Propidium monoazide combined with qPCR to differentiate live and dead conidia of Neofabraea actinidiae

Authors

  • Lucia Ramos Plant & Food Research
  • I.P. Shamini Pushparajah Plant & Food Research
  • M. Shahjahan Kabir Plant & Food Research
  • Bethan E. Parry Plant & Food Research
  • Kerry R. Everett Plant & Food Research

DOI:

https://doi.org/10.30843/nzpp.2018.71.218

Abstract

Neofabraea actinidiae can occasionally cause post-harvest rot in kiwifruit. Quantitative polymerase chain reaction (qPCR) analysis represents a feasible and accurate option for identifying and quantifying this rot but is limited because qPCR results do not differentiate live and dead conidia. Propidium monoazide (PMA) is a photoreactive dye that penetrates into the damaged cell-wall membranes of dead conidia binding to the DNA and thus suppressing its amplification by qPCR. A commercial kit containing PMA was trialled for differentiating between live and dead N. actinidiae conidia. The most suitable conditions were 1 μM PMA with 10 min light emitting diode (LED) exposure, and could clearly distinguish high concentrations of live from similar concentrations of dead conidia when tested separately and as a mixture. Low concentrations of live N. actinidiae conidia could be distinguished from dead ones when tested separately, but not as a mixture. Additional work is needed to optimise the effectiveness of the PMA binding and apply this concept in the orchard.

Published

2018-07-30

How to Cite

Ramos, Lucia, I.P. Shamini Pushparajah, M. Shahjahan Kabir, Bethan E. Parry, and Kerry R. Everett. “Propidium Monoazide Combined With QPCR to Differentiate Live and Dead Conidia of Neofabraea Actinidiae”. New Zealand Plant Protection 71 (July 30, 2018): 348. Accessed February 22, 2024. https://journal.nzpps.org/index.php/nzpp/article/view/218.

Issue

Section

Poster Abstracts

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