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Research Article

Development of 34 New Microsatellite Markers from Actinidia arguta: Intra- and Interspecies Genetic Analysis

Plant Breeding and Biotechnology 2013;1(2):137-147.
Published online: June 30, 2013

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*Corresponding author: Kyung-Ho Ma, khma@korea.kr, Tel: +82-31-294-6029, Fax: +82-31-294-6029
• Received: June 15, 2013   • Revised: June 21, 2013   • Accepted: June 23, 2013

Copyright © 2013 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/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Development of 34 New Microsatellite Markers from Actinidia arguta: Intra- and Interspecies Genetic Analysis
Plant Breed. Biotech.. 2013;1(2):137-147.   Published online June 30, 2013
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Development of 34 New Microsatellite Markers from Actinidia arguta: Intra- and Interspecies Genetic Analysis
Plant Breed. Biotech.. 2013;1(2):137-147.   Published online June 30, 2013
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Development of 34 New Microsatellite Markers from Actinidia arguta: Intra- and Interspecies Genetic Analysis
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Fig. 1 Phylogenetic dendrogram of the varieties and species in Actinidia based on SSR amplification profiles.
Development of 34 New Microsatellite Markers from Actinidia arguta: Intra- and Interspecies Genetic Analysis

List of Actinidia species used in this study.

Taxon Number of accessions Variety Geographic location
A. arguta (Sieb. and Zucc.) Planch. ex Miq. var. arguta 40 63. K5-1-22 73. K5-3-6 83. K5-4-12 93. K5-10-1 Korea
64. K5-2-3 74. K5-3-7 84. K5-5-1 94. K5-10-2
65. K5-2-7 75. K5-3-8 85. K5-5-8 95. K5-10-3
66. K5-2-8 76. K5-3-12 86. K5-5-12 96. K5-10-5
67. K5-2-10 77. K5-3-13 87. K5-5-14 97. K5-10-8
68. K5-2-13 78. K5-3-18 88. K5-6-10 98. K5-11-1
69. K5-2-16 79. K5-4-3 89. K5-6-12 99. K5-11-2
70. K5-2-18 80. K5-4-7 90. K5-9-1 100. K5-11-3
71. K5-2-22 81. K5-4-8 91. K5-9-4 101. K5-11-8
72. K5-3-3 82. K5-4-10 92. K5-9-8 102. K5-12-6

A. chinensis Planch. var. chinensis 13 14. SKK3 20. SKK10 104. Red-Kiwi 106. Jesi-Gold Korea
16. SKK5 21. SKK11, 105. Hort16A
17. SKK7 (Gracies) 23. SKK13, 108. Yeo-San-Hyang
19. SKK9 (Chin Mei) 62. Jeju-Gold 115. Guem-Goi

A. chinensis Planch. var. chinensis 6 12. SKK1 30. S13 China
13. SKK2 15. SKK4 18. SKK8 (Tomuri) 117. SKK12-2

A. deliciosa (A. Chev.) C.F. Liang and A.R. Ferguson var. deliciosa 25 1. Abbott 7. Hyang-Rok 11. Yellow Queen (First Emperor) 45. SKK 60 New Zealand, Japan, China
2. Bruno 8. Golden King 38. SKK22 (Jinkui) 47. SKK80 (Red Princess)
3. Hayward 9. Golden Yellow 40. SKK33 (Wha-Pyung 2 Ho) 116. Jin-Mi
4. Monty 10. Sensation Apple 42. SKK43 (Seo-Hyang)
5. Matua 37. SKK18 (Chieftain) 43. SKK44 110. M51-2
6. Tomuri 39. SKK23 46. SKK 61
34. Sun-WuKong 41. SKK34 (Mi-Lyang 2 Ho) 109. M51-1

A. eriantha Benth. 3 50. S10 51. S20 103. Bi-Dan China, Korea

A. arguta var. purpurea (Rehd.) C.F. Liang 1 24. S3 China

A. kolomikta (Maxim. and Rupr.) Maxim. 1 25. S4 China

A. macrosperma C.F. Liang 1 27. S7 China

A. rufa (Sieb. and Zucc.) Planch. ex Miq. 1 61. Seom-Darae Korea

Efficiency of the procedure adopted for the identification of microsatellite markers in A. arguta.

Procedural step Number (percentage)
Sequenced clones 762
Redundant clones 35 (4.6%)
Unique clones 727 (95.4%)
SSR clones 565 (74.1%)
Truncated clones (5′ or 3′ end) 162 (21.3%)
Fewer than 12 nucleotides 263 (34.5%)
Primer pairs designed 140 (18.4%)
Polymorphic loci 34 (24.3%)z)

z)percentage based on the designed primer pairs.

Frequency and type of di-, tri- and ≥ tetra-nucleotide repeats isolated from the A. arguta microsatellite-enriched library.

Repeat unit Repeat class Number of microsatellite loci Polymorphic loci


Number Percentage Number Percentage


Di-nucleotide AC/CA/TG/GT 238 23.7% 8 17.4%
AG/GA/TC/CT 603 60.1% 30 65.2%
AT/TA 22 2.2%
CG/GC 1 0.1%


Total 864 86.1%


Tri-nucleotide AAC/ACA/CAA 15 1.5%
AAG/AGA/GAA 44 4.4% 1 2.2%
AAT/ATA/TAA 4 0.4%
ACC/CCA/CAC 3 0.3%
AGC/CGA/GAC 4 0.4%
AGG/GGA/GAG 14 1.4% 2 4.3%
ATC/TCA/CAT 1 0.1%
CCG/CGC/GCC 2 0.2%


Total 87 8.7%


≥ Tetra-nucleotide 53 5.8% 5 10.9%


Total repeat motifs 1004 46

Characteristics of 34 polymorphic microsatellite loci for Actinidia species.

Marker GeneBank accession Repeat motif Primer sequence Size (bp) Ta (°C) A MAF He Ho PIC
GB-AA-005 FJ647762 (CT)17 F: AGTTGTGCATCCAAAGGC
R: CAGTGGGGTGAAGAACGA
192–218 57 12 0.32 0.73 0.78 0.76
GB-AA-012 FJ647763 (TG)13 F: TCACAACACTCATTTCGGC
R: ATCCGCTTCCTTAGCTGC
154–176 58 10 0.49 0.47 0.71 0.68
GB-AA-015 FJ647764 (AG)19 F: CCTGGTCGTTCAGGGAAT
R: ATGGCATTTGTTGCCTTG
267–293 58 14 0.24 0.89 0.83 0.83
GB-AA-017 FJ647765 (CT)6 F: AAAGTGTGAGCACGTGACAA
R: TGAGAGAGAGAGGTGGCG
170–182 58 2 0.54 0.00 0.50 0.37
GB-AA-018 FJ647766 (CA)8, (GGA)4 F: ACCATGGCACAGATGGAA
R: TCCAGTGCCTTTTTAAGCC
147–171 58 11 0.23 0.75 0.81 0.80
GB-AA-024 FJ647767 (GA)19 F: AGGAGACCCAACAGGAACA
R: AATTCGGGTCACCACACA
155–181 58 12 0.40 0.49 0.75 0.74
GB-AA-054 FJ647768 (GA)15 F: ACCAAAAACCACCTGCCT
R: TGAACCCGTATTCGCATC
186–214 58 15 0.23 0.87 0.85 0.84
GB-AA-065 FJ647769 (GA)11(GAA)23 F: ATTGAAGCCCCCATTGAG
R: CCAAGGAGGGCATTTAGG
212–238 58 11 0.43 0.60 0.74 0.72
GB-AA-069 FJ647770 (GGGA)6(GA)8 F: CGTTCTCCTTCGACCCTT
R: CCGTTACCTTGTCCAATCC
196–258 58 11 0.43 0.67 0.74 0.71
GB-AA-080 FJ647771 (GA)15 F: CCAATCAACAAGATGCACG
R: TGGGAGGTTGAAACTGGA
179–213 58 11 0.26 1.00 0.82 0.80
GB-AA-084 FJ647772 (GA)15 F: CATTCGAACCAACGCAAT
R: AGTCGGAGCTGGGAGAAG
233–259 58 12 0.22 0.96 0.83 0.82
GB-AA-088 FJ647773 (CT)7 F: TCTGGTTTGTTTTCCACCA
R: GGTTGAGTTCCATTCCCG
177–197 57 8 0.45 0.56 0.69 0.67
GB-AA-091 FJ647774 (CT)19 F: TGACTTAAGGGCGACCAA
R: GGAAATCGCTCATGGACA
201–223 58 8 0.38 0.62 0.74 0.69
GB-AA-094 FJ647775 (AC)15 F: ACAGGGGAACATCAGTGC
R: GTGGGATATAACCGGGGA
208–220 57 6 0.38 0.41 0.68 0.63
GB-AA-096 FJ647776 (GA)6, (GA)3 F: TTGGTACACAAGACGCCC
R: GAAATTTCTTCACCCGCC
229–235 58 3 0.52 0.01 0.50 0.39
GB-AA-302 FJ647777 (AG)13 F: TCGGTGATAAATGGCAGG
R: GGCTCCTTGACAGCACAG
269–311 58 11 0.43 0.31 0.74 0.72
GB-AA-303 FJ647778 (CT)15 F: TGGGGCATTTATGCCTAT
R: TCCAACCTTCTTGGCTCA
156–258 57 12 0.33 0.35 0.75 0.73
GB-AA-304 FJ647779 (AG)11 F: CAGTCCACAAAATGGGTCA
R: GGTGAACACCCCCAAAGT
202–364 58 13 0.41 0.15 0.67 0.64
GB-AA-308 FJ647780 (CCCT)4, (GGGT)2, (GGGT)2 F:TAGTGCTGCGAAGGAGGA
R: ATTGCACCATTCTGTGGC
265–304 58 8 0.32 0.36 0.77 0.75
GB-AA-331 FJ647781 (GA)15, (CAGA)4 F: CCATCTTTTTGTGCCTTTG
R: TTGTTGGTATCATGCCCC
259–303 57 13 0.28 0.59 0.83 0.80
GB-AA-333 FJ647782 (TG)12 F: AAGTTCATTCCACGCACG
R: ATTGCATTTGAGCCGCTA
242–276 58 6 0.49 1.00 0.67 0.61
GB-AA-337 FJ647783 (GA)10, (AC)6 F: TTCTCTGCGCGTTCTCTC
R: AGCCTCAACCAAGAAGGG
290–332 58 10 0.36 0.53 0.74 0.72
GB-AA-340 FJ647784 (CCT)5 F: AAGGAATTCGCCCTCAAA
R: GGCTGACAAGAAGCGATG
287–296 58 4 0.65 0.68 0.49 0.45
GB-AA-342 FJ647785 (GA)2, (GA)4 F: AAAATTCCAATTCCCCCA
R: CATTTCGGAATCCCCTTT
261–293 57 13 0.35 0.59 0.81 0.80
GB-AA-343 FJ647786 (AG)14 F: TGCTTCTTCGGTCATGCT
R: CACCATTTCGAACCCAGA
232–262 58 5 0.44 0.67 0.70 0.64
GB-AA-356 FJ647787 (AC)12 F: CCCGACTTCCAAGTCTCC
R: CTCCGGATGCCCTTTATC
251–287 58 16 0.25 0.70 0.85 0.83
GB-AA-366 FJ647788 (GA)7 F: CAGCTCCAAGGGCTATGA
R: TGTTCCCACTACCGCAAC
241–247 57 5 0.51 0.09 0.53 0.43
GB-AA-369 FJ647789 (CT)16(CA)17 F: TGATCCACAACGTCATCAA
R: GGGCACGCTAGACACACC
185–215 58 13 0.32 0.31 0.76 0.74
GB-AA-370 FJ647790 (GA)16 F: GGGAATTGGTGAGTGGGT
R: ATAGCCCAAACCGTTGGT
229–273 58 16 0.57 0.77 0.61 0.63
GB-AA-372 FJ647791 (TC)16 F: GGACTTCGGTCACCCTTC
R: CATCCAAAAACACCTCGG
149–215 57 10 0.51 0.59 0.71 0.67
GB-AA-374 FJ647792 (CT)17 F: GAACGAATCAGGAATCGAAA
R: TGAAGTGTAATAAAAGACTTCGCA
153–171 58 9 0.32 0.68 0.78 0.74
GB-AA-380 FJ647793 (CT)16 F: GGCAAACCTACACCCTCA
R: TTTTCTCGCTCCTCGTGA
296–326 57 9 0.57 0.02 0.56 0.52
GB-AA-393 FJ647794 (GA)7 F: CAAGCAGTGAAGATGCTTACC
R: CAGCTCAGGGGTCGACTA
118–216 57 20 0.22 0.68 0.87 0.88
GB-AA-398 FJ647795 (GA)11 F: CGGGAATGTGAAATCCTTT
R: CCAATTGCTTGGGAGTGA
233–267 57 10 0.40 0.65 0.75 0.72

 Mean 10.3 0.39 0.55 0.72 0.69

*Loci with significant deviations from Hardy-Weinberg equilibrium.

Ta, annealing temperature; MAF, major allele frequency; A, number of alleles; He, expected heterozygosity; Ho, observed heterozygosity; PIC, polymorphism information content.

Table 1 List of Actinidia species used in this study.
Table 2 Efficiency of the procedure adopted for the identification of microsatellite markers in A. arguta.

percentage based on the designed primer pairs.

Table 3 Frequency and type of di-, tri- and ≥ tetra-nucleotide repeats isolated from the A. arguta microsatellite-enriched library.
Table 4 Characteristics of 34 polymorphic microsatellite loci for Actinidia species.

Loci with significant deviations from Hardy-Weinberg equilibrium.

Ta, annealing temperature; MAF, major allele frequency; A, number of alleles; He, expected heterozygosity; Ho, observed heterozygosity; PIC, polymorphism information content.