大豆子粒蛋白亚基变异种质的鉴定与筛选

doi:10.16035/j.issn.1001-7283.2018.03.007

摘要/Abstract

摘要：

本研究以大豆优良品种中品661的298份甲基磺酸乙酯（EMS）诱变株系和610份大豆品种（系）为试验材料,利用聚丙烯酰胺凝胶电泳（SDS-PAGE）技术,分离大豆种子蛋白并计算各蛋白亚基的相对含量及11S/7S值。研究结果表明,突变群体与自然群体中不同亚基的变异系数差异极显著,但均以β亚基变异范围最大。另外,突变群体的α、β和酸性蛋白亚基的变异范围均大于自然群体。相关性分析显示：11S球蛋白与7S球蛋白含量呈极显著负相关;11S/7S值与蛋白含量相关性不显著,11S/7S值与11S和7S球蛋白各组成亚基均呈显著相关。此外,本研究还筛选出亚基明显变异材料6份,其中Ax亚基突变体尚未见报道,11S/7S值大于3的材料10份,蛋白含量大于48%的材料7份。本研究鉴定和筛选的种子蛋白变异种质为大豆品质相关基因发掘和品种改良提供了材料基础。

关键词: 大豆, 蛋白亚基, 11S/7S, 突变体, SDS-PAGE

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

张明俊,李忠峰,于莉莉,王俊,邱丽娟. 大豆子粒蛋白亚基变异种质的鉴定与筛选[J]. 作物杂志, 2018 (3): 44–50

Mingjun Zhang,Zhongfeng Li,Lili Yu,Jun Wang,Lijuan Qiu. Identification and Screening of Protein Subunit Variation Germplasm from Both Mutants and Natural Population in Soybean[J]. Crops, 2018(3): 44–50

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参考文献 27

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蛋白亚基 Protein subunit	变异幅度Range			平均值±标准差Mean±SD			变异系数CV
蛋白亚基 Protein subunit	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

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

	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

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