Utilization of single nucleotide polymorphism-based allele-specific PCR to Identify shiikuwasha (citrus depressa hayata) and calamondin (Citrus madurensis Lour.) in processed juice

Ayumi Musou-Yahada, Ken-Ichi Honjoh, Kenta Yamamoto, Takahisa Miyamoto, Hideaki Ohta

Research output: Contribution to journalArticle

Abstract

To develop a method for the identification of shiikuwasha (Citrus depressa Hayata) and calamondin (Citrus madurensis Lour.), trnL-trnF and trnT-trnL intergenic spacer regions of their chloroplast DNA were amplified using PCR and the nucleotide sequences were determined. In each region, a single nucleotide polymorphism (SNP) site specific to the respective citrus species (shiikuwasha and calamondin) was found. For species discrimination using PCR, two forward primers containing the allele-specific SNP site at the 3’-end and a mismatched nucleotide at the 3 rd base from the 3’-end were designed. The allele-specific forward primers specific to shiikuwasha and calamondin were respectively designated CiDeLF-F and CiMaTL-F. To confirm the specificity of the designed primers, PCR was carried out with DNA prepared from citrus peel or hand-squeezed juice as the template. Results showed that shiikuwasha and calamondin fruits and juices were identifiable by PCR using the allele-specific primers. Furthermore, this allele-specific PCR method can be applied to industrially processed and concentrated juice by amplifying DNA in advance.

Original languageEnglish
Pages (from-to)19-27
Number of pages9
JournalFood Science and Technology Research
Volume25
Issue number1
DOIs
Publication statusPublished - Jan 1 2019

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Food Science
  • Chemical Engineering(all)
  • Industrial and Manufacturing Engineering
  • Marketing

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