Crops ›› 2018, Vol. 34 ›› Issue (6): 53-57.doi: 10.16035/j.issn.1001-7283.2018.06.009

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Genetic Diversity of Flax Germplasm Resources in Inner Mongolia

Yi Liuxi1,Siqin Bateer1,Gao Fengyun1,Zhou Yu1,Jia Xiaoyun1,Chen Mingzhe2,Zhang Hui1,Jia Haibin3   

  1. 1 Biotechnology research center of the Inner Mongolia Academy of Agriculture and Husbandry Sciences, Huhhot 010031, Inner Mongolia, China
    2 Xilinguole Meng Institute of Agricultural Sciences, Xilinhaote 026000, Inner Mongolia, China
    3 Wulanchabu Academy of Agriculture and Animal Husbandry, Jining 012000, Inner Mongolia, China
  • Received:2018-04-18 Revised:2018-08-23 Online:2018-12-15 Published:2018-12-06

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Abstract:

In order to further understand the genetic diversity and genetic relationship of Inner Mongolia flax local varieties, the genetic diversity of 23 flax local varieties was analyzed using SRAP markers. The optimum SRAP PCR system contained 0.3μL of forward and reverse primer (10μmol/L), 9.0μL of 2×Taq Master Mix and 40ng template DNA in 25μL. 219 bands were produced by the 18 pairs of primers with an average of 12.17 bands for each pair of primers. 136 polymorphic bands were obtained and the polymorphic band proportion was 62.10%. The number of observed allele was 2.00, with 1.59 effective alleles for each locus. The average polymorphic information content was 0.61. The effective allele number (Ne), Shannon’s information index (I), and Nei’s genetic similarity coefficient (H) were 1.5630, 0.6715 and 0.438, respectively. When H was 0.55, the population was divided into 2 groups, when H was 0.38, the first population was dividid into 3 sub-groups. It could be concluded that the Inner Mongolia flax local varieties possess abundant genetic diversity and the flax varieties cultivated in the same area are closely related.

Key words: Flax, Inner Mongolia, Genetic diversity, SRAP

Table 1

Name and code of 23 flax cultivars (lines)"

编号Code 品种（系）Cultivar (Line)
1 H919
2 H920
3 H921
4 H922
5 内亚油一号Neiyayouyihao
6 临河17号Linhe No.17
7 集宁1号Jining No.1
8 集宁2号Jining No.2
9 轮选3号Lunxuan No.3
10 轮选2号Lunxuan No.2
11 轮选1号Lunxuan No.1
编号Code 品种（系）Cultivar (Line)
12 内蒙红Neimenghong
13 内亚2号Neiya No.2
14 CH-89
15 内亚5号Neiya No.5
16 内亚6号Neiya No.6
17 内亚7号Neiya No.7
18 华德小胡麻Huadexiaohuma
19 华德大粒高杆Huadedaligaogan
20 多伦小胡麻Duolunxiaohuma
21 多伦大胡麻Duolundahuma
22 后旗普通胡麻Houqiputonghuma
23 乌拉特中旗3号Wulatezhongqi No.3

Table 2

SRAP primer sequences used in the experiment"

正向引物Forward primer 反向引物Reverse primer
名称Name 序列Sequence (5'→3') 名称Name 序列Sequence (5'→3')
M1 5′-TGAGTCCAAACCGGCAT-3′ E1 5′-GACTGCGTACGAATTGTA-3′
M2 5′-TGAGTCCAAACCGGACA-3′ E2 5′-GACTGCGTACGAATTCGA-3′
M3 5′-TGAGTCCAAACCGGAAG-3′ E3 5′-GACTGCGTACGAATTGTC-3′
M4 5′-TGAGTCCAAACCGGATG-3′ E4 5′-GACTGCGTACGAATTCAA-3′
M5 5′-TGAGTCCAAACCGGTGC-3′ E5 5′-GACTGCGTACGAATTGTC-3′
M6 5′-TGAGTCCAAACCGGAAT-3′ E6 5′-GACTGCGTACGAATTCTA-3′
M7 5′-TGAGTCCAAACCGGTGC-3′ E7 5′-GACTGCGTACGAATTCGA-3′
M8 5′-TGAGTCCAAACCGGACG-3′ E8 5′-GACTGCGTACGAATTTGA-3′
M9 5′-TGAGTCCAAACCGGTCC-3′ E9 5′-GACTGCGTACGAATTGTA-3′
M10 5′-GACTGCGTACGAATTACT-3′ E10 5′-GACTGCGTACGAATTAGC-3′
M11 5′-TGAGTCCAAACCGGAAC-3′ E11 5′-GACTGCGTACGAATTACT-3′
M12 5′-GACTGCGTACGAATTCGA-3′ E12 5′-GACTGCGTACGAATTAAT-3′

Table 3

Orthogonal design L16(4)3 of reaction system"

编号
Code		 引物量(μL)
Primer quantity		 2×Taq (μL)
Master Mix		 DNA模板量(ng)
DNA template quantity
1 0.1 7.0 25
2 0.1 9.0 30
3 0.1 12.0 35
4 0.1 15.0 40
5 0.2 7.0 30
6 0.2 9.0 25
7 0.2 12.0 40
8 0.2 15.0 35
9 0.3 7.0 35
10 0.3 9.0 40
11 0.3 12.0 25
12 0.3 15.0 30
13 0.5 7.0 40
14 0.5 9.0 35
15 0.5 12.0 30
16 0.5 15.0 25

Fig.1

Amplification of SRAP-PCR system under orthogonal design M, 1500bp DNA marker; 1~16, Orthogonal test combination"

Table 4

Amplification of flax with the 18 pairs of SRAP primer"

引物名称
Primer name		 总位点数
Total loci		 多态性位点数Polymorphic loci 多态性位点百分率(%)
Percentage of polymorphic loci		 有效等位基因数
Effective alleles		 PIC
M9E11 12.00 7.00 58.33 1.61 0.59
M1E5 11.00 8.00 72.73 1.71 0.63
M8E2 13.00 9.00 69.23 1.62 0.58
M10E9 14.00 10.00 71.43 1.73 0.67
M6E11 10.00 7.00 70.00 1.49 0.65
M1E9 12.00 8.00 66.67 1.60 0.70
M5E2 13.00 8.00 61.54 1.66 0.54
M7E4 16.00 9.00 56.25 1.55 0.62
M4E4 12.00 7.00 58.33 1.53 0.66
M4E10 11.00 6.00 54.55 1.51 0.62
M3E5 14.00 8.00 57.14 1.69 0.57
M9E6 15.00 9.00 60.00 1.57 0.64
M10E3 13.00 7.00 53.85 1.44 0.61
M4E7 11.00 8.00 72.73 1.65 0.68
M5E6 10.00 5.00 50.00 1.49 0.51
M3E3 9.00 6.00 66.67 1.67 0.58
M10E6 11.00 7.00 63.64 1.55 0.55
M1E3 12.00 7.00 58.33 1.59 0.61
总计Total 219.00 136.00 1 121.42 28.66 11.01
平均值Average 12.17 7.56 62.30 1.59 0.61

Fig.2

Clustering of 23 flax cultivars with SRAP markers A, First group of cultivars; B, Second group of cultivars; Ⅰ, Ⅱ, Ⅲ, Represent the 3 sub-groups; 1~23 corresponding materials as table 1"

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