Construction of EMS Mutagenesis Mung Bean Mutant Library and Phenotypic Analysis

doi:10.11924/j.issn.1000-6850.casb19020066

Abstract

Abstract:

To find out the optimal concentration and treatment time of ethyl methyl sulfonate (EMS) for mung bean mutant library construction, ‘Wankelü 3’ was used as material, 16 treatment combinations including 4 EMS concentrations (0.6%, 1.0%, 1.4% and 1.8%) and 4 kinds of treatment time (6, 10, 14 and 18 h) were employed to treat the seeds, and the semi-lethal dose was selected for mutant library construction. The results showed that the optimal condition for ‘Wankelü 3’ mutagenesis was 1.0%/10 h. 6000 mature seeds of ‘Wankelü 3’ were treated under this optimal condition, 998 M₁ lines were obtained. Traits such as leaf color, leaf shape, plant height, flower morphology and flowering time of M₂ generation plants were investigated. A total of 324 mutants were obtained, and the mutation frequency was 2.97%. Among these mutants, leaf color mutation and dwarf were the dominated variation types, accounting for 1.10 % and 0.46%, respectively. Mutation sites of 4 mutant materials were determined by genome re-sequencing, the result showed that their base mutation frequency was 1/37.78 kb. These results also demonstrated that using EMS mutagenesis method, abundant phenotypic variant materials could be obtained, which could be used in genetic improvement and genome functional analysis for mung bean.

Key words: mung bean, ethyl methane sulfonate (EMS), mutant library, phenotype, sequencing

CLC Number:

S322

Ye Weijun, Yang Yong, Zhang Liya, Tian Dongfeng, Zhang Lingling, Zhou Bin. Construction of EMS Mutagenesis Mung Bean Mutant Library and Phenotypic Analysis[J]. Chinese Agricultural Science Bulletin, 2020, 36(17): 36-41.

Figures/Tables 4

References 30

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性状	变异类型	突变体株数	突变频率/%
叶片	叶色	120	1.10
叶片	叶型	32	0.29
茎	高秆	12	0.11
	矮秆	50	0.46
	畸形茎	21	0.19
	茎色	3	0.03
花	畸形花	4	0.04
花期	早花	19	0.17
花期	晚花	25	0.23
育性	半不育	32	0.29
育性	全不育	15	0.14
荚	长荚	6	0.06
荚	短荚	25	0.23
合计		364	3.34
复合性状突变		40	0.37
实际数量		324	2.97

性状	变异类型	突变体株数	突变频率/%
叶片	叶色	120	1.10
叶片	叶型	32	0.29
茎	高秆	12	0.11
	矮秆	50	0.46
	畸形茎	21	0.19
	茎色	3	0.03
花	畸形花	4	0.04
花期	早花	19	0.17
花期	晚花	25	0.23
育性	半不育	32	0.29
育性	全不育	15	0.14
荚	长荚	6	0.06
荚	短荚	25	0.23
合计		364	3.34
复合性状突变		40	0.37
实际数量		324	2.97

编号	测序深度/×	测序总碱基数/bp	突变碱基数/bp	突变频率/1/kb
ME1	18.33	7883350874	161372	48.85
ME2	20.15	8666094933	156116	55.51
ME3	18.20	7827440585	157308	49.76
ME4	17.85	7664010506	161813	47.36
平均	18.63	8010224225	159152	50.37

编号	测序深度/×	测序总碱基数/bp	突变碱基数/bp	突变频率/1/kb
ME1	18.33	7883350874	161372	48.85
ME2	20.15	8666094933	156116	55.51
ME3	18.20	7827440585	157308	49.76
ME4	17.85	7664010506	161813	47.36
平均	18.63	8010224225	159152	50.37