A Rapid and Reliable Propidium Monoazide Polymerase Chain Reaction for Detecting Viable <i>Pseudomonas syringae</i> pv. <i>actinidiae</i>
Yi Luo | Wenfei Liao | Yue Li | Wen Chen | Sen Zhong | Cuiping Wu | Kaikai Yao | Rui Yang | Miaomiao Ma | Guoshu Gong
<i>Pseudomonas syringae</i> pv. <i>actinidiae</i> (Psa) is responsible for causing kiwifruit canker disease. The detection of Psa is commonly carried out using normal PCR and culture-based isolation. However, normal PCR does not differentiate between live and dead cells, potentially resulting in the incorrect estimation of the amount of infectious substance in a sample. Such an incorrect estimation could result in unnecessary phytosanitary strategies and control measures. This study attempts to establish a specific assay for detecting only live Psa bacterial cells. To achieve this, a pair of strain-specific primers designed from <i>HopZ3</i> effector were used, and the traditional PCR method was assessed using a nucleic acid-binding dye (propidium monoazide—PMA), establishing a PMA–PCR system and conditions for detecting live Psa in this study. Sensitivity tests showed a detection limit of 10 cfu/mL and 1 pg/μL. This method was also tested in diseased kiwifruit tissues and can be seen as a rapid and dependable replacement to PCR methods for detecting only those infective kiwifruit materials with viable Psa.
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