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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (7): 1095-1102.doi: 10.3724/SP.J.1006.2018.01095

• RESEARCH NOTES • Previous Articles    

Development and Validation of Molecular Marker for Protein Content in Tetraploid Potato Tuber

Hong-Bo SHAN,Jia-Wen SHI,Ying SHI()   

  1. College of Agronomy, Northeast Agricultural University, Harbin 150030, Heilongjiang, China
  • Received:2018-01-16 Accepted:2018-03-26 Online:2018-07-10 Published:2018-04-27
  • Contact: Ying SHI E-mail:yshi@neau.edu.cn
  • Supported by:
    This study was supported by the National Key Research and Development Programs (2017YFD0101906).

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Abstract:

Protein content is one of the most important quality traits in potato. In this study, SCAR8-107, a protein content linked molecular marker, was developed using Atlantic, Dingshu 1, and their F1 segregation population consisting of 173 individuals by reduced-representation sequencing and bulked segregant analysis. A further testing of marker SCAR8-107 was performed using 74 high protein content individuals, 42 low protein content individuals and some tetraploid varieties, showing the selection accuracy of 90.51% and 72.97% between marker test and phenotyping in segregation population and tetraploid varieties respectively. Pearson’s correlation analysis also showed that the marker SCAR8-107 was significantly correlated to protein content in potato. The SCAR8-107 marker could be used in marker-assisted selection.

Key words: potato protein, marker development, BSA mixing pool sequencing, marker-assisted selection

Table1

Information of potato varieties"

序号
Code		 品种
Variety		 蛋白含量
Protein content (%)		 序号
Code		 品种
Variety		 蛋白含量
Protein content (%)
1 -37 1.18 28 Shlin-6 1.54
2 东农305 Dongnong 305 1.39 29 坝薯10号 Bashu 10 1.50
3 东农308 Dongnong 308 1.45 30 BC 1.45
4 东农310 Dongnong 310 2.21 31 大200305-32 Da 200305-32 1.50
5 东农311 Dongnong 311 1.17 32 大200410-19 Da 200410-19 2.15
6 东农312 Dongnong 312 1.68 33 大地3号 Dadi 3 1.53
7 东农313 Dongnong 313 1.26 34 定薯3号 Dingshu 3 2.01
8 东农314 Dongnong 314 1.68 35 东农03-1 Dongnong 03-1 1.44
9 东农316 Dongnong 316 1.44 36 费乌瑞它 Favourite 1.69
10 东农317 Dongnong 317 1.63 37 H9900-1 1.58
11 Allora 1.56 38 H99-9 2.12
12 B190 1.59 39 克新12 Kexin 12 2.06
13 B192 1.41 40 克新22 Kexin 22 1.98
14 B200 1.64 41 克新27 Kexin 27 1.68
15 B201 1.71 42 希森3号 Xisen 3 1.95
16 Carlar 2.30 43 夏坡蒂 Shepody 2.55
17 CIP39048 1.33 44 鑫科1号 Xinke 1 1.51
18 D412 1.45 45 兴佳2号 Xingjia 2 1.47
19 DESIREE 1.22 46 延薯3号 Yanshu 3 1.79
20 FW5 1.75 47 延薯4号 Yanshu 4 2.20
21 Gala 1.53 48 延薯6号 Yanshu 6 1.47
22 K200410-137 2.32 49 延薯7号 Yanshu 7 1.51
23 NY128 1.02 50 延薯8号 Yanshu 8 0.94
24 Q12 1.59 51 延薯97-8 Yanshu 97-8 1.71
25 早大白 Zaodabai 1.37 52 云薯201 Yunshu 201 1.18
26 中薯3号 Zhongshu 3 1.90 53 云薯301 Yunshu 301 2.09
27 中薯5号 Zhongshu 5 0.94 54 早50 Zao 50 1.59

Fig. 1

Distribution of group protein content"

Table 2

Determination of group protein content"

蛋白含量
Protein content (%)		 总数
Total number		 极端材料数量
Number of extreme individuals
极高蛋白 Extremely high protein >2.5 18 25 (>2.4)
高蛋白 High protein 2.0-2.5 56
中蛋白 Medium protein 1.5-2.0 57
中低蛋白 Medium-low protein 1.0-1.5 39
极低蛋白 Extremely low protein <1.0 3 25 (<1.3)

Table 3

Primer information"

引物名称
Primer name		 正向引物
Forward primer (5'-3')		 反向引物
Reverse primer (5'-3')		 退火温度
Anneal temp. (°C)		 产物大小
Product size (bp)
chr8-100 CTGTTGGTCCGTGGTCTGG CGAGAGAGTGAATGGGCTAAGG 58 798
chr8-101 TCGCACAAGAACAACCACAATC ATACATTCAGCCATTCAGACTTCG 59 474
chr8-102 CAACAAAGTCTGAATGGCTGAATG AGGCTATGCTGCTAAGGATGC 59 488
chr8-103 TGAGAGGTCATCGTCTGGTTG TGGCTGTGGTTTAAGAAGTTGG 59 717
chr8-104 GGATGGCTCGTGATTGTCTTG TTTACTTCATTGGGTCGTTCGTC 59 699
chr8-105 TTCAGCGAGGACATAGCACAG CTCACTACACTCTTGGCACATTG 58 892
chr8-106 ACTCATAAGCATCCTCATTCTCAG CTAACCTCCTTCACTCTTCTATCC 60 569
chr8-107 GATGTCTAATACTCCGTCAAGGTG TCAAAGCATTACAACAACAAAGGG 58 724
chr8-108 CGCCAAGCCTCCAACTACTG ACTTTGAATCCTCTTTCCAACTCC 58 651
chr8-109 TTACCGTCACATTACATTTCACAG CTCCCATACTCTCTTCTCTAAACC 58 544
chr8-110 ATGGGATGTGTCAAGGGTCTC CCTCTCACTTCACTCTTCTCAAC 59 772
chr8-111 TCAATCCACGGACACAGACC GCCTAAACCCTCAACTCAAACG 58 515
chr8-112 CAAAAGTGATGGACAAAGTGACG CCTATCTCCCTCTGCTGTTCC 58 807
chr8-113 CACCTGAATGGACGGATATAAGC TGGGAGTGATAAGTCTGGTATGG 58 686
chr8-114 ATCAAGAAGAGAACGAGCAAGAC CAGACCCACCACAACATCAATC 58 671
chr8-115 GCTATCCTGCTATTCTCCGAAG TCTGACCATCACGACATCTCC 59 461
chr8-116 AACATCTGACTGTCTATGAAGCC GAGCACCGATATGAGGGAGAG 61 733
chr8-117 CTTACCGCTTCAGGCTTTCTC TTGATAGTCCTCAGGCATAAACG 58 752
chr8-118 CTGGCTTATGAACTCGGTGTC ATTCCTTGCTGTGTCCTTTGC 58 575
chr8-119 GGTTCAATTAAAGGTGTGCTGTG ATGAGGTGAGGACAATGCTAGG 58 745
chr8-120 TCAACAAAGACCCACATCACAAG TCACCTTCACGAGCCACTTC 58 686
chr8-121 TGCCGCATTATTGTTGTTATTCG GCCGCTCTCAGTTCTCTCC 59 484
chr8-89 TCAGATCACGCATAAATCGGTTC TGGACATAAACACGGGACTTAGG 59 406
chr8-90 TTGCTTCCGTGAAAGTGTTATGG TCATCAGTGGCTTGGCTTACC 60 748
chr8-91 CCTCCAGAGCCACCACTTAC TTGAACTTCCAATGCTACCATCC 58 626
chr8-92 GAGGGTGGAAAGTCGTGTTATG AATGAATGTGGTGGTGGATGC 58 831
chr8-93 GAGGGTGGAAAGTCGTGTTATG TGGATGCTCTTAATTGGCTATGG 58 817
chr8-94 AGGTGTAAGTGTAGGTGTAGGTG GCGAGCATCTTGGAGTTAGC 58 464
chr8-95 ACAAGGTGAATTGGAAGATGCTC TGGAAGGACAATCTGCTACGAC 58 578
chr8-96 GCGATGCTGCTATATGGTTCAC CTCCTCACGAATCTCCTCTGC 58 453
chr8-97 CATCCTCCGCCACCCAAAG CAGAATCAGCAGCAGCATCC 58 835
chr8-98 GGAGGAATTTGCGTGGGAAAG GCCGTATGTTGTGAAGGAAGC 58 705
chr8-99 CTCATCAGCATCACACTTCACTC AAGGCAAGGGTAAAGGCAAATC 58 571

Fig. 2

PCR products of some primers in parents Dingshu 1 and AtlanticPrimer number: 8-106, 8-107, 8-108, 8-109, 8-110. The first and second lanes of each pair of primers are Dingshu 1 and Atlantic."

Fig. 3

PCR amplification results of primers 8-107 in partial extremely high protein and extremely low protein progeniesP1: low protein parent Dingshu 1; P2: high protein parent Atlantic; numbers 39, 46, 49, 50, and 51 are extremely high protein progeny materials; numbers 32, 43, 45, 53, and 54 are extremely low protein progeny materials."

Fig. 4

PCR amplification results of SCAR8-107 marks in partial high protein (A) and low protein (B) progeniesP1: low protein parent Dingshu 1; P2: high protein parent Atlantic; code of low protein content and high protein content progenies."

Table 4

Identification of phenotype for protein content and statistics of marker test results in partial samples of F1"

标记检测
Marker test		 样本总数
Total number of samples		 表型
Phenotype		 样本数
Number of samples		 百分比
Percentage (%)
阳性
Positive		 70 高蛋白含量 High protein content 67 95.71
低蛋白含量 Low protein content 3
阴性
Negative		 46 高蛋白含量 High protein content 8 82.61
低蛋白含量 Low protein content 38

Table 5

Identification of protein phenotype and statistics of marker test results in partial varieties"

标记检测
Marker test		 样本总数
Total number of samples		 表型
Phenotype		 样本数
Number of samples		 百分比
Percentage (%)
阳性
Positive		 14 高蛋白含量 High protein content 10 71.42
低蛋白含量 Low protein content 4
阴性
Negative		 23 高蛋白含量 High protein content 6 73.91
低蛋白含量 Low protein content 17

Fig. 5

PCR amplification results of SCAR8-107 marks in partial high protein (A) and low protein (B) varietiesP1: low protein parent Dingshu 1; P2: high protein parent Atlantic; 1-12: Shepody, K200410-137, Carlora, Dongnong 310, Yanshu 4, Da 200410-19, Hu H 99-9, Yunshu 301, Kexin 12, Dingshu 3, Kexin 22, Xisen 3; 13-24: Dongnong 316, B192, Dongnong 305, Zaodabai, CIP39048, Dongnong 313, DESIREE, Yunshu 201, Dongnong 311, NY128, Zhongshu 5, Yanshu 8."

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