Please suggest some good media compositions that could induce endophytic fungi to produce secondary metabolites of rich medicinal value.
In very general terms, since there are no details which fungi are the producers, some kind of stress, nutritional or environmental could be beneficial for secondary metabolite formation.
Please clarify whether you want to work on fungal mycelium or fruiting bodies. And another thing an important question arises as any particular protein or carbohydrate may or may not present in the cultured mycelium which may be present in the fruiting bodies collected directly from any area. So first you decide then proceed.
Yeast extract Sucrose broth is better one as I used. The fermented broth gives lots of peaks when compared to others. PDB supplemented with 0.5% soytone was used by many workers to produce taxol and taxanes..
PDA, PDA+Y,Corn meal agar are the best media for growing spores of endophytic fungi.
You have to standardize media based on your target compound, I do not know any available standard methods
I have used modified media, such as a PDA agar base, supplemented with plant extract
I used semisolid medium agro industrial wastes (ananas, grapes, passion fruit) and rice.
You may contact Dr. Gary Strobel for his expert advice!
cheers,
shenoy
First try with basic complex of PD with salt solution and then for better growth try synthetic media for further culture.
First of all , you should use a simple medium (PDA or Czapek is OK) for isolation of the endophyte from plant. Then when you have obtained a pure culture, you'll go on with identification of the strain. For sporulation maybe you need near-UV light.
For production of metabolites, generally I use malt extract+ peptone+glucose+agar (MPGA; 20-2-20-13 g/L respectively). To increase metabolite production I had good results also increasing the glucose amount until 90-100 g/L.
I use agar CYA (Czapek Yeast Autolysate Agar) and have good results.
If you are looking for metabolites constitutively produced by your strain, in my opinion the best medium is Czapek-Dox, which contains sucrose only as a C source. However, a number of fungal species are unable to grow on this substrate, so it may not work for your strain/s.
use a defined media like Czapek-Dox where you can also standardize composition specific for the fungi by varying / adding / deleting components.
Hi, You could try some of the media that Charles Bacon used/developed for secondary metabolite production in the clavicipitaceous endophytes. These include M104T, SM medium and another defined medium for Balansia alkaloid production. I'm attaching a pdf of a chapter Charles Bacon and I wrote a few years ago with this media included. These media were designed to maximize ergot alkaloid production but they may work for other secondary metabolites. The chapter also includes for other media or techniques that have been useful over the years with the clavicipitaceous endophytes. I think the metabolite production will increase once the endophytes go into the lag phase and the endophytes stop producing hyphal/cell mass and begin to put resources into secondary metabolite production.
Good luck with it.
Yours,
Jim White
Rutgers University
Yes, start with the classic complex media pre-mixes such as PDA/PDB, Czapek-Dox and so on. very soon though, you will need more defined media so that you can separate out the effect of various added substrates e.g. the carbon source(s). I have used the following medium with much success from various fungi including an endophytic Fusarium sp. You can use and cite the medium here (note that the fungus it was used for was Aspergillus; the Fusarium paper is being prepared):
http://clemkuek.com/papers/kuek1991.pdf
You can substitute the carbon source with anything which you think is worth trying - from glucose to more complex oligomers.
Clem
Hello, While I am generally considered to be one of the top ethnomycologists in the world, I do have a Power Point program on my website of Endophytic and mycorhrizal fungi with Dr. Prakitsin Sihanonth, also a member here. The original Power Point Program was used by Dr. Sihanonth of Chulalongkorn University with Hawksworth at a conference. Since I have not visited the site in a while I am not really sure where it might be at on my site, but it should read as John's Power Point maybe in the general Index or in All Things Shroomy or in the Articles1 page, maybe in my articles section at my site or in the species Identification page which has several thousand Psilocybian mushroom identification photos and descriptions.
I will provide a link to my site and then look for the actual slide presentation. If you send to me an email address to [email protected], I can send you the actual slide that only covered the Endophytic Fungi you asked about.
http://www.mushroomjohn.org
http://www.mushroomjohn.org/articles1.htm
http://www.mushroomjohn.org/species.htm
And for the actual one I used which is different than the one I will email you if you like the original which covers Mycorhrizal and Endophytic Fungi.
http://www.mushroomjohn.org/johnsslideshow1.htm
Remember I converted the original for my presentation with Dr. Sihanonth's work at Chulalongkorn and that of Dr. Hawksworth's original one without Psilocybe and other species . John. I hope that is helpful
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