A rapid and enhanced DNA detection method for crop cultivar discrimination

J Biotechnol. 2014 Sep 20:185:57-62. doi: 10.1016/j.jbiotec.2014.06.013. Epub 2014 Jun 19.

Abstract

In many crops species, the development of a rapid and precise cultivar discrimination system has been required for plant breeding and patent protection of plant cultivars and agricultural products. Here, we successfully evaluated strawberry cultivars via a novel method, namely, the single tag hybridization (STH) chromatographic printed array strip (PAS) using the PCR products of eight genomic regions. In a previous study, we showed that genotyping of eight genomic regions derived from FaRE1 retrotransposon insertion site enabled to discriminate 32 strawberry cultivars precisely, however, this method required agarose/acrylamide gel electrophoresis, thus has the difficulty for practical application. In contrast, novel DNA detection method in this study has some great advantages over standard DNA detection methods, including agarose/acrylamide gel electrophoresis, because it produces signals for DNA detection with dramatically higher sensitivity in a shorter time without any preparation or staining of a gel. Moreover, this method enables the visualization of multiplex signals simultaneously in a single reaction using several independent amplification products. We expect that this novel method will become a rapid and convenient cultivar screening assay for practical purposes, and will be widely applied to various situations, including laboratory research, and on-site inspection of plant cultivars and agricultural products.

Keywords: Cultivar discrimination; Multiplex PCR; Practical application; Retrotransposon; Strawberry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agriculture / methods*
  • Chromatography / methods
  • Crops, Agricultural / genetics*
  • DNA Primers / genetics
  • DNA, Plant / isolation & purification*
  • Electrophoresis, Agar Gel
  • Fragaria / genetics*
  • Nucleic Acid Hybridization / methods
  • Sensitivity and Specificity
  • Species Specificity

Substances

  • DNA Primers
  • DNA, Plant