한국내 양귀비과 식물의 DNA barcode 데이터베이스 구축 및 종(種) 식별 Marker 발굴
Constructing a DNA Barcode Database and Identification of Marker Nucleotides for Forensic Discrimination of Opium Poppy in South Korea
2
조회
0
다운로드
3
인용
0
참고문헌
키워드
초록
본 연구에서는 수집된 양귀비과(Papaveraceae family) 식물 60개체의 게놈 DNA (genomic DNA)에서 확보한 7개 범용성 DNA 바코드 구간(ITS, rbcL, trnL-trnF, trnL-intron, psbA-trnH, 18S rRNA, matK) 의 염기서열을 분석하여 종 식별 기술 개발에 활용 가능 한 게놈 DNA 염기서열 정보 기반의 바코드 데이터베이스(DB)를 제공하였다. 7개 바코드 중에서 가장 낮은 염기변이를 보인 3개의 바코드를 제외한 4개의 바코드 구간, ITS (0.042), psbA-trnH (0.025), trnL-trnF와 matK (0.015)는 높은 수준의 종간변이와 염기치환율을 나타냈다. 계통수 분석결과, 비규제종인 P. orientale 과 높은 유연관계를 나타낸 18S rRNA 를제외한 4개의 바코드 구간에서, 60개체의 양귀비과 시료 모두 규제종인 P. somniferum-P. setigerum과 높은 유연관계를 나타냈으며, 비마약 식물 종과는 뚜렷하게 독립된 형태의 분계조를 형성하는 것을 확인하였다. 또한, 본 연구에서 처음으로 종 단위 감별이 가능한 14개의 SNPs 와 9개의 P. bracteatum 특이적 염기치환을 발견하였으며, 이러한 염기서열의 특이성은 종식별에 이용가능 할 것으로 판단된다. 이러한 결과는 4개의 DNA 바코드 구간이 규제 양귀비 종을 감별하는데 활용 가능함을 나타낸다. 따라서 본 연구에서지속적인 염기서열 기반DB 구축의 중요성을 확인하였으며, 이와같은 유전체 정보들은 규제 양비귀 종 식별에 대한 이해를 돕는데 유용한 자료가 될 것이다.
Abstract
In this study, we deciphered the sequences of seven universal DNA barcodes, ITS, rbcL, trnL-trnF, trnL-intron, psbA-trnH, 18S rRNA and matK regions in 60 seized samples and provides a barcoding database based on genomic DNA sequences, will be useful to develop a powerful tool for species-specific identification. Among the seven DNA barcodes, ITS (0.042), psbA-trnH (0.025), trnL-trnF, and matK (0.015) exhibited both higher levels of interspecific distance and higher rates of nucleotide substitution than those of four DNA barcodes. Phylogenetic analysis confirmed that all tested samples were genetically distinct from non-drug type Papaver species and clustered into groups constituting P. somniferum or P. setigerum, while the 18S rRNA sequences were also closely related to licit Papaver species, including the P. orientale. Furthermore, fourteen SNPs and nine P. bracteatum- specific nucleotide substitutions in four DNA barcodes were the first identified by comparing the genome sequences and it can also be used for illicit Papaver species identification. Thus, the four regions can be used as reliable barcodes for identifying illicit Papaver species. Taken together, our work highlights the importance of continuously constructing the genome database, and it facilitates a better understanding of forensic discrimination for opium poppy.
참고문헌 (28)
- 정종길, "한국산 양귀비과 식물에 관한 본초학적 연구", 대한본초학회지, 31(5), pp.63~69 (2016), 대한본초학회
- PA Egan, "Flora of Nepal Papaveraceae" (2011), Royal Botanic Gardens
- W Kukula-Koch, "The elevation of {LC}-ESIQ-TOF-MS response in the analysis of isoquinoline alkaloids from some Papaveraceae and Berberidaceae representatives", J Anal Methods Chem, 2017, pp.1~9 (2017)
- S Choe, "Species identification of Papaver by metabolite profiling", Forensic Sci Int, 10, pp.51~60 (2011)
- (약칭: 마약류관리법), , 2019
- WJ Kress, "Use of DNA barcodes to identify flowering plants", Proc Natl Acad Sci USA, 102, pp.8369~8374 (2005)
- RS Purty, "DNA barcoding:an effective technique in molecular taxonomy", Austin J Biotechnol Bioeng, 3(1), pp.1059~ (2016)
- PDN Hebert, "Biological identification through DNA barcodes", Proceedings of the Royal Society of London. Weries B: Biological Sciences, 270, pp.313~321 (2003)
- WJ Kress, "DNA barcodes:genes, genomics, and bioinformatics", Proc Natl Acad Sci USA, 105, pp.2761~2762 (2008)
- I Meusnier, "A universal DNA mini-barcode for biodiversity analysis", BMC Genomics, 9(1), pp.214~217 (2008)
- CBOL Plant Working Group, "A DNA barcode for land plants", Proc Natl Acad Sci USA, 106, pp.12794~12797 (2009)
- EJ Lee, "An assessment of the utility of universal and specific genetic markers for opium poppy identification", J Forensic Sci, 55, pp.1202~1208 (2010)
- S Choe, "Genetic and chemical components analysis of Papaver setigerum naturalized in Korea", Forensic Sci Int, 222, pp.387~393 (2012)
- EJ Lee, "Exploiting expressed sequence tag database for the development and characterization of gene-derived simple sequence repeat markers in the opium poppy (Papaver somniferum L.) for forensic applications", J Forensic Sci, 56, pp.1131~1135 (2011)
- V Mičianová, "Forensic application of ESTderived STR markers in opium poppy", Biologia (Poland), 72(6), pp.587~594 (2017)
- K Tamura, "MEGA5: Molecular Evolutionary Genetics Analysis using maximum likelihood distance, and maximum parsimony methods", Mol Biol Evol, 28, pp.2731~2739 (2011)
- M Kimura, "A simple method for estimating evolutionary rate of base substitutions through comparative studies of nucleotide sequences", J Mol Evol, 16(2), pp.111~120 (1980)
- J Felsenstein, "Confidence limits on phylogenies : an approach using the bootstrap", Evolution, 39, pp.783~791 (1985)
- TS White, "Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics", pp.315~322 (1990), Academic Press, Inc
- Adrienne E Ng, "Identification and Individualization of Lophophora using DNA Analysis of the trnL", trnF Region and rbcL Gene, 2015, pp.61(S1), S26~S229
- P Taberlet, "Universal primers for amplification of three noncoding regions of chloroplast DNA", Plant Mol Bio, 17, pp.1105~1109 (1991)
- T Sang, "Chloroplast DNA phylogeny, reticulate evolution, and biogeography of Paeonia (Paeoniaceae)", Am J Bot, 84, pp.1120~1136 (1997)
- JA Tate, "Paraphyly of Tarasa(Malvaceae)and diverse origins of the polyploid species", Systematic Botany, 28, pp.723~737 (2003)
- DE Soltis, "Angiosperm Phylogeny Inferred from 18S Ribosomal DNA Sequences", Ann Mo Bo Gard, 84(1), pp.1~49 (1997)
- F Rollo, "Otzi’s last meals: DNA analysis of the intestinal content of the Neolithic glacier mummy from the Alps", Proc Natl Acad Sci USA, 99(20), pp.12594~12599 (2002)
- P Cuénoud, "Molecular phylogenetics of Caryophyllales based on nuclear 18S rDNA and plastid rbcL, atpB and matK DNA sequences", Am J Bot, 89, pp.132~144 (2002)
- M Aragane M, "Rapid identification of a narcotic plant Papaver bracteatum using flow cytometry", J Nat Med, 68(4), pp.677~685 (2014)