Crops ›› 2018, Vol. 34 ›› Issue (3): 44-50.doi: 10.16035/j.issn.1001-7283.2018.03.007

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Identification and Screening of Protein Subunit Variation Germplasm from Both Mutants and Natural Population in Soybean

Zhang Mingjun1,2,Li Zhongfeng2,Yu Lili2,Wang Jun1,Qiu Lijuan1,2   

  1. 1 College of Agronomy, Yangtze University, Jingzhou 434020, Hubei, China
    2 Institute of Crop Science, Chinese Academy of Agricultural Sciences/The National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI)/Key Open Laboratory of Crop Germplasm Resources & Biotechnology, Ministry of Agriculture, Beijing 100081, China;
  • Received:2018-04-27 Revised:2018-05-20 Online:2018-06-20 Published:2018-06-20

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Abstract:

In this study, soybean seed storage protein from 298 EMS mutant families derived from Zhongpin 661 and 610 varieties (lines), was separated by polyacrylamide gel electrophoresis (SDS-PAGE), repectively, and then the Quantity One (V4.62) software was used to quantify the relative content of protein subunits and the 11S/7S value. The results showed that significant difference for coefficient of variation (CV) of different subunits was observed between mutants and natural population, and the variation range of alpha, beta and acidic protein subunits in mutants was all larger than that in natural population, and notably the β subunit displayed the largest CV in both mutants and natural population. Furthermore, correlation analysis showed that the relative content of 11S glycinin and 7S β-conglycinin were negatively correlated, the 11S/7S value and each subunit of 11S glycinin and 7S β-conglycinin were significantly correlated as well, while the correlation was not significant for crude protein content and 11S/7S value. Also some elite germplasm was identified, among which, six displayed obvious variation in protein subunits, and a novel subunit mutant designated as Ax was also identified. Besides, ten showed higher 11S/7S value than 3, and seven exhibited higher protein content than 48%. These seed protein variants here we identified will provide important material basis for the related gene mining and genetic improvement of seed protein quality in soybean.

Key words: Soybean, Protein subunits, 11S/7S, Mutant, SDS-PAGE

Fig.1

SDS-PAGE profile for soybean seed storage protein"

Fig.2

Optical density scanning of the Zhongpin 661 seed protein"

Fig.3

The frequency distribution of the relative content of soybean globulin 11S and 7S in both mutant and natural population"

Table 1

Variation analysis of soybean seed protein subunits content in natural and mutant population %"

蛋白亚基
Protein
subunit		 变异幅度Range 平均值±标准差Mean±SD 变异系数CV
自然群体
Natural
population		 突变群体
Mutant
population		 中品661
Zhongpin 661		 自然群体
Natural
population		 突变群体
Mutant
population		 中品661
Zhongpin 661		 自然群体
Natural
population		 突变群体
Mutant
population		 中品661
Zhongpin 661
Lox 0.01~27.09 3.22~26.40 5.53~13.06 7.68±2.59 8.84±3.26 9.53±2.42 33.72 36.93 25.43
α' 0.86~23.53 2.11~21.05 4.01~10.39 8.34±2.41 8.63±2.78 8.37±1.96 28.89 32.22 23.36
α 7.36~36.60 4.47~35.58 9.40~18.65 15.53±3.91 14.36±4.08 12.55±2.32 25.18 28.45 18.49
β 5.94~38.73 3.61~45.77 6.53~13.36 12.55±3.76 10.93±5.04 10.42±2.10 29.99 46.11 20.16
A3 1.37~12.21 0.40~10.30 1.74~8.85 5.58±1.53 5.13±2.25 5.67±1.80 27.50 43.78 31.74
Acidic 9.01~39.43 5.30~37.79 19.69~36.98 20.38±4.30 22.33±6.82 25.42±5.26 22.57 30.52 20.71
Basic 17.95~63.40 14.00~44.93 20.54~33.24 29.60±4.92 29.51±5.38 28.03±3.63 16.61 18.22 12.94
7S 20.84~56.92 13.25~60.39 25.53~35.60 36.42±4.54 33.99±5.09 31.35±2.63 12.46 18.50 8.39
11S 38.11~75.01 33.00~74.06 52.54~67.79 55.90±5.09 57.04±7.47 59.12±4.04 10.00 13.10 6.83
11S/7S 66.95~341.97 54.65~351.38 147.60~265.55 157.47±34.22 179.35±51.13 190.80±28.94 21.73 28.51 15.17

Fig.4

The frequency distribution of the ratio of 11S/7S in both natural and mutant population"

Table 2

Analysis of soybean seed protein content in mutants and natural population %"

材料
Material		 变异幅度
Range		 平均值
Mean		 标准差
SD		 变异系数
CV
自然群体Natural population 33.29~48.50 40.51 2.32 5.73
中品661 EMS突变体Mutant 35.57~49.04 44.36 2.01 4.54

Table 3

Correlation analysis of the relative content of soybean protein subunits between natural and mutant population"

α′ α β A3 Acidic Basic 7S 11S 11S/7S Protein
α′ -1 -0.12* -0.05 -0.11 -0.29** -0.05 -0.30** -0.26** -0.36** -0.03
α -0.33** -1 -0.08 -0.30** -0.28** -0.23** -0.47** -0.32** -0.51** -0.14*
β -0.27** -0.25** -1 -0.06 -0.33** -0.45** -0.74** -0.58** -0.59** -0.04
A3 -0.02 -0.21** -0.08 -1 -0.36** -0.31** -0.27** -0.24** -0.38** -0.03
Acidic -0.02 -0.03 -0.21** -0.37** -1 -0.01 -0.56** -0.75** -0.69** -0.01
Basic -0.20** -0.24** -0.40** -0.05 -0.28** -1 -0.52** -0.60** -0.53** -0.04
7S -0.14** -0.39** -0.73** -0.24** -0.24** -0.52** -1 -0.77** -0.93** -0.10
11S -0.16** -0.36** -0.57** -0.25** -0.40** -0.59** -0.80** -1 -0.85** -0.03
11S/7S -0.11* -0.44** -0.63** -0.27** -0.30** -0.58** -0.94** -0.92** -1 -0.09
Protein -0.11* -0.07 -0.08 -0.08 -0.14* -0.13* -0.09 -0.04 -0.10 -1

Table 4

The list of screened elite soybean germplasm"

名称Name 统一编号ID number 类型Type 来源Origin 11S/7S值11S/7S ratios 蛋白含量Content of protein (%)
16M4704 — α′缺失 突变群体 2.36 41.30
16M4960 — α′低 突变群体 1.78 44.01
16M647 — β低 突变群体 1.90 44.12
16M3980 — A3低 突变群体 2.16 44.54
汾豆5号Fendou 5 ZDD24776 Ax高 自然群体 1.29 45.17
吉育83 Jiyu 83 ZDD24461 11/7S高 自然群体 3.42 37.70
新大豆9号Xindadou 9 ZDD24674 11/7S高 自然群体 3.36 38.20
徐豆13 Xudou 13 ZDD24780 11/7S高 自然群体 3.36 40.00
北豆26 Beidou 26 ZDD24367 11/7S高 自然群体 3.31 38.50
16M2748 ZDD25880 11/7S高 突变群体 3.12 42.14
16M2768 — 11/7S高 突变群体 3.04 45.64
16M2784 — 11/7S高 突变群体 3.51 43.01
16M3127 — 11/7S高 突变群体 3.10 41.18
16M3178 — 11/7S高 突变群体 3.19 43.58
16M5083 — 11/7S高 突变群体 3.15 42.56
PSB368 WDD02748 高蛋白 自然群体 2.17 48.21
晋豆38 Jindou 38 ZDD24712 高蛋白 自然群体 1.83 48.50
16M2223 — 高蛋白 突变群体 1.51 48.43
16M2352 — 高蛋白 突变群体 1.39 48.62
16M2788 — 高蛋白 突变群体 2.61 49.04
16M3637 — 高蛋白 突变群体 2.01 48.30
16NF5437 — 高蛋白 突变群体 1.14 48.30
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