Susann,

bacteria grow or do not grow on different sources of carbon, nitrogen, vitamins, etc. - thus, it easy to analyse the data.

Your endophitic fungus probably will grow on most of chemicals you test, but will differ in rate of growth, morphology of cells, sporulation (if it has spores), production of secondary metabolites, etc. - mostly quantitative traits. So, you must be prepeared for complex analysis

Good luck!

I suggest you a paper entitled “Trichoderma virens, a Plant Beneficial Fungus, Enhances Biomass Production and Promotes Lateral Root Growth through an Auxin-Dependent Mechanism in Arabidopsis by Hexon Angel Contreras-Cornejo, Lourdes Macı´as-Rodrı´guez, Carlos Corte´s-Penagos, and Jose´ Lo´pez-Bucio Plant Physiology, March 2009, Vol. 149, pp. 1579–1592.”

They made use of 0.2 x MS medium for working out Effects of T. virens and T. atroviride inoculation on the growth of Arabidopsis seedlings.I suppose that helps you. Good Luck

I suggest you a paper entitled “Trichoderma virens, a Plant Beneficial Fungus, Enhances Biomass Production and Promotes Lateral Root Growth through an Auxin-Dependent Mechanism in Arabidopsis by Hexon Angel Contreras-Cornejo, Lourdes Macı´as-Rodrı´guez, Carlos Corte´s-Penagos, and Jose´ Lo´pez-Bucio Plant Physiology, March 2009, Vol. 149, pp. 1579–1592.”

They made use of 0.2 x MS medium for working out Effects of T. virens and T. atroviride inoculation on the growth of Arabidopsis seedlings.I suppose that helps you. Good Luck

attached here is the suggested paper

Yes, you can follow the papers on rep-PCR technique for charaterization of Bacteria and Fungi.

We work with Azospirillum and Trichoderma, which put them to grow in liquid culture medium supplemented with tryptophan. HPLC determined the amount of acetic acid produced indole. Also measure other metabolites (Kyn, anthranilic acid, Pyr, TRM and IAM) involved in the synthesis of this compound. The methodology for the design and apply Azospirillum also in Trichoderma.

good luck.

Hernández-Mendoza J. L., Quiroz-Velázquez J. D., Díaz-Franco A., García-Olivares J. G., Bustamante-Dávila A. J. and Gill-Langarica H. R. 2012. Detection of metabolites in Flor de Mayo common beans (Phaseolus vulgaris L.) and their response to inoculation with Trichoderma harzianum. African Journal of Biotechnology. Vol. 11(55): 11767-11771.

Hernández Mendoza, J. Luis; Quiroz Velásquez J.Dicarlo; Moreno Medina R. y Mayek Pérez, N. 2008. Biosíntesis de ácido antranílico y ácido indolacético a partir de triptofano en una cepa de Azospirillum brasilense nativa de Tamaulipas, México. Rev AIA. 12(1): 57-67.

A other sugestion

Hernández-Mendoza, J. Luis, Moreno-Medina, Víctor Ricardo, Quiroz-Velásquez, Jesús DiCarlo, García-Olivares, Jesús Gerardo, Mayek-Pérez, Netzahualcóyotl. 2010. Efecto de diferentes concentraciones de ácido antranílico en el crecimiento del maíz. REVISTA COLOMBIANA DE BIOTECNOLOGIA. 12(1): 57-63.

There are numerous articles on production of fungi, especially Trichoderma spp. on solid substrates. The choice of substrate depends on the enzymes you would like the fungus to produce. The better the choice the better the results you are looking for.However, bacteria grow best in liquid medium.

The experiment you referred could be an incidence of symbiosis. Some bacteria and fungus survive through mechanism of symbiosis, particularly the endophytic fungus. Please test whether the bacterial strain that you are trying has the that relationship. Very often this relationship is linked to genetic makeup of the fungus and the bacteria.

Upananda

This is to my earlier suggestion made. I am referring the MS media used in the paper can be utilized for characterizing the plant growth promoting activity

Trichoderma virens, a Plant Beneficial Fungus, Enhances Biomass Production and Promotes Lateral Root Growth through an Auxin-Dependent Mechanism in Arabidopsis by Hexon Angel Contreras-Cornejo, Lourdes Macı´as-Rodrı´guez, Carlos Corte´s-Penagos, and Jose´ Lo´pez-Bucio Plant Physiology, March 2009, Vol. 149, pp. 1579–1592.”

They made use of 0.2 x MS medium for working out Effects of T. virens and T. atroviride inoculation on the growth of Arabidopsis seedlings.I suppose that helps you. Good Luck

Anil Kotasthane

One important suggestion is that before to test the ability of your isolate/s to promote plant growth you can select an inoculation method of the microrganism (p.e.: seed inoculation, seedling inoculation, roots inoculation, soil inoculation, etc). This method needs to replicate the environmental conditions in which the plant-microbe interaction happens in nature. This helps that the plant growth promotion ability of this/these microrganism/s can be then applied in comercial growing conditions.

Have nice experiments!

Thank you all for your suggestions.

I just wanted to add some more background to make my question more specific.

The fungus I study actually does promote the growth of plants in vitro, but does not so in vivo. The whole thing is a mutualistic relationship and my theory is that the plant benefits under certain conditions from the interaction with this fungus.

Right now I am interested in the question whether the fungus is able to solubilize certain minerals from the soil to make it available to the plant (biofertilization).

(I have come across the paper from Trichoderma but that deals with hormones and pathways I have looked at already briefly, if time allows I will look at that later in greater detail.)

For bacteria it seems to me the procedures to test what causes growth promotion are established and widely used. So hence my question if it would be adviseable to adapt those methods to the fungus. So far I haven´t really come across a "standard procedure" (if that exists) to test a fungus in terms of biofertilization.

The seeds, particularly corn, have studied, that starting germination releases compounds as tryptophan and the Azospirillum and Trichoderma can metabolize to this compound for synthesize the IAA.

I suggest making inoculations and determined by HPLC metabolites are produced. These tests can be done in sterile petri dish water to retrieve the germination medium.

Also make inoculations and to make observations in scanning electron microscopy and analyze the behavior of the organism and determine the level of interaction

Yes I suppose the basic supportive media changes which is suitable for fungi, characterization method remains the same as of bacteria.

Good that you explained it further. Now my guess that symbiotic relationship exists is correct. But you observe that only it in vitro. It implies it will operate under certain conditions. these conditions does not exists in the environment you carried out your experiment. These conditions , very often, might be concentration of ions of other minerals which retard the forward chemical reaction. Then you have either add a stimulant or change the environmental variables (i.e. Temp., RH, etc.).

Solubility of certain chemical often vary with concentration of ions which give the same effect of the ion under consideration. Or it may give rise to disassociation of some compounds. Ion concentration of minerals may lead you to a method to develop a hypothesis on solubility of plan nutrients (bio fertilization) and the concentration of chemical/hormone released by the fungus under different environment variables (i.e., temperature, relative humidity etc.). I believe RH is a potential variable to manipulate.

Endophytic fungi known to produce bio active compounds like growth promoting substances. We have shown Colletotrichum sp produced IAA in culture. You can test method adopted for bacteria to fungus. Prepare PD media with a precursor for IAA it will work.

If you would like to read about bio-fertilization, I suggest : Implications of Rhizosphere-competence of Trichoderma harzianum in Canadian Journal of Microbiology 1988.

These experiments were done in vivo as well as in vitro. There were statistically significant differences in treated and control plants

You need review some published research work, like this one http://onlinelibrary.wiley.com/doi/10.1002/jobm.201200579/pdf

Carlos, I am also interested in this subject. Would it be possible for you to add the pdf? Now I can not open it...

Thanks for your help.

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