I am working on the Isolation of Endophytes from Seaweeds. I have surface-sterilized my samples using Sodium hypochlorite, and Ethanol. Then washed several times with sterile distilled water and plated on Potato Dextrose Agar with Streptomycin. After 10 days of Incubation, Fungal growth occurs on the media. Now, How do I confirm that the grown fungus is an Endophyte? Are there any specific protocols available for confirmation? Kindly suggest your opinion.
If you have properly surface sterilized your sample, then there is very less chance that your fungus is not an endophyte. To confirm the sterilization procedure, you can try different concentration of ethanol and sodium hypochlorite and also you can change the dipping time too. I think your are proceeding in a correct way. All the best
The best and sure shot way to confirm that you're only isolating endophytes is to spread 100microlitre aliquot of water from the last wash that you gave to your samples. That way if there is a growth of fungal colonies on PDA plate where you spread the aliquot of wash water, you will know that surface fungi are still there and surface sterilization has not been done properly. Hope this helps.
doi.org/10.1002/jobm.202000035 you can view this method paper
Moreover our lab specifically works on endophytic actinobacteria, you can refer those too..doi.org/10.3389/fmicb.2020.00318
You can inoculate the plant with a suitable endophytic fungus and left the plant for several weeks. After that make a cross section from the inoculated plant to see and follow the growth and the development of the endophytic fungus by staining the tissue with lactophenol and methylene blue stain.
Hi Dr Jeyamanikandan Venkatachalam . I hope the following link could help: https://www.scielo.br/j/aib/a/9BsgGZjz5TMsQcCckHBQGPf/?format=pdf&lang=en
Also, see the following link that might help: Article Isolation and characterization of endophytic bacteria isolat...
Also may the following link could help: https://link.springer.com/content/pdf/10.1007%2F978-3-319-90484-9_25.pdf
You can try
1)the colonization of endophytic fungus in host.
2) confocal microscopy of original Host.
3) SEM analysis of Original Host TS.