Selective detection of viable seed-borne Acidovorax citrulli by real-time PCR with propidium monoazide View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2016-12

AUTHORS

Qian Tian, Jian-Jun Feng, Jie Hu, Wen-Jun Zhao

ABSTRACT

In recent years, use of the DNA-intercalating dye propidium monoazide (PMA) in real-time PCR has been reported as a novel method to detect viable bacteria in different types of samples, such as food, environmental, and microbiological samples. In this study, viable cells of Acidovorax citrulli, the causal agent of bacterial seedling blight and fruit blotch, were selectively detected and differentiated from dead cells by real-time fluorescent polymerase chain reaction amplification after the bacterial solution was treated with the DNA-binding dye PMA. The primers and TaqMan probe were based on the A. citrulli genome (Aave_1909, Gene ID: 4669443) and were highly specific for A. citrulli. The detection threshold of this assay was 103 colony-forming units per mL (CFU/mL) in pure cell suspensions containing viable and dead cells and infected watermelon seeds. Application of this assay enables the selective detection of viable cells of A. citrulli and facilitates monitoring of the pathogen in watermelon and melon seeds. More... »

PAGES

35457

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/srep35457

DOI

http://dx.doi.org/10.1038/srep35457

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1001530203

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/27739469


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