I have many isolates of Pyrenophora teres and i want to know the fsp for them, is ITS region sequencing give me a good results for that.
Bette to include more regions like 1Ealfa, and other genes will give you better results
Hi Oadi,
Due to the high similarity among P. graminea, P. teres f. teres and P. teres f. maculata, ITS is not reliable to differentiate amongst these pathogens. Housekeeping genes such as Btub, GPD or Actin are not suitable, either. EF1 might be similar (I have never tried).
You should use specific PCR primers. There are different primer sets available for PG, PTT and PTM. It is always useful to include PG as control.
FOR PG:
Taylor EJA, Stevens EA, Bates JA et al. (2001) Rapid-cycle PCR detection of Pyrenophora graminea from barley seed. Plant Pathology 50:347–355.
FOR PTT and PTM:
Williams KJ, Smyl C, Lichon A, Wong KY, Wallwork H. (2001) Development and use of an assay based on the polymerase chain reaction that differentiates the pathogens causing spot form and net form of net blotch of barley. Australasia Plant Pathology 30:37–44.
Lu S, Platz GJ, Edwards MC, Friesen TL. Mating type locus-specific polymerase chain reaction markers for differentiation of Pyrenophora teres f. teres and P. teres f. maculata, the causal agents of barley net blotch. Phytopathology. 2010 Dec;100(12):1298-306. doi: 10.1094/PHYTO-05-10-0135.
Leisova L, Kucera L, Minarikova V, Ovesna J. 2005. AFLP-based PCR markers that differentiate spot and net forms of Pyrenophora teres. Plant Pathology, 54 66–73 Doi: 10.1111/j.1365-3059.2004.01117.x
Hope this helps you.
cheers,
József
see Lu et al. 2013 EJPP for primers for targeting the mating type genes for P. teres forms.
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