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

研究成果: ジャーナルへの寄稿学術誌査読

1 被引用数 (Scopus)

抄録

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.

本文言語英語
ページ(範囲)19-27
ページ数9
ジャーナルFood Science and Technology Research
25
1
DOI
出版ステータス出版済み - 2019

!!!All Science Journal Classification (ASJC) codes

  • バイオテクノロジー
  • 食品科学
  • 化学工学(全般)
  • 産業および生産工学
  • マーケティング

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