you can lose some activities.
But you can also restore and Activity...
it's important which kind of activity do you mean. Activity which is dedicated to this microorganism, or it's inducible activity . I need more details.
Dear Mr. Karen,
Oh I see, thankyou for explaining :)
I mean the secondary metabolite which has biological activity (antimicrobial, antifungal, or etc). I heard some researchers lost activity of their isolate, it's estimated because of several subculture or because separation from "their friends" in nature.
Now, im just starting my endophyte fermentation in small scale, 100 mL.
But, when i do my mix culture i didnt make a starter. Maybe, in my next fermentation i should to make a starter right ?
Im sorry for many questions, because it is my first time doing research in endophyte :D
Yes, The starter always better. What concern secondary metabolites synthesis it's depends from substrate in nutrient media. If you know the composition of nutrient media for your cultures, you can use it for grow cultures and then you can check your activities by classical microbiological methods.
Thank you Mr. Karen
Im sorry, do you have recommended papers/references for my research ?
In response to your question about loss of activity due to sub-culturing. This is a real problem. Whatever environmental pressures the organisms experience in the natural environment are removed in the lab. Two ways to prevent this is to try and keep a frozen or spore sample that you can go back to if loss of activity occurs. You can also attempt to passage the culture through more natural conditions to select for the trait you are looking for.
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