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

Previous Articles     Next Articles

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

RichHTML

8

PDF (PC)

132

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
[1] Thanh V H, Shibasaki K . Major proteins of soybean seeds. A straight forward fractionation and their characterization. Journal of Agricultural & Food Chemistry, 1976,24(6):1117-1121.
doi: 10.1021/jf60208a030 pmid: 1033950
[2] Tsukada Y, Kitamura K, Harada K , et al. Genetic analysis of subunits of two major storage proteins (β-conglycinin and glycinin) in soybean seeds. Scandinavian Cardiovascular Journal, 1986,36(4):390-400.
[3] Paek N C, Imsande J, Shoemaker R C , et al. Nutritional control of soybean seed storage protein. Crop Science, 1997,37(37):498-503.
doi: 10.2135/cropsci1997.0011183X003700020031x
[4] Koshiyama I . Chemical and physical properties of a 7S protein in soybean globulins. Cereal Chemistry, 1968,45(9):394-404.
[5] Staswick P E, Hermodson M A, Nielsen N C . Identification of the acidic and basic subunit complexes of glycinin. Journal of Biological Chemistry, 1981,256(16):8752-8755.
doi: 10.1016/S0167-6911(81)80039-4 pmid: 7263683
[6] Kitamura K, Kaizuma N . Mutant strains with low level of subunits of 7S globulin in soybean (Glycine max L.) seed. Japanese Journal of Breeding, 2008,31(4):353-359.
doi: 10.1270/jsbbs1951.31.353
[7] Shimada S, Shimada H, Takada Y , et al. New soybean cultivar "Yumeminori" with low allergenicity and high content of 11S. Bulletin of the National Agricultural Research Center for Tohoku Region, 2004.
[8] Takahashi K, Mizuno Y, Yumoto S , et al. Inheritance of the α-subunit deficiency of β-conglycinin in soybean (Glycine max L.) line induced by γ-ray irradiation. Breeding Science, 1996,46(3):251-255.
[9] Yagasaki K, Kalzuma N, Kitamura K . Inheritance of glycinin subunits and characterization of glycinin molecules lacking the subunits in soybean (Glycine max L.). Breeding Science, 1996,46(1):11-15.
[10] Kiamura K . Genetic improvement of nutritional and food processing quality in soybean. Japan Agricultural Research Quarterly, 1995,29(12):926-931.
doi: 10.6013/jbrewsocjapan1988.89.926
[11] 刘春, 王显生, 张占琴 , 等. 大豆种子贮藏蛋白亚基含量变异种质的筛选与创制. 湖南农业大学学报(自然科学版), 2008,34(3):249-255.
[12] 杨玛丽 . 大豆理化诱变突变体库的构建及大豆单链特异性核酸酶基因的克隆. 南京:南京农业大学, 2007.
[13] 关荣霞, 常汝镇, 邱丽娟 , 等. 栽培大豆蛋白亚基11S/7S组成及过敏蛋白缺失分析. 作物学报, 2004,30(11):1076-1079.
doi: 10.3321/j.issn:0496-3490.2004.11.002
[14] 郭顺堂, 孟岩, 张雪梅 , 等. 中国大豆蛋白亚基构成分析与缺失部分亚基的特异大豆品种的筛选. 作物学报, 2006,32(8):1130-1134.
[15] 宋波, 蓝岚, 田福东 , 等. 大豆7S球蛋白α'亚基缺失及(α'+α)亚基双缺失品系的回交转育. 作物学报, 2012,38(12):2297-2305.
doi: 10.3724/SP.J.1006.2012.02297
[16] 刘珊珊, 滕卫丽, 姜自芹 , 等. 大豆7S球蛋白α-亚基缺失型种质创新. 作物学报, 2010,36(8):1409-1413.
doi: 10.3724/SP.J.1006.2010.01409
[17] Hayashi M, Nishioka M, Kitamura K , et al. Identification of AFLP markers tightly linked to the gene for deficiency of the 7S globulin in soybean seed and characterization of abnormal phenotypes involved in the mutation. Breeding Science, 2000,50(2):123-129.
[18] Liu S, Kazuyoshi O, Dong C , et al. Genetic diversity of soybean (Glycine max L.) 7S globulin protein subunits. Genetic Resources & Crop Evolution, 2006,53(6):1209-1219.
doi: 10.1007/s10722-005-2434-y
[19] Li Z F, Guo Y, Ou L , et al. Identification of the dwarf gene GmDW1 in soybean (Glycine max L.) by combining mapping-by-sequencing and linkage analysis. Theoretical & Applied Genetics, 2018,131(5):1001-1016.
[20] Poysa V, Yu L W . Effect of soy protein subunit composition on tofu quality. Food Research International, 2006,39(3):309-317.
doi: 10.1016/j.foodres.2005.08.003
[21] 姜莹, 吴娴静, 董德坤 , 等. 浙江省大豆种质资源蛋白亚基构成分析. 浙江农业学报, 2010,22(4):403-407.
[22] 王丽侠, 郭顺堂, 付翠真 , 等. 大豆种子贮藏蛋白11S与7S组分的研究. 中国粮油学报, 2004,19(4):54-59.
[23] 徐豹, 林忠平 . 野生大豆(G. soja)种子贮藏蛋白组份11S/7S的研究. 作物学报, 1990,16(3):235-241.
[24] 胡超, 黄丽华, 李文哲 . 大豆球蛋白11S/7S比值对大豆蛋白功能性的影响. 中国粮油学报, 2004,19(1):40-42.
[25] 卢为国, 王树峰, 李卫东 , 等. 大豆籽粒贮藏蛋白11S/7S比值与生态因子相关关系的研究. 中国农业科学, 2005,38(5):1059-1064.
[26] 王燕平, 李贵全, 郭数进 , 等. 山西不同生态型大豆种质资源蛋白亚基的变异. 生态学报, 2011,31(1):203-211.
[27] 王义谅, 袁洪伟, 高山 . 快中子和EMS诱发大豆高蛋白和高油分突变的研究. 辽宁农业科学, 1980(6):1-8.
[1] Yun Zhao,Cailong Xu,Xu Yang,Suzhen Li,Jing Zhou,Jicun Li,Tianfu Han,Cunxiang Wu. Effects of Sowing Methods on Seedling Stand and Production Profit of Summer Soybean under Wheat-Soybean System [J]. Crops, 2018, 34(4): 114-120.
[2] Li Zhang,Zantang Li,Shiyin Wang,Yanchao Ma,Yang Dongfang,Xueyong Li,Jiang Xu. Physiological and Genetic Analysis of Rice Mutant osnad1 Defective in Nitrogen Absorption [J]. Crops, 2018, 34(3): 68-76.
[3] Jiani Zhu,Huiping Dai,Shuhe Wei,Genliang Jia,Dejing Chen,Jinjin Pei,Qing Zhang,Long Qiang. Effects of Applying Zn on the Growth and Zn Accumulation in Soybean at Flowering Stage [J]. Crops, 2018, 34(1): 152-155.
[4] Tianle Ma,Jianxin Zhang. Effects of Different Multiple Cropping Methods on Dry Matter Accumulation, Distribution and Yield of Summer Soybean [J]. Crops, 2018, 34(1): 156-159.
[5] Linjian Dai,Wu Chen,Tian Zhou,Jun Zhong. Selection Mutant of Resistant to Black Shank on Tobacco by Anther Culture [J]. Crops, 2018, 34(1): 66-70.
[6] Lina Li,Longguo Jin,Chuanxiao Xie,Changlin Liu. Determining Blind Samples of Transgenic Maize and Transgenic Soybean [J]. Crops, 2017, 33(6): 37-44.
[7] Ying Jiang,Yufeng Sun,Dongmei Pan,Qiuzhi Li. EMS Mutant Screening and RAPD Analysis on Industrial Hemp (Cannabis sativa L.) [J]. Crops, 2017, 33(6): 50-54.
[8] Xuechao Zhou,Surong Ding,Yunshan Wei,Yanfang Zhou,Xue Wei,Rina Na,Feng Li. Evaluation of Adaptability of Different Vegetable Soybean Cultivars (Lines) in Chifeng Area [J]. Crops, 2017, 33(3): 44-48.
[9] Guoyong Ren,Wei Li,Lifeng Zhang,Caijie Wang,Haiying Dai,Jinlong Wang,Ran Xu,Yanwei Zhang. The Resistance to Soybean Cyst Nematode Race 1 of Transgenic Soybean with hrf2 Encoding HarpinXooc [J]. Crops, 2017, 33(3): 49-53.
[10] Xuli Zhang,Baolong Xing,Guimei Wang,Lili Yin. Effects of Planting Density on Agronomic Traits, Economic Traits and Yield of Soybean in North of Shanxi Province [J]. Crops, 2017, 33(3): 127-131.
[11] Xiting Zhang,Liwei Cao,Shucai Lü,Guoxing Chen,Yongji Wang,Shuhan Yu,Zhenping Gong. Effects of Bulk Density on Nitrogen Absorption and Yield of Soybean on Black Soil [J]. Crops, 2017, 33(3): 132-137.
[12] Yixin Tian,Fengju Gao. The Response of Growth and Dry Matter Accumulation and Distribution of High Protein Soybean to Plant Density [J]. Crops, 2017, 33(2): 121-125.
[13] Li Yan,Qiang Yang,Yupeng Shao,Dandan Li,Zhikun Wang,Wenbin Li. Cloning and Sequence Analysis of GmWRI1a Gene Promoter in Soybean [J]. Crops, 2017, 33(2): 51-58.
[14] Yu Gao,Yanchao Liu,Shusen Shi,Juan Cui,Jinfeng Xiong. Review on Soybean Thrips in China [J]. Crops, 2017, 33(1): 8-13.
[15] Haiyan Li,Deli Cai,Jingsheng Chen,Yuxi Duan,Lijie Chen,Yingyu Shang. The Influence of Resistant and Susceptible Soybean Germplasm on Growth Dynamic Changes of Soybean Cyst Nematode Race 3（SCN3） [J]. Crops, 2017, 33(1): 144-149.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Guangcai Zhao,Xuhong Chang,Demei Wang,Zhiqiang Tao,Yanjie Wang,Yushuang Yang,Yingjie Zhu. General Situation and Development of Wheat Production[J]. Crops, 2018, 34(4): 1 -7 .
[2] Baoquan Quan,Dongmei Bai,Yuexia Tian,Yunyun Xue. Effects of Different Leaf-Peg Ratio on Photosynthesis and Yield of Peanut[J]. Crops, 2018, 34(4): 102 -105 .
[3] Xuefang Huang,Mingjing Huang,Huatao Liu,Cong Zhao,Juanling Wang. Effects of Annual Precipitation and Population Density on Tiller-Earing and Yield of Zhangzagu 5 under Film Mulching and Hole Sowing[J]. Crops, 2018, 34(4): 106 -113 .
[4] Wenhui Huang, Hui Wang, Desheng Mei. Research Progress on Lodging Resistance of Crops[J]. Crops, 2018, 34(4): 13 -19 .
[5] Yun Zhao,Cailong Xu,Xu Yang,Suzhen Li,Jing Zhou,Jicun Li,Tianfu Han,Cunxiang Wu. Effects of Sowing Methods on Seedling Stand and Production Profit of Summer Soybean under Wheat-Soybean System[J]. Crops, 2018, 34(4): 114 -120 .
[6] Mei Lu,Min Sun,Aixia Ren,Miaomiao Lei,Lingzhu Xue,Zhiqiang Gao. Effects of Spraying Foliar Fertilizers on Dryland Wheat Growth and the Correlation with Yield Formation[J]. Crops, 2018, 34(4): 121 -125 .
[7] Xiaofei Wang,Haijun Xu,Mengqiao Guo,Yu Xiao,Xinyu Cheng,Shuxia Liu,Xiangjun Guan,Yaokun Wu,Weihua Zhao,Guojiang Wei. Effects of Sowing Date, Density and Fertilizer Utilization Rate on the Yield of Oilseed Perilla frutescens in Cold Area[J]. Crops, 2018, 34(4): 126 -130 .
[8] Pengjin Zhu,Xinhua Pang,Chun Liang,Qinliang Tan,Lin Yan,Quanguang Zhou,Kewei Ou. Effects of Cold Stress on Reactive Oxygen Metabolism and Antioxidant Enzyme Activities of Sugarcane Seedlings[J]. Crops, 2018, 34(4): 131 -137 .
[9] Jie Gao,Qingfeng Li,Qiu Peng,Xiaoyan Jiao,Jinsong Wang. Effects of Different Nutrient Combinations on Plant Production and Nitrogen, Phosphorus and Potassium Utilization Characteristics in Waxy Sorghum[J]. Crops, 2018, 34(4): 138 -142 .
[10] Na Shang,Zhongxu Yang,Qiuzhi Li,Huihui Yin,Shihong Wang,Haitao Li,Tong Li,Han Zhang. Response of Cotton with Vegetative Branches to Plant Density in the Western of Shandong Province[J]. Crops, 2018, 34(4): 143 -148 .