Enhancement of fungal DNA templates and PCR amplification yield by three types of nanoparticles
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Department of Biology, Science and Humanities College, Alquwayiyah, Shaqra University, Saudi Arabia
Plant Pathology Research Institute, Agricultural Research Centre (ARC), 12619, Giza, Egypt
College of Biotechnology, Misr University for Science and Technology, P.O. Box: 77, 6th of October City, Egypt
Submission date: 2017-08-21
Acceptance date: 2017-12-04
Corresponding author
Tahsin Shoala
College of Biotechnology, Misr University for Science and Technology, P.O. Box: 77, 6th of October City, Egypt
Journal of Plant Protection Research 2018;58(1):66-72
Nanodiagonastic methods in plant pathology are used for enhancing detection and identification of different plant pathogens and toxigenic fungi. Improvement of the specificity and efficiency of the polymerase chain reaction (PCR) by using some nanoparticles is emerging as a new area of research. In the current research, silver, zinc, and gold nanoparticles were used to increase the yield of DNA for two plant pathogenic fungi including soil-borne fungus Rhizoctonia solani and toxigenic fungus Alternaria alternata. Gold nanoparticles combined with zinc and silver nanoparticles enhanced both DNA yield and PCR products compared to DNA extraction methods with ALB buffer, sodium dodecyl sulfate, ALBfree from protinase K, ZnNPs and AgNPs. Also, by using ZnNPs and AgNPs the DNA yield was enhanced and the sensitivity of random amplified polymorphic DNA (RAPD) PCR products was increased. Application of nanomaterials in the PCR reaction could increase or decrease the PCR product according to the type of applied nanometal and the type of DNA template. Additions of AuNPs to PCR mix increased both sensitivity and specificity for PCR products of the tested fungi. Thus, the use of these highly stable, commercially available and inexpensive inorganic nano reagents open new opportunities for improving the specificity and sensitivity of PCR amplicon, which is the most important standard method in molecular plant pathology and mycotoxicology.
The authors have declared that no conflict of interests exist.
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