The cultural features of fungal isolates are not sufficient to confirm the identification. Can you show me the microscopic photo of Morphology of the isolates to see the conidiophore and confirm a genera.

Dear Dr. Jawad, 

Since you can't see any sporulation, I suggest that you use DNA approach by sequencing the ITS region of genomic DNA. This approach will help you identifying the identity of your fungus. Here is an example:

Best wishes and regards

Simon

Methodology article Open Access

Fungal-specific PCR primers developed for analysis of the ITS

region of environmental DNA extracts

Kendall J Martin*1 and Paul T Rygiewicz2

Address: 1Dynamac Corporation, National Health and Environmental Effects Research Laboratory, Corvallis, OR USA and 2USEPA National

Health and Environmental Effects Research Laboratory, Corvallis, OR, USA

Email: Kendall J Martin* - [email protected]; Paul T Rygiewicz - [email protected]

* Corresponding author

PCRprimersbasidiomycetesascomycetesInternal Transcribed Spacer (ITS)X/T rapid extraction method

Abstract

Background: The Internal Transcribed Spacer (ITS) regions of fungal ribosomal DNA (rDNA) are

highly variable sequences of great importance in distinguishing fungal species by PCR analysis.

Previously published PCR primers available for amplifying these sequences from environmental

samples provide varying degrees of success at discriminating against plant DNA while maintaining

a broad range of compatibility. Typically, it has been necessary to use multiple primer sets to

accommodate the range of fungi under study, potentially creating artificial distinctions for fungal

sequences that amplify with more than one primer set.

Results: Numerous sequences for PCR primers were tested to develop PCR assays with a wide

range of fungal compatibility and high discrimination from plant DNA. A nested set of 4 primers

was developed that reflected these criteria and performed well amplifying ITS regions of fungal

rDNA. Primers in the 5.8S sequence were also developed that would permit separate

amplifications of ITS1 and ITS2. A range of basidiomycete fruiting bodies and ascomycete cultures

were analyzed with the nested set of primers and Restriction Fragment Length Polymorphism

(RFLP) fingerprinting to demonstrate the specificity of the assay. Single ectomycorrhizal root tips

were similarly analyzed. These primers have also been successfully applied to Quantitative PCR

(QPCR), Length Heterogeneity PCR (LH-PCR) and Terminal Restriction Fragment Length

Polymorphism (T-RFLP) analyses of fungi. A set of wide-range plant-specific primers were

developed at positions corresponding to one pair of the fungal primers. These were used to verify

that the host plant DNA was not being amplified with the fungal primers.

Conclusion: These plant primers have been successfully applied to PCR-RFLP analyses of forest

plant tissues from above- and below-ground samples and work well at distinguishing a selection of

plants to the species level. The complete set of primers was developed with an emphasis on

discrimination between plant and fungal sequences and should be particularly useful for studies of

fungi where samples also contain high levels of background plant DNA, such as verifying

ectomycorrhizal morphotypes or characterizing phylosphere communities.

Published: 18 May 2005

BMC Microbiology 2005, 5:28 doi:10.1186/1471-2180-5-28

Received: 30 November 2004

Ac

Thank you very much my friend Dr. Simon Francis Shamoun, you are right, but I try to recognize it at morphological level, for sure I will use the molecular technique to identify this fungus.

Dear Saida Messgo Moumene, thank you for you comment, I will present the fungal structure soon. 

Dearest friend Dr. Jawad,

Of course continue your efforts in recognizing the fungus at morphological level. Having said that, I suggest placing the cultures under near ultraviolet light. Or try to grow the fungus in poor media (other than routine media such as PDA or MEA). Also try to amend your media with with reed extract and use light/dark lighting alternative .. Just few of my own thoughts.. I wish you all tthe best in your research..

Best wishes and regards..\

Simon.. 

My dear friend Dr. Simon, you are usually, kind and generous with presenting very useful idea. I did amend the medium with reed extract (dry and fresh), also I will use another media with different circumstances... Thank you again  

Dear Jawad,

I recommend you some papers about occuerence of microsopic fungi on Phragmites australis in Poland (one unfortunatelly some in Polish, but names of fungi are in English), maybe this will help you to find what species of fungi is yours.

http://bomax.botany.pl/pubs/data/article_pdf?id=2246

https://pbsociety.org.pl/journals/index.php/aa/article/view/aa.2005.062/1586

http://agro.icm.edu.pl/agro/element/bwmeta1.element.agro-8869d5c9-d5af-4dd8-bf30-c0867daad462

http://www.progress.plantprotection.pl/download.php?ma_id=948

Regards

Thanks Dr. Łukasz Łopusiewicz, that is very kind of you, I will look at them, hoping useful for this purposes

Dear & Simon, Saida and Łukasz

I attached new photos related to the blight reed fungus, Thank you