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記事

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.

元の言語英語
ページ(範囲)19-27
ページ数9
ジャーナルFood Science and Technology Research
25
発行部数1
DOI
出版物ステータス出版済み - 1 1 2019

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Citrus madurensis
Citrus
Nucleotides
Polymorphism
single nucleotide polymorphism
Single Nucleotide Polymorphism
juices
Alleles
alleles
Polymerase Chain Reaction
DNA
Chloroplast DNA
citrus peels
Fruits
Intergenic DNA
intergenic DNA
chloroplast DNA
hands
Hand
nucleotides

All Science Journal Classification (ASJC) codes

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

これを引用

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title = "Utilization of single nucleotide polymorphism-based allele-specific PCR to Identify shiikuwasha (citrus depressa hayata) and calamondin (Citrus madurensis Lour.) in processed juice",
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.",
author = "Ayumi Musou-Yahada and Ken-Ichi Honjoh and Kenta Yamamoto and Takahisa Miyamoto and Hideaki Ohta",
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AU - Musou-Yahada, Ayumi

AU - Honjoh, Ken-Ichi

AU - Yamamoto, Kenta

AU - Miyamoto, Takahisa

AU - Ohta, Hideaki

PY - 2019/1/1

Y1 - 2019/1/1

N2 - 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.

AB - 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.

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