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An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species

The present study utilizes polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using partial plastid rbcL and mitochondrial trnC–trnP gene sequences to distinguish the six representative Pyropia species produced via mariculture in Korea. The rbcL, trnC, and trnP se...

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Detalles Bibliográficos
Autores principales: Kim, Yonguk, Choi, Sung-Je, Choi, Chulyung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6149700/
https://www.ncbi.nlm.nih.gov/pubmed/29292786
http://dx.doi.org/10.3390/molecules22122182
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author Kim, Yonguk
Choi, Sung-Je
Choi, Chulyung
author_facet Kim, Yonguk
Choi, Sung-Je
Choi, Chulyung
author_sort Kim, Yonguk
collection PubMed
description The present study utilizes polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using partial plastid rbcL and mitochondrial trnC–trnP gene sequences to distinguish the six representative Pyropia species produced via mariculture in Korea. The rbcL, trnC, and trnP sequences of 15 Pyropia species from the NCBI database were aligned to determine specific restriction enzyme sites of the six Pyropia species. To confirm the presence of restriction sites of eight enzymes, PCR amplicons were digested as follows: a 556 bp fragment within the rbcL region of chloroplast DNA was confirmed in P. yezoensis using BglI, whereas Tth111I, AvaII, BsrI, and BsaAI enzymes produced fragments of 664, 271, 600, and 510 bp, respectively, from the rps11–trnG region of mitochondrial DNA in P. seriata, P. dentata, P. suborbiculata, and P. haitanensis. In the case of P. pseudolinearis, HindIII, SacII, and SphI enzymes each had two cleavage sites, at positions 174 and 825, 788 and 211, and 397 and 602 bp, respectively. All six species were successfully distinguished using these eight restriction enzymes. Therefore, we propose that PCR-RFLP analysis is an efficient tool for the potential use of distinguishing between the six Pyropia species cultivated via mariculture in Korea.
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spelling pubmed-61497002018-11-13 An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species Kim, Yonguk Choi, Sung-Je Choi, Chulyung Molecules Technical Note The present study utilizes polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis using partial plastid rbcL and mitochondrial trnC–trnP gene sequences to distinguish the six representative Pyropia species produced via mariculture in Korea. The rbcL, trnC, and trnP sequences of 15 Pyropia species from the NCBI database were aligned to determine specific restriction enzyme sites of the six Pyropia species. To confirm the presence of restriction sites of eight enzymes, PCR amplicons were digested as follows: a 556 bp fragment within the rbcL region of chloroplast DNA was confirmed in P. yezoensis using BglI, whereas Tth111I, AvaII, BsrI, and BsaAI enzymes produced fragments of 664, 271, 600, and 510 bp, respectively, from the rps11–trnG region of mitochondrial DNA in P. seriata, P. dentata, P. suborbiculata, and P. haitanensis. In the case of P. pseudolinearis, HindIII, SacII, and SphI enzymes each had two cleavage sites, at positions 174 and 825, 788 and 211, and 397 and 602 bp, respectively. All six species were successfully distinguished using these eight restriction enzymes. Therefore, we propose that PCR-RFLP analysis is an efficient tool for the potential use of distinguishing between the six Pyropia species cultivated via mariculture in Korea. MDPI 2017-12-08 /pmc/articles/PMC6149700/ /pubmed/29292786 http://dx.doi.org/10.3390/molecules22122182 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Technical Note
Kim, Yonguk
Choi, Sung-Je
Choi, Chulyung
An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title_full An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title_fullStr An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title_full_unstemmed An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title_short An Efficient PCR-RFLP Method for the Rapid Identification of Korean Pyropia Species
title_sort efficient pcr-rflp method for the rapid identification of korean pyropia species
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6149700/
https://www.ncbi.nlm.nih.gov/pubmed/29292786
http://dx.doi.org/10.3390/molecules22122182
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