Construction and Phenotypic Variation Analysis of Ningchun No. 4 Wheat EMS Mutant Library

doi:10.13560/j.cnki.biotech.bull.1985.2025-0027

Abstract

Abstract:

Objective To accelerate the creation of high-quality germplasm resources for saline wheat and to enrich genetic diversity, we constructed Ningchun No. 4 EMS mutant library. Method In this study, 10 000 seeds of the wheat variety Ningchun No. 4 were treated with 0.5% ethyl methanesulfonate (EMS) solution, and a library of M₁ generation mutants was constructed and statistically analyzed for the mutation rates of their phenotypic traits. In addition, the M₁ generation material was subjected to targeted screening with 250 mmol/L NaCl solution to obtain salt-tolerant mutants, and the mutants were phenotyped as well as measured for morphological indices, substance accumulation, and physiological indices. Result The 3 709 plants survived in M₁ generation with a survival rate of 37.09%, of which 1 226 had phenotypic mutant traits with a mutation rate of 33.05%. The 1 226 plants were rich in mutation types, containing 151 mutations for leaf traits, 212 mutations for spike traits, 161 mutations for tillers, 133 mutations for plant height, 130 mutations for fertility and fecundity, and 75 mutations for other traits, with mutation rates of 13.89%, 5.72%, 4.34%, 3.59%, 3.50%, and 2.02%, respectively. After salt stress treatment, Ningchun No. 4 showed yellowing and wilting of leaves, significant differences in seedling height, maximum root length, number of primordial roots, aboveground fresh weight, root fresh weight, aboveground dry weight, and root-crown ratio compared to the mutant, while root dry weight showed no significant change. In terms of physiological indices, malondialdehyde (MDA) content and cell membrane permeability of the mutants were significantly lower than those of the wild type, proline content, peroxidase (POD) and catalase (CAT) activities were significantly higher than those of the wild type, and there was no significant difference in the soluble sugar content. Conclusion The types of mutants produced by EMS mutagenesis from Ningchun No. 4 are abundant, providing rich germplasm resources for rapid localization of wheat genes for excellent traits and study of their functions.

Key words: wheat, Ningchun No. 4, EMS mutagenesis, mutant library construction, statistical analysis of phenotypic variation

DOU Fei-fei, REN Yu-zhao, WANG Shi-lei, LIU Chun-ying, WANG Xiao-dong, WANG Zhao-yi, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng. Construction and Phenotypic Variation Analysis of Ningchun No. 4 Wheat EMS Mutant Library[J]. Biotechnology Bulletin, 2025, 41(8): 92-101.

Figures/Tables 14

References 31

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EMS浓度 EMS concentration （%）	发芽率 Germination rate （%）	相对致死率 Relative lethality rate （%）
0 （CK）	92.0a	0
0.3	54.2b	41.1
0.5	45.7c	50.3
0.7	35.6d	61.3

EMS浓度 EMS concentration （%）	发芽率 Germination rate （%）	相对致死率 Relative lethality rate （%）
0 （CK）	92.0a	0
0.3	54.2b	41.1
0.5	45.7c	50.3
0.7	35.6d	61.3

性状 Trait	表型 Phenotype	突变数量 Number of mutations	突变比率 Mutation ratio （%）
叶部 Leaf	间绿 Mesophyllous green	36	0.97
	单边失绿带 Single-sided loss of green tap	98	2.64
	两侧失绿带 Loss of green belt on both sides	22	0.59
	半边失绿 Lose one half of one’s head	7	0.19
	中间失绿带 Lose the center green zone	54	1.46
	叶变黄 Lose color	206	5.55
	卷叶 Leafroll	92	2.48
穗 Spike	双头穗 Double-headed spikelet	42	1.13
	三头穗 Three-headed spikelet	16	0.43
	弯穗 Bent spikelet	47	1.27
	密穗 Dense spikelet	2	0.05
	不育穗 Sterile spikelet	3	0.08
	畸形穗 Distorted spike	30	0.81
	短穗 Short spikelet	72	1.94
分蘖 Tiller	独秆 Solitary culm	96	2.59
	寡蘖 Few tiller	49	1.32
	多蘖 Multiple tiller	16	0.43
株高 Plant height	高秆 Tall stalks	12	0.32
株高 Plant height	矮秆 Short stalks	121	3.26
育性和生育期 Fertility and maturity	早熟 Early-maturingt	27	0.73
	晚熟 Late-maturing	53	1.43
	不育 Sterile	50	1.35
其他 Others	叶环腐斑 Leaf ring rot	23	0.62
	叶黄斑 Leaf yellow spot	20	0.54
	叶白斑 Leaf white spot	21	0.57
	叶白线 White lines on the leaf	9	0.24
	叶黑线 Black lines on the leaf	2	0.05
合计 Total		1 226	33.05

性状 Trait	表型 Phenotype	突变数量 Number of mutations	突变比率 Mutation ratio （%）
叶部 Leaf	间绿 Mesophyllous green	36	0.97
	单边失绿带 Single-sided loss of green tap	98	2.64
	两侧失绿带 Loss of green belt on both sides	22	0.59
	半边失绿 Lose one half of one’s head	7	0.19
	中间失绿带 Lose the center green zone	54	1.46
	叶变黄 Lose color	206	5.55
	卷叶 Leafroll	92	2.48
穗 Spike	双头穗 Double-headed spikelet	42	1.13
	三头穗 Three-headed spikelet	16	0.43
	弯穗 Bent spikelet	47	1.27
	密穗 Dense spikelet	2	0.05
	不育穗 Sterile spikelet	3	0.08
	畸形穗 Distorted spike	30	0.81
	短穗 Short spikelet	72	1.94
分蘖 Tiller	独秆 Solitary culm	96	2.59
	寡蘖 Few tiller	49	1.32
	多蘖 Multiple tiller	16	0.43
株高 Plant height	高秆 Tall stalks	12	0.32
株高 Plant height	矮秆 Short stalks	121	3.26
育性和生育期 Fertility and maturity	早熟 Early-maturingt	27	0.73
	晚熟 Late-maturing	53	1.43
	不育 Sterile	50	1.35
其他 Others	叶环腐斑 Leaf ring rot	23	0.62
	叶黄斑 Leaf yellow spot	20	0.54
	叶白斑 Leaf white spot	21	0.57
	叶白线 White lines on the leaf	9	0.24
	叶黑线 Black lines on the leaf	2	0.05
合计 Total		1 226	33.05

品种 Material	NaCl （mmol/L）	苗高 Seedling height （cm）	最大根长 Maximum root length （cm）	初生根数 Number of initial roots
宁春4号 Ningchun No. 4	0	14.12a	8.92a	5.47a
宁春4号 Ningchun No. 4	250	15.16b	9.96b	5.62b
突变体 Mutant	0	14.12a	9.77c	5.46a
突变体 Mutant	250	18.44c	12.34d	5.74c