Location: Agricultural Systems Research Unit
Title: Detection of Pyrenophora teres in infested plant tissues by PCR Authors
Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: May 11, 2010
Publication Date: June 1, 2010
Citation: Lartey, R.T., Caesar, T., Caesar, A.J., Hanson, S.L., Sainju, U.M. 2010. Detection of Pyrenophora teres in infested plant tissues by PCR. American Phytopathological Society Annual Meeting. 100(6)S68. Technical Abstract: Net blotch of barley, a commonly occurring foliar disease is caused by Pyrenophora teres Drechs. The disease is characterized by small circular elliptical spots which enlarge to the typical narrow netlike pattern. Lesions in mature plants appear similar to spot blotch of Cochliobolus sativus, both of which occur extensively in the Northern Great Plains of the US. Conidia and ascospores from infested straw are probably the most important sources of primary inoculum. To speed up detection and identification of P. teres in infected hosts, a rapid PCR technique using Extract-N-Amp Plant PCR Kit (Sigma-Aldrich) was developed. Bypassing the standard DNA extraction, leaf disks from diseased tissues, uninfected barley leaves and P. teres pure culture controls were first homogenized in extraction solution and diluted with the solution provided in the kit. Aliquots of the homogenate were added to PCR reaction and subjected to amplification using the newly developed PTACTIN, a P. teres actin and ITS primers. Sizes of amplicons from diseases leaves which were resolved on agarose gel, correlated with amplicon from the control pure cultures. The amplicons were purified from the gel and sequenced. Sequences alignment confirmed the detection of P. teres. The technique will enhance rapid detection of P. teres in diseased barley and alternate hosts, including asymptomatic plants as well as verification of efficacy of management of net blotch.