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Research Article

Authentication of Golden-Berry P. ginseng Cultivar ‘Gumpoong’ from a Landrace ‘Hwangsook’ Based on Pooling Method Using Chloroplast-Derived Markers

Ho Jun Joh1, Nam-Hoon Kim1, Murukarthick Jayakodi1, Woojong Jang1, Jee Young Park1, Young Chang Kim2, Jun-Gyo In3, Tae-Jin Yang1,4,*
Plant Breeding and Biotechnology 2017;5(1):16-24.
Published online: March 1, 2017

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*Corresponding author: Tae-Jin Yang, tjyang@snu.ac.kr, Tel: +82-2-880-4547, Fax: +82-2-873-2056
• Received: February 1, 2017   • Revised: February 7, 2017   • Accepted: February 8, 2017

Copyright © 2017 The Korean Society of Breeding Science

This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Authentication of Golden-Berry P. ginseng Cultivar ‘Gumpoong’ from a Landrace ‘Hwangsook’ Based on Pooling Method Using Chloroplast-Derived Markers
Plant Breed. Biotech.. 2017;5(1):16-24.   Published online March 1, 2017
DOI: https://doi.org/10.9787/PBB.2017.5.1.16
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Authentication of Golden-Berry P. ginseng Cultivar ‘Gumpoong’ from a Landrace ‘Hwangsook’ Based on Pooling Method Using Chloroplast-Derived Markers
Plant Breed. Biotech.. 2017;5(1):16-24.   Published online March 1, 2017
DOI: https://doi.org/10.9787/PBB.2017.5.1.16
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Authentication of Golden-Berry P. ginseng Cultivar ‘Gumpoong’ from a Landrace ‘Hwangsook’ Based on Pooling Method Using Chloroplast-Derived Markers
Image Image Image Image Image
Fig. 1 Distinct characteristics of berry and stem of two P. ginseng landraces. (A, B) Red berry and violet stem of P. ginseng landrace Jakyung. (C, D) Golden berry and green stem of P. ginseng landrace Hwangsook (HS). Most of registered P. ginseng cultivars have developed from Jakyung except for P. ginseng cultivars Gumpoong (GU) developed from HS. GU shows very similar morphologies including berry color to HS.
Fig. 2 Location of InDel and SNP variations between chloroplast genome sequences of GU and HS. The intra-species polymorphic sites were identified among 11 ginseng cultivars in our previous study (Kim et al. 2015), of which five (a–e) showed polymorphisms (two InDels as color-filled triangles and three SNPs as empty triangles) between GU and HS. Among the five polymorphism sites, one InDel and two SNPs (indicated by a, b and c) were used to design makers in this study. a, pgcpd02; b, pgcp_ccsA; c, pgycf1. pgcp_ccsA marker was newly developed in this study. Polymorphic site c was duplicated in two inverted repeats (IRA and IRB).
Fig. 3 Diagram of dCAPS marker pgcp_ccsA for discriminating GU and HS sequence. Nucleotide in red box represents single nucleotide polymorphism (SNP) in ccsA gene region between GU and HS. Nucleotide in blue box represent designed nucleotide mismatch for SmaI cutting. This dCAPS marker is developed to digest only GU sequence. After PCR amplification and SmaI cutting, PCR product of GU is cleaved into two fragments of 199 bp and 24 bp, while product of HS is not cleaved and remains to be 223 bp. These size differences can easily be detected through electrophoresis.
Fig. 4 Application of three molecular markers to individual plants in GU and HS population. A. Marker analyses of individual plants in GU population. B. Marker analyses of individual plants in HS population. Numbers (1–32) indicate plant nos. as described in Materials and Methods. G and H indicate authentication controls for GU and HS, respectively. Electrophoresis was performed using 1% agarose gel for pgcpd02 and pgcp_ccsA marker, and 3% agarose gel for pgycf1 marker. M represents 100-bp DNA ladder.
Fig. 5 Bulked population analysis of GU and HS with three molecular marker. GU and HS represent authentication controls, respectively. GU-P and HS-P represent the 32 plant bulked DNA template sample of GU and HS population, respectively. Electrophoresis was performed using 1% agarose gel for pgcpd02 and pgcp_ccsA marker, and 3% agarose gel for pgycf1 marker. M represents 100-bp DNA ladder.

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

Authentication of Golden-Berry P. ginseng Cultivar ‘Gumpoong’ from a Landrace ‘Hwangsook’ Based on Pooling Method Using Chloroplast-Derived Markers

Polymorphic sites and marker information in chloroplast genomes of GU and HS.

Type Position Nucleotide position GU / HS Polymorphisms GU / HS Primer ID Product size (bp) GU/HS Restriction enzyme
InDel ycf1 111,305 / 111,304 57×4 / 57×3 pgycf1z) 729/672
InDel rps16 intron 5,473 (C)9 / (C)8
SNP rpoC2 21,345 / 21,344 T / C pgcpd02z) 177/200 ScaI
SNP ccsA 117,427 / 117,369 G / A pgcp_ccsAy) 199/223 SmaI
SNP ycf1 127,069 A / T

Polymorphic sites and marker information were from Kim et al. (2015).

z)Developed in Kim et al. (2015).

y)Developed in this study.

The genotyping data of individual plant in GU and HS population using three markers.

Primer ID Gumpoong (GU) Hwangsook (HS)
No. of GU type No. of HS type No. of GU type No. of HS type
pgcpd02 32 0 12 20
pgcp_ccsA 32 0 12 20
pgycf1 32 0 20 12
Table 1 Polymorphic sites and marker information in chloroplast genomes of GU and HS.

Polymorphic sites and marker information were from Kim et al. (2015).

Developed in Kim et al. (2015).

Developed in this study.

Table 2 The genotyping data of individual plant in GU and HS population using three markers.

Table 1

Table 2

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