Crops ›› 2016, Vol. 32 ›› Issue (2): 43-49.doi: 10.16035/j.issn.1001-7283.2016.02.008

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Development of a Set of 7S and 11S Multi-Subunit-Deficient Mutants with Chinese Soybean Genetic Background

Guo Bowen,Zhao Xue,Wei Xiaoshuang,Han Yanjing,Liang Qiangfei,Song Bo,Liu Shanshan   

  1. Key Laboratory of Soybean Biology in Chinese Ministry of Education,Northeast Agricultural University,Harbin 150030,Heilongjiang,China
  • Received:2015-12-23 Revised:2016-03-15 Online:2016-04-15 Published:2018-08-26
  • Contact: Shanshan Liu

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Abstract:

By different breeding methods, i.e. backcross, three-way cross combined pedigree selection, a novel set of multi-subunit-deficiency lines was developed by using a multi-subunit-deficiency breeding material RiB as subunit-deficiency donor parent, and Chinese cultivar Dongnong 47 as a genetic background. By sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis, seven band-deficiency types characterized by:(α+11S groupⅡa)-null, (α′+11S groupⅡa)-null, [(α′+α)+11S groupⅡa,Ⅱb]-null, [(α′+α)+11S groupⅡa]-null, [(α′+α)+11S groupⅡb+X1X2]-null, [(α′+α)+11S groupⅡb]-null and (α′+11S groupⅠ,Ⅱa)-null were detected. An comprehensive evaluation of the agronomic performance, amino acid composition and amino acid content analysis revealed that, compared with control, all 7 mutant genotypes showed a generally higher content of amino acids composition. Total protein content in these mutant seeds was higher than that of recurrent parent Dongnong 47, and arginine (Arg) content was especially enriched. The content of total amino acids, 17 amino acid compositions and sulfer-containing amino acids of novel line G2-2-3, which lacked (α+11S groupⅡa)-subunits, were all significantly higher than that of Dongnong 47, and its Arg content was 7.27mg/g higher than Dongnong 47. These results indicate that novel multi-subunit-null lines lacking allergen-subunits appear to contain markedly increased amino acid content and modified storage protein subunit composition. This set of multi-subunit-deficiency variants with Chinese soybean genetic background would be expected to be favorable material for soybean engineering breeding of modifying globulin composition and subunit composition projects.

Key words: Soybean, Storage protein, Chinese genetic background, Multi-subunit-deficient mutants, New germplasm

Fig.1

SDS-PAGE patterns of protein obtained form soybean lines have different 7S and 11S subunit deficiency composition."

Fig.2

The representative plants of 7S and 11S multi-subunit-null soybean lines"

Table 1

Protein, oil contents and performance of major agronomics of 7S and 11S multi-subunit-null soybean lines"

品系
Lines		 亚基表现
Subunit composition		 世代
Generation		 叶型
Blade profile		 花色
Flower color		 熟期(d)
Growth period		 株高(cm)
Plant height		 百粒重(g)
100-seed weight		 蛋白(%)
Protein		 油份(%)
Fat
东农47 7S、11S亚基表现正常 亲本 尖叶 白色 115 78.20±2.1680b 20.39±0.8862bc 40.92±0.6458cd 22.38±0.1643a
绥农10 7S、11S亚基表现正常 三交亲本 尖叶 白色 120 93.00±3.3912a 18.54±1.0896cd 41.02±0.8228cd 21.40±0.2646c
G2-2-3 (α+11S groupⅡa)-缺失型 三交F8 中间型 白色 132 72.00±3.9370bc 16.96±0.8961d 43.96±0.8143a 20.20±0.5568d
BC5-84 (αˊ+11S groupⅡa)-缺失型 BC1F6 尖叶 白色 130 68.00±3.5355c 22.23±1.5341ab 41.36±1.4293bcd 21.80±0.1899bc
BC5-148-1 [(α+αˊ)+11S groupⅡa、Ⅱb]-缺失型 BC1F5 尖叶 白、紫 138 76.00±5.2440b 21.31±0.3938b 40.72±1.2598cd 21.98±0.4712ab
BC5-94 [(α+αˊ)+11S groupⅡa]-缺失型 BC1F6 尖叶 白色 128 67.00±2.4495c 23.37±1.1481a 41.10±0.4848cd 21.98±0.3033ab
BC5-11-1 [(α+αˊ)+11S groupⅡb+X1X2]-缺失型 BC1F5 尖叶 白色 140 69.00±7.6812c 22.37±1.9067ab 42.80±1.1180ab 21.74±0.3209bc
BC5-11-2 [(α+αˊ)+11S groupⅡb]-缺失型 BC1F5 尖叶 白色 138 72.80±6.4962bc 18.94±2.2821cd 41.60±1.9634bc 21.74±0.5505bc
C4832 (αˊ+11S groupⅠ、Ⅱa)-缺失型 BC3F4 尖叶 白色 110 72.00±4.3589bc 17.86±1.7196d 39.84±0.8264d 22.08±0.2388ab

Table 2

Amino acid and free amino acid composition in soybean of Dongnong47、G2-2-3、BC5-84 and BC5-94"

氨基酸含量（mg/g）Amino acids 游离氨基酸含量（mg/g）Free amino acids
东农47
Dongnong47		 G2-2-3 BC5-84 BC5-94 东农47
Dongnong47		 G2-2-3 BC5-84 BC5-94
必需氨基酸Essential amino acids
Thr 13.20±0.10c 16.10±0.20a 15.30±0.26b 16.03±0.15a 0.2340±0.0030b 0.3330±0.0547a 0.2673±0.0673ab 0.3377±0.0321a
Val 14.90±0.30c 18.00±0.17a 16.70±0.46b 17.63±0.15a 0.1155±0.0015b 0.1153±0.0032b 0.1350±0.0151a 0.1307±0.0106ab
Met 3.70±0.10c 5.17±0.06a 4.47±0.06b 4.50±0.17b 0.0715±0.0025a 0.0720±0.0017a 0.0760±0.0122a 0.0713±0.0031a
Ile 14.50±0.30c 18.17±0.15a 17.40±0.36b 17.50±0.20b 0.0995±0.0025a 0.0763±0.0015b 0.1037±0.0107a 0.0987±0.0040a
Leu 27.00±0.30c 31.33±0.23a 29.53±0.55b 29.40±0.20b 0.1860±0.0060a 0.1090±0.0044c 0.1720±0.0128a 0.1537±0.0070b
Phe 18.17±0.21c 21.20±0.17a 19.67±0.32b 19.60±0.00b 0.1890±0.0010c 0.2107±0.0029bc 0.2340±0.0161ab 0.2517±0.0318a
Lys 22.83±0.21d 26.37±0.21a 24.47±0.42b 23.53±0.12c 0.2365±0.0085b 0.2600±0.0036ab 0.2947±0.0410a 0.2723±0.0144ab
非必需氨基酸Non-essential amino acids
Asp 38.20±0.79c 46.47±0.76a 42.73±0.81b 42.80±0.35b 0.6475±0.0475c 0.9000±0.0376b 0.9130±0.0467b 1.0103±0.0572a
Ser 17.93±0.31c 20.47±0.42a 19.17±0.32b 19.13±0.15b 0.0915±0.0015b 0.1193±0.0145ab 0.1117±0.0239ab 0.1307±0.0111a
Glu 60.40±1.05c 66.53±0.40a 61.90±0.87b 57.87±0.45d 0.4185±0.0465c 0.7137±0.0862b 0.8237±0.0723b 1.0900±0.0244a
Gly 14.67±0.6d 16.73±0.06a 15.43±0.31c 16.23±0.12b 0.0535±0.0005b 0.0860±0.0964a 0.0863±0.0257a 0.0897±0.0096a
Ala 14.60±0.20c 16.40±0.00a 15.73±0.40b 16.13±0.12ab 0.1290±0.0020c 0.1403±0.0133c 0.1993±0.0121b 0.2397±0.0153a
Cys 5.30±0.10b 5.80±0.17a 5.30±0.10b 5.37±0.15b 0.1670±0.0020c 0.2317±0.0168a 0.2007±0.0025b 0.1797±0.0032c
Tyr 11.30±0.17b 12.00±0.10a 10.97±0.32b 11.30±0.00b 0.1110±0.0030b 0.1540±0.0017a 0.1667±0.0341a 0.1570±0.0210a
His 8.87±0.12d 12.43±0.23a 10.40±0.17c 11.03±0.12b 0.0995±0.0115b 0.3560±0.0821a 0.3380±0.0389a 0.3247±0.0355a
Arg 23.27±0.35c 35.77±1.70a 31.87±0.91b 30.87±0.99b 1.1735±0.0445c 8.4473±1.8440a 5.8207±0.3857b 6.4193±0.7376b
Pro 18.33±0.45d 20.73±0.25a 19.57±0.21b 18.93±0.25c 0.1665±0.0005b 0.2050±0.0053a 0.1887±0.0194ab 0.1800±0.0131b
TAA 327.17±4.15c 389.67±4.71a 360.60±6.15b 357.87±1.25b 4.1895±0.1045c 12.5297±2.1430a 10.1320±0.6706b 11.1370±0.8595ab
含硫氨基酸Sulfur-cotaining amino acids
Met+Cys 9.00±0.20c 10.97±0.15a 9.77±0.12b 9.87±0.25b 0.2385±0.0045c 0.3037±0.0152a 0.2767±0.0100b 0.2510±0.0062c
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