Home
Print _CMN_EMAIL_ALT

Enhancement of fungal DNA templates and PCR amplification yield by three types of nanoparticles

 

Fahad A. Al-Dhabaan1, Heba Yousef2, Tahsin Shoala3*, Jumana Shaheen3, Yousra El Sawi3, Tasneem Farag2

 

1Department of Biology, Science and Humanities College, Alquwayiyah, Shaqra University, Saudi Arabia

2Plant Pathology Research Institute, Agricultural Research Centre (ARC), 12619, Giza, Egypt 3College of Biotechnology, Misr University for Science and Technology, P.O. Box: 77, 6th of October City, Egypt

 

*Corresponding address:

This e-mail address is being protected from spam bots, you need JavaScript enabled to view it

 

Received: August 21, 2017

Accepted: December 4, 2017

 

Abstract

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.

 

Key words: Alternaria alternata, DNA extraction, metal nanoparticles, nano-PCR, Rhizoctonia solani

 

 

 
Editor in Chief
This e-mail address is being protected from spam bots, you need JavaScript enabled to view it

Managing Editors
This e-mail address is being protected from spam bots, you need JavaScript enabled to view it

This e-mail address is being protected from spam bots, you need JavaScript enabled to view it
Current issue cover
Who's Online
We have 1 guest online