Hi All,
Rhizoctonia solani is a soilborne pathogen, which do not produce spores in lab. R.solani is multinuclear and the transformation is quite difficult. We can not perform gene knock down or RNAi in R.solani.
Therefore, if we obtain some differential expressed genes during the development progress of R.solani, how to design a suitable experiment protocol in a follow-up research?
Do you have any good ideas?
Thank you!
Canwei
Could you try Host-Induced Gene Silencing (HIGS) using a tractable plant host? This has worked with rust fungi.
Rs AG3 (solanaceous hosts) can form homokaryotic basidiospores. Transform heterokaryotic mycelia/hyphal fragments/protoplasts, select on high levels of selection agent, induce basidia and plate single (homokaryotic?) basidiospores on selection media.
The genetic and phenotypic expression of Rhizoctonia can be affected by hypovirulence. This hypovirulence resultisfrom the infection with suspected RNA virus which knocks out the capacity to Rhizoctonia to be pathogenic and its saprophytic competitivenss. You might like to literature search that topic area to see if there something potentially useful for your investigation..
Hi Stephen,
Thank you for your reply.
Host-Induced Gene Silencing (HIGS) is a good idea. We will have a try.
But Rs AG1 is difficut to produce basidiospores. It is diffrent form Rs AG3.
The critical problem is that we failed to transformc by hyphal fragments or protoplasts methods. Althought we have tried for more than 2 years.
We can find some transformants generate on regenetation media, but they can't grow on selection media amend with Hygromycin.
Can we study some Rs AG1 gene function in another model fungal species, eg, Saccharomyces cerevisiae, by a Heterologous expression method?
Is it workable?
Regards,
Canwei
Perhaps Rs cDNAs could be tested in Saccharomyces; or, maybe in a tractable basidiomycete like Ustilago or Coprinus.
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