Normal PDA is ok.

You can as well refer to Plant pathological methods of fungi and bacteria.

With regards to the isolates, I have recently isolated some of the cucumber but proximity is the main challenge

thank you Osabutey Samuel

Thank you Kouadri Mohamed El Amine

We have isolated Fusarium oxysporum from clinical and environmental materials on sevearl media, such as potato dextrose agar, Sabouraud dextrose agar and oat meal agar.

We have published two papers on oculomycosis in animals due to Fusarium solani and both papers can be easily downloaded from the Research Gate.

Thank you sir, I will go through it. Mahendra Pal

Dear, Navya,

I have worked with isolation of Fusarium oxysporum for a long time, and see that Potato Dextrose Agar (PDA) is very suitable medium to isolate this fungus from soil , infected roots, seeds or plant crowns (The contact between roots and stems).

With Regards

The medium PDA consists of the following:

Potato 200 gm

Dextrose 20 gm

Agar 20 gm

Distilled water 1L

Autoclaved at 121 C and 15 pounds/Inch2 for about 30 minutes and adding suitable antibiotic such as Chloramphenicol or any other antibiotic to prevent the growth of bacteria. Then it poured in sterile Petri dishes.

Please take a look at my attached research which titled: Isolation and identification of Fusarium species from different types of soils at Basra Governorate and their pathogenicity testing. Muthanna Journal of Agriculture Science, VOL.(7), NO.(1), 2019: 83-91.

Also, you can use Carnation Leaves Agar (CLA) for Fusarium spp., if you want the fungus to produce sporodochia successfully.

Some other media like CMA, OME, MEA and SDA can be used also, but PDA stays the best one for good isolation of most species of the fungus Fusarium.

If you want to isolate Fusarium oxysporum f. sp. cucumerinum, you should take roots of cucumber which infected with fusarium wilt and apply the following items:

1 - Washing these roots with tap water carefully to remove dirts and cutting them

to about 1 cm for each portion

2 - Sterilizing them with sodium hypochlorite 10% (from the trade solution) for 2-3

minutes

3 - Drying them with filter papers

4 - Culturing them on sterile PDA with a rate of 5 portions per each plate circularly

5 - Incubating the plates in incubator for 4-7 days.

Dear, Navya Botlagunta,

I think you can't isolate the fungus F. oxysporum f.sp. cucumerinum from the leaves of cucumber. Therefore, I suggest you must not use the leaves for isolation the fungus. You can isolate the fungus from soil, roots, seeds and some times stems, but not from leaves.

With greetings.

Thank you so much, sir. you are giving so much information with so much patience sir. I will definitely try it, sir. Yehya A. Salih

You are most welcome dear Navya. So, I hope you have more information from these different comments and apply them in the laboratory to get a good result in your work.

With kind regards.

The fungus Fusarium oxysporum can be isolated from soil, plant roots, plant leaves and some times from spoilage food. So, the methods of isolation the fungus from theses sources are approximately same.

The best technique for getting consistent morphology for Fusarium indication is carnation leaves which are sterilized in polypropylene gas and planted on water agar.

The Fusarium laboratory of Pennsylvania State had much expertise in teaching the techniques for dealing with Fusarium species.

Standard consistent techniques are a very good start.

For looking at various Fusarium species the use of seed isolation is convenient.

From surface disinfected seeds a wide range of Fusarium species can be represresented.

These seed isolates are internally seedborne and can represent potentials of a wide variety of pathogenic neutral and even beneficial reactions.

For isolating the cucumber F. oxysporum the best strategy is to go to disease plants and isolate from the stems and seeds. The stem isolation from a plant showing discoloration of the vascular tissue is performed by immersing the stem pieces in alcohol flaming them and plating on potato dextrose agar.

Confirm the morphology and genomic confirmation nice also. Complete Koch postulates and reisolate the pathogen upon inoculation and incubation.

Fusarium on cucumber gives characteristic vascular wilt and growing of the xyem tissue.