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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (11): 1631-1639.doi: 10.3724/SP.J.1006.2018.01631

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Genome-wide Screening and Evaluation of SNP Core Loci for Identification of Upland Cotton Varieties

Guo-Zhong ZHU,Fang ZHANG,Jie FU,Le-Chen LI,Er-Li NIU,Wang-Zhen GUO()   

  1. State Key Laboratory of Crop Genetics & Germplasm Enhancement / Hybrid Cotton R&D Engineering Research Center (the Ministry of Education) / Nanjing Agricultural University, Nanjing 210095, Jiangsu, China
  • Received:2018-02-25 Accepted:2018-08-20 Online:2018-11-12 Published:2018-09-04
  • Contact: Wang-Zhen GUO E-mail:moelab@njau.edu.cn
  • Supported by:
    This study was supported by the National Key R&D Program for Crop Breeding(2017YFD0102000);Jiangsu Collaborative Innovation Center for Modern Crop Production Project(No.10)

Abstract:

Utilizing the genome-wide SNP information to screen the core SNP loci may provide an accurate and efficient method for the identification of upland cotton varieties. Using the CottonSNP80K array, SNP genotyping was performed within 326 upland cotton accessions. Then, the SNP loci were annotated with TM-1 genomic sequence of Gossypium hirsutum (AD1) genome NBI v1.1 Upland cotton of Nanjing Agricultural University as reference sequence. Statistical analysis of all loci in CottonSNP80K showed that the call rate of 93.85% loci (72 990 in 77 774) was more than 99%, and 61 595 (79.20%) SNPs were polymorphic loci among the tested upland cotton accessions. Among them, minor allele frequency (MAF) of 76.32% (47 009) loci was greater than 0.1. Based on call frequency for each locus > 0.99; loci with polymorphism; MAF > 0.2; heterozygosity rate < 0.05; SNP density with ~400 kb/SNP in each chromosome, we obtained 4857 high-quality core SNP loci. The characteristic statistics of the core SNP loci combination showed that the average call rate was nearly 100%; the average MAF was 0.34; and the average heterozygosity was 0.02. Using these core SNPs, more than 99% of the materials could be identified accurately and effectively. In addition, the identification results of core SNP loci showed extremely significant linear correlation with that of CottonSNP80K. Taken together, a core combination containing 4857 SNP loci for fingerprint identification of upland cotton varieties is constructed, which can accurately identify the purity and reality of modern upland cotton varieties.

Key words: DNA array, fingerprint, SNP, core loci, Upland cotton

Fig. 1

Characteristic statistics of the SNPs on CottonSNP80K arrayThe abscissa represents statistical SNP characteristic parameters, involved in loci call frequency, minor allele frequency (MAF), heterozygosity and distance between adjacent SNPs. The ordinate represents the number of SNPs."

Table 1

Chromosome distribution of the core SNPs for fingerprint analysis"

染色体
Chr.
染色体长度
Chr. length
(Mb)
标记数Number of SNPs 分布密度
Density
(kb/SNP)
染色体
Chr.
染色体长度
Chr. length
(Mb)
标记数Number of SNPs 分布密度
Density
(kb/SNP)
A01 99.9 250 399.5 D01 61.5 153 401.7
A02 83.4 207 403.1 D02 67.3 169 398.1
A03 100.3 248 404.3 D03 46.7 118 395.7
A04 62.9 156 403.3 D04 51.5 128 402.0
A05 92.0 237 388.4 D05 61.9 156 397.0
A06 103.2 255 404.6 D06 64.3 164 392.0
A07 78.3 197 397.2 D07 55.3 138 400.8
A08 103.6 258 401.7 D08 65.9 164 401.8
A09 75.0 183 409.8 D09 51.0 128 398.4
A10 100.9 256 394.0 D10 63.4 156 406.2
A11 93.3 242 385.6 D11 66.1 169 391.1
A12 87.5 215 406.9 D12 59.1 150 394.1
A13 80.0 204 392.0 D13 60.5 156 388.0
Total 1160.2 2908 399.0 Total 774.4 1949 397.3

Table 2

Characteristic statistics of the core SNPs for fingerprint analysis"

类型
Type
最大值
Max.
最小值
Min.
平均值
Mean
检出率 Call frequency 1.00 0.99 1.00
最小等位基因频率 MAF 0.50 0.20 0.34
杂合率 Heterozygosity 0.05 0.00 0.02
多态信息含量 PIC 0.50 0.32 0.44

Fig. 2

Correlation analysis of genetic distance of the 312 upland cotton accessions The abscissa represents the genetic distance between the materials calculated by the total loci in CottonSNP80K. The ordinate represents the genetic distance between the materials calculated by the core loci."

Table 3

Detection of polymorphism between the near-isogenic lines of upland cotton using the core SNP loci combination"

材料
Accession
多态位点(总位点/核心位点)
No. of polymorphic loci
(total loci/core loci)
多态率(总位点/核心位点)
Polymorphic rate
(total loci/core loci)
新乡小吉和新乡小吉无绒无絮突变体
Xinxiangxiaoji linted-fuzzless vs. Xinxiangxiaoji lintless-fuzzless
12529/1215 0.161/0.250
徐州142和徐州142无绒无絮突变体
Xuzhou-142 vs. Xuzhou-142 lintless-fuzzless
14808/1488 0.190/0.306
7235和7235纤维突变体
7235 vs. 7235 mutant
19451/2546 0.250/0.524
TM-1和SL1-7-1突变体
TM-1 vs. SL1-7-1
20859/2188 0.268/0.450
TM-1和 MD-17突变体
TM-1 vs. MD-17
23814/2367 0.306/0.487
TM-1和显性光子N1突变体
TM-1 vs. N1 (dominant naked seed)
17678/2170 0.227/0.447
TM-1和隐性光子n2突变体
TM-1 vs. n2 (recessive naked seed)
19996/2446 0.257/0.503
TM-1和im突变体
TM-1 vs. im (immature fiber)
16333/3116 0.210/0.642

Table 4

Consistency analysis between array genotyping and SNP-PCR for the selected SNP loci"

SNP 染色体
Chr.
位置
Position (bp)
验证不一致数目
Number of mismatches
引物
Primer (5'-3')
TM3086 A01 86363489 1 F: CTTGATTTTCAATTCAACCAAAAAACTCCG
R: AAAATACATGGACGCAACTATCTGATGCATAATTT
TM4824 A02 45647806 0 F: TTGAATGAGTCTCATTTTATCAATTGACCTTTATTA
R: TCATAAGGTAATTTGGCAAGTAATTTTGAAAAAGTC
TM7830 A03 89740533 1 F: AAGCCTATATTTTGCCAACTTACCTTGTGCAG
R: ATTAAGTTAAAAAGTGCCACATGTCACAAAACCAT
TM9574 A04 51844539 0 F: GGCCCTAATCTTAGGCATAGTTTACCCCAAATAA
R: CTTAGGGTAGACGGATGGAGGGATGAGA
TM19508 A07 21138995 0 F: TGCGAGGGATTAAGTTTAGCTTGGAAAGC
R: CCATTAGTTGACAATCTTCAAGAACCTTACGAAAA
TM27528 A08 63064554 0 F: GCTCCCATAATTTTCTTTCACTTCGAGACG
R: AAGAGAAAGGTGAAAATTTTAAGTACAAAAGGTTGA
TM27748 A08 65768181 0 F: GGCTTTTACTGATTTACATTTTGAACAATGTTAAAG
R: GGCTGAATGCTCCTTTACCTTATTTATGGGTT
TM33913 A10 4141534 2 F: GCTCCCTTAAACCCGCAACCTTAGTCA
R: AGAAAGTTGTTTCGGCTGGTATTTTGAGGTT
TM53506 D03 2560865 0 F: ATATTTTCTTTTTCTTTTTTCGATAAGATAGAGCAA
R: CCAAGGTTAGTTAACTAGAAAAACTGACCGAATCAA
TM58088 D05 33818451 0 F: CTCGTATGTCACAAGACCATCAAAATTTAAATGAA
R: TGTATCCCCATTTTGAACACCCACATG
TM69807 D08 63425972 0 F: TTTGTCAATTCTTACCAATTATACCTTCCTACCAAG
R: TATGTTTTGAAAATGCAACACCTATAGAAAAAGCAA
TM72841 D09 47573336 0 F: CAGGAATCATTGGGGTTGGACAAGC
R: TATGCTTGACTAAGGGATGCATTATCATACATTGTT
TM75830 D11 13710651 0 F: CGCGTCGTAATCGAGTTTCACCGA
R: GGTGTCCGGTCCGGTGTGCTTA
TM80004 D13 163793 0 F: GTGGCCACTGAAGAGCAAGCTTAATGA
R: ATGCATGTTCAAGCTTCTTGGTAATGTCATAATC

Fig. 3

Verification on SNPs from array genotyping by SNP-PCR analysisM: DNA marker; 1: TM-1; 2: 7235; 3: J02-508 (7-50); 4: DPL16; 5: Emian 21; 6: Emian 23; 7: Guoxinmian 9; 8: Handan 885; 9: Heishanmian 1; 10: Ji 122; 11: Jummian 1; 12: Shannongmian 8; 13: Shanxi W1; 14: Shanxi W8; 15: Stoneville 2B; 16: Simian 3; 17: Wanmian 17; 18: Xinluzao 32; 19: Xinluzao 7; 20: Xinluzhong 26; 21: Xinluzhong 35; 22: Yumian 15; 23: Zhongmiansuo 12; 24: Zhongmiansuo 41."

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