오이풀, 흰오이풀, 긴오이풀의 NGS 기반 유전체 서열의 완전 해독 및차세대 염기서열 재분석으로 탐색된 SNP 기반 HRM 분자표지 개발
Development of HRM Markers Based on Identification of SNPs from Next-Generation Sequencing of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link
심미옥, 장지훈, 정호경, 황태연, 김선영, 조현우,
소속 상세정보
심미옥 ( Sim Mi-Ok )
한국한의약진흥원 한약자원본부
장지훈 ( Jang Ji-Hun )
한국한의약진흥원 한약자원본부
정호경 ( Jung Ho-Kyung )
한국한의약진흥원 한약자원본부
황태연 ( Hwang Tae-Yeon )
한국한의약진흥원 한약자원본부
김선영 ( Kim Sun-Young )
한국한의약진흥원 한약자원본부
조현우 ( Cho Hyun-Woo )
한국한의약진흥원 한약자원본부
Abstract
Objective : To establish a reliable tool between for the distinction of original plants of Sanguisorbae Radix, we analyzed the complete chloroplast genome sequence of Sanguisorbae Radix and identified single nucleotide polymorphisms (SNPs).
Materials and methods : The chloroplast genome sequence of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link obtained using next-generation sequencing technology were described and compared with those of other species to develop specific markers. Candidate genetic markers were identified to distinguish species from the chloroplast sequences of each species using Modified Phred Phrap Consed and CLC Genomics Workbench programs.
Results : The structure of the chloroplast genome of each sample that had been assembled and verified was circular, and the length was about 155 kbp. Through comparative analysis of the chloroplast sequences, we found 220 nucleotides, 158 SNPs, and 62 Indel (insertion and/or deletion), to distinguish Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link. Finally, 15 specific SNP genetic markers were selected for the verification at positions. Avaliable primers for the dried herb, which is used as medicine, were used to develop the PCR amplification product of Sanguisorbae Radix to assess the applicability of PCR analysis.
Conclusion : In this study, we found that Fendel-qPCR analysis based on the chloroplast DNA sequences can be an efficient tool for discrimination of Sanguisorba officinalis, Sanguisorba tenuifolia f. alba (Trautv. & Mey.) Kitam and Sanguisorba tenuifolia Fisch. ex Link.
키워드
Sanguisorbae Radix; Genetic Marker; Chloroplast Genome; Single Nucleotide Polymorphism(SNP)
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