A Review of Conventional PCR Assays for the Detection of Selected Phytopathogens of Wheat

Abstract

Infection of phyllosphere (stems, leaves, husks, and grains) by pathogenic fungi reduces the wheat yield and grain quality. Detection of the main wheat pathogenic fungi provides information about species composition and allows effective and targeted plant treatment. Since conventional procedures for the detection of these organisms are unreliable and time consuming, diagnostic DNA-based methods are required. Nucleic acid amplification technologies are independent of the morphological and biochemical characteristics of fungi. Microorganisms do not need to be cultured. Therefore, a number of PCR-based methodologies have been developed for the identification of key pathogenic fungi, such as Fusarium spp., Puccinia spp., Zymoseptoria tritici, Parastagonospora nodorum, Blumeria graminis f. sp. tritici, and Pyrenophora tritici-repentis. This article reviews frequently used DNA regions for fungus identification and discusses already known PCR assays for detection of the aforementioned wheat pathogens. We demonstrate that PCR-based wheat pathogen identification assays require further research. In particular, the number of diagnostic tests for Fusarium graminearum, Puccinia spp., and P. tritici-repentis are insufficient.

The Development of Diagnostics Tools and Techniques in the Isolation and Detection of Fungal Pathogens

“…Invasive fungal infections (IFIs) can be detected even at an early stage of development; this confers more sensitivity to the non-culture method [1][2][3][4][5][6][7][8][9][10][11]. The principle, just as in bacterial serology is the detection of antibodies or antigen in the serum.…”

“…This brought a boost to the identification of human diseases. Of course their reliability, sensitivity and specificity put them before the traditional methods [1][2][3][4][5][6][7][8][9][10][11][12].…”

“…Also the choice of the buffer used might hinder the homogenization of cells. According to many fungal pathologists, CTAB or guanidinium thiocyanate based buffer is most appropriate [2][3][4][5][6][7][8][9][10]. In response to the need of attaining pure form of DNA, there are number of procedures that can assure the quality and quantity of DNA.…”

“…Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is used in conjunction with the broad-range PCR that targets rDNA fragment. This enables the identification of pathogens from the specimens of patients and also methylprednisolone vials [2][3][4][5][6][7][8]. MALDI has shown increasing positive results for the rapid diagnosing Candida spp.…”

“…The Vitek MS system is an example of the newer version and it makes the method user friendly [2][3][4][5][6][7][8][9][10].…”