Trichoderma no infected banana

To get isolation of Trichoderma I suggest using Trichoderma selective media. I would use fresh root sections which throughly washed of all soil and then surface disinfected with 10% bleach 0.5% sodium hypochlorite 4 minutes 1 minute 70% ethyl alchohol and 2 minutes rinse sterile distilled water. let the tissue dry under aseptic hood. Use root sections of 0.5 cm in length. 

The Trichoderma-selective medium (TSM) developed and used in this work consisted of the following components (g/1 distilled water): MgSO 4 9 7 H2 O, 0.2; K2 HPO4,

0.9; KC1, 0.15; NH4 NO3, 1.0; glucose, 3.0; chloramphenicol (Chloromycetin, Sigma

Chemical Co., USA), 0.25; p-dimethylaminobenzenediazo sodium sulfonate (Dexon

60% w.p., Farbenfabrik Bayer A.G., Germany), 0.3; pentachloronitrobenzene (Terraclor

75 w.p., Olin Chemicals, USA), 0.2; rose-bengal (tetrachlorotetradiodofluorescein,

BDH Chemicals Ltd., England), 0.15; agar (Difco Laboratories, USA), 20. See Chet Plant Pathologist from Israel. 

Use 100 to 400 sections to get a percentage of incidence.Trichoderma is an endophyte at low incidence. 

The ability of Trichoderma to counteract the prevalent root pathogens make them very attractive for biocontrol. Trichoderma harzianum is of much interest because it has been shown to stimulate the defensive metabolism of host plants. Trichoderma is also interesting for its hyperparastism. 

good luck in your project and keep us posted. P

Yes, Trichoderma is as endophyte , and it able to induce the plant defences of banana

Hay,

My opinion is that you cant get a direct concentration but comparing with other experiment treatments you would get a relative concentration in the roots. First you have to isolate the Trichoderma population from the banana roots. For that try culturing root tissues (make sure you get the same portion of the tissues from other roots. Second try a broth culture (mostly PDA, Selective media - if the broth commercially not available you'll have to make it from a recipe) to filter most of the bacteria. Third you will see Trichoderma colony after 4- 5 days. Net you can dilute the broth culture with the colony in a beaker with standard volume of sterile water (make sure to follow the same volume for others too) (you can dilute and filter it with a clean cloth). Finally you can use the hemocytometre to count the spores in a ml of innoculum from the beaker. Your count will be spores/ml. As the volume in the beaker was same in other samples and size of the tissue was same in all you can compare this concentration with other roots. 

Cheers

Thank you Dr. Laith Al-Ani, Dr. Paul Reed, and Surantha Salgadoe for your valuable suggestions.

It will help me to solve my problems and misunderstandings.

Thanks a lot! -R-

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Welcome