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Research Article

A Glimpse of Panax ginseng Genome Structure Revealed from Ten BAC Clone Sequences Obtained by SMRT Sequencing Platform

Plant Breeding and Biotechnology 2017;5(1):25-35.
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 13, 2017   • Accepted: February 13, 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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A Glimpse of Panax ginseng Genome Structure Revealed from Ten BAC Clone Sequences Obtained by SMRT Sequencing Platform
Plant Breed. Biotech.. 2017;5(1):25-35.   Published online March 1, 2017
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A Glimpse of Panax ginseng Genome Structure Revealed from Ten BAC Clone Sequences Obtained by SMRT Sequencing Platform
Plant Breed. Biotech.. 2017;5(1):25-35.   Published online March 1, 2017
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A Glimpse of Panax ginseng Genome Structure Revealed from Ten BAC Clone Sequences Obtained by SMRT Sequencing Platform
Image Image Image
Fig. 1 Sequence analysis of ten BAC clones in ginseng. Horizontal black bars represent ten BAC sequences. Identified genome components were shown above the black bar and depicted according to their position on the BAC sequences. The description of each components is shown at the bottom right. Graphs under the black bars represent the depth distribution of 30 Gbp WGS read mapping (approximately 10× coverage). RD indicate the read depth plot for each nucleotide position using 10× WGS.
Fig. 2 Structure of newly identified long terminal repeat retrotransposons (LTR-RTs) in ginseng BAC sequences. Vertical bars on both ends represent the target site duplication (TSD). The mark (+ or −) next to the transposable elements ID indicates the direction on the BAC sequence. Identified internal domain, AP (aspartic protease), CH (chromodomain), GAG (capsid protein), INT (integrase), RH (RNase H), RT (reverse transcriptase) and Zn (zinc knuckle), were depicted according to their position in the internal regions. The dotted box region of PgDel5 are estimated structure because it was truncated in the BAC clone sequence. The omitted region of PgSire1 indicates area lost by external factors.
Fig. 3 The FISH analysis of PgDel2 (green signals) and PgDel5 (red signals) on somatic metaphase chromosomes. Bar, 5 μm.
A Glimpse of Panax ginseng Genome Structure Revealed from Ten BAC Clone Sequences Obtained by SMRT Sequencing Platform

Summary of sequence statistics.

Read size Sequencing chemistry

P4-C2 P6-C4


Total read length (bp) Number of reads Average read length (bp) Total read length (bp) Number of reads Average read length (bp)
~5 kb 7,107,7597 33,924 2,095 160,132,596 66,268 2,416
5 kb~10 kb 58,026,397 8,263 7,022 164,293,017 22,992 7,145
~10 kb 32,183,323 2,549 12,625 161,018,656 11,584 13,900
Total 161,287,317 44,736 3,605 485,444,269 100,844 4,814

Assembly statistics of ten BAC clones.

BAC ID Assembled length (bp) Number of contigs Non-repetitive region (bp, <50 mapping depth) GenBank Accession number

P4-C2 P6-C4 Final
8L14 173,429 5 1 1 11,843 KY513616
9P08 139,247 1 1 1 7,290 KY513618
6B09 131,717 2 1 1 25,544 KY513612
8G13 123,350 1 1 1 902 KY513615
5N01 117,096 2 1 1 90 KY513611
8P22z) 109,113 1 - 1 232 KY513617
8C22 107,984 1 1 1 61,448 KY513614
5B21 100,349 1 1 1 196 KY513610
10M13z) 97,432 1 - 1 2,611 KY513619
7P20 63,647 2 2 2 9,707 KY513613

z)Two BAC clones, 8P22 and 10M13, were not included in the second sequencing run using P6-P4 chemistry.

Annotation summary of identified eleven genes in BAC sequences.

Gene annotation based on BLASTP searches BAC ID Position (bp) # of exon Accession no. (E-value)
Cellulose synthase A catalytic subunit 3 [UDP-forming] 9P08 215–9,773 14 XP_017226278.1 (0.0)
Calcium-dependent protein kinase 28 6B09 1–5,383 (partial) 9 XP_017251289.1 (0.0)
Uncharacterized protein 6B09 18,057–19,415 3 XP_017251291.1 (8e-115)
Transformation/transcription domain-associated protein 6B09 21,538–59,489 35 XP_017217620.1 (0.0)
Transformation/transcription domain-associated protein 6B09 94,283–120,564 35 XP_017217620.1 (0.0)
Acyl carrier protein 1 10M13 1,127–3,315 4 XP_017257036.1 (1e-55)
Superoxide dismutase 10M13 33,367–36,315 7 O22668.1 (1e-100)
Bifunctional 3-dehydroquinate dehydratase/shikimate dehydrogenase 10M13 44,804–50,967 10 XP_017220070.1 (0.0)
Uncharacterized protein 10M13 55,540–58,599 1 XP_017218710.1 (0.0)
Uncharacterized protein 7P20 2,245–7,217 5 XP_017238997.1 (0.0)
Protein FAR1-related sequence 5-like 7P20 53,287–55,512 2 XP_015866013.1 (0.0)

List of novel LTR retrotransposons identified in BAC sequences.

Type BAC ID Position (bp) TSDz) Length / LTR length (bp) Tsy) Tvx) Ts/Tv Kw) Insertion timev) (MYA)
gDel4 8G13 75,443–76,688, 77,213–87,016 GCAAC 11,050 Left 2,542 21 15 1.4 0.014 0.57
Right 2,551
PgDel5 8P22 1–10,587 CAAGC 10,587 Left 1,332 5 8 0.63 0.010 0.41
Right 3,522
PgDel6 8P22 20,535–32,786 GCGCT 12,252 Left 3,195 60 53 1.13 0.037 1.51
Right 3,195
PgTork2 8L14 140,265–148,881 GCAAC 8,615 Left 1,544 13 6 2.17 0.012 0.49
Right 1,544
PgSire1 7P20 32,485–35,143, 39,282–46,836 AAAGG 10,214 Left 1,558 41 14 2.93 0.037 1.51
Right 1,554
PgSire2 10M13 65,051–65,495, 65,765–71,239 CCAGT 5,920 Left 251 8 3 2.67 0.045 1.81
Right 253

z)Target site duplication.

y)Number of transition mutations.

x)Number of transversion mutations.

w)Kimura’s distance.

v)Insertion times were estimated by adopting the substitution rate of 1.22 × 10−8.

Proportion of LTR-RTs in the ginseng genome calculated from repeat masking and WGS read mapping.

Type Proportion in BACs (%) Expected proportion in genome (%)
Ty3/Gypsy 46.88 28.66
PgDel 34.53 24.04
  PgDel1 24.17 20.0
  PgDel2 0.72 0.93
  PgDel3 4.07 1.17
  PgDel4 1.08 0.37
  PgDel5 1.61 0.51
  PgDel6 2.88 1.06
PgTat 11.13 4.1
  PgTat1 9.83 3.85
  PgTat2 1.3 0.25
PgAthila 1.22 0.52
Ty1/Copia 5.09 5.1
PgTork 2.32 1.28
  PgTork1 0.9 0.72
  PgTork2 1.12 0.56
PgSire 2.9 1.2
  PgSire1 2.25 0.92
  PgSire2 0.65 0.28
PgOryco 0.17 0.04
Degenerated LTR-RTs 8.57 2.58
Total 60.54 36.34
Table 1 Summary of sequence statistics.
Table 2 Assembly statistics of ten BAC clones.

Two BAC clones, 8P22 and 10M13, were not included in the second sequencing run using P6-P4 chemistry.

Table 3 Annotation summary of identified eleven genes in BAC sequences.
Table 4 List of novel LTR retrotransposons identified in BAC sequences.

Target site duplication.

Number of transition mutations.

Number of transversion mutations.

Kimura’s distance.

Insertion times were estimated by adopting the substitution rate of 1.22 × 10−8.

Table 5 Proportion of LTR-RTs in the ginseng genome calculated from repeat masking and WGS read mapping.