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基本情報 |
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氏名 |
山本 健太 |
氏名(カナ) |
ヤマモト ケンタ |
氏名(英語) |
YAMAMOTO KENTA |
所属 |
中村学園大学 栄養科学部 フード・マネジメント学科 |
職名 |
講師 |
Utilization of Single Nucleotide Polymorphism-based Allele-specific PCR to Identify Shiikuwasha (Citrus depressa Hayata) and Calamondin (Citrus madurensis Lour.) in Processed Juice
Musou-Yahada, Ayumi Honjoh, Ken-ichi Yamamoto, Kenta Miyamoto, Takahisa Ohta, Hideaki
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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 3rd 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.
FOOD SCIENCE AND TECHNOLOGY RESEARCH
JAPANESE SOC FOOD SCI & TECHNOLOGY
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